Sample records for acanthocephalan pomphorhynchus laevis

  1. Paratenic hosts as regular transmission route in the acanthocephalan Pomphorhynchus laevis: potential implications for food webs

    NASA Astrophysics Data System (ADS)

    Médoc, Vincent; Rigaud, Thierry; Motreuil, Sébastien; Perrot-Minnot, Marie-Jeanne; Bollache, Loïc

    2011-10-01

    Although trophically transmitted parasites are recognized to strongly influence food-web dynamics through their ability to manipulate host phenotype, our knowledge of their host spectrum is often imperfect. This is particularly true for the facultative paratenic hosts, which receive little interest. We investigated the occurrence and significance both in terms of ecology and evolution of paratenic hosts in the life cycle of the fish acanthocephalan Pomphorhynchus laevis. This freshwater parasite uses amphipods as intermediate hosts and cyprinids and salmonids as definitive hosts. Within a cohort of parasite larvae, usually reported in amphipod intermediate hosts, more than 90% were actually hosted by small-sized fish. We demonstrated experimentally, using one of these fish, that they get infected through the consumption of parasitized amphipods and contribute to the parasite's transmission to a definitive host, hence confirming their paratenic host status. A better knowledge of paratenic host spectrums could help us to understand the fine tuning of transmission strategies, to better estimate parasite biomass, and could improve our perception of parasite subwebs in terms of host-parasite and predator-parasite links.

  2. Comparison of morphological characters in Irish and English populations of the acanthocephalan Pomphorhynchus laevis (Müller, 1776).

    PubMed

    O'Mahony, E M; Kennedy, C R; Holland, C V

    2004-10-01

    Pomphorhynchus laevis is believed on ecological evidence to exist as three strains in the British Isles. However, the strains have never been shown to be capable of being distinguished using morphological characters. A morphological comparison was made between a sample of P. laevis from Salmo trutta in L. Feeagh in the west of Ireland and a sample from Leuciscus cephalus in R. Culm in the south of England. The length and width of the trunk, neck, bulb, proboscis and hooks were measured. The number of hooks per row, the number of rows and the positions of the stoutest and longest hooks were also recorded. A Principal Components Analysis based on the morphological measurements confirmed the separation of the two populations and showed that two characters successfully identified the populations: the position of the stoutest hook and the ratio of numbers of anterior to posterior hooks.

  3. Does fish reproduction and metabolic activity influence metal levels in fish intestinal parasites, acanthocephalans, during fish spawning and post-spawning period?

    PubMed

    Filipović Marijić, Vlatka; Vardić Smrzlić, Irena; Raspor, Biserka

    2014-10-01

    Application of fish intestinal parasites, acanthocephalans, as bioindicators in metal exposure assessment usually involves estimation of their metal levels and bioconcentration factors. Metal levels in parasite final host, fishes, are influenced by fish physiology but there is no data for acanthocephalan metal levels. Gastrointestinal Zn, Fe, Mn, Cd, Ag levels in European chub (Squalius cephalus L.) from the Sava River were significantly higher during chub spawning (April/May) compared to the post-spawning period (September). In acanthocephalans (Pomphorhynchus laevis and Acanthocephalus anguillae) significantly higher metal levels during chub spawning were observed only for Zn in P. laevis. Bioconcentration factors were twice as high for Fe, Mn, Ag, Pb in the post-spawning period, probably as a consequence of lower gastrointestinal metal levels in fish rather than metal exposure. Therefore, bioconcentration factors should be interpreted with caution, due to their possible variability in relation to fish physiology. In addition, gastrointestinal Cu, Cd and Pb levels were lower in infected than uninfected chub, indicating that metal variability in fishes might be affected by the presence of acanthocephalans. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Enteric neuromodulators and mucus discharge in a fish infected with the intestinal helminth Pomphorhynchus laevis.

    PubMed

    Bosi, Giampaolo; Shinn, Andrew Paul; Giari, Luisa; Sayyaf Dezfuli, Bahram

    2015-07-08

    In vertebrates, the presence of enteric worms can induce structural changes to the alimentary canal impacting on the neuroendocrine system, altering the proper functioning of the gastrointestinal tract and affecting the occurrence and relative density of endocrine cells (ECs). This account represents the first immunohistochemistry and ultrastructure-based study which documents the intimate relationship between the intestinal mucous cells and ECs in a fish-helminth system, investigating the potential effects of enteric neuromodulators on gut mucus secretion/discharge. A modified dual immunohisto- and histochemical staining technique was applied on intestinal sections from both infected and uninfected fish. Sections were incubated in antisera to a range of neuromodulators (i.e. leu-enkephalin, met-enkephalin, galanin and serotonin) and the glycoconjugate histochemistry of the mucous cells was determined using a subsequent alcian blue - periodic acid Schiff staining step. Dual fluorescent staining on sections prepared for confocal laser scanning microscopy and transmission electron microscopy were also used to document the relationship between ECs and mucous cells. From a total of 26 specimens of Squalius cephalus sampled from the River Paglia, 16 (i.e. 62 %) specimens were found to harbour an infection of the acanthocephalan Pomphorhynchus laevis (average intensity of infection 9.2 ± 0.8 parasites host(-1), mean ± standard error). When acanthocephalans were present, the numbers of mucous cells (most notably those containing acidic or mixed glycoconjugates) and ECs secreting leu-enkephalin, met-enkephalin, galanin, serotonin were significantly higher than those seen on sections from uninfected fish. The relationship between met-enkephalin-like or serotonin-like ECs and lectin DBA positive mucous cells was demonstrated through a dual fluorescent staining. The presence of tight connections and desmosomes between mucous and ECs in transmission electron

  5. Infection with an acanthocephalan manipulates an amphipod's reaction to a fish predator's odours.

    PubMed

    Baldauf, Sebastian A; Thünken, Timo; Frommen, Joachim G; Bakker, Theo C M; Heupel, Oliver; Kullmann, Harald

    2007-01-01

    Many parasites with complex life cycles increase the chances of reaching a final host by adapting strategies to manipulate their intermediate host's appearance, condition or behaviour. The acanthocephalan parasite Pomphorhynchus laevis uses freshwater amphipods as intermediate hosts before reaching sexual maturity in predatory fish. We performed a series of choice experiments with infected and uninfected Gammarus pulex in order to distinguish between the effects of visual and olfactory predator cues on parasite-induced changes in host behaviour. When both visual and olfactory cues, as well as only olfactory cues were offered, infected and uninfected G. pulex showed significantly different preferences for the predator or the non-predator side. Uninfected individuals significantly avoided predator odours while infected individuals significantly preferred the side with predator odours. When only visual contact with a predator was allowed, infected and uninfected gammarids behaved similarly and had no significant preference. Thus, we believe we show for the first time that P. laevis increases its chance to reach a final host by olfactory-triggered manipulation of the anti-predator behaviour of its intermediate host.

  6. Conflict between co-occurring manipulative parasites? An experimental study of the joint influence of two acanthocephalan parasites on the behaviour of Gammarus pulex.

    PubMed

    Cezilly, F; Gregoire, A; Bertin, A

    2000-06-01

    When two parasite species are manipulators and have different definitive hosts, there is a potential for conflict between them. Selection may then exist for either avoiding hosts infected with conflicting parasites, or for hijacking, i.e. competitive processes to gain control of the intermediate host. The evidence for both phenomena depends largely on the study of the relative competitive abilities of parasites within their common intermediate host. We studied the effects of simultaneous infection by a fish acanthocephalan parasite, Pomphorhynchus laevis, and a bird acanthocephalan parasite, Polymorphus minutus, on the behaviour of their common intermediate host, the amphipod Gammarus pulex. We compared the reaction to light and vertical distribution of individuals infected with both parasites to those of individuals harbouring a single parasite species and uninfected ones under controlled conditions. Compared to uninfected gammarids that were photophobic and tended to remain at the bottom of the water column, P. laevis-infected gammarids were attracted to light, whereas P. minutus-infected individuals showed a modified vertical distribution and were swimming closer to the water surface. The effects of both P. laevis and P. minutus appeared to be dependent only on their presence, not on their intensity. Depending on the behavioural trait under study, however, the outcome of the antagonism between P. laevis and P. minutus differed. The vertical distribution of gammarids harbouring both parasites was half-way between those of P. laevis- and P. minutus-infected individuals, whereas P. laevis was able to induce altered reaction to light even in the presence of P. minutus. We discuss our results in relation to the occurrence of active avoidance or hijacking between conflicting manipulative parasites and provide some recommendations for future research.

  7. Response of the gut neuroendocrine system of Leuciscus cephalus (L.) to the presence of Pomphorhynchus laevis Müller, 1776 (Acanthocephala).

    PubMed

    Bosi, G; Domeneghini, C; Arrighi, S; Giari, L; Simoni, E; Dezfuli, B S

    2005-04-01

    Immunohistochemical tests were applied to sections of intestine of uninfected and Pomphorhynchus laevis Muller-infected chub, Leuciscus cephalus (L.) using 15 different antisera. Nerve cell bodies and fibres immunoreactive (IR) to the anti-bombesin, -Cholecystokinin-8 (CCK-8), -galanin, -Gastrin-Releasing Peptide (-GRP), -Nitric Oxide Synthase (-NOS), -Substance P (-SP), and -Vasoactive Intestinal Peptide (-VIP) sera were observed in the myenteric plexus of uninfected chub. The density of nerve components immunoreactive to these antisera was high in the intestine of the infected fish, especially near the site of attachment. Moreover, numerous nerve fibres, immunoreactive to anti-bombesin, -GRP, -galanin, -SP, and -VIP sera, were encountered in the connective tissue capsule surrounding the bulb and proboscis of P. laevis. The occurrence of P. laevis in the chub gut significantly increased the number of endocrine cells per intestinal fold immunoreactive to galanin, met-enkephalin and leu-enkephalin antisera. CCK-8, Neuropeptide Y and glucagon-like immunoreactive cells were less numerous in the intestine of infected chub. A large number of cells in the tunica propria-submucosa of L. cephalus infected with P. laevis were immunoreactive to anti-serotonin and -leu-enkephalin sera.

  8. ITS rDNA sequences of Pomphorhynchus laevis (Zoega in Müller, 1776) and P. lucyi Williams and Rogers, 1984 (Acanthocephala: Palaeacanthocephala).

    PubMed

    Král'ová-Hromadová, Iva; Tietz, David F; Shinn, Andrew P; Spakulová, Marta

    2003-10-01

    The internal transcribed spacers (ITS-1 and ITS-2) of the ribosomal RNA gene of Pomphorhynchus laevis (Zoega in Müller, 1776) (Acanthocephala) isolated from various fish species across Central and Southern Europe were compared with those of P. lucyi Williams and Rogers, 1984 collected from the largemouth bass Micropterus salmonoides Boulenger from the USA. The nucleotide sequences of ITS regions of P. laevis from minnows Phoxinus phoxinus (L.) and chub Leuciscus cephalus (L.) from two distant localities in the Slovak Republic were found to be 100% identical. The ITS-1 and ITS-2 of P. laevis from chub from the Czech Republic and Italy were also mutually identical, but significantly different from Slovak worms (88.7% identity for ITS-1, 91.3% identity for ITS-2). A fifth sample collected from Barbus tyberinus Bonaparte from Italy was very similar to the sympatric Italian isolate from chub, possessing four nucleotide substitutions in ITS-1 (98.4% identity). The ITS rDNA sequences of P. lucyi differed significantly from those of P. laevis; the values of identity were 51.8-56.1% for ITS-1 and 63.1-65.3% for ITS-2, and were significantly higher than the range of P. laevis within-species variability. The results based on the ITS sequences confirmed the occurrence of strains in P. laevis from Continental Europe which are well defined by molecules but reveal only slight differences in their morphology.

  9. Nematode eel parasite found inside acanthocephalan cysts--a "Trojan horse" strategy?

    PubMed

    Emde, Sebastian; Rueckert, Sonja; Kochmann, Judith; Knopf, Klaus; Sures, Bernd; Klimpel, Sven

    2014-11-18

    The invasive eel parasite Anguillicoloides crassus (syn. Anguillicola crassus) is considered one of the major causes for the decline of the European eel (Anguilla anguilla) panmictic population. It impairs the swim bladder function and reduces swimming performance of its host. The life cycle of this parasite involves different intermediate and paratenic hosts. Despite an efficient immune system of the paratenic fish hosts acting against infections with A. crassus, levels of parasitized eels remain high in European river systems. Recently, the round goby Neogobius melanostomus (Gobiidae) has become dominant in many rivers in Europe and is still spreading at a rapid pace. This highly invasive species might potentially act as an important, so far neglected paratenic fish host for A. crassus. Based on own observations and earlier single sightings of A. crassus in N. melanostomus, 60 fresh individuals of N. melanostomus were caught in the Rhine River and examined to assess the infection levels with metazoan parasites, especially A. crassus. Glycerin preparations were used for parasite identification. The parasite most frequently found in N. melanostomus was the acanthocephalan Pomphorhynchus sp. (subadult stage) which occurred mainly encysted in the mesenteries and liver. Every third gobiid (P = 31.7%) was infected by A. crassus larvae (L3) which exclusively occurred inside the acanthocephalan cysts. No intact or degenerated larvae of A. crassus were detected elsewhere in the goby, neither in the body cavity and mesenteries nor in other organs. Affected cysts contained the acanthocephalan larvae and 1-12 (mI =3) living A. crassus larvae. Additionally, encysted larvae of the nematode Raphidascaris acus were detected in the gobies, but only in the body cavity and not inside the acanthocephalan cysts. Based on our observations, we suggest that A. crassus might actively bypass the immune response of N. melanostomus by invading the cysts of acanthocephalan parasites of the

  10. Suckers and other bursal structures of Pomphorhynchus bulbocolli and Acanthocephalus dirus (Acanthocephala).

    PubMed

    Doyle, L R; Gleason, L N

    1991-06-01

    Accessory copulatory structures in the bursa of Pomphorhynchus bulbocolli include 2 suckers on the inner surface and bursal rays. A stylet was present in the penis of this species. Two suckers were present in the bursa of Acanthocephalus dirus, but bursal rays and a stylet in the penis were not observed. This is the first report of suckers present in the bursas of acanthocephalans. Copulatory cement caps and holes surrounded by a ring approximating the inner bursal diameter and not covered by a cap were present on the sides of some P. bulbocolli, indicating the possibility of hypodermic insemination in this species.

  11. Morphological and molecular data for a new species of Pomphorhynchus Monticelli, 1905 (Acanthocephala: Pomphorhynchidae) in the Mexican redhorse Moxostoma austrinum Bean (Cypriniformes: Catostomidae) in central Mexico.

    PubMed

    García-Varela, Martín; Mendoza-Garfias, Berenit; Choudhury, Anindo; Pérez-Ponce de León, Gerardo

    2017-11-01

    Pomphorhynchus purhepechus n. sp. is described from the intestine of the Mexican redhorse Moxostoma austrinum Bean (Catostomidae) in central Mexico. The new species can be distinguished from the other seven described species of Pomphorhynchus Monticelli, 1905 in the Americas by a subspherical proboscis and 14 longitudinal rows with 16-18 hooks each; the third and the fourth row of hooks are alternately longest. Sequences of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene and the large subunit (LSU) rDNA (including the domains D2-D3) were used to corroborate the morphological distinction between the new species and Pomphorhynchus bulbocolli Linkins in Van Cleave, 1919, a species widely distributed in several freshwater fish species across Canada, USA, and Mexico. The genetic divergence estimated between the new species and the isolates of P. bulbocolli ranged between 13 and 14% for cox1, and between 0.6 and 0.8% for LSU. Maximum likelihood and Bayesian inference analyses of each dataset showed that the isolates of P. bulbocolli parasitising freshwater fishes from three families, the Catostomidae, Cyprinidae and Centrarchidae, represent a separate lineage, and that the acanthocephalans collected from two localities in central Mexico comprise an independent lineage. In addition, our analysis of the genetic variation of P. bulbocolli demonstrates that individuals of this acanthocephalan from different host species are conspecific. Finally, the distribution, host-association, and phylogenetic relationship of the new species, when placed in the context of the region's geological history, suggest that both host and parasite underwent speciation after their ancestors became isolated in Central Mexico.

  12. Species associations among larval helminths in an amphipod intermediate host.

    PubMed

    Dezfuli, B S; Giari, L; Poulin, R

    2000-10-01

    Larval helminths that share the same intermediate host may or may not also share the same definitive hosts. If one or more of these helminth species can manipulate the phenotype of the intermediate host, there can be great advantages or severe costs for other helminths resulting from co-occurring with a manipulator, depending on whether they have the same definitive host or not. Among 2372 specimens of the amphipod Echinogammarus stammeri collected from the river Brenta, northern Italy, there was a positive association between two acanthocephalan species with the same fish definitive hosts, the relatively common Pomphorhynchus laevis and the much less prevalent Acanthocephalus clavula. The number of cystacanths of P. laevis per infected amphipod, which ranged from one to five, did not influence the likelihood that the amphipod would also host A. clavula. A third acanthocephalan species, Polymorphus minutus,which matures in birds, showed no association with either of the two other species. These results show that associations among helminth species in intermediate hosts are not random, and are instead the product of selection favouring certain pathways of transmission.

  13. The endohelminth fauna of barbel ( Barbus barbus) correlates with water quality of the Danube River in Bulgaria.

    PubMed

    Nachev, M; Sures, B

    2009-04-01

    Infection of barbel with 10 species of metazoan parasites including 3 trematodes, 3 acanthocephalans and 4 nematodes was observed in fish collected from 3 localities in the Bulgarian part of the river Danube between summer 2004 and summer 2007. The composition as well as the diversity characteristics of the parasite communities were analysed seasonally and showed a clear correlation with the composition of the invertebrate fauna and water quality. The most prevalent species was the acanthocephalan Pomphorhynchus laevis, which was also the dominant species of the intestinal component communities at all sampling sites. The second most frequent parasite at all Danube localities was Rhabdochona hellichi, which occurred in significantly higher numbers at the less polluted sites. Overall, the diversity of helminth communities increased with decreasing levels of nutrients and pollutants at all sampling sites. Therefore, the composition and diversity of parasite communities may be used to characterize ecosystem health and integrity.

  14. The effects of parasite age and intensity on variability in acanthocephalan-induced behavioural manipulation.

    PubMed

    Franceschi, Nathalie; Bauer, Alexandre; Bollache, Loïc; Rigaud, Thierry

    2008-08-01

    Numerous parasites with complex life cycles are able to manipulate the behaviour of their intermediate host in a way that increases their trophic transmission to the definitive host. Pomphorhynchus laevis, an acanthocephalan parasite, is known to reverse the phototactic behaviour of its amphipod intermediate host, Gammarus pulex, leading to an increased predation by fish hosts. However, levels of behavioural manipulation exhibited by naturally-infected gammarids are extremely variable, with some individuals being strongly manipulated whilst others are almost not affected by infection. To investigate parasite age and parasite intensity as potential sources of this variation, we carried out controlled experimental infections on gammarids using parasites from two different populations. We first determined that parasite intensity increased with exposure dose, but found no relationship between infection and host mortality. Repeated measures confirmed that the parasite alters host behaviour only when it reaches the cystacanth stage which is infective for the definitive host. They also revealed, we believe for the first time, that the older the cystacanth, the more it manipulates its host. The age of the parasite is therefore a major source of variation in parasite manipulation. The number of parasites within a host was also a source of variation. Manipulation was higher in hosts infected by two parasites than in singly infected ones, but above this intensity, manipulation did not increase. Since the development time of the parasite was also different according to parasite intensity (it was longer in doubly infected hosts than in singly infected ones, but did not increase more in multi-infected hosts), individual parasite fitness could depend on the compromise between development time and manipulation efficiency. Finally, the two parasite populations tested induced slightly different degrees of behavioural manipulation.

  15. The Meristogram: a neglected tool for acanthocephalan systematics

    PubMed Central

    2016-01-01

    Abstract Background The hooks of the acanthocephalan proboscis exhibit serial variation in size and shape. The Meristogram was developed by Huffman and Bullock (1975) to provide a graphical representation of this positional variation in hook morphology. Initial studies demonstrated the ability of the Meristogram to discriminate species within the genera Echinorhynchus and Pomphorhynchus (Huffman and Bullock 1975, Huffman and Nickol 1978, Gleason and Huffman 1981). However, the reliability of the method for taxonomic work was questioned by Shostak et al. (1986) after they found intra-specific variation in two Echinorhynchus species. Uncertainty about the usefulness of the Meristogram and the absence of a readily available software implementation of the algorithm, might explain why this abstract proboscis character has yet to be adopted by acanthocephalan systematists. New information The Meristogram algorithm was implemented in the R language and a simple graphical user interface created to facilitate ease of use (the software is freely available from https://github.com/WaylandM/meristogram). The accuracy of the algorithm's formula for calculating hook cross-sectional area was validated by data collected using a digitizing tablet. Meristograms were created from data in public respositories for eight Echinorhynchus taxa: E. bothniensis, E. 'bothniensis', E. gadi spp. A, B and I, E. brayi, E. salmonis and E. truttae. In this preliminary analysis, the meristogram differentiated E. bothniensis, E. brayi, E. gadi sp. B, E. salmonis and E. truttae from each other, and from the remaining taxa in this study, but independent data will be required for validation. Sample sizes for E. 'bothniensis' and E. gadi spp. A and I were too small to identify diagnostic features with any degree of confidence. Meristogram differences among the sibling species of the E. gadi and E. bothniensis groups suggest that the 'intra-specfic' variation in meristogram previously reported for some

  16. A minimalist macroparasite diversity in the round goby of the Upper Rhine reduced to an exotic acanthocephalan lineage.

    PubMed

    David, Gwendoline M; Staentzel, Cybill; Schlumberger, Olivier; Perrot-Minnot, Marie-Jeanne; Beisel, Jean-Nicolas; Hardion, Laurent

    2017-12-12

    The round goby, Neogobius melanostomus, is a Ponto-Caspian fish considered as an invasive species in a wide range of aquatic ecosystems. To understand the role that parasites may play in its successful invasion across Western Europe, we investigated the parasitic diversity of the round goby along its invasion corridor, from the Danube to the Upper Rhine rivers, using data from literature and a molecular barcoding approach, respectively. Among 1666 parasites extracted from 179 gobies of the Upper Rhine, all of the 248 parasites barcoded on the c oxidase subunit I gene were identified as Pomphorhynchus laevis. This lack of macroparasite diversity was interpreted as a loss of parasites along its invasion corridor without spillback compensation. The genetic diversity of P. laevis was represented by 33 haplotypes corresponding to a haplotype diversity of 0·65 ± 0·032, but a weak nucleotide diversity of 0·0018 ± 0·00015. Eight of these haplotypes were found in 88·4% of the 248 parasites. These haplotypes belong to a single lineage so far restricted to the Danube, Vistula and Volga rivers (Eastern Europe). This result underlines the exotic status of this Ponto-Caspian lineage in the Upper Rhine, putatively disseminated by the round goby along its invasion corridor.

  17. Invasive Ponto-Caspian Amphipods and Fish Increase the Distribution Range of the Acanthocephalan Pomphorhynchus tereticollis in the River Rhine

    PubMed Central

    Emde, Sebastian; Rueckert, Sonja; Palm, Harry W.; Klimpel, Sven

    2012-01-01

    Non-indigenous species that become invasive are one of the main drivers of biodiversity loss worldwide. In various freshwater systems in Europe, populations of native amphipods and fish are progressively displaced by highly adaptive non-indigenous species that can perform explosive range extensions. A total of 40 Ponto-Caspian round gobies Neogobius melanostomus from the Rhine River near Düsseldorf, North Rhine-Westphalia, Germany, were examined for metazoan parasites and feeding ecology. Three metazoan parasite species were found: two Nematoda and one Acanthocephala. The two Nematoda, Raphidascaris acus and Paracuaria adunca, had a low prevalence of 2.5%. The Acanthocephala, Pomphorhynchus tereticollis, was the predominant parasite species, reaching a level of 90.0% prevalence in the larval stage, correlated with fish size. In addition, four invasive amphipod species, Corophium curvispinum (435 specimens), Dikerogammarus villosus (5,454), Echinogammarus trichiatus (2,695) and Orchestia cavimana (1,448) were trapped at the sampling site. Only D. villosus was infected with P. tereticollis at a prevalence of 0.04%. The invasive goby N. melanostomus mainly preys on these non-indigenous amphipods, and may have replaced native amphipods in the transmission of P. tereticollis into the vertebrate paratenic host. This study gives insight into a potential parasite-host system that consists mainly of invasive species, such as the Ponto-Caspian fish and amphipods in the Rhine. We discuss prospective distribution and migration pathways of non-indigenous vertebrate (round goby) and invertebrates (amphipods) under special consideration of parasite dispersal. PMID:23300895

  18. Evolutionary anatomy of the muscular apparatus involved in the anchoring of Acanthocephala to the intestinal wall of their vertebrate hosts.

    PubMed

    Herlyn, Holger; Taraschewski, Horst

    2017-04-01

    Different conceptions exist regarding structure, function, and evolution of the muscles that move the acanthocephalan presoma, including the proboscis, i.e., the usually hooked hold-fast anchoring these endoparasites to the intestinal wall of their vertebrate definitive hosts. In order to clarify the unresolved issues, we carried out a light microscopic analysis of series of semi-thin sections and whole mounts representing the three traditional acanthocephalan classes: Archiacanthocephala (Macracanthorhynchus hirudinaceus), Eoacanthocephala (Paratenuisentis ambiguus, Tenuisentis niloticus), and Palaeacanthocephala (Acanthocephalus anguillae, Echinorhynchus truttae, Pomphorhynchus laevis, Corynosoma sp.). Combining our data with published light, transmission electron, and scanning electron microscopic data, we demonstrate that receptacle protrusor and proboscis receptacle in Archi- and Eoacanthocephala are homologous to the outer and inner wall of the proboscis receptacle in Palaeacanthocephala. Besides the proboscis receptacle and a "surrounding muscle," the last common ancestor of Acanthocephala presumably possessed a proboscis retractor, receptacle retractor, neck retractor (continuous with lemnisci compressors), and retinacula. These muscles most probably evolved in the acanthocephalan stem line. Moreover, the last common ancestor of Acanthocephala presumably possessed only a single layer of muscular cords under the presomal tegument while the metasomal body wall had circular and longitudinal strands. Two lateral receptacle flexors (also lateral receptacle protrusors), an apical muscle plate (surrounding one or two apical sensory organs), a midventral longitudinal muscle, and the differentiation of longitudinal body wall musculature at the base of the proboscis probably emerged within Archiacanthocephala. All muscles have a common organization principle: a peripheral layer of contractile filaments encloses the cytoplasm.

  19. The discovery of acanthocephalans parasitizing chaetognaths.

    PubMed

    Lozano-Cobo, Horacio; Gómez-Gutiérrez, Jaime; Franco-Gordo, Carmen; Prado-Rosas, María Del Carmen Gómez Del

    2017-06-01

    A comprehensive literature review shows that 12 types of pathogens, micropredators and parasites are reported to interact with chaetognaths, mostly digenean trematodes, cestodes and nematodes larval stages. Through analysis of 78,152 chaetognaths from a monthly zooplankton time series (Jan 1996-Dec 1998) collected in the Mexican Central Pacific twelve acanthocephalan larvae parasitizing chaetognaths were discovered. This is the first record of an acanthocephalan parasitizing chaetognaths, raising to 13 the types of symbionts known to interact with them (excluding predators). Cystacanth larval specimens of Corynosoma sp. (Polymorphidae) were observed parasitizing the head, trunk and caudal cavities of three of the eight chaetognath species inhabiting this tropical coastal region (Flaccisagitta enflata, Flaccisagitta hexaptera and Zonosagitta bedoti). Because Corynosoma sp. parasitized chaetognaths during different months and years (Jan-Feb 1996, Mar and Jul 1997, Jan, Jun, Aug-Sep 1998) and because the total length of these cystacanths varied between 165-480 µm, suggesting growth inside the hosts, we conclude that chaetognaths are intermediate hosts of Corynosoma sp. The twelve parasitized chaetognaths were juveniles (without gonads) or immature adults (none in reproductively mature stage IV); therefore Corynosoma may have a negative influence on host reproduction. Marine crustaceans (amphipods, decapods, copepods, mysids and euphausiids) and fishes are common intermediate or paratenic hosts of acanthocephalans. Fish, sea birds and marine mammals are definitive hosts for marine Corynosoma species. The present discovery implies that acanthocephalans are transmitted trophically through different intermediate hosts (crustaceans, chaetognaths and/or fish); thus chaetognaths can also be part of the marine acanthocephalan life cycle.

  20. Understanding trophic interactions in host-parasite associations using stable isotopes of carbon and nitrogen.

    PubMed

    Nachev, Milen; Jochmann, Maik A; Walter, Friederike; Wolbert, J Benjamin; Schulte, S Marcel; Schmidt, Torsten C; Sures, Bernd

    2017-02-17

    Stable isotope analysis of carbon and nitrogen can deliver insights into trophic interactions between organisms. While many studies on free-living organisms are available, the number of those focusing on trophic interactions between hosts and their associated parasites still remains scarce. In some cases information about taxa (e.g. acanthocephalans) is completely missing. Additionally, available data revealed different and occasionally contrasting patterns, depending on the parasite's taxonomic position and its degree of development, which is most probably determined by its feeding strategy (absorption of nutrients through the tegument versus active feeding) and its localization in the host. Using stable isotope analysis of carbon and nitrogen we provided first data on the trophic position of an acanthocephalan species with respect to its fish host. Barbels (Barbus barbus) infected only with adult acanthocephalans Pomphorhynchus laevis as well as fish co-infected with the larval (L4) nematodes Eustrongylides sp. from host body cavity were investigated in order to determine the factors shaping host-parasite trophic interactions. Fish were collected in different seasons, to study also potential isotopic shifts over time, whereas barbels with single infection were obtained in summer and co-infected ones in autumn. Acanthocephalans as absorptive feeders showed lower isotope discrimination values of δ 15 N than the fish host. Results obtained for the acanthocephalans were in line with other parasitic taxa (e.g. cestodes), which exhibit a similar feeding strategy. We assumed that they feed mainly on metabolites, which were reprocessed by the host and are therefore isotopically lighter. In contrast, the nematodes were enriched in the heavier isotope δ 15 N with respect to their host and the acanthocephalans, respectively. As active feeders they feed on tissues and blood in the body cavity of the host and thus showed isotope discrimination patterns resembling those of

  1. A comparative analysis of adult body size and its correlates in acanthocephalan parasites.

    PubMed

    Poulin, Robert; Wise, Megan; Moore, Janice

    2003-07-30

    Adult acanthocephalan body sizes vary interspecifically over more than two orders of magnitude; yet, despite its importance for our understanding of the coevolutionary links between hosts and parasites, this variation remains unexplained. Here, we used a comparative analysis to investigate how final adult sizes and relative increments in size following establishment in the definitive host are influenced by three potential determinants of acanthocephalan sizes: initial (cystacanth) size at infection, host body mass, and the thermal regime experienced during growth, i.e. whether the definitive host is an ectotherm or an endotherm. Relative growth from the cystacanth stage to the adult stage ranged from twofold to more than 10,000-fold across acanthocephalan species, averaging just over 100-fold. However, this relative increment in size did not correlate with host mass, and did not differ between acanthocephalan species using ectothermic hosts and those growing in endothermic hosts. In contrast, final acanthocephalan adult sizes correlated positively with host mass, and after correction for host mass, final adult sizes were higher in species parasitising endotherms than in those found in ectotherms. The relationship between host mass and acanthocephalan adult size practically disappears, however, once phylogenetic influences are taken into account. Positive relationships between adult acanthocephalan size, cystacanth size and egg size indicate that a given relative size is a feature of an acanthocephalan species at all stages of its life cycle. These relationships also suggest that adult size is to some extent determined by cystacanth size, and that the characteristics of the definitive host are not the sole determinants of parasite life history traits.

  2. [BIODIVERSITY OF ACANTHOCEPHALANS (ACANTHOCEPHALA) IN FRESHWATER FISHES OF ASIATIC SUB-ARCTIC REGION].

    PubMed

    Atrashkevich, G I; Mikhailova, E I; Orlovskaya, O M; Pospekhov, V V

    2016-01-01

    The analysis of taxonomical and ecological diversity of acanthocephalans in fishes of Asiatic sub-Arctic region freshwaters, summarizing changes in modern views on species composition, life cycles, and ecology of background groups of these parasites is given. A priority role of studies provided by O. N. Bauer and his scientific school in organization and development of these aspects of acanthocephalology is demonstrated. Special attention is paid to the assessment of acanthocephalan biodiversity of the genus Neoechinorhynchus, the background group of freshwater fish parasites of the Asiatic sub-Arctic region, and an original key for their species is given. The distribution of acanthocephalans of the genus Acanthocephalus in northeastern Asia is analyzed and prospective study of this parasite group, evolutionary associated with freshwater isopods of the genus Asellus as intermediate hosts, is outlined. The absence of documented evidences on intermediate hosts of other background parasites of freshwater fishes in the region, acanthocephalans of the genus Metechinorhynchus, is revealed. It is assumed that subsequent taxonomic revisions based both on morphological and molecular genetic studies are necessary for the reliable revealing of species composition in each genus of the background acanthocephalans from freshwater fishes of Northern Asia. Theoretical significance of the study of acanthocephalan life cycles and revealing their natural intermediate hosts for the reliable estimation of structural and functional organization of their host-parasite systems in different parts of the range is substantiated and the possibility of the distribution of taxonomic conclusions in new territories is analyzed. A brief annotated taxonomical list of freshwater acanthocephalans of the Asiatic sub-Arctic region is given.

  3. The complete mitochondrial genome of Pallisentis celatus (Acanthocephala) with phylogenetic analysis of acanthocephalans and rotifers.

    PubMed

    Pan, Ting Shuang; Nie, Pin

    2013-07-01

    Acanthocephalans are a small group of obligate endoparasites. They and rotifers are recently placed in a group called Syndermata. However, phylogenetic relationships within classes of acanthocephalans, and between them and rotifers, have not been well resolved, possibly due to the lack of molecular data suitable for such analysis. In this study, the mitochondrial (mt) genome was sequenced from Pallisentis celatus (Van Cleave, 1928), an acanthocephalan in the class Eoacanthocephala, an intestinal parasite of rice-field eel, Monopterus albus (Zuiew, 1793), in China. The complete mt genome sequence of P. celatus is 13 855 bp long, containing 36 genes including 12 protein-coding genes, 22 transfer RNAs (tRNAs) and 2 ribosomal RNAs (rRNAs) as reported for other acanthocephalan species. All genes are encoded on the same strand and in the same direction. Phylogenetic analysis indicated that acanthocephalans are closely related with a clade containing bdelloids, which then correlates with the clade containing monogononts. The class Eoacanthocephala, containing P. celatus and Paratenuisentis ambiguus (Van Cleave, 1921) was closely related to the Palaeacanthocephala. It is thus indicated that acanthocephalans may be just clustered among groups of rotifers. However, the resolving of phylogenetic relationship among all classes of acanthocephalans and between them and rotifers may require further sampling and more molecular data.

  4. [Infection of the pacific saury Cololabis saira by acanthocephalans in the Kuril Islands area].

    PubMed

    Gordeev, I I; Grigorov, I V; Afanasyev, P K

    2017-01-01

    The Pacific saury Cololabis saira (Brevoort, 1856) is one of the important target species of commercial fisheries. Food manufacturers and consumers encounter problems due to the infection of the saury by acanthocephalans, which are quite difficult to clean out completely during on-board catch processing. Infection of C. saira was not studied on a regular basis, therefore, our knowledge about the parasites of saury is fragmentary. This paper contains infection indices (only acanthocephalans) of the Pacific saury caught in the Kuril Islands area (Russian Exclusive Economic Zone) in 2015.

  5. First paleoparasitological record of acanthocephalan eggs from Northwestern Patagonia (Late Holocene, Argentina).

    PubMed

    Beltrame, María Ornela; Fernández, Fernando Julián; Sardella, Norma Haydeé

    2015-06-01

    Eggs representative of an acanthocephalan were found in an ancient fragment of raptor pellet, probably belonged to the barn owl, Tyto alba, from the archeological site named "Epullán Chica cave." This site is a cave located at the southern of Neuquén Province, Patagonia, Argentina. The fragment of pellet was found in a layer with charcoals dated at 1980±80 years B.P. A total of 56 eggs were found. Eggs were brown colored and thick-shelled, and presented four membranes, the outer lightly sculpted. The embryos presented hooks in one extremity. Measurements ranged from 87.5 to 107.5μm long and 50 to 57.5μm wide. Eggs were very well-preserved, and were identified as belonged to Class Archiacanthocephala, Order Oligacanthorhynchida, Family Oligacanthorhynchidae, probably Macracanthorhynchus Travassos, 1917, or an unidentified species. This is the first report of small mammal acanthocephalans from ancient material worldwide. Copyright © 2015 Elsevier B.V. All rights reserved.

  6. Acanthocephalans of the genus Centrorhynchus (Palaeacanthocephala: Centrorhynchidae) of birds of prey (Falconiformes) and owls (Strigiformes) in Slovakia.

    PubMed

    Komorová, P; Špakulová, M; Hurníková, Z; Uhrín, M

    2015-06-01

    Three species of thorny-headed worms of the genus Centrorhynchus were found to parasitize birds of prey and owls in the territory of the Slovakia during the years 2012-2014. Out of 286 examined bird individuals belonging to 23 species, only Buteo buteo, Buteo rufinus, Falco tinnunculus (Falconiformes), Asio otus, Strix aluco, Strix uralensis and Tyto alba (Strigiformes) were infected by acanthocephalans. All the bird species except for S. aluco represent new host records for Slovakia. The most prevalent acanthocephalan Centrorhynchus aluconis was detected in all 15 examined birds of non-migratory Ural owl S. uralensis (P = 100%); however, it was found occasionally also in two individuals of the tawny owl S. aluco (P = 20%), one long-eared owl A. otus (P = 7.7%), one barn owl T. alba (P = 33.3%) and the common buzzard B. buteo (P = 0.8%). Two other thorny-headed worms occurred exclusively in Falconiformes in raw or mixed infections: Centrorhynchus buteonis was found in 11 individuals of B. buteo (P = 9.2%), and two birds (B. buteo and B. rufinus) were parasitized simultaneously by C. buteonis and the species Centrorhynchus globocaudatus. Moreover, the latest, relatively rare acanthocephalan was found alone in two common kestrels F. tinnunculus (P = 2.7%). Regarding intensity of infection, it ranged from a single female of C. buteonis, C. globocaudatus or C. aluconis per host (four cases) to a maximum of 82 C. aluconis per an Ural owl. The difference in acanthocephalan species spectrum between birds of prey and owls in Slovakia was apparent.

  7. Xenopus laevis and Emerging Amphibian Pathogens in Chile.

    PubMed

    Soto-Azat, Claudio; Peñafiel-Ricaurte, Alexandra; Price, Stephen J; Sallaberry-Pincheira, Nicole; García, María Pía; Alvarado-Rybak, Mario; Cunningham, Andrew A

    2016-12-01

    Amphibians face an extinction crisis with no precedence. Two emerging infectious diseases, ranaviral disease caused by viruses within the genus Ranavirus and chytridiomycosis due to Batrachochytrium dendrobatidis (Bd), have been linked with amphibian mass mortalities and population declines in many regions of the globe. The African clawed frog (Xenopus laevis) has been indicated as a vector for the spread of these pathogens. Since the 1970s, this species has been invasive in central Chile. We collected X. laevis and dead native amphibians in Chile between 2011 and 2013. We conducted post-mortem examinations and molecular tests for Ranavirus and Bd. Eight of 187 individuals (4.3 %) tested positive for Ranavirus: seven X. laevis and a giant Chilean frog (Calyptocephallela gayi). All positive cases were from the original area of X. laevis invasion. Bd was found to be more prevalent (14.4 %) and widespread than Ranavirus, and all X. laevis Bd-positive animals presented low to moderate levels of infection. Sequencing of a partial Ranavirus gene revealed 100 % sequence identity with Frog Virus 3. This is the first report of Ranavirus in Chile, and these preliminary results are consistent with a role for X. laevis as an infection reservoir for both Ranavirus and Bd.

  8. Distribution of the Hawaiian ghost crab, Ocypode laevis Dana

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fellows, D.P.

    1975-01-01

    The presence of a small breeding population of Ocypode laevis at Enewetak Atoll, Marshall Islands, is reported, and morphological and behavioral comparisons are made with the Hawaiian O. laevis. Previous distribution records for the species are discussed and corrected.

  9. Acanthocephalan fish parasites (Rhadinorhynchidae Lühe, 1912) as potential biomarkers: Molecular-chemical screening by pyrolysis-field ionization mass spectrometry

    NASA Astrophysics Data System (ADS)

    Kleinertz, S.; Eckhardt, K.-U.; Theisen, S.; Palm, H. W.; Leinweber, P.

    2016-07-01

    The present study represents the first molecular-chemical screening by pyrolysis-field ionization mass spectrometry applied on fish parasites. A total of 71 fishes from Balinese fish markets, 36 Auxis rochei (Risso, 1810) and 35 A. thazard (Lacepède, 1800), were studied for their acanthocephalan parasites. This is the first record of Rhadinorhynchus zhukovi in Balinese waters, Indonesia, and we describe for the first time A. rochei and A. thazard as R. zhukovi hosts. Using this method, small scale variations within the chemical compounds of acanthocephalans could be detected. Using this methodology it will be possible to generate additional, pollutant specific information from aquatic habitats in future with the potential of a new bioindicator application for parasite/host origin and/or environmental pollution.

  10. Acanthocephalan parasites of slimy sculpin, Cottus cognatus, and Ninespine Stickleback, Pungitius pungitius, from Lake Michigan, U.S.A.

    USGS Publications Warehouse

    Muzzall, Patrick M.; Lima, Michael; Gentile, Alex; Gunn, Jacob; Jones, Amanda; Morrison, Jamie; French, John R. P.

    2012-01-01

    In total, 288 slimy sculpins, Cottus cognatus, were collected in September 2003 from 6 Lake Michigan, U.S.A., ports, along with 220 ninespine sticklebacks, Pungitius pungitius, from 3 ports. The ports included Waukegan, Illinois; Port Washington (PW) and Sturgeon Bay (SB), Wisconsin; and Manistique (MS), Frankfort (FF), Ludington (LD), and Saugatuck, Michigan. Echinorhynchus salmonis infected sculpins from 6 ports, Acanthocephalus dirus infected sculpins from 4 ports, and Neoechinorhynchus pungitius infected sculpins from 3 ports. Echinorhynchus salmonis infected significantly more sculpins at PW and at FF than at MS and LD. There were several significant differences in the intensities and abundances of E. salmonis among ports. Acanthocephalus dirus significantly infected more sculpins and had significantly higher abundances at FF than at PW, MS, and LD. Echinorhynchus salmonis, A. dirus, and N. pungitius infected sticklebacks from SB, MS, and FF. Neoechinorhynchus pungitius significantly infected more sculpins and more sticklebacks, and it had significantly higher abundances at MS than at FF. Neoechinorhynchus pungitius was the most common acanthocephalan in C. cognatus and P. pungitius at MS. These acanthocephalan species infecting C. cognatus and P. pungitius corresponded in their occurrence to those organisms that serve as their intermediate hosts found in the stomachs of both fish species. Potential changes in the diet of C. cognatus played a role in significant differences found for E. salmonis and N. pungitius at MS. One of these acanthocephalan species was always the most numerous helminth species found in the digestive tracts of P. pungitius and C. cognatus from these Lake Michigan ports.

  11. Xenopus laevis oocyte maturation is affected by metal chlorides.

    PubMed

    Marin, Matthieu; Slaby, Sylvain; Marchand, Guillaume; Demuynck, Sylvain; Friscourt, Noémie; Gelaude, Armance; Lemière, Sébastien; Bodart, Jean-François

    2015-08-01

    Few studies have been conducted using Xenopus laevis germ cells as oocytes, though these cells offer many advantages allowing both electrophysiological studies and morphological examination. Our aim was to investigate the effects of metal (cadmium, lead, cobalt and zinc) exposures using cell biology approaches. First, cell survival was evaluated with both phenotypical and electrophysiological approaches. Secondly, the effect of metals on oocyte maturation was assessed with morphological observations and electrophysiological recordings. From survival experiments, our results showed that metal chlorides did not affect cell morphology but strongly depolarized X. laevis oocyte resting potential. In addition, cadmium chloride was able to inhibit progesterone-induced oocyte maturation. By contrast, zinc, but also to a lesser extent cadmium, cobalt and lead, were able to enhance spontaneous oocyte maturation in the absence of progesterone stimulation. Finally, electrophysiological recordings revealed that some metal chlorides (lead, cadmium) exposures could disturb calcium signaling in X. laevis oocyte by modifying calcium-activated chloride currents. Our results demonstrated the high sensitivity of X. laevis oocytes toward exogenous metals such as lead and cadmium. In addition, the cellular events recorded might have a predictive value of effects occurring later on the ability of oocytes to be fertilized. Together, these results suggest a potential use of this cellular lab model as a tool for ecotoxicological assessment of contaminated fresh waters. Copyright © 2015 Elsevier Ltd. All rights reserved.

  12. Getting What Is Served? Feeding Ecology Influencing Parasite-Host Interactions in Invasive Round Goby Neogobius melanostomus

    PubMed Central

    Emde, Sebastian; Kochmann, Judith; Kuhn, Thomas; Plath, Martin; Klimpel, Sven

    2014-01-01

    Freshwater ecosystems are increasingly impacted by alien invasive species which have the potential to alter various ecological interactions like predator-prey and host-parasite relationships. Here, we simultaneously examined predator-prey interactions and parasitization patterns of the highly invasive round goby (Neogobius melanostomus) in the rivers Rhine and Main in Germany. A total of 350 N. melanostomus were sampled between June and October 2011. Gut content analysis revealed a broad prey spectrum, partly reflecting temporal and local differences in prey availability. For the major food type (amphipods), species compositions were determined. Amphipod fauna consisted entirely of non-native species and was dominated by Dikerogammarus villosus in the Main and Echinogammarus trichiatus in the Rhine. However, the availability of amphipod species in the field did not reflect their relative abundance in gut contents of N. melanostomus. Only two metazoan parasites, the nematode Raphidascaris acus and the acanthocephalan Pomphorhynchus sp., were isolated from N. melanostomus in all months, whereas unionid glochidia were only detected in June and October in fish from the Main. To analyse infection pathways, we examined 17,356 amphipods and found Pomphorhynchus sp. larvae only in D. villosus in the river Rhine at a prevalence of 0.15%. Dikerogammarus villosus represented the most important amphipod prey for N. melanostomus in both rivers but parasite intensities differed between rivers, suggesting that final hosts (large predatory fishes) may influence host-parasite dynamics of N. melanostomus in its introduced range. PMID:25338158

  13. Population genetic structure of the acanthocephalan Acanthosentis cheni in anadromous, freshwater, and landlocked stocks of its fish host, Coilia nasus.

    PubMed

    Song, Rui; Li, Wen X; Wu, Shan G; Zou, Hong; Wang, Gui T

    2014-04-01

    The acanthocephalan Acanthosentis cheni was found in anadromous, freshwater, and landlocked stocks of its fish host, Coilia nasus. To examine the genetic variations of the acanthocephalan among the 3 populations with the adaptation of the host to the freshwater, the genetic structure of the helminth was investigated in anadromous (Zhoushan and Chongming islands, and Anqing), freshwater (Anqing, Ezhou, and Poyang Lake), and landlocked (Tian'ezhou Reserve) populations by sequencing intergenic transcribed spacers (ITS) of the ribosomal RNA coding genes. Low Fst values and high gene flow were found among the 7 populations (Fst = 0.0135, P = 0.2723; Nm = 36.48) and the 3 ecotypes of Acanthosentis cheni (Fst = 0.0178, P = 0.1044; Nm = 27.67). On the other hand, significant genetic differentiation of the C. nasus host populations was detected between the upstream and downstream areas of Xiaogu Mountain (Fst = 0.1961, P = 0.0030; Nm = 2.05), which is the farthest location of spawning migration for C. nasus . However, the migration break of the fish host appeared not to cause significant genetic differentiation of A. cheni populations between the upper and lower reaches of Xiaogu Mountain. Other factors might promote genetic exchange of A. cheni populations such as dispersal of the intermediate host by flooding or other fish species serving as the definitive or paratenic hosts. In Anqing, nucleotide diversity of the acanthocephalan was highest in the freshwater population (0.0038) and lower in the anadromous population (0.0026). This suggested that new mutations may have occurred in the freshwater A. cheni population in Anqing when adapting to a freshwater environment.

  14. A quantitative adverse outcome pathway model for thyroid axis disruption in Xenopus laevis tadpoles

    EPA Science Inventory

    The development of Xenopus laevis tadpoles is tightly controlled by the thyroid hormones tetraiodothyronine (T4) and triiodothyronine (T3). Toxicity testing efforts have shown that several compounds interfere with development in X. laevis tadpoles by disrupting the thyroid axis a...

  15. Effect of acanthocephalan parasites on hiding behaviour in two species of shore crabs.

    PubMed

    Latham, A D M; Poulin, R

    2002-12-01

    The effect of acanthocephalan parasites (Profilicollis spp.) on the hiding behaviour during low tide of two species of shore crabs (intermediate hosts), Macrophthalmus hirtipes (Brachyura: Ocypodidae) and Hemigrapsus crenulatus (Brachyura: Grapsidae), was examined at Blueskin Bay, South Island, New Zealand. Exposed M. hirtipes were found to have significantly higher infection levels than did hidden conspecifics. This pattern was not observed for H. crenulatus. Mean cystacanth numbers were found to be considerably higher in M. hirtipes than H. crenulatus. Crabs exposed at low tide are at a greater risk of predation by definitive shorebird hosts than are hidden conspecifics. Preferential manipulation of one intermediate host species over another could influence diversity within ecosystems.

  16. XENOPUS LAEVIS: A CULTURING AND REARING PROTOCOL

    EPA Science Inventory

    Xenopus laevis are used extensively here at MED-Duluth as a model for assessing development toxicity to xenobiotics. As a result, a culturing system has been developed that provides eggs and tadpoles of consistent high quality for use by researchers at the facility. The methods ...

  17. On four species of echinorhynchid acanthocephalans from marine fish in Halong Bay, Vietnam, including the description of three new species and a key to the species of Gorgorhynchus.

    PubMed

    Amin, Omar M; Van Ha, Nguyen

    2011-09-01

    Four species of echinorhynchid acanthocephalans were collected from marine fish off Cat Ba Island, Halong Bay, Gulf of Tonkin, Vietnam, in the spring of 2009. Acanthocephalus halongensis n. sp. (Echinorhynchidae) from the redtail scad, Decapterus kurroides Bleeker 1855 (Carangidae), has a unique proboscis armature with a spiniform basal hook with lateral root and an incomplete receptacle wall posteriorly. Gorgorhynchus tonkinensis n. sp. (Rhadinorhynchidae) also from D. kurroides, has long, slender, winding lemnisci, many epidermal nuclei, and a narrow anterior trunk with a shoulder armed with 20 circles of tightly packed spines, the posterior four circles of which have abruptly larger spines than those in the anterior circles. Neorhadinorhynchus atypicalis n. sp. (Cavisomidae) from the rabbitfish, Siganus fuscescens (Houttuyn 1782) (Siganidae), has the largest number of proboscis hooks per row, testes wider than long, and four clustered cement glands. Micracanthorhynchica kuwaitensis Amin and Sey 1996 (Rhadinorhynchidae) from the spottail needlefish Strongylura strongylura (van Hasselt 1823) (Belonidae) was similar to specimens originally described from the Arabian Gulf off the Kuwaiti coast. These acanthocephalans were collected in small numbers but stood out as uniquely and considerably different from their closest relatives to warrant their reporting. All species of acanthocephalans and their host and geographic distribution are described, and a key to the species of Gorgorhynchus is provided.

  18. Xenopus laevis in Developmental and Molecular Biology.

    ERIC Educational Resources Information Center

    Dawid, Igor B.; Sargent, Thomas D.

    1988-01-01

    Discusses the advantages of Xenopus laevis as an experimental animal in the study of embryogenesis in vertebrates. Summarizes the contributions of this system to the analysis of ribosomal and 5S RNA genes, and the diverse and highly productive applications of the oocyte injection technology. (RT)

  19. Unequal contribution of native South African phylogeographic lineages to the invasion of the African clawed frog, Xenopus laevis, in Europe

    PubMed Central

    Courant, Julien; Herrel, Anthony; Rebelo, Rui; Rödder, Dennis; Measey, G. John; Backeljau, Thierry

    2016-01-01

    Due to both deliberate and accidental introductions, invasive African Clawed Frog (Xenopus laevis) populations have become established worldwide. In this study, we investigate the geographic origins of invasive X. laevis populations in France and Portugal using the phylogeographic structure of X. laevis in its native South African range. In total, 80 individuals from the whole area known to be invaded in France and Portugal were analysed for two mitochondrial and three nuclear genes, allowing a comparison with 185 specimens from the native range. Our results show that native phylogeographic lineages have contributed differently to invasive European X. laevis populations. In Portugal, genetic and historical data suggest a single colonization event involving a small number of individuals from the south-western Cape region in South Africa. In contrast, French invasive X. laevis encompass two distinct native phylogeographic lineages, i.e., one from the south-western Cape region and one from the northern regions of South Africa. The French X. laevis population is the first example of a X. laevis invasion involving multiple lineages. Moreover, the lack of population structure based on nuclear DNA suggests a potential role for admixture within the invasive French population. PMID:26855879

  20. Transcriptome analysis identifies genes involved in sex determination and development of Xenopus laevis gonads.

    PubMed

    Piprek, Rafal P; Damulewicz, Milena; Kloc, Malgorzata; Kubiak, Jacek Z

    Development of the gonads is a complex process, which starts with a period of undifferentiated, bipotential gonads. During this period the expression of sex-determining genes is initiated. Sex determination is a process triggering differentiation of the gonads into the testis or ovary. Sex determination period is followed by sexual differentiation, i.e. appearance of the first testis- and ovary-specific features. In Xenopus laevis W-linked DM-domain gene (DM-W) had been described as a master determinant of the gonadal female sex. However, the data on the expression and function of other genes participating in gonad development in X. laevis, and in anurans, in general, are very limited. We applied microarray technique to analyze the expression pattern of a subset of X. laevis genes previously identified to be involved in gonad development in several vertebrate species. We also analyzed the localization and the expression level of proteins encoded by these genes in developing X. laevis gonads. These analyses pointed to the set of genes differentially expressed in developing testes and ovaries. Gata4, Sox9, Dmrt1, Amh, Fgf9, Ptgds, Pdgf, Fshr, and Cyp17a1 expression was upregulated in developing testes, while DM-W, Fst, Foxl2, and Cyp19a1 were upregulated in developing ovaries. We discuss the possible roles of these genes in development of X. laevis gonads. Copyright © 2018 International Society of Differentiation. Published by Elsevier B.V. All rights reserved.

  1. Zebrafish transgenic constructs label specific neurons in Xenopus laevis spinal cord and identify frog V0v spinal neurons.

    PubMed

    Juárez-Morales, José L; Martinez-De Luna, Reyna I; Zuber, Michael E; Roberts, Alan; Lewis, Katharine E

    2017-09-01

    A correctly functioning spinal cord is crucial for locomotion and communication between body and brain but there are fundamental gaps in our knowledge of how spinal neuronal circuitry is established and functions. To understand the genetic program that regulates specification and functions of this circuitry, we need to connect neuronal molecular phenotypes with physiological analyses. Studies using Xenopus laevis tadpoles have increased our understanding of spinal cord neuronal physiology and function, particularly in locomotor circuitry. However, the X. laevis tetraploid genome and long generation time make it difficult to investigate how neurons are specified. The opacity of X. laevis embryos also makes it hard to connect functional classes of neurons and the genes that they express. We demonstrate here that Tol2 transgenic constructs using zebrafish enhancers that drive expression in specific zebrafish spinal neurons label equivalent neurons in X. laevis and that the incorporation of a Gal4:UAS amplification cassette enables cells to be observed in live X. laevis tadpoles. This technique should enable the molecular phenotypes, morphologies and physiologies of distinct X. laevis spinal neurons to be examined together in vivo. We have used an islet1 enhancer to label Rohon-Beard sensory neurons and evx enhancers to identify V0v neurons, for the first time, in X. laevis spinal cord. Our work demonstrates the homology of spinal cord circuitry in zebrafish and X. laevis, suggesting that future work could combine their relative strengths to elucidate a more complete picture of how vertebrate spinal cord neurons are specified, and function to generate behavior. © 2017 Wiley Periodicals, Inc. Develop Neurobiol 77: 1007-1020, 2017. © 2017 Wiley Periodicals, Inc.

  2. Pattern formation in early embryogenesis of Xenopus laevis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mglinets, V.A.

    1995-07-01

    Establishment of egg polarity, separation of germ layers, and the appearance of animal-vegetal, dorsoventral, and anteroposterior axes in Xenopus laevis embryos are considered. The control of these processes by gene coding for growth factors, protooncogens, and homeobox-containing genes is also been reviewed.

  3. Effects of nutritional factors on the growth and heterotrophic eicosapentaenoic acid production of diatom Nitzschia laevis

    NASA Astrophysics Data System (ADS)

    Cao, Xiaohong; Li, Songyao; Wang, Chunling; Lu, Meifang

    2008-08-01

    The effects of several nutritional factors on the growth and eicosapentaenoic acid (EPA) production of diatom Nitzschia laevis were studied. 4 LDM (quadrupled concentration of the nutrient salt) was the optimal concentration of nutrient salt for the growth and EPA production of N. laevis. The growth of N. laevis was inhibited when the glucose concentration was either lower than 10 gL-1 or higher than 15 gL-1. Both sodium nitrate and urea were good nitrogen sources for the growth and EPA production, while ammonium chloride seriously decreased the dry cell weight (DW) and the EPA content. Silicate seriously influenced the growth of N. laevis. The maximum DW of 2.34 gL-1 was obtained in the presence of 150 mgL-1 Na2SiO3·9H2O. The EPA content remained almost the same when the silicate concentration was lower than 150 mgL-1; however, higher silicate concentrations resulted in a steady decrease of EPA content. Low medium salinity (⩽29) did not seem to influence the DW of N. laevis, and high salinity resulted in a decrease of DW. The highest EPA content (4.08%) and yield (110 mgL-1) were observed at the salinity of 36 and 29, respectively.

  4. Cloning of an origin of DNA replication of Xenopus laevis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Watanabe, S.; Taylor, J.H.

    1980-09-01

    DNA fragments of Xenopus laevis, the African frog, were cloned in the EcoRI site of the Eschrichia coli plasmid pACYC189 and tested for ability to initiate and complete replication of the recombinant plasmid when injected into unfertilized eggs of X. laevis. After measurement of the (/sup 3/H)-thymidine incorporation per egg for a number of recombinant plasmids, pSW14 and pSW9, which respectively contain a small segment (550 base pairs) and several kilobases of frog DNA, were selected for more extensive analysis. In spite of the small size of th segment in pSW14, it incorporates in 2 hr at least 3 timesmore » as much labeled thymidine as either pSW9 or the vector alone. To determine the number of replications of pSW14, a novel method was employed. The results showed that about 50% of the labeled, supercoiled DNA recovered from eggs after 4 hr was sensitive to EcoRI digestion, which indicates that most of the DNA that incorporated (/sup 3/H)thymidine had replicated twice during the 4 hr in the unfertilized eggs of X. laevis. We conclude the pSW14 has a functional origin in the Xenopus DNA segment.« less

  5. Patterns of trunk spine growth in two congeneric species of acanthocephalan: investment in attachment may differ between sexes and species.

    PubMed

    Hernández-Orts, Jesús S; Timi, Juan T; Raga, Juan A; García-Varela, M; Crespo, Enrique A; Aznar, Francisco J

    2012-06-01

    Acanthocephalans have evolved a hooked proboscis and some taxa have trunk spines to attach to their definitive hosts. These structures are generated before being used, thus a key question is how investment in attachment could optimally be allocated through the ontogeny. The number and arrangement of hooks and spines are never modified in the definitive host, but it is unclear whether these structures grow during adult development. A comparison of the size of trunk spines between cystacanths and adults of Corynosoma cetaceum and C. australe indicated that spines grow in both species, but only in females, which also had significantly larger spines than males. This sexual dimorphism did not result from pure allometry because the body of females was smaller, and did not grow more than that of males. However, having a longer lifespan, females would need to withstand the extreme flow conditions prevailing in marine mammals for longer, inducing different investment and development schedules for spines. Patterns of spine growth also differed between species: fore-trunk spines grew in both species, but hind-trunk spines did only in C. cetaceum. In conclusion, investment strategies on attachment may differ, not only between congeneric species of acanthocephalan, but also between sexes of the same species.

  6. Sequencing and analysis of 10967 full-length cDNA clones from Xenopus laevis and Xenopus tropicalis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Morin, R D; Chang, E; Petrescu, A

    2005-10-31

    Sequencing of full-insert clones from full-length cDNA libraries from both Xenopus laevis and Xenopus tropicalis has been ongoing as part of the Xenopus Gene Collection initiative. Here we present an analysis of 10967 clones (8049 from X. laevis and 2918 from X. tropicalis). The clone set contains 2013 orthologs between X. laevis and X. tropicalis as well as 1795 paralog pairs within X. laevis. 1199 are in-paralogs, believed to have resulted from an allotetraploidization event approximately 30 million years ago, and the remaining 546 are likely out-paralogs that have resulted from more ancient gene duplications, prior to the divergence betweenmore » the two species. We do not detect any evidence for positive selection by the Yang and Nielsen maximum likelihood method of approximating d{sub N}/d{sub S}. However, d{sub N}/d{sub S} for X. laevis in-paralogs is elevated relative to X. tropicalis orthologs. This difference is highly significant, and indicates an overall relaxation of selective pressures on duplicated gene pairs. Within both groups of paralogs, we found evidence of subfunctionalization, manifested as differential expression of paralogous genes among tissues, as measured by EST information from public resources. We have observed, as expected, a higher instance of subfunctionalization in out-paralogs relative to in-paralogs.« less

  7. A life cycle database for parasitic acanthocephalans, cestodes, and nematodes

    USGS Publications Warehouse

    Benesh, Daniel P.; Lafferty, Kevin D.; Kuris, Armand

    2017-01-01

    Parasitologists have worked out many complex life cycles over the last ~150 years, yet there have been few efforts to synthesize this information to facilitate comparisons among taxa. Most existing host-parasite databases focus on particular host taxa, do not distinguish final from intermediate hosts, and lack parasite life-history information. We summarized the known life cycles of trophically transmitted parasitic acanthocephalans, cestodes, and nematodes. For 973 parasite species, we gathered information from the literature on the hosts infected at each stage of the parasite life cycle (8510 host-parasite species associations), what parasite stage is in each host, and whether parasites need to infect certain hosts to complete the life cycle. We also collected life-history data for these parasites at each life cycle stage, including 2313 development time measurements and 7660 body size measurements. The result is the most comprehensive data summary available for these parasite taxa. In addition to identifying gaps in our knowledge of parasite life cycles, these data can be used to test hypotheses about life cycle evolution, host specificity, parasite life-history strategies, and the roles of parasites in food webs.

  8. Prolonged in vivo imaging of Xenopus laevis.

    PubMed

    Hamilton, Paul W; Henry, Jonathan J

    2014-08-01

    While live imaging of embryonic development over long periods of time is a well established method for embryos of the frog Xenopus laevis, once development has progressed to the swimming stages, continuous live imaging becomes more challenging because the tadpoles must be immobilized. Current imaging techniques for these advanced stages generally require bringing the tadpoles in and out of anesthesia for short imaging sessions at selected time points, severely limiting the resolution of the data. Here we demonstrate that creating a constant flow of diluted tricaine methanesulfonate (MS-222) over a tadpole greatly improves their survival under anesthesia. Based on this result, we describe a new method for imaging stage 48 to 65 X. laevis, by circulating the anesthetic using a peristaltic pump. This supports the animal during continuous live imaging sessions for at least 48 hr. The addition of a stable optical window allows for high quality imaging through the anesthetic solution. This automated imaging system provides for the first time a method for continuous observations of developmental and regenerative processes in advanced stages of Xenopus over 2 days. Developmental Dynamics 243:1011-1019, 2014. © 2014 Wiley Periodicals, Inc. © 2014 Wiley Periodicals, Inc.

  9. The first record of the slender sunfish Ranzania laevis from the Red Sea.

    PubMed

    Abu El-Regal, M A; El-Moselhy, K

    2013-11-01

    A female specimen of the slender sunfish Ranzania laevis of 600 mm total length was recorded for the first time from the Red Sea after being stranded on a shallow sandy bay at Hurghada beach (27° 06' 16″ N; 33° 50' 01″ E) on 13 May 2012. Ranzania laevis is believed to have migrated from the Indian Ocean as the nearest area where it was found is coastal waters of Oman. © 2013 The Fisheries Society of the British Isles.

  10. Expression and hypophysiotropic actions of corticotropin-releasing factor in Xenopus laevis.

    PubMed

    Boorse, Graham C; Denver, Robert J

    2004-07-01

    Members of the corticotropin-releasing factor (CRF) family of peptides play pivotal roles in the regulation of neuroendocrine, autonomic, and behavioral responses to physical and emotional stress. In amphibian tadpoles, CRF-like peptides stimulate both thyroid and interrenal (adrenal) hormone secretion, and can thereby modulate the rate of metamorphosis. To better understand the regulation of expression and actions of CRF in amphibians we developed a homologous radioimmunoassay (RIA) for Xenopus laevis CRF (xCRF). We validated this RIA and tissue extraction procedure for the measurement of brain CRF content in tadpoles and juveniles. We show that the CRF-binding protein, which is highly expressed in X. laevis brain, is largely removed by acid extraction and does not interfere in the RIA. We analyzed CRF peptide content in five microdissected brain regions in prometamorphic tadpoles and juveniles. CRF was detected throughout the brain, consistent with its role as both a hypophysiotropin and a neurotransmitter/neuromodulator. CRF content was highest in the region of the preoptic area (POa) and increased in all brain regions after metamorphosis. Exposure to 4h of handling/shaking stress resulted in increased CRF peptide content in the POa in juvenile frogs. Injections of xCRF into prometamorphic tadpoles increased whole body corticosterone and thyroxine content, thus supporting findings in other anuran species that this peptide functions as both a corticotropin- and a thyrotropin (TSH)-releasing factor. Furthermore, treatment of cultured tadpole pituitaries with xCRF (100nM for 24h) resulted in increased medium content, but decreased pituitary content of TSHbeta-immunoreactivity. Our results support the view that CRF functions as a stress neuropeptide in X. laevis as in other vertebrates. Furthermore, we provide evidence for a dual hypophysiotropic action of CRF on the thyroid and interrenal axes in X. laevis as has been shown previously in other amphibian species.

  11. Regulation of Xenopus laevis DNA topoisomerase I activity by phosphorylation in vitro

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kaiserman, H.B.; Ingebritsen, T.S.; Benbow, R.M.

    1988-05-03

    DNA topoisomerase I has been purified to electrophoretic homogeneity from ovaries of the frog Xenopus laevis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the most purified fraction revealed a single major band at 110 kDa and less abundant minor bands centered at 62 kDa. Incubation of the most purified fraction with immobilized calf intestinal alkaline phosphatase abolished all DNA topoisomerase enzymatic activity in a time-dependent reaction. Treatment of the dephosphorylated X. laevis DNA topoisomerase I with a X. laevis casein kinase type II activity and ATP restored DNA topoisomerase activity to a level higher than that observed in the most purifiedmore » fraction. In vitro labeling experiments which employed the most purified DNA topoisomerase I fraction, (..gamma..-/sup 32/P)ATP, and the casein kinase type II enzyme showed that both the 110- and 62-kDa bands became phosphorylated in approximately molar proportions. Phosphoamino acid analysis showed that only serine residues became phosphorylated. Phosphorylation was accompanied by an increase in DNA topoisomerase activity in vitro. Dephosphorylation of DNA topoisomerase I appears to block formation of the initial enzyme-substrate complex on the basis of the failure of the dephosphorylated enzyme to nick DNA in the presence of camptothecin. The authors conclude that X. laevis DNA topoisomerase I is partially phosphorylated as isolated and that this phosphorylation is essential for expression of enzymatic activity in vitro. On the basis of the ability of the casein kinase type II activity to reactivate dephosphorylated DNA topoisomerase I, they speculate that this kinase may contribute to the physiological regulation of DNA topoisomerase I activity.« less

  12. Colonisation and extinction in relation to competition and resource partitioning in acanthocephalans of freshwater fishes of the British Isles.

    PubMed

    Lyndon, A R; Kennedy, C R

    2001-01-01

    This paper challenges two paradigms long held in relation to the ecology of parasites in freshwater systems: (1) autogenic species are poorer colonisers than allogenic ones; and (2) parasites with direct life cycles are more successful colonisers than those with complex life cycles. Using new and existing data for Acanthocephala in freshwater fish from the British Isles, it is suggested that all six species present have been able to colonise and persist successfully, in spite of the supposed limitations of their autogenic life-style. It is proposed that these parasites have overcome these limitations by a variety of means, which apply equally to all species considered. Foremost among these is the utilisation of a migratory fish host as either a preferred or a suitable host in their life cycle, allowing colonisation of new areas and rescue effects in established areas, whilst equally important is the use of a common and widespread crustacean as the intermediate host. In addition, all six species appear to exhibit resource partitioning by host at either or both the larval and adult stages, thus reducing the potential for competition and further facilitating colonisation and survival. This hypothesis is supported by data from previous studies both on acanthocephalans from Europe and North America and on other autogenic parasites. It also provides an explanation for the apparently atypical host utilisation patterns of some acanthocephalan species in areas on the edge of their distributions, notably in Ireland.

  13. Metabolism of ribosomal proteins microinjected into the oocytes of Xenopus laevis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Tsurugi, K.; Motizuki, M.; Mitsui, K.

    1988-01-01

    When the total proteins from Xenopus laevis 60 S ribosomal subunits (TP60) were /sup 3/H-labeled in vitro and injected back into X. laevis oocytes, most /sup 3/H-TP60 are integrated into the cytoplasmic 60 S subunits via the nucleus during 16 h of incubation. In the oocytes whose rRNA synthesis is inhibited, /sup 3/H-TP60 are rapidly degraded with a half-life of 2-3 h. This degradation ceased as soon as rRNA synthesis was resumed, suggesting that ribosomal proteins unassociated with nascent rRNA are unstable in the oocytes. The degradation of /sup 3/H-TP60 in the absence of RNA synthesis was inhibited by iodoacetamide,more » a cysteine protease inhibitor, resulting in the accumulation of /sup 3/H-TP60 in the nucleus reaching about a threefold concentration in the cytoplasm. Considering the results with enucleated oocytes, we suggest that the X. laevis nucleus has a limited capacity to accumulate ribosomal proteins in an active manner but that those ribosomal proteins accumulated in excess over rRNA synthesis are degraded by a cysteine protease in the nucleus. By contrast, ribosomal proteins from Escherichia coli only equilibrate between the nucleus and the cytoplasm and are degraded by serine protease(s) in the cytoplasm without being integrated in the form of ribosomes in the nucleus.« less

  14. Phenolic sodium sulphates of Frankenia laevis L.

    PubMed

    Hussein, S A M

    2004-04-01

    Four new phenolic anionic conjugates have been isolated from the whole plant aqueous alcohol extract of Frankenia laevis L. Their structures were established, mainly on the basis of ESI-MS, 1D and 2D NMR spectroscopic evidence, as gallic acid-3-methyl ether-5-sodium sulphate, acetophenone-4-methyl ether-2-sodium sulphate, ellagic acid-3,3'-dimethyl ether-4,4'-di-sodium sulphate and ellagic acid-3-methyl ether-4-sodium sulphate.

  15. Retention of duplicated ITAM-containing transmembrane signaling subunits in the tetraploid amphibian species Xenopus laevis

    PubMed Central

    Guselnikov, S.V.; Grayfer, L.; De Jesús Andino, F.; Rogozin, I.B.; Robert, J.; Taranin, A.V.

    2015-01-01

    The ITAM-bearing transmembrane signaling subunits (TSS) are indispensable components of activating leukocyte receptor complexes. The TSS-encoding genes map to paralogous chromosomal regions, which are thought to arise from ancient genome tetraploidization(s). To assess a possible role of tetraploidization in the TSS evolution, we studied TSS and other functionally linked genes in the amphibian species Xenopus laevis whose genome was duplicated about 40 MYR ago. We found that X. laevis has retained a duplicated set of sixteen TSS genes, all except one being transcribed. Furthermore, duplicated TCRα loci and genes encoding TSS-coupling protein kinases have also been retained. No clear evidence for functional divergence of the TSS paralogs was obtained from gene expression and sequence analyses. We suggest that the main factor of maintenance of duplicated TSS genes in X. laevis was a protein dosage effect and that this effect might have facilitated the TSS set expansion in early vertebrates. PMID:26170006

  16. Effects of tributyltin on metamorphosis and gonadal differentiation of Xenopus laevis at environmentally relevant concentrations.

    PubMed

    Shi, Huahong; Zhu, Pan; Guo, Suzhen

    2014-05-01

    Tributyltin (TBT), a well known endocrine disruptor, has high teratogenicity to embryos of amphibian (Xenopus tropicalis). An amphibian metamorphosis assay (AMA) and a complete AMA (CAMA) were conducted for TBT. In AMA, the body weight, the snout-to-vent length and the hind limb length of X. laevis tadpoles were decreased in tributyltin chloride (TBTCl; 12.5-200 ng/L) treatment groups after 7 days exposure. TBT greatly retarded the development of tadpoles, decreased the number of follicle and induced thyroid follicle cell hyperplasia after 19 days exposure. In CAMA, 10 and 100 ng/L TBTCl led to various malformations of gonad, including intersex, segmental aplasia and multiple ovary cavities of X. laevis following exposure from stages 46 to stage 66. The sex ratio was male-biased in TBT treatment groups. These results suggest that TBT delayed the metamorphosis, inhibited the growth of tadpoles and disrupted the gonadal differentiation of X. laevis at environmentally relevant concentrations.

  17. Spermatozoon structure and motility in the anuran Lepidobatrachus laevis.

    PubMed

    Waggener, W L; Carroll, E J

    1998-02-01

    Synthetic human gonadotropin releasing hormone (GnRH) injections were used for induction of spermatozoon release followed by cloacal lavage or mechanical stimulation of sperm release in Lepidobatrachus laevis. Light microscopic observations of Lepidobatrachus laevis spermatozoa indicated an acrosomal segment with a length of 4.1 microm delineated by an indentation, a nuclear region of 12.6 microm in length and a midpiece of 0.87 microm in length. The tail was 54.9 microm long by 1.35 microm wide with two lateral axial fibers and a central undulating membrane. At the electron microscopic level, the unusual tail had two complete axonemes that emanated from the distal centriole. The tail also contained two axial fibers 77 nm in diameter medial to the axonemes and was connected by an undulating membrane. An unusual accessory cell adherent to the head of the spermatozoon was noted in freshly obtained suspensions of spermatozoa. Spermatozoa with the accessory cell were motile and a subsequent loss of motility was correlated with the shedding of the accessory cell.

  18. CONCENTRATION DEPENDENT ACCUMULATION OF [3H]-DELTAMETHRIN IN XENOPUS LAEVIS OOCYTES.

    EPA Science Inventory

    Pyrethroid insecticides such as deltamethrin have been demonstrated to target and disrupt voltage-sensitive sodium channels (VSSCs). VSSCs were expressed in Xenopus laevis oocytes and used to study the effects of deltamethrin on VSSCs. This study evaluated the amount of deltameth...

  19. Biochemical study of prolactin binding sites in Xenopus laevis brain and choroid plexus

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Muccioli, G.; Guardabassi, A.; Pattono, P.

    1990-03-01

    The occurrence of prolactin binding sites in some brain structures (telencephalon, ventral hypothalamus, myelencephalon, hypophysis, and choroid plexus) from Xenopus laevis (anuran amphibian) was studied by the in vitro biochemical technique. The higher binding values were obtained at the level of the choroid plexus and above all of the hypothalamus. On the bases of hormonal specificity and high affinity, these binding sites are very similar to those of prolactin receptors of classical target tissues as well as of those described by us in other structures from Xenopus. To our knowledge, the present results provide the first demonstration of the occurrencemore » of prolactin specific binding sites in Xenopus laevis choroid plexus cells.« less

  20. Swimming kinematics and respiratory behaviour of Xenopus laevis larvae raised in altered gravity.

    PubMed

    Fejtek, M; Souza, K; Neff, A; Wassersug, R

    1998-06-01

    We examined the respiratory behaviours and swimming kinematics of Xenopus laevis tadpoles hatched in microgravity (Space Shuttle), simulated microgravity (clinostat) and hypergravity (3 g centrifuge). All observations were made in the normal 1 g environment. Previous research has shown that X. laevis raised in microgravity exhibit abnormalities in their lungs and vestibular system upon return to 1 g. The tadpoles raised in true microgravity exhibited a significantly lower tailbeat frequency than onboard 1 g centrifuge controls on the day of landing (day0), but this behaviour normalized within 9 days. The two groups did not differ significantly in buccal pumping rates. Altered buoyancy in the space-flight microgravity tadpoles was indicated by an increased swimming angle on the day after landing (day1). Tadpoles raised in simulated microgravity differed to a greater extent in swimming behaviours from their 1 g controls. The tadpoles raised in hypergravity showed no substantive effects on the development of swimming or respiratory behaviours, except swimming angle. Together, these results show that microgravity has a transient effect on the development of locomotion in X. laevis tadpoles, most notably on swimming angle, indicative of stunted lung development. On the basis of the behaviours we studied, there is no indication of neuromuscular retardation in amphibians associated with embryogenesis in microgravity.

  1. Swimming kinematics and respiratory behaviour of Xenopus laevis larvae raised in altered gravity

    NASA Technical Reports Server (NTRS)

    Fejtek, M.; Souza, K.; Neff, A.; Wassersug, R.

    1998-01-01

    We examined the respiratory behaviours and swimming kinematics of Xenopus laevis tadpoles hatched in microgravity (Space Shuttle), simulated microgravity (clinostat) and hypergravity (3 g centrifuge). All observations were made in the normal 1 g environment. Previous research has shown that X. laevis raised in microgravity exhibit abnormalities in their lungs and vestibular system upon return to 1 g. The tadpoles raised in true microgravity exhibited a significantly lower tailbeat frequency than onboard 1 g centrifuge controls on the day of landing (day0), but this behaviour normalized within 9 days. The two groups did not differ significantly in buccal pumping rates. Altered buoyancy in the space-flight microgravity tadpoles was indicated by an increased swimming angle on the day after landing (day1). Tadpoles raised in simulated microgravity differed to a greater extent in swimming behaviours from their 1 g controls. The tadpoles raised in hypergravity showed no substantive effects on the development of swimming or respiratory behaviours, except swimming angle. Together, these results show that microgravity has a transient effect on the development of locomotion in X. laevis tadpoles, most notably on swimming angle, indicative of stunted lung development. On the basis of the behaviours we studied, there is no indication of neuromuscular retardation in amphibians associated with embryogenesis in microgravity.

  2. An environmentally relevant endocrine-disrupting antiandrogen, vinclozolin, affects calling behavior of male Xenopus laevis.

    PubMed

    Hoffmann, Frauke; Kloas, Werner

    2010-09-01

    Vinclozolin (VIN) is an antiandrogenic model substance as well as a common fungicide that can affect the endocrine system of vertebrates. The objective of this study was to investigate how VIN affects mate calling behavior of South African clawed frogs (Xenopus laevis) and whether it is effective at environmentally relevant concentrations. Male X. laevis were injected with human chorionic gonadotropin (hCG) to stimulate their androgen-controlled mate calling behavior and were treated with VIN at concentrations of 10(-6), 10(-8) and 10(-10)M. VIN at 10(-6)M reduced calling activity. Furthermore, the vocalization composition of VIN-treated X. laevis was altered. The call types advertisement calls and chirping are uttered by reproductively active males, whereas the call types growling, ticking, and rasping indicate a sexually unaroused state of a male. VIN at any of the tested concentrations led to a decrease in utterance of calls, which indicate a sexually aroused state of the males, and an increase in relative proportions of calls, indicating a sexually unaroused state of the males. Additionally, the mean duration of clicks and the number of accentuated clicks during the advertisement calls decreased at all concentrations of VIN. No significant differences were observed in any other temporal or spectral calling parameters between the treatments. This study illustrates that exposure to the antiandrogen VIN might result in a reduced reproductive success by altering mate calling behavior of X. laevis. Moreover, it suggests that the behavioral parameters examined in this study can be used as sensitive biomarkers for detecting antiandrogenic endocrine disrupting compounds in amphibians. Copyright (c) 2010 Elsevier Inc. All rights reserved.

  3. Cortical Isolation from Xenopus laevis Oocytes and Eggs.

    PubMed

    Sive, Hazel L; Grainger, Robert M; Harland, Richard M

    2007-06-01

    INTRODUCTIONIn Xenopus laevis, the cortex is the layer of gelatinous cytoplasm that lies just below the plasma membrane of the egg. Rotation of the cortex relative to the deeper cytoplasm soon after fertilization is intimately linked to normal dorsal axis specification. The cortex can be dissected from the egg to analyze its composition and activity or to clone associated RNAs. This protocol describes a procedure for isolating the vegetal cortex of the fertilized egg.

  4. Parasites of the African clawed frog, Xenopus laevis, in southern California, U.S.A

    USGS Publications Warehouse

    Kuperman, Boris I.; Matey, Victoria E.; Fisher, Richard N.; Ervin, Edward L.; Warburton, Manna L.; Bakhireva, Ludmila; Lehman, Cynthia A.

    2004-01-01

    A total of 230 feral African clawed frogs, Xenopus laevis, from 3 localities in southern California were examined for parasites. The following species were found: 3 species of Protozoa, Nyctotherussp., Balantidium xenopodis, Protoopalina xenopodus; 2 species of Monogenea, Protopolystoma xenopodis, Gyrdicotylus gallieni; 1 species of Digenea, Clinostomum sp. (as metacercariae); 1 species of Cestoda, Cephalochlamys namaquensis; 2 species of Nematoda, Contracaecum sp. (as larvae), Eustrongylides sp. (as larvae); and 1 species of Acanthocephala, Acanthocephalus sp. (as cystacanth). Of these, the protozoans P. xenopodus and B. xenopodis, both monogeneans, and the cestode have an African origin. Contracaecum sp., Eustrongylides sp., and Acanthocephalus sp. have not been previously reported from X. laevis.

  5. Vocal communication between male Xenopus laevis.

    PubMed

    Tobias, Martha L; Barnard, Candace; O'Hagan, Robert; Horng, Sam H; Rand, Masha; Kelley, Darcy B

    2004-02-01

    This study focuses on the role of male-male vocal communication in the reproductive repertoire of the South African clawed frog, Xenopus laevis . Six male and two female call types were recorded from native ponds in the environs of Cape Town, South Africa. These include all call types previously recorded in the laboratory as well as one previously unidentified male call: chirping. The amount of calling and the number of call types increased as the breeding season progressed. Laboratory recordings indicated that all six male call types were directed to males; three of these were directed to both sexes and three were directed exclusively to males. Both female call types were directed exclusively to males. The predominant call type, in both field and laboratory recordings, was the male advertisement call. Sexual state affected male vocal behaviour. Male pairs in which at least one male was sexually active (gonadotropin injected) produced all call types, whereas pairs of uninjected males rarely called. Some call types were strongly associated with a specific behaviour and others were not. Clasped males always growled and clasping males typically produced amplectant calls or chirps; males not engaged in clasping most frequently advertised. The amount of advertising produced by one male was profoundly affected by the presence of another male. Pairing two sexually active males resulted in suppression of advertisement calling in one; suppression was released when males were isolated after pairing. Vocal dominance was achieved even in the absence of physical contact (clasping). We suggest that X. laevis males gain a reproductive advantage by competing for advertisement privileges and by vocally suppressing neighbouring males.

  6. Dehydration triggers differential microRNA expression in Xenopus laevis brain.

    PubMed

    Luu, Bryan E; Storey, Kenneth B

    2015-11-15

    African clawed frogs, Xenopus laevis, although primarily aquatic, have a high tolerance for dehydration, being capable of withstanding the loss of up to 32-35% of total water body water. Recent studies have shown that microRNAs play a role in the response to dehydration by the liver, kidney and ventral skin of X. laevis. MicroRNAs act by modulating the expression of mRNA transcripts, thereby affecting diverse biochemical pathways. In this study, 43 microRNAs were assessed in frog brains comparing control and dehydrated (31.2±0.83% of total body water lost) conditions. MicroRNAs of interest were measured using a modified protocol which employs polyadenylation of microRNAs prior to reverse transcription and qPCR. Twelve microRNAs that showed a significant decrease in expression (to 41-77% of control levels) in brains from dehydrated frogs (xla-miR-15a, -150, -181a, -191, -211, -218, -219b, -30c, -30e, -31, -34a, and -34b) were identified. Genomic analysis showed that the sequences of these dehydration-responsive microRNAs were highly conserved as compared with the comparable microRNAs of mice (91-100%). Suppression of these microRNAs implies that translation of the mRNA transcripts under their control could be enhanced in response to dehydration. Bioinformatic analysis using the DIANA miRPath program (v.2.0) predicted the top two KEGG pathways that these microRNAs collectively regulate: 1. Axon guidance, and 2. Long-term potentiation. Previous studies indicated that suppression of these microRNAs promotes neuroprotective pathways by increasing the expression of brain-derived neurotrophic factor and activating anti-apoptotic pathways. This suggests that similar actions may be triggered in X. laevis brains as a protective response to dehydration. Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.

  7. Susceptibility of early life stages of Xenopus laevis to cadmium

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Herkovits, J.; Perez-Coll, C.S.; Cardellini, P.

    1997-02-01

    The susceptibility of Xenopus laevis to cadmium during different stages of development was evaluated by exposing embryos to cadmium concentrations ranging from 0.1 to 10 mg Cd{sup 2+}/L for 24, 48, and 72 h and assessing lethality and malformations. Susceptibility increased from the two blastomeres stage (stage 2) to stage 40, in which the 24-h LC100 was 1.13 mg Cd{sup 2+}/L, and resistance increased from this stage onward. Malformations occurred at all developmental stages evaluated, the most common being reduced size, incurvated axis, underdeveloped or abnormally developed fin, microcephaly, and microphtalmy. Scanning electron microscopy revealed changes in the ectodermal surfacemore » ranging from slightly vaulted cells to a severe reduction in the number of ciliated cells as the concentration of cadmium increased. The intraspecific variation evaluated in embryos (from four sets of parents) at seven developmental stages, expressed as the coefficient of variation of the LC100, ranged from 10 to 112% and reflects the capacity of Xenopus laevis to adapt to changing environmental conditions at different embryonic stages.« less

  8. How does the Xenopus laevis embryonic cell cycle avoid spatial chaos?

    PubMed Central

    Gelens, Lendert; Huang, Kerwyn Casey; Ferrell, James E.

    2015-01-01

    Summary Theoretical studies have shown that a deterministic biochemical oscillator can become chaotic when operating over a sufficiently large volume, and have suggested that the Xenopus laevis cell cycle oscillator operates close to such a chaotic regime. To experimentally test this hypothesis, we decreased the speed of the post-fertilization calcium wave, which had been predicted to generate chaos. However, cell divisions were found to develop normally and eggs developed into normal tadpoles. Motivated by these experiments, we carried out modeling studies to understand the prerequisites for the predicted spatial chaos. We showed that this type of spatial chaos requires oscillatory reaction dynamics with short pulse duration, and postulated that the mitotic exit in Xenopus laevis is likely slow enough to avoid chaos. In systems with shorter pulses, chaos may be an important hazard, as in cardiac arrhythmias, or a useful feature, as in the pigmentation of certain mollusk shells. PMID:26212326

  9. Description of a New Quadrigyrid Acanthocephalan from Kashmir, with Notes on Metal Analysis and Histopathology, and a Key to Species of the Subgenus Acanthosentis from the Indian Subcontinent.

    PubMed

    Amin, Omar M; Heckmann, Richard A; Zargar, Ummer Rashid

    2017-10-01

    Acanthogyrus (Acanthosentis) kashmirensis n. sp. is described from recently collected acanthocephalan specimens in the Jhelum River in northern Kashmir that are conspecific with Neoechinorhynchus kashmirensis Fotedar and Dhar, 1977 originally described in a Ph.D. thesis in 1972 from 4 species of cyprinid fishes: Tor tor Hamilton, Bangana diplostoma (Heckel) (syn. Labeo diplostoma Heckel), Labeo rohita Hamilton, and Ptychobarbus sp. Steindachner. The poor unpublished diagnosis was followed by 1 uninformative abstract in a scientific meeting in 1977. The acanthocephalan was later designated as invalid because of the lack of a formal published description and absence of information on deposited type or voucher specimens. Recent collections of specimens of the same species were made from 2 other cyprinid species of cyprinid fishes, Schizothorax plagiostomus Heckel and Schizothorax labiatus (McClelland) from the Sandran River, a tributary of the Jhelum River, in southern Kashmir. It is now possible to provide a full description of these specimens and reassign them in the subgenus Acanthosentis Verma and Datta, 1929 based on the finding of circles of vestigial spines at the anterior end of the trunk of male and female specimens. These vestigial spines are barely visible and easy to miss with optical microscopy. The new species is also characterized by having (1) a para-receptacle structure in males and females, (2) unique double Saefftigen's pouches, (3) large round single-nucleated cells in the proboscis, and (4) the lemnisci being either equal or distinctly unequal with no intermediate states. A key to the species of Acanthosentis of the Indian subcontinent is provided. Histopathological sections show extensive damage to the host intestine with subsequent blood loss, cell necrosis, and attempted encapsulation. Results of the energy dispersive X-ray analysis (EDAX) study show hollow hooks high in sulfur but with limited calcium ions. Hooks of most acanthocephalans

  10. Sequencing and analysis of 10,967 full-length cDNA clones from Xenopus laevis and Xenopus tropicalis reveals post-tetraploidization transcriptome remodeling

    PubMed Central

    Morin, Ryan D.; Chang, Elbert; Petrescu, Anca; Liao, Nancy; Griffith, Malachi; Kirkpatrick, Robert; Butterfield, Yaron S.; Young, Alice C.; Stott, Jeffrey; Barber, Sarah; Babakaiff, Ryan; Dickson, Mark C.; Matsuo, Corey; Wong, David; Yang, George S.; Smailus, Duane E.; Wetherby, Keith D.; Kwong, Peggy N.; Grimwood, Jane; Brinkley, Charles P.; Brown-John, Mabel; Reddix-Dugue, Natalie D.; Mayo, Michael; Schmutz, Jeremy; Beland, Jaclyn; Park, Morgan; Gibson, Susan; Olson, Teika; Bouffard, Gerard G.; Tsai, Miranda; Featherstone, Ruth; Chand, Steve; Siddiqui, Asim S.; Jang, Wonhee; Lee, Ed; Klein, Steven L.; Blakesley, Robert W.; Zeeberg, Barry R.; Narasimhan, Sudarshan; Weinstein, John N.; Pennacchio, Christa Prange; Myers, Richard M.; Green, Eric D.; Wagner, Lukas; Gerhard, Daniela S.; Marra, Marco A.; Jones, Steven J.M.; Holt, Robert A.

    2006-01-01

    Sequencing of full-insert clones from full-length cDNA libraries from both Xenopus laevis and Xenopus tropicalis has been ongoing as part of the Xenopus Gene Collection Initiative. Here we present 10,967 full ORF verified cDNA clones (8049 from X. laevis and 2918 from X. tropicalis) as a community resource. Because the genome of X. laevis, but not X. tropicalis, has undergone allotetraploidization, comparison of coding sequences from these two clawed (pipid) frogs provides a unique angle for exploring the molecular evolution of duplicate genes. Within our clone set, we have identified 445 gene trios, each comprised of an allotetraploidization-derived X. laevis gene pair and their shared X. tropicalis ortholog. Pairwise dN/dS, comparisons within trios show strong evidence for purifying selection acting on all three members. However, dN/dS ratios between X. laevis gene pairs are elevated relative to their X. tropicalis ortholog. This difference is highly significant and indicates an overall relaxation of selective pressures on duplicated gene pairs. We have found that the paralogs that have been lost since the tetraploidization event are enriched for several molecular functions, but have found no such enrichment in the extant paralogs. Approximately 14% of the paralogous pairs analyzed here also show differential expression indicative of subfunctionalization. PMID:16672307

  11. Effects of 17α-trenbolone and melengestrol acetate on Xenopus laevis growth, development, and survival.

    PubMed

    Finch, Bryson E; Blackwell, Brett R; Faust, Derek R; Wooten, Kimberly J; Maul, Jonathan D; Cox, Stephen B; Smith, Philip N

    2013-02-01

    The synthetic growth-promoting hormones trenbolone and melengestrol acetate have been detected in the environment near beef cattle feedlots and are reportedly transported via wind-borne particulate matter. Therefore, movement of synthetic hormones from beef cattle feedlots to water bodies via particulate matter is possible. Our objective was to evaluate potential effects of 17α-trenbolone (17α-TB), melengestrol acetate (MGA), and combinations of both on growth, development, and survival of Xenopus laevis larvae. On post-hatch day 2 (stage 33/34), X. laevis larvae were exposed to three nominal concentrations of 17α-TB (10, 100, and 500 ng/L), MGA (1, 10, and 100 ng/L), a combination of both (1/10, 10/100, and 100/500 ng/L MGA/17α-TB), frog embryo teratogenesis assay-Xenopus medium, or a solvent control. Significant increases in all X. laevis growth metrics were observed among larvae in the 1 ng/L MGA + 10 ng/L 17α-TB and 10 ng/L MGA + 100 ng/L 17α-TB treatments. Stage of development was increased among larvae in the 1 ng/L MGA + 10 ng/L 17α-TB treatment group and significantly decreased among those in the 500 ng/L 17α-TB treatment. Total body mass and snout-vent length of X. laevis larvae were significantly reduced in the 100 ng/L MGA and 100 ng/L MGA + 500 ng/L 17α-TB treatment groups. Larvae exposed to 500 ng/L 17α-TB had decreased total body mass, snout-vent length, and total length. In general, growth measurements decreased with increasing concentration of MGA, 17α-TB, or a combination of both. Survival among all treatments was not significantly different from controls. Amphibians exposed to MGA and 17α-TB in the environment may experience alterations in growth and development.

  12. Probing the Xenopus laevis inner ear transcriptome for biological function

    PubMed Central

    2012-01-01

    Background The senses of hearing and balance depend upon mechanoreception, a process that originates in the inner ear and shares features across species. Amphibians have been widely used for physiological studies of mechanotransduction by sensory hair cells. In contrast, much less is known of the genetic basis of auditory and vestibular function in this class of animals. Among amphibians, the genus Xenopus is a well-characterized genetic and developmental model that offers unique opportunities for inner ear research because of the amphibian capacity for tissue and organ regeneration. For these reasons, we implemented a functional genomics approach as a means to undertake a large-scale analysis of the Xenopus laevis inner ear transcriptome through microarray analysis. Results Microarray analysis uncovered genes within the X. laevis inner ear transcriptome associated with inner ear function and impairment in other organisms, thereby supporting the inclusion of Xenopus in cross-species genetic studies of the inner ear. The use of gene categories (inner ear tissue; deafness; ion channels; ion transporters; transcription factors) facilitated the assignment of functional significance to probe set identifiers. We enhanced the biological relevance of our microarray data by using a variety of curation approaches to increase the annotation of the Affymetrix GeneChip® Xenopus laevis Genome array. In addition, annotation analysis revealed the prevalence of inner ear transcripts represented by probe set identifiers that lack functional characterization. Conclusions We identified an abundance of targets for genetic analysis of auditory and vestibular function. The orthologues to human genes with known inner ear function and the highly expressed transcripts that lack annotation are particularly interesting candidates for future analyses. We used informatics approaches to impart biologically relevant information to the Xenopus inner ear transcriptome, thereby addressing the

  13. From mammals back to birds: Host-switch of the acanthocephalan Corynosoma australe from pinnipeds to the Magellanic penguin Spheniscus magellanicus

    PubMed Central

    Brandão, Martha; Georgieva, Simona; Raga, Juan Antonio; Crespo, Enrique Alberto; Luque, José Luis

    2017-01-01

    Trophically-transmitted parasites are regularly exposed to potential new hosts through food web interactions. Successful colonization, or switching, to novel hosts, occur readily when ‘donor’ and ‘target’ hosts are phylogenetically related, whereas switching between distantly related hosts is rare and may result from stochastic factors (i.e. rare favourable mutations). This study investigates a host-switching event between a marine acanthocephalan specific to pinnipeds that is apparently able to reproduce in Magellanic penguins Spheniscus magellanicus from Brazil. Detailed analysis of morphological and morphometrical data from acanthocephalans from penguins indicates that they belong to Corynosoma australe Johnston, 1937. Partial fragments of the 28S rRNA and mitochondrial cox1 genes were amplified from isolates from penguins and two pinniped species (i.e. South American sea lion Otaria flavescens and South American fur seal Arctocephalus australis) to confirm this identification. Infection parameters clearly differ between penguins and the two pinniped species, which were significantly lower in S. magellanicus. The sex ratio of C. australe also differed between penguins and pinnipeds; in S. magellanicus was strongly biased against males, while in pinnipeds it was close to 1:1. Females of C. australe from O. flavescens were smaller than those from S. magellanicus and A. australis. However, fecundity (i.e. the proportion of fully developed eggs) was lower and more variable in females collected from S. magellanicus. At first glance, the occurrence of reproductive individuals of C. australe in Magellanic penguins could be interpreted as an adaptive colonization of a novel avian host through favourable mutations. However, it could also be considered, perhaps more likely, as an example of ecological fitting through the use of a plesimorphic (host) resource, since the ancestors of Corynosoma infected aquatic birds. PMID:28981550

  14. From mammals back to birds: Host-switch of the acanthocephalan Corynosoma australe from pinnipeds to the Magellanic penguin Spheniscus magellanicus.

    PubMed

    Hernández-Orts, Jesús Servando; Brandão, Martha; Georgieva, Simona; Raga, Juan Antonio; Crespo, Enrique Alberto; Luque, José Luis; Aznar, Francisco Javier

    2017-01-01

    Trophically-transmitted parasites are regularly exposed to potential new hosts through food web interactions. Successful colonization, or switching, to novel hosts, occur readily when 'donor' and 'target' hosts are phylogenetically related, whereas switching between distantly related hosts is rare and may result from stochastic factors (i.e. rare favourable mutations). This study investigates a host-switching event between a marine acanthocephalan specific to pinnipeds that is apparently able to reproduce in Magellanic penguins Spheniscus magellanicus from Brazil. Detailed analysis of morphological and morphometrical data from acanthocephalans from penguins indicates that they belong to Corynosoma australe Johnston, 1937. Partial fragments of the 28S rRNA and mitochondrial cox1 genes were amplified from isolates from penguins and two pinniped species (i.e. South American sea lion Otaria flavescens and South American fur seal Arctocephalus australis) to confirm this identification. Infection parameters clearly differ between penguins and the two pinniped species, which were significantly lower in S. magellanicus. The sex ratio of C. australe also differed between penguins and pinnipeds; in S. magellanicus was strongly biased against males, while in pinnipeds it was close to 1:1. Females of C. australe from O. flavescens were smaller than those from S. magellanicus and A. australis. However, fecundity (i.e. the proportion of fully developed eggs) was lower and more variable in females collected from S. magellanicus. At first glance, the occurrence of reproductive individuals of C. australe in Magellanic penguins could be interpreted as an adaptive colonization of a novel avian host through favourable mutations. However, it could also be considered, perhaps more likely, as an example of ecological fitting through the use of a plesimorphic (host) resource, since the ancestors of Corynosoma infected aquatic birds.

  15. THYROID AXIS INHIBITION IN XENOPUS LAEVIS: DEVELOPMENT OF AN AMPHIBIAN-BASED SCREENING ASSAY

    EPA Science Inventory

    In response to the initial EDSTAC recommendations, research was conducted on the development of a Xenopus laevis based tail resorption assay for evaluating thyroid axis disruption. These experiments highlighted key limitations associated with relying on tail resorption as a measu...

  16. Localization, structure and polymorphism of two paralogous Xenopus laevis mitochondrial malate dehydrogenase genes.

    PubMed

    Tlapakova, Tereza; Krylov, Vladimir; Macha, Jaroslav

    2005-01-01

    Two paralogous mitochondrial malate dehydrogenase 2 (Mdh2) genes of Xenopus laevis have been cloned and sequenced, revealing 95% identity. Fluorescence in-situ hybridization (FISH) combined with tyramide amplification discriminates both genes; Mdh2a was localized into chromosome q3 and Mdh2b into chromosome q8. One kb cDNA probes detect both genes with 85% accuracy. The remaining signals were on the paralogous counterpart. Introns interrupt coding sequences at the same nucleotide as defined for mouse. Restriction polymorphism has been detected in the first intron of Mdh2a, while the individual variability in intron 6 of Mdh2b gene is represented by an insertion of incomplete retrotransposon L1Xl. Rates of nucleotide substitutions indicate that both genes are under similar evolutionary constraints. X. laevis Mdh2 genes can be used as markers for physical mapping and linkage analysis.

  17. Morphological and molecular evidence on the existence of a single estuarine and rocky intertidal acanthocephalan species of Profilicollis Meyer, 1931 (Acanthocephala: Polymorphidae) along the Atlantic and Pacific coasts of southern South America.

    PubMed

    Rodríguez, Sara M; Diaz, Julia I; D'Elía, Guillermo

    2017-05-01

    Profilicollis chasmagnathi Holcman-Spector, Mañé-Garzón & Dei-Cas, 1977 (Acanthocephala: Polymorphidae) has been reported to parasitise different grapsid species as intermediate hosts along the South Atlantic shores, i.e. Cyrtograpsus angulatus (Dana) and Neohelice granulata (Dana) in Uruguay and Cyrtograpsus altimanus (Rathbun) in Argentina. Larvae of a similar acanthocephalan described as Profilicollis antarcticus Zdzitowiecki, 1985 were recorded in the crab Hemigrapsus crenulatus (Milne-Edwards) from an estuarine habitat on the Southeast Pacific shore in Chile. Earlier studies have questioned the specific assignation of the Chilean estuarine populations of Profilicollis Meyer, 1931. The aim of this study was to re-examine the identification of these acanthocephalans by means of morphological and molecular analyses of cystacanths of Profilicollis spp. gathered from C. angulatus, N. granulata, C. altimanus and H. crenulatus. Our analyses showed that a single species of Profilicollis, P. chasmagnathi, parasitises these four crab species. The assessment of specimens from the South Shetlands Islands, the type-locality of P. antarcticus, is needed before formally proposing that P. antarcticus is a junior subjective synonym of P. chasmagnathi.

  18. Population structure of the African Clawed Frog (Xenopus laevis) in maize-growing areas with atrazine application versus non-maize-growing areas in South Africa

    USGS Publications Warehouse

    Du Preez, L.H.; Solomon, K.R.; Carr, J.A.; Giesy, J.P.; Gross, T.S.; Kendall, R.J.; Smith, E.E.; Van Der Kraak, G. L.; Weldon, C.

    2005-01-01

    The herbicide atrazine has been suggested to cause gonadal deformities in frogs and could possibly impact on reproduction. Since the early 1960s, atrazine has been used in large amounts in maize production areas of South Africa. These areas overlap with populations of the African Clawed Frog (Xenopus laevis) that has a wide distribution in southern Africa and is found in most water-bodies including those where atrazine residues are detected. The aim of this study was to compare various attributes of individual- and population-level responses of X. laevis from maize-growing and non-maize-growing areas. Xenopus laevis were studied in three reference and five maize-growing sites. Sex ratio, snout-vent length, body-mass and age profiles were found to be similar for populations in maize-growing and non-maize-growing areas. Our mark-recapture data indicated that all sites had robust populations. There were no significant relationships between exposure to atrazine and any of the parameters investigated in populations of X. laevis.

  19. Impacts of Climate Change on the Global Invasion Potential of the African Clawed Frog Xenopus laevis

    PubMed Central

    Ihlow, Flora; Courant, Julien; Secondi, Jean; Herrel, Anthony; Rebelo, Rui; Measey, G. John; Lillo, Francesco; De Villiers, F. André; Vogt, Solveig; De Busschere, Charlotte; Backeljau, Thierry; Rödder, Dennis

    2016-01-01

    By altering or eliminating delicate ecological relationships, non-indigenous species are considered a major threat to biodiversity, as well as a driver of environmental change. Global climate change affects ecosystems and ecological communities, leading to changes in the phenology, geographic ranges, or population abundance of several species. Thus, predicting the impacts of global climate change on the current and future distribution of invasive species is an important subject in macroecological studies. The African clawed frog (Xenopus laevis), native to South Africa, possesses a strong invasion potential and populations have become established in numerous countries across four continents. The global invasion potential of X. laevis was assessed using correlative species distribution models (SDMs). SDMs were computed based on a comprehensive set of occurrence records covering South Africa, North America, South America and Europe and a set of nine environmental predictors. Models were built using both a maximum entropy model and an ensemble approach integrating eight algorithms. The future occurrence probabilities for X. laevis were subsequently computed using bioclimatic variables for 2070 following four different IPCC scenarios. Despite minor differences between the statistical approaches, both SDMs predict the future potential distribution of X. laevis, on a global scale, to decrease across all climate change scenarios. On a continental scale, both SDMs predict decreasing potential distributions in the species’ native range in South Africa, as well as in the invaded areas in North and South America, and in Australia where the species has not been introduced. In contrast, both SDMs predict the potential range size to expand in Europe. Our results suggest that all probability classes will be equally affected by climate change. New regional conditions may promote new invasions or the spread of established invasive populations, especially in France and Great

  20. Impacts of Climate Change on the Global Invasion Potential of the African Clawed Frog Xenopus laevis.

    PubMed

    Ihlow, Flora; Courant, Julien; Secondi, Jean; Herrel, Anthony; Rebelo, Rui; Measey, G John; Lillo, Francesco; De Villiers, F André; Vogt, Solveig; De Busschere, Charlotte; Backeljau, Thierry; Rödder, Dennis

    2016-01-01

    By altering or eliminating delicate ecological relationships, non-indigenous species are considered a major threat to biodiversity, as well as a driver of environmental change. Global climate change affects ecosystems and ecological communities, leading to changes in the phenology, geographic ranges, or population abundance of several species. Thus, predicting the impacts of global climate change on the current and future distribution of invasive species is an important subject in macroecological studies. The African clawed frog (Xenopus laevis), native to South Africa, possesses a strong invasion potential and populations have become established in numerous countries across four continents. The global invasion potential of X. laevis was assessed using correlative species distribution models (SDMs). SDMs were computed based on a comprehensive set of occurrence records covering South Africa, North America, South America and Europe and a set of nine environmental predictors. Models were built using both a maximum entropy model and an ensemble approach integrating eight algorithms. The future occurrence probabilities for X. laevis were subsequently computed using bioclimatic variables for 2070 following four different IPCC scenarios. Despite minor differences between the statistical approaches, both SDMs predict the future potential distribution of X. laevis, on a global scale, to decrease across all climate change scenarios. On a continental scale, both SDMs predict decreasing potential distributions in the species' native range in South Africa, as well as in the invaded areas in North and South America, and in Australia where the species has not been introduced. In contrast, both SDMs predict the potential range size to expand in Europe. Our results suggest that all probability classes will be equally affected by climate change. New regional conditions may promote new invasions or the spread of established invasive populations, especially in France and Great Britain.

  1. PHENOBARBITAL AFFECTS THYROID HISTOLOGY AND LARVAL DEVELOPMENT IN THE AFRICAN CLAWED FROG XENOPUS LAEVIS

    EPA Science Inventory

    The abstract highlights our recent study to explore endocrine disrupting effects of phenobarbital in the African clawed frog, Xenopus laevis. In mammals, this chemical is known to induce the biotransforming enzyme UDP-glucuronosyltransferase (UDPGT) resulting in increased thyroid...

  2. Gene expression analysis of the ovary of hybrid females of Xenopus laevis and X. muelleri

    PubMed Central

    2008-01-01

    Background Interspecific hybrids of frogs of the genus Xenopus result in sterile hybrid males and fertile hybrid females. Previous work has demonstrated a dramatic asymmetrical pattern of misexpression in hybrid males compared to the two parental species with relatively few genes misexpressed in comparisons of hybrids and the maternal species (X. laevis) and dramatically more genes misexpressed in hybrids compared to the paternal species (X. muelleri). In this work, we examine the gene expression pattern in hybrid females of X. laevis × X. muelleri to determine if this asymmetrical pattern of expression also occurs in hybrid females. Results We find a similar pattern of asymmetry in expression compared to males in that there were more genes differentially expressed between hybrids and X. muelleri compared to hybrids and X. laevis. We also found a dramatic increase in the number of misexpressed genes with hybrid females having about 20 times more genes misexpressed in ovaries compared to testes of hybrid males and therefore the match between phenotype and expression pattern is not supported. Conclusion We discuss these intriguing findings in the context of reproductive isolation and suggest that divergence in female expression may be involved in sterility of hybrid males due to the inherent sensitivity of spermatogenesis as defined by the faster male evolution hypothesis for Haldane's rule. PMID:18331635

  3. Glider and Vision: two new families of miniature inverted-repeat transposable elements in Xenopus laevis genome.

    PubMed

    Lepetit, D; Pasquet, S; Olive, M; Thézé, N; Thiébaud, P

    2000-01-01

    We have characterised from Xenopus laevis two new short interspersed repetitive elements, we have named Glider and Vision, that belong to the family of miniature inverted-repeat transposable elements (MITEs). Glider was first characterised in an intronic region of the alpha-tropomyosin (alpha-TM) gene and database search has revealed the presence of this element in 10 other Xenopus laevis genes. Glider elements are about 150 bp long and for some of them, their terminal inverted repeats are flanked by potential target-site duplications. Evidence for the mobility of Glider element has been provided by the presence/absence of one element at corresponding location in duplicated alpha-TM genes. Vision element has been identified in the promoter region of the cyclin dependant kinase 2 gene (cdk2) where it is boxed in a Glider element. Vision is 284bp long and is framed by 14-bp terminal inverted repeats that are flanked by 7-bp direct repeats. We have estimated that there are about 20,000 and 300 copies of Glider and Vision respectively scattered throughout the Xenopus laevis genome. Every MITEs elements but two described in our study are found either in 5' or in 3' regulatory regions of genes suggesting a potential role in gene regulation.

  4. Isolation and Expression Profile of the Ca2+-Activated Chloride Channel-like Membrane Protein 6 Gene in Xenopus laevis

    PubMed Central

    Lee, Ra Mi; Ryu, Rae Hyung; Jeong, Seong Won; Oh, Soo Jin; Huang, Hue; Han, Jin Soo; Lee, Chi Ho; Lee, C. Justin; Jan, Lily Yeh

    2011-01-01

    To clone the first anion channel from Xenopus laevis (X. laevis), we isolated a calcium-activated chloride channel (CLCA)-like membrane protein 6 gene (CMP6) in X. laevis. As a first step in gene isolation, an expressed sequence tags database was screened to find the partial cDNA fragment. A putative partial cDNA sequence was obtained by comparison with rat CLCAs identified in our laboratory. First stranded cDNA was synthesized by reverse transcription polymerase-chain reaction (RT-PCR) using a specific primer designed for the target cDNA. Repeating the 5' and 3' rapid amplification of cDNA ends, full-length cDNA was constructed from the cDNA pool. The full-length CMP6 cDNA completed via 5'- and 3'-RACE was 2,940 bp long and had an open reading frame (ORF) of 940 amino acids. The predicted 940 polypeptides have four major transmembrane domains and showed about 50% identity with that of rat brain CLCAs in our previously published data. Semi-quantification analysis revealed that CMP6 was most abundantly expressed in small intestine, colon and liver. However, all tissues except small intestine, colon and liver had undetectable levels. This result became more credible after we did real-time PCR quantification for the target gene. In view of all CLCA studies focused on human or murine channels, this finding suggests a hypothetical protein as an ion channel, an X. laevis CLCA. PMID:21826170

  5. Exploring the Underlying Mechanisms of the Xenopus laevis Embryonic Cell Cycle.

    PubMed

    Zhang, Kun; Wang, Jin

    2018-05-31

    The cell cycle is an indispensable process in proliferation and development. Despite significant efforts, global quantification and physical understanding are still challenging. In this study, we explored the mechanisms of the Xenopus laevis embryonic cell cycle by quantifying the underlying landscape and flux. We uncovered the Mexican hat landscape of the Xenopus laevis embryonic cell cycle with several local basins and barriers on the oscillation path. The local basins characterize the different phases of the Xenopus laevis embryonic cell cycle, and the local barriers represent the checkpoints. The checkpoint mechanism of the cell cycle is revealed by the landscape basins and barriers. While landscape shape determines the stabilities of the states on the oscillation path, the curl flux force determines the stability of the cell cycle flow. Replication is fundamental for biology of living cells. We quantify the input energy (through the entropy production) as the thermodynamic requirement for initiation and sustainability of single cell life (cell cycle). Furthermore, we also quantify curl flux originated from the input energy as the dynamical requirement for the emergence of a new stable phase (cell cycle). This can provide a new quantitative insight for the origin of single cell life. In fact, the curl flux originated from the energy input or nutrition supply determines the speed and guarantees the progression of the cell cycle. The speed of the cell cycle is a hallmark of cancer. We characterized the quality of the cell cycle by the coherence time and found it is supported by the flux and energy cost. We are also able to quantify the degree of time irreversibility by the cross correlation function forward and backward in time from the stochastic traces in the simulation or experiments, providing a way for the quantification of the time irreversibility and the flux. Through global sensitivity analysis upon landscape and flux, we can identify the key elements for

  6. Developing Xenopus Laevis as a Model to Screen Drugs for Fragile X Syndrome

    DTIC Science & Technology

    2013-10-01

    several candidate treatments for Fragile X Syndrome have gone to clinical trials. Though promising, no treatment has yet been approved. This sad ...Xenopus laevis tadpoles. J Comp Neurol 520, 401-433. Dong, W., Lee, R.H., Xu, H., Yang, S., Pratt, K.G., Cao, V., Song , Y.K., Nurmikko, A., and

  7. Extracellular Ca2+ Is Required for Fertilization in the African Clawed Frog, Xenopus laevis

    PubMed Central

    Duray, Alexis M.; Tembo, Maiwase; Beleny, David O.; Napolitano, Marc A.; Sauer, Monica L.; Wisner, Bennett W.

    2017-01-01

    Background The necessity of extracellular Ca2+ for fertilization and early embryonic development in the African clawed frog, Xenopus laevis, is controversial. Ca2+ entry into X. laevis sperm is reportedly required for the acrosome reaction, yet fertilization and embryonic development have been documented to occur in high concentrations of the Ca2+ chelator BAPTA. Here we sought to resolve this controversy. Methodology/principal finding Using the appearance of cleavage furrows as an indicator of embryonic development, we found that X. laevis eggs inseminated in a solution lacking added divalent cations developed normally. By contrast, eggs inseminated in millimolar concentrations of BAPTA or EGTA failed to develop. Transferring embryos to varying solutions after sperm addition, we found that extracellular Ca2+ is specifically required for events occurring within the first 30 minutes after sperm addition, but not after. We found that the fluorescently stained sperm were not able to penetrate the envelope of eggs inseminated in high BAPTA, whereas several had penetrated the vitelline envelope of eggs inseminated without a Ca2+ chelator, or with BAPTA and saturating CaCl2. Together these results indicate that fertilization does not occur in high concentrations of Ca2+ chelators. Finally, we found that the jelly coat includes >5 mM of readily diffusible Ca2+. Conclusions/Significance Taken together, these data are consistent with requirement of extracellular Ca2+ for fertilization. Based on our findings, we hypothesize that the jelly coat surrounding the egg acts as a reserve of readily available Ca2+ ions to foster fertilization in changing extracellular milieu. PMID:28114360

  8. METAMORPHIC INHIBITION OF XENOPUS LAEVIS BY SODIUM PERCHLORATE: EFFECTS ON DEVELOPMENT AND THYROID HISTOLOGY

    EPA Science Inventory

    The perchlorate anion inhibits thyroid hormone (TH) synthesis via inhibition of the sodium-iodide symporter. It is, therefore, a good model chemical to aid in the development of a bioassay to screen chemicals for effects on thyroid function. Xenopus laevis larvae were exposed to ...

  9. Assessment of laryngeal muscle and testicular cell types in Xenopus laevis (Anura Pipidae) inhabiting maize and non-maize growing areas of South Africa

    USGS Publications Warehouse

    Smith, E.E.; Du Preez, L.H.; Gentles, A.; Solomon, K.R.; Tandler, B.; Carr, J.A.; Van Der Kraak, G. L.; Kendall, R.J.; Giesy, J.P.; Gross, T.S.

    2005-01-01

    We tested the hypothesis that adult African clawed frogs (Xenopus laevis) inhabiting water bodies in maize-growing areas (MGA) of South Africa would exhibit differences in testicular structure compared to frogs from water bodies in non-maize-growing areas (NMGA) in the same locale. Adults of both sexes were collected during the autumn of 2002 in South Africa, and stereological analytical techniques were used to quantify the distribution of testicular cell types. In addition, total laryngeal mass was used as a gauge of secondary sex differences in animals from MGA and NMGA study sites. Evaluation of the total laryngeal mass revealed that there were no statistically significant differences between X. laevis of the same sex from the NMGA and MGA sites. Mean percent fractional-volume values for seminiferous tubule distribution of testicular cell types of mature X. laevis, ranged from 3-4% for spermatogonia, 26-28% for spermatocytes, 54-57% for spermatozoa, and 14-15% for other cells types. The mean percent volume for blood vessels ranged from 0.3-0.4%. These values did not differ significantly between frogs from NMGA and MGA areas. Collectively, these data demonstrated no differences in gonadal and laryngeal development in X. laevis collected in South Africa from MGA and NMGA areas and that there is little evidence for an effect of agricultural chemicals used in maize production functioning as endocrine disrupters in this species. Screening of X. laevis testes revealed a small incidence of Stage 1 testicular oocytes in adult male frogs collected from the NMGA (3%) and MGA (2%).

  10. Effects of Cylindrospermopsis raciborskii strains (Woloszynska, 1912) Senayya & Subba Raju on the mobility of Daphnia laevis (Cladocera, Daphniidae).

    PubMed

    Restani, G C; Fonseca, A L

    2014-02-01

    Cylindrospermopsis raciborskii is a cyanobacterium distributed worldwide that is known to produce cyanotoxins. Some of the Brazilian strains can produce saxitoxins (STXs), which are classified as neurotoxins and can paralyze cladocerans .Daphnia laevis is a cladoceran with a wide distribution in the Americas and has been studied as a possible test-organism in toxicity bioassays. The present work tested the acute effect on D laevis mobility when fed a saxitoxin-producing (STX and neoSTX) C. raciborskii strain, CYRF-01, and compared the results with the effects of a non-toxic strain (NPCS-1). Neonates (6-24 hours after birth) were exposed to concentrations of C. raciborskii varying from 102 to 106 cells·mL-1 of each strain for up to three hours. The cladocerans were then transferred to a medium without toxic filaments for 24 hours. Only the organisms exposed to the STX-producing strain showed signs of the immobilization of swimming movements, confirming the effects of the toxins. There was a linear correlation between the time required to induce stopping the swimming movement, with a shorter time to needed to induce immobilization at a higher the concentration; this correlation was inverse to the time required to recover the swimming movements (longer at higher concentrations, p < 0.1). D. laevis is a tropical and subtropical species with great potential for use in toxicity tests for the detection of STXs, despite being native to and found in a great array of freshwater bodies. This is the first assay testing STX-producing and non-producing C. raciborskii strains on D. laevis, species that are both found in Brazilian ecosystems.

  11. Development of a New Decision Tree to Rapidly Screen Chemical Estrogenic Activities of Xenopus laevis.

    PubMed

    Wang, Ting; Li, Weiying; Zheng, Xiaofeng; Lin, Zhifen; Kong, Deyang

    2014-02-01

    During the last past decades, there is an increasing number of studies about estrogenic activities of the environmental pollutants on amphibians and many determination methods have been proposed. However, these determination methods are time-consuming and expensive, and a rapid and simple method to screen and test the chemicals for estrogenic activities to amphibians is therefore imperative. Herein is proposed a new decision tree formulated not only with physicochemical parameters but also a biological parameter that was successfully used to screen estrogenic activities of the chemicals on amphibians. The biological parameter, CDOCKER interaction energy (Ebinding ) between chemicals and the target proteins was calculated based on the method of molecular docking, and it was used to revise the decision tree formulated by Hong only with physicochemical parameters for screening estrogenic activity of chemicals in rat. According to the correlation between Ebinding of rat and Xenopus laevis, a new decision tree for estrogenic activities in Xenopus laevis is finally proposed. Then it was validated by using the randomly 8 chemicals which can be frequently exposed to Xenopus laevis, and the agreement between the results from the new decision tree and the ones from experiments is generally satisfactory. Consequently, the new decision tree can be used to screen the estrogenic activities of the chemicals, and combinational use of the Ebinding and classical physicochemical parameters can greatly improves Hong's decision tree. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  12. Effects of depleted uranium on survival, growth, and metamorphosis in the african clawed frog (Xenopus laevis)

    USGS Publications Warehouse

    Mitchell, S.E.; Caldwell, C.A.; Gonzales, G.; Gould, W.R.; Arimoto, R.

    2005-01-01

    Embryos (stage 8-47, Nieuwkoop and Faber) of the African clawed frog (Xenopus laevis) were subjected to water-borne depleted uranium (DU) concentrations that ranged from 4.8 to 77.7 mg/Lusing an acute 96-h frog embryo teratogenesis assay-Xenopus (FETAX). In a chronic 64-d assay, X. laevis (from embryo through metamorphosis; stages 8-66) were subjected to concentrations of DU that ranged from 6.2 to 54.3 mg/L Our results indicate DU is a non teratogenic metal. No effects on mortality, malformations, or growth were observed in the 96-h FETAX with concentrations of DU that ranged from 4.8 to 77.7 mg/L From stage 8 to stage 47, X. laevis tadpoles do not actively feed and the gills are not well developed. Thus, uptake of DU was reduced despite exposure to elevated concentrations. The 64-d assay resulted in no concentration response for either mortality or malformations; however, a delay in metamorphosis was observed in tadpoles subjected to elevated DU concentrations (from 13.1 to 54.3 mg/L) compared to tadpoles in both the well-water control and reference. The delay in metamorphosis was likely due to increasing body burden of DU that ranged from 0.98 to 2.82 mg/kg. Copyright?? Taylor & Francis Inc.

  13. Competitive growth, energy allocation, and host modification in the acanthocephalan Acanthocephalus dirus: field data.

    PubMed

    Caddigan, Sara C; Pfenning, Alaina C; Sparkes, Timothy C

    2017-01-01

    The acanthocephalan Acanthocephalus dirus is a trophically transmitted parasite that modifies both the physiology and behavior of its intermediate host (isopod) prior to transmission to its definitive host (fish). Infected isopods often contain multiple A. dirus individuals and we examined the relationships between host sharing, body size, energy content, and host modification to determine if host sharing was costly and if these costs could influence the modification of host behavior (mating behavior). Using field-based measures of parasite energy content (glycogen, lipid) and parasite body size (volume), we showed that host sharing was costly in terms of energy content but not in terms of body size. Analysis of the predictors of host behavior revealed that energy content, and body size, were not predictors of host behavior. Of the variables examined, parasite intensity was the only predictor of host behavior. Hosts that contained more parasites were less likely to be modified (i.e., less likely to undergo mating suppression). We suggest that intraspecific competition influenced parasite energy content and that the costs associated with competition are likely to shape the strategy of growth and energy allocation adopted by the parasites. These costs did not appear to have a direct effect on the modification of host mating behavior.

  14. The Finding of Pacific Transvenid Acanthocephalan in the Arabian Gulf, with the Description of Paratrajectura longcementglandatus N. Gen., N. Sp. from Perciform Fishes and Emendation of Transvenidae.

    PubMed

    Amin, Omar M; Heckmann, Richard A; Ali, Atheer H

    2018-02-01

    The acanthocephalan Paratrajectura longcementglandatus n. gen., n. sp. (Transvenidae) is described from specimens of 2 perciform fish species, Nemipterus japonicus Bloch (Nemipteridae) and Otolithes ruber Bloch and Schneider, collected in the marine territorial waters of Iraq and Iran in the Arabian Gulf. Metal analysis of hook tip, middle, and base is also described using energy disruptive analysis for X-ray. The new genus is distinguished from the closely related genus Trajectura Pichelin and Cribb, 2001 described from wrasses (Labridae) (Perciformes) in the Pacific off Australia and Japan by having a proboscis with apical epidermal cone, long rhadinorhynchid-like tubular cement glands, relatively short and lobulated lemnisci, all proboscis hooks with prominent roots, females with subterminal gonopore and a rounded projection on the antero-dorsal end of the trunk, and males with elongate pre-equatorial testes reaching proboscis receptacle. In Trajectura, the proboscis lacks apical epidermal cone, the cement glands are pyriform or ovoid, the lemnisci are digitiform and considerably longer than the receptacle, the posterior proboscis hooks are rootless, the females have prominent finger-like trunk projection and terminal gonopore, and males with equatorial testes that may not be elongate and may be distant from the receptacle. The importance of cement glands in the diagnosis of genera and families in acanthocephalan taxonomy is stressed. Other features especially the type and arrangement of hooks on the proboscis, but not hook roots, are comparable in the 2 genera. Diagnosis of the family Transvenidae is emended.

  15. Dehydration mediated microRNA response in the African clawed frog Xenopus laevis.

    PubMed

    Wu, Cheng-Wei; Biggar, Kyle K; Storey, Kenneth B

    2013-10-25

    Exposure to various environmental stresses induces metabolic rate depression in many animal species, an adaptation that conserves energy until the environment is again conducive to normal life. The African clawed frog, Xenopus laevis, is periodically subjected to arid summers in South Africa, and utilizes entry into the hypometabolic state of estivation as a mechanism of long term survival. During estivation, frogs must typically deal with substantial dehydration as their ponds dry out and X. laevis can endure >30% loss of its body water. We hypothesize that microRNAs play a vital role in establishing a reversible hypometabolic state and responding to dehydration stress that is associated with amphibian estivation. The present study analyzes the effects of whole body dehydration on microRNA expression in three tissues of X. laevis. Compared to controls, levels of miR-1, miR-125b, and miR-16-1 decreased to 37±6, 64±8, and 80±4% of control levels during dehydration in liver. By contrast, miR-210, miR-34a and miR-21 were significantly elevated by 3.05±0.45, 2.11±0.08, and 1.36±0.05-fold, respectively, in the liver. In kidney tissue, miR-29b, miR-21, and miR-203 were elevated by 1.40±0.09, 1.31±0.05, and 2.17±0.31-fold, respectively, in response to dehydration whereas miR-203 and miR-34a were elevated in ventral skin by 1.35±0.05 and 1.74±0.12-fold, respectively. Bioinformatic analysis of the differentially expressed microRNAs suggests that these are mainly involved in two processes: (1) expression of solute carrier proteins, and (2) regulation of mitogen-activated protein kinase signaling. This study is the first report that shows a tissue specific mode of microRNA expression during amphibian dehydration, providing evidence for microRNAs as crucial regulators of metabolic depression. © 2013 Elsevier B.V. All rights reserved.

  16. Tissue Distribution of Enrofloxacin in African Clawed Frogs (Xenopus laevis) after Intramuscular and Subcutaneous Administration

    PubMed Central

    Felt, Stephen; Papich, Mark G; Howard, Antwain; Long, Tyler; McKeon, Gabriel; Torreilles, Stéphanie; Green, Sherril

    2013-01-01

    As part of an enrofloxacin pharmacokinetic study, concentrations of enrofloxacin and ciprofloxacin (metabolite) were measured in various tissues (brain, heart, kidney, liver, lung, and spleen) collected from treated (subcutaneous delivery, n = 3; intramuscular delivery, n = 3; untreated controls, n = 2) adult female Xenopus laevis by using HPLC. Enrofloxacin was rapidly absorbed after administration by either route and readily diffused into all sampled tissues. Enrofloxacin and ciprofloxacin were present in the tissue samples collected at 8 h. The highest average tissue concentrations for enrofloxacin were found in kidney, with the lowest concentrations in liver. Ciprofloxacin tissue concentrations paralleled but were always lower than those of enrofloxacin for all time points and tissues except brain and kidney. These results, together with previously published pharmacokinetic data and known minimal inhibitory concentrations of common pathogenic bacteria, provide a strong evidence-based rationale for choosing enrofloxacin to treat infectious diseases in X. laevis. PMID:23562103

  17. Tissue distribution of enrofloxacin in African clawed frogs (Xenopus laevis) after intramuscular and subcutaneous administration.

    PubMed

    Felt, Stephen; Papich, Mark G; Howard, Antwain; Long, Tyler; McKeon, Gabriel; Torreilles, Stéphanie; Green, Sherril

    2013-03-01

    As part of an enrofloxacin pharmacokinetic study, concentrations of enrofloxacin and ciprofloxacin (metabolite) were measured in various tissues (brain, heart, kidney, liver, lung, and spleen) collected from treated (subcutaneous delivery, n = 3; intramuscular delivery, n = 3; untreated controls, n = 2) adult female Xenopus laevis by using HPLC. Enrofloxacin was rapidly absorbed after administration by either route and readily diffused into all sampled tissues. Enrofloxacin and ciprofloxacin were present in the tissue samples collected at 8 h. The highest average tissue concentrations for enrofloxacin were found in kidney, with the lowest concentrations in liver. Ciprofloxacin tissue concentrations paralleled but were always lower than those of enrofloxacin for all time points and tissues except brain and kidney. These results, together with previously published pharmacokinetic data and known minimal inhibitory concentrations of common pathogenic bacteria, provide a strong evidence-based rationale for choosing enrofloxacin to treat infectious diseases in X. laevis.

  18. Expression of mammalian beta-adrenergic receptors in Xenopus laevis oocytes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bahouth, S.W.; Malbon, C.C.

    1987-05-01

    Xenopus laevis oocytes are a useful transcription and expression system for DNA and RNA, respectively. Total cellular RNA was extracted from mouse lymphoma S49 cells and poly(A)/sup +/mRNA prepared by affinity chromatography of RNA on oligo(dT) cellulose. The membranes of S49 cells contain beta-adrenergic receptors that display pharmacological characteristics of beta/sub 2/-subtype. Xenopus laevis oocytes were injected with 50 ng of mRNA/oocyte. Expression of beta-adrenergic receptors in oocytes incubated for 30 hr after microinjection was assessed in membranes by radioligand binding using (/sup 3/H) dihydroalprenolol. The injected oocytes displayed 0.34 fmol receptor/oocyte as compared to 0.02 fmol receptor/oocyte in themore » control oocytes. The affinity of beta-adrenergic receptors in injected oocytes for this radioligand was 2 nM, a value similar to the affinity of beta-adrenergic receptors for DHA in S49 cell membranes. The potency of beta-adrenergic agonists in competing for DHA binding to oocytes membranes was isoproterenol > epinephrine > norepineprine, indicating that the expressed beta-adrenergic receptors were of the beta/sub 2/-subtype. The K/sub I/ of these agonists for the beta-adrenergic receptor in oocyte membranes was 0.03, 0.15 and 1.2 ..mu..M, respectively. The role of post-translational modification in dictating receptor subtype is analyzed using mRNA of beta/sub 1/- as well as beta/sub 2/-adrenergic receptors.« less

  19. Three-dimensional reconstruction of the cranial and anterior spinal nerves in early tadpoles of Xenopus laevis (Pipidae, Anura).

    PubMed

    Naumann, Benjamin; Olsson, Lennart

    2018-04-01

    Xenopus laevis is one of the most widely used model organism in neurobiology. It is therefore surprising, that no detailed and complete description of the cranial nerves exists for this species. Using classical histological sectioning in combination with fluorescent whole mount antibody staining and micro-computed tomography we prepared a detailed innervation map and a freely-rotatable three-dimensional (3D) model of the cranial nerves and anterior-most spinal nerves of early X. laevis tadpoles. Our results confirm earlier descriptions of the pre-otic cranial nerves and present the first detailed description of the post-otic cranial nerves. Tracing the innervation, we found two previously undescribed head muscles (the processo-articularis and diaphragmatico-branchialis muscles) in X. laevis. Data on the cranial nerve morphology of tadpoles are scarce, and only one other species (Discoglossus pictus) has been described in great detail. A comparison of Xenopus and Discoglossus reveals a relatively conserved pattern of the post-otic and a more variable morphology of the pre-otic cranial nerves. Furthermore, the innervation map and the 3D models presented here can serve as an easily accessible basis to identify alterations of the innervation produced by experimental studies such as genetic gain- and loss of function experiments. © 2017 Wiley Periodicals, Inc.

  20. A Novel Trypsin Inhibitor-Like Cysteine-Rich Peptide from the Frog Lepidobatrachus laevis Containing Proteinase-Inhibiting Activity.

    PubMed

    Wang, Yu-Wei; Tan, Ji-Min; Du, Can-Wei; Luan, Ning; Yan, Xiu-Wen; Lai, Ren; Lu, Qiu-Min

    2015-08-01

    Various bio-active substances in amphibian skins play important roles in survival of the amphibians. Many protease inhibitor peptides have been identified from amphibian skins, which are supposed to negatively modulate the activity of proteases to avoid premature degradation or release of skin peptides, or to inhibit extracellular proteases produced by invading bacteria. However, there is no information on the proteinase inhibitors from the frog Lepidobatrachus laevis which is unique in South America. In this work, a cDNA encoding a novel trypsin inhibitor-like (TIL) cysteine-rich peptide was identified from the skin cDNA library of L. laevis. The 240-bp coding region encodes an 80-amino acid residue precursor protein containing 10 half-cysteines. By sequence comparison and signal peptide prediction, the precursor was predicted to release a 55-amino acid mature peptide with amino acid sequence, IRCPKDKIYKFCGSPCPPSCKDLTPNCIAVCKKGCFCRDGTVDNNHGKCVKKENC. The mature peptide was named LL-TIL. LL-TIL shares significant domain similarity with the peptides from the TIL supper family. Antimicrobial and trypsin-inhibitory abilities of recombinant LL-TIL were tested. Recombinant LL-TIL showed no antimicrobial activity, while it had trypsin-inhibiting activity with a Ki of 16.5178 μM. These results suggested there was TIL peptide with proteinase-inhibiting activity in the skin of frog L. laevis. To the best of our knowledge, this is the first report of TIL peptide from frog skin.

  1. Comparative toxicity of methidathion and glyphosate on early life stages of three amphibian species: Pelophylax ridibundus, Pseudepidalea viridis, and Xenopus laevis.

    PubMed

    Güngördü, Abbas

    2013-09-15

    The assessments of pesticide toxicity on nontarget organisms have largely been focused on the determination of median lethal concentration (LC50) values using single/laboratory species. Although useful, these studies cannot describe the biochemical mechanisms of toxicity and also cannot explain the effects of pesticides on natural species. In this study, the toxic effects of glyphosate and methidathion were evaluated comparatively on early developmental stages of 3 anurans-2 natural (Pelophylax ridibundus, Pseudepidalea viridis) and 1 laboratory species (Xenopus laevis). The 96-h LC50 values for methidathion and glyphosate were determined as 25.7-19.6 mg active ingredient (AI)/L for P. viridis, 27.4-22.7 mg AI/L for P. ridibundus, and 15.3-5.05 mg AI/L for X. laevis tadpoles. Furthermore, as early signs of intoxication, glutathione S-transferase (GST), acetylcholinesterase (AChE), carboxylesterase (CaE), glutathione reductase, lactate dehydrogenase, and aspartate aminotrasferase were assayed in 4-day-old tadpoles after 96-h pesticide exposure. The GST induction after 3.2mg AI/L methidathion exposure was determined to be 173%, 83%, and 38% of control, and the AChE inhibition for the same dose was determined to be 86%, 96%, and 30% of control for P. ridibundus, P. viridis, and X. laevis, respectively. Unlike the application of methidathion, all enzyme activities showed statistically significant increases on glyphosate exposure compared to controls. However, these increases in enzyme activities were not shown to be parallel with the increase of concentration. The levels of increases of GST and AChE were determined to be 111% and 31% for P. ridibundus, 13% and 51% for P. viridis, and 15% and 36% for X. laevis after 3.2mg AI/L glyphosate exposure, respectively. The findings of the study suggest that the most sensitive species to pesticide exposure is X. laevis. The selected biomarker enzymes AChE, CaE, and GST are useful in understanding the toxic mechanisms of these

  2. THYROID AXIS INHIBITION IN XENOPUS LAEVIS: DEVELOPMENT OF AN AMPHIBIAN-BASED SCREENING ASSAY FOR THYROID DISRUPTION

    EPA Science Inventory

    In response to the initial EDSTAC recommendations, research was conducted on the development of a Xenopus laevis based tail resorption assay for evaluating thyroid axis disruption. These experiments highlighted key limitations associated with reliance on tail resorption as a meas...

  3. Identification of a candidate CD5 homologue in the amphibian Xenopus laevis.

    PubMed

    Jürgens, J B; Gartland, L A; Du Pasquier, L; Horton, J D; Göbel, T W; Cooper, M D

    1995-11-01

    We identified a novel T cell Ag in the South African clawed toad (Xenopus laevis) by a mAb designated 2B1. This Ag is present in relatively high levels on most thymocytes, approximately 65% of splenocytes, 55% of PBL, and 65% of intestinal lymphocytes, but is rarely seen on IgM+ B cells in any of these tissues. Lymphocytes bearing the 2B1 Ag proliferate in response to stimulation with Con A or PHA, whereas the 2B1- lymphocytes are reactive to LPS. Biochemical analysis indicates that this Ag is a differentially phosphorylated glycoprotein of 71 to 82 kDa. The protein core of 64 kDa bears both N- and O-linked carbohydrate side chains. The amino-terminal protein sequence of the 2B1 Ag shares significant homology with both the macrophage scavenger receptor type 1 motif and the mammalian CD5/CD6 family. The biochemical characteristics and cellular distribution of the 2B1 Ag suggest that it represents the CD5 homologue in X. laevis. While T cells constitutively express this highly conserved molecule, Xenopus B cells acquire the CD5 homologue only when they are stimulated in the presence of T cells.

  4. RMND5 from Xenopus laevis is an E3 ubiquitin-ligase and functions in early embryonic forebrain development.

    PubMed

    Pfirrmann, Thorsten; Villavicencio-Lorini, Pablo; Subudhi, Abinash K; Menssen, Ruth; Wolf, Dieter H; Hollemann, Thomas

    2015-01-01

    In Saccharomyces cerevisiae the Gid-complex functions as an ubiquitin-ligase complex that regulates the metabolic switch between glycolysis and gluconeogenesis. In higher organisms six conserved Gid proteins form the CTLH protein-complex with unknown function. Here we show that Rmnd5, the Gid2 orthologue from Xenopus laevis, is an ubiquitin-ligase embedded in a high molecular weight complex. Expression of rmnd5 is strongest in neuronal ectoderm, prospective brain, eyes and ciliated cells of the skin and its suppression results in malformations of the fore- and midbrain. We therefore suggest that Xenopus laevis Rmnd5, as a subunit of the CTLH complex, is a ubiquitin-ligase targeting an unknown factor for polyubiquitination and subsequent proteasomal degradation for proper fore- and midbrain development.

  5. Survival fraction and phenotype alterations of Xenopus laevis embryos at 3 Gy, 150 kV X-ray irradiation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Carotenuto, Rosa; Tussellino, Margherita; Mettivier, Giovanni

    To determine the radiosensitivity of Xenopus laevis embryos, aquatic organism model, for toxicity studies utilizing X-rays at acute high dose levels, by analysing its survival fraction and phenotype alterations under one-exposure integral dose. We used the standard Frog Embryo Teratogenesis Assay Xenopus test during the early stages of X. laevis development. The embryos were harvested until st. 46 when they were irradiated. The radiation effects were checked daily for a week and the survival, malformations and growth inhibition were assessed. Sibling tadpoles as control organisms were used. Statistical analysis was performed to assess the extent of any damage. Irradiation was performedmore » with an X-ray tube operated at 150 kV. The tube containing the tadpoles was exposed to an air kerma of 3 Gy as measured in air with an in-beam ionization chamber. After one week, survival fraction of irradiated embryos was 58%, while for control embryos it was 81%. Hence, irradiation with 150 kV, 3 Gy X-rays produced a 23% decrease of survival in regard to unirradiated embryos. The 70% of the irradiated embryos showed an altered distribution of the skin pigmentation, in particular on the dorsal area and in the olfactory pits, where the pigment concentration increased by a factor 2. In conclusion exposure of X. laevis to 3 Gy, 150 kV X-rays induced a reduction of embryos survival and a significant modification of pigmentation. The authors think that X. laevis embryos, at st 46, is a suitable biological model for large scale investigations on the effects of ionizing radiation.« less

  6. A bio-inspired swellable microneedle adhesive for mechanical interlocking with tissue

    NASA Astrophysics Data System (ADS)

    Yang, Seung Yun; O'Cearbhaill, Eoin D.; Sisk, Geoffroy C.; Park, Kyeng Min; Cho, Woo Kyung; Villiger, Martin; Bouma, Brett E.; Pomahac, Bohdan; Karp, Jeffrey M.

    2013-04-01

    Achieving significant adhesion to soft tissues while minimizing tissue damage poses a considerable clinical challenge. Chemical-based adhesives require tissue-specific reactive chemistry, typically inducing a significant inflammatory response. Staples are fraught with limitations including high-localized tissue stress and increased risk of infection, and nerve and blood vessel damage. Here inspired by the endoparasite Pomphorhynchus laevis, which swells its proboscis to attach to its host’s intestinal wall, we have developed a biphasic microneedle array that mechanically interlocks with tissue through swellable microneedle tips, achieving ~3.5-fold increase in adhesion strength compared with staples in skin graft fixation, and removal force of ~4.5 N cm-2 from intestinal mucosal tissue. Comprising a poly(styrene)-block-poly(acrylic acid) swellable tip and non-swellable polystyrene core, conical microneedles penetrate tissue with minimal insertion force and depth, yet high adhesion strength in their swollen state. Uniquely, this design provides universal soft tissue adhesion with minimal damage, less traumatic removal, reduced risk of infection and delivery of bioactive therapeutics.

  7. A Bio-Inspired Swellable Microneedle Adhesive for Mechanical Interlocking with Tissue

    PubMed Central

    Yang, Seung Yun; O'Cearbhaill, Eoin D.; Sisk, Geoffroy C.; Park, Kyeng Min; Cho, Woo Kyung; Villiger, Martin; Bouma, Brett E.; Pomahac, Bohdan; Karp, Jeffrey M.

    2013-01-01

    Achieving significant adhesion to soft tissues while minimizing tissue damage poses a considerable clinical challenge. Chemical-based adhesives require tissue-specific reactive chemistry, typically inducing a significant inflammatory response. Staples are fraught with limitations including high-localized tissue stress and increased risk of infection, and nerve and blood vessel damage. Here, inspired by the endoparasite Pomphorhynchus laevis which swells its proboscis to attach to its host’s intestinal wall, we have developed a biphasic microneedle array that mechanically interlocks with tissue through swellable microneedle tips, achieving ~ 3.5 fold increase in adhesion strength compared to staples in skin graft fixation, and removal force of ~ 4.5 N/cm2 from intestinal mucosal tissue. Comprising a poly(styrene)-block-poly(acrylic acid) swellable tip and non-swellable polystyrene core, conical microneedles penetrate tissue with minimal insertion force and depth, yet high adhesion strength in their swollen state. Uniquely, this design provides universal soft tissue adhesion with minimal damage, less traumatic removal, reduced risk of infection and delivery of bioactive therapeutics. PMID:23591869

  8. Root iron uptake efficiency of Ulmus laevis and U. minor and their distribution in soils of the Iberian Peninsula

    PubMed Central

    Venturas, Martin; Fernández, Victoria; Nadal, Paloma; Guzmán, Paula; Lucena, Juan J.; Gil, Luis

    2014-01-01

    The calcifuge and calcicole character of wild plants has been related to nutrient availability shortages, including iron (Fe)-deficiency. Surprisingly, just a few studies examined the relation between root Fe uptake and plant distribution in different soil types. We assessed the root Fe acquisition efficiency of two Ulmus species with calcareous (Ulmus minor) and siliceous (U. laevis) soil distribution patterns in the Iberian Peninsula. Seedlings of both elm species were grown hydroponically with different Fe concentrations during 6 weeks. Plant physiological responses to Fe-limiting conditions were evaluated as were the ferric reductase activity and proton (H+) extrusion capacity of the roots. Iron deprived elm seedlings of both species were stunted and suffered severe Fe-chlorosis symptoms. After Fe re-supply leaf chlorophyll concentrations rose according to species-dependent patterns. While U. minor leaves and seedlings re-greened evenly, U. laevis did so along the nerves of new growing leaves. U. minor had a higher root ferric reductase activity and H+-extrusion capability than U. laevis and maintained a better nutrient balance when grown under Fe-limiting conditions. The two elm species were found to have different Fe acquisition efficiencies which may be related to their natural distribution in calcareous and siliceous soils of the Iberian Peninsula. PMID:24723927

  9. Accelerated Gene Evolution and Subfunctionalization in thePseudotetraploid Frog Xenopus Laevis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hellsten, Uffe; Khokha, Mustafa K.; Grammar, Timothy C.

    2007-03-01

    Ancient whole genome duplications have been implicated in the vertebrate and teleost radiations, and in the emergence of diverse angiosperm lineages, but the evolutionary response to such a perturbation is still poorly understood. The African clawed frog Xenopus laevis experienced a relatively recent tetraploidization {approx} 40 million years ago. Analysis of the considerable amount of EST sequence available for this species together with the genome sequence of the related diploid Xenopus tropicalis provides a unique opportunity to study the genomic response to whole genome duplication.

  10. Subcellular Metabolite and Lipid Analysis of Xenopus laevis Eggs by LAESI Mass Spectrometry

    PubMed Central

    Reschke, Brent R.; Henderson, Holly D.; Powell, Matthew J.; Moody, Sally A.; Vertes, Akos

    2014-01-01

    Xenopus laevis eggs are used as a biological model system for studying fertilization and early embryonic development in vertebrates. Most methods used for their molecular analysis require elaborate sample preparation including separate protocols for the water soluble and lipid components. In this study, laser ablation electrospray ionization (LAESI), an ambient ionization technique, was used for direct mass spectrometric analysis of X. laevis eggs and early stage embryos up to five cleavage cycles. Single unfertilized and fertilized eggs, their animal and vegetal poles, and embryos through the 32-cell stage were analyzed. Fifty two small metabolite ions, including glutathione, GABA and amino acids, as well as numerous lipids including 14 fatty acids, 13 lysophosphatidylcholines, 36 phosphatidylcholines and 29 triacylglycerols were putatively identified. Additionally, some proteins, for example thymosin β4 (Xen), were also detected. On the subcellular level, the lipid profiles were found to differ between the animal and vegetal poles of the eggs. Radial profiling revealed profound compositional differences between the jelly coat vitelline/plasma membrane and egg cytoplasm. Changes in the metabolic profile of the egg following fertilization, e.g., the decline of polyamine content with the development of the embryo were observed using LAESI-MS. This approach enables the exploration of metabolic and lipid changes during the early stages of embryogenesis. PMID:25506922

  11. Subcellular metabolite and lipid analysis of Xenopus laevis eggs by LAESI mass spectrometry.

    PubMed

    Shrestha, Bindesh; Sripadi, Prabhakar; Reschke, Brent R; Henderson, Holly D; Powell, Matthew J; Moody, Sally A; Vertes, Akos

    2014-01-01

    Xenopus laevis eggs are used as a biological model system for studying fertilization and early embryonic development in vertebrates. Most methods used for their molecular analysis require elaborate sample preparation including separate protocols for the water soluble and lipid components. In this study, laser ablation electrospray ionization (LAESI), an ambient ionization technique, was used for direct mass spectrometric analysis of X. laevis eggs and early stage embryos up to five cleavage cycles. Single unfertilized and fertilized eggs, their animal and vegetal poles, and embryos through the 32-cell stage were analyzed. Fifty two small metabolite ions, including glutathione, GABA and amino acids, as well as numerous lipids including 14 fatty acids, 13 lysophosphatidylcholines, 36 phosphatidylcholines and 29 triacylglycerols were putatively identified. Additionally, some proteins, for example thymosin β4 (Xen), were also detected. On the subcellular level, the lipid profiles were found to differ between the animal and vegetal poles of the eggs. Radial profiling revealed profound compositional differences between the jelly coat vitelline/plasma membrane and egg cytoplasm. Changes in the metabolic profile of the egg following fertilization, e.g., the decline of polyamine content with the development of the embryo were observed using LAESI-MS. This approach enables the exploration of metabolic and lipid changes during the early stages of embryogenesis.

  12. Effect of metal ions on the activity of casein kinase II from Xenopus laevis.

    PubMed

    Gatica, M; Hinrichs, M V; Jedlicki, A; Allende, C C; Allende, J E

    1993-01-04

    Casein kinase II purified from the nuclei of Xenopus laevis oocytes as well as the recombinant alpha and beta subunits of the X. laevis CKII, produced in E. coli from the cloned cDNA genes, were tested with different divalent metal ions. The enzyme from both sources was active with either Mg2+, Mn2+, or Co2+. Optimal concentrations were 7-10 mM for Mg2+, 0.5-0.7 mM for Mn2+ and 1-2 mM for Co2+. In the presence of Mn2+ or Co2+ the enzyme used GTP more efficiently than ATP as a phosphate donor while the reverse was true in the presence of Mg2+. The apparent Km values for both nucleotide triphosphates were greatly decreased in the presence of Mn2+ as compared with Mg2+. Addition of Zn2+ (above 150 microM) to an assay containing the optimal Mg2+ ion concentration caused strong inhibition of both holoenzyme and alpha subunit. Inhibition of the holoenzyme by 400 microM Ni2+ could be reversed by high concentrations of Mg2+ but no reversal of this inhibition was observed with the alpha subunit.

  13. Regulation of the insulin-Akt signaling pathway and glycolysis during dehydration stress in the African clawed frog Xenopus laevis.

    PubMed

    Wu, Cheng-Wei; Tessier, Shannon N; Storey, Kenneth B

    2017-12-01

    Estivation is an adaptive stress response utilized by some amphibians during periods of drought in the summer season. In this study, we examine the regulation of the insulin signaling cascade and glycolysis pathway in the African clawed frog Xenopus laevis during the dehydration stress induced state of estivation. We show that in the brain and heart of X. laevis, dehydration reduces the phosphorylation of the insulin growth factor-1 receptor (IGF-1R), and this is followed by similar reductions in the phosphorylation of the Akt and mechanistic target of rapamycin (mTOR) kinase. Interestingly, phosphorylation levels of IGF-1R and mTOR were not affected in the kidney, and phosphorylation levels of P70S6K and the ribosomal S6 protein were elevated during dehydration stress. Animals under estivation are also susceptible to periods of hypoxia, suggesting that glycolysis may also be affected. We observed that protein levels of many glycolytic enzymes remained unchanged during dehydration; however, the hypoxia response factor-1 alpha (HIF-1α) protein was elevated by greater than twofold in the heart during dehydration. Overall, we provide evidence that shows that the insulin signaling pathway in X. laevis is regulated in a tissue-specific manner during dehydration stress and suggests an important role for this signaling cascade in mediating the estivation response.

  14. Anxa4 Genes are Expressed in Distinct Organ Systems in Xenopus laevis and tropicalis But are Functionally Conserved

    PubMed Central

    Massé, Karine L; Collins, Robert J; Bhamra, Surinder; Seville, Rachel A

    2007-01-01

    Anxa4 belongs to the multigenic annexin family of proteins which are characterized by their ability to interact with membranes in a calcium-dependent manner. Defined as a marker for polarized epithelial cells, Anxa4 is believed to be involved in many cellular processes but its functions in vivo are still poorly understood. Previously, we cloned Xanx4 in Xenopus laevis (now referred to as anxa4a) and demonstrated its role during organogenesis of the pronephros, providing the first evidence of a specific function for this protein during the development of a vertebrate. Here, we describe the strict conservation of protein sequence and functional domains of anxa4 during vertebrate evolution. We also identify the paralog of anxa4a, anxa4b and show its specific temporal and spatial expression pattern is different from anxa4a. We show that anxa4 orthologs in X. laevis and tropicalis display expression domains in different organ systems. Whilst the anxa4a gene is mainly expressed in the kidney, Xt anxa4 is expressed in the liver. Finally, we demonstrate Xt anxa4 and anxa4a can display conserved function during kidney organogenesis, despite the fact that Xt anxa4 transcripts are not expressed in this domain. This study highlights the divergence of expression of homologous genes during Xenopus evolution and raises the potential problems of using X. tropicalis promoters in X. laevis. PMID:19279706

  15. Acanthocephalans from fishes and amphibians in Vietnam, with descriptions of five new species

    PubMed Central

    2014-01-01

    Eight species of acanthocephalans are reported, and five are new. Specimens of Neoechinorhynchus (Hebesoma) manubrianus Amin, Ha & Ha, 2011 were similar to the original description. Neoechinorhynchus (Hebesoma) spiramuscularis n. sp. (Neoechinorhynchidae), from Xenocypris davidi, has a unique proboscis receptacle wrapped in a spiral muscular layer, and an undulating flask-shaped lemnisci, as well as double para-receptacle structures. Heterosentis mongcai n. sp. (Arhythmacanthidae), from Acreichthys sp., has a small fusiform trunk with an unarmed cone and anterior trunk spines, and a proboscis with two circles of rooted apical hooks and 3–4 circles of rooted posterior spines as well as a para-receptacle-like structure at the posterior end. The poorly known Filisoma indicum Van Cleave, 1928 is fully described and illustrated for the first time. Acanthocephalus parallelcementglandatus n. sp. (Echinorhynchidae), from Clarias batrachus, is distinguished from other species of Acanthocephalus by its small fusiform trunk and parallel tubular cement glands. Pseudoacanthocephalus coniformis n. sp. (Echinorhynchidae), from Hylarana sp., is distinguished from other species by having an anterior trunk collar and staggered prominent filiform cement glands, among other features. Cathayacanthus spinitruncatus n. sp. (Rhadinorhynchidae), from Leiognathus equulus, is distinguished from the only two known species of the genus by having a very long and slender proboscis with more than 50 hooks per row and a totally spined trunk. The generic diagnosis of Cathayacanthus Golvan, 1969 is emended. Rhadinorhynchus johnstoni Golvan, 1969 (Rhadinorhynchidae) perfectly fits the only complete description of that species from the Fiji Islands. PMID:25331738

  16. Acanthocephalans from fishes and amphibians in Vietnam, with descriptions of five new species.

    PubMed

    Amin, Omar Mohamed; Heckmann, Richard Anderson; Van Ha, Nguyen

    2014-01-01

    Eight species of acanthocephalans are reported, and five are new. Specimens of Neoechinorhynchus (Hebesoma) manubrianus Amin, Ha & Ha, 2011 were similar to the original description. Neoechinorhynchus (Hebesoma) spiramuscularis n. sp. (Neoechinorhynchidae), from Xenocypris davidi, has a unique proboscis receptacle wrapped in a spiral muscular layer, and an undulating flask-shaped lemnisci, as well as double para-receptacle structures. Heterosentis mongcai n. sp. (Arhythmacanthidae), from Acreichthys sp., has a small fusiform trunk with an unarmed cone and anterior trunk spines, and a proboscis with two circles of rooted apical hooks and 3-4 circles of rooted posterior spines as well as a para-receptacle-like structure at the posterior end. The poorly known Filisoma indicum Van Cleave, 1928 is fully described and illustrated for the first time. Acanthocephalus parallelcementglandatus n. sp. (Echinorhynchidae), from Clarias batrachus, is distinguished from other species of Acanthocephalus by its small fusiform trunk and parallel tubular cement glands. Pseudoacanthocephalus coniformis n. sp. (Echinorhynchidae), from Hylarana sp., is distinguished from other species by having an anterior trunk collar and staggered prominent filiform cement glands, among other features. Cathayacanthus spinitruncatus n. sp. (Rhadinorhynchidae), from Leiognathus equulus, is distinguished from the only two known species of the genus by having a very long and slender proboscis with more than 50 hooks per row and a totally spined trunk. The generic diagnosis of Cathayacanthus Golvan, 1969 is emended. Rhadinorhynchus johnstoni Golvan, 1969 (Rhadinorhynchidae) perfectly fits the only complete description of that species from the Fiji Islands. © O.M. Amin et al., published by EDP Sciences, 2014.

  17. CONCENTRATION DEPENDENT ACCUMULATION OF [3H]-DELTAMETHRIN IN SODIUM CHANNEL N AV1.2 EXPRESSING XENOPUS LAEVIS OOCYTES.

    EPA Science Inventory

    Disruption of neuronal voltage-sensitive sodium channels (VSSCs) by pyrethroid insecticides such as deltamethrin (DLT) has been widely studied using Xenopus laevis oocytes transfected with VSSC. However, the extent of pyrethroid accumulation in VSSC-expressing oocytes is unknown....

  18. Purification of Xenopus laevis mitochondrial RNA polymerase and identification of a dissociable factor required for specific transcription

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Bogenhagen, D.F.; Insdorf, N.F.

    1988-07-01

    The Xenopus laevis mitochondrial RNA (mtRNA) polymerase was purified to near homogeneity with an overall yield approaching 50%. The major polypeptides in the final fraction were a doublet of proteins of approximately 140 kilodaltons that copurified with the mtRNA polymerase activity. It appeared likely that the smaller polypeptide is a breakdown product of the larger one. The highly purified polymerase was active in nonspecific transcription but required a dissociable factor for specific transcription of X. laevis mtDNA. The factor could be resolved from mtRNA polymerase by hydrophobic chromatography and had a sedimentation coefficient of 3.0 S. The transcription factor elutedmore » from both the hydrophobic column and a Mono Q anion-exchange column as a single symmetrical peak. The mtRNA polymerase and this factor together are necessary and sufficient for active transcription from four promoters located in a noncoding region of the mtDNA genome between the gene for tRNA/sup Phe/ and the displacement loop.« less

  19. Functional and structural effects of amyloid-β aggregate on Xenopus laevis oocytes.

    PubMed

    Parodi, Jorge; Ochoa-de la Paz, Lenin; Miledi, Ricardo; Martínez-Torres, Ataúlfo

    2012-10-01

    Xenopus laevis oocytes exposed to amyloid-β aggregate generated oscillatory electric activity (blips) that was recorded by two-microelectrode voltage-clamp. The cells exhibited a series of "spontaneous" blips ranging in amplitude from 3.8 ± 0.9 nA at the beginning of the recordings to 6.8 ± 1.7 nA after 15 min of exposure to 1 μM aggregate. These blips were similar in amplitude to those induced by the channel-forming antimicrobial agents amphotericin B (7.8 ± 1.2 nA) and gramicidin (6.3 ± 1.1 nA). The amyloid aggregate-induced currents were abolished when extracellular Ca(2+) was removed from the bathing solution, suggesting a central role for this cation in generating the spontaneous electric activity. The amyloid aggregate also affected the Ca(2+)-dependent Cl(-) currents of oocytes, as shown by increased amplitude of the transient-outward chloride current (T(out)) and the serum-activated, oscillatory Cl(-) currents. Electron microcopy revealed that amyloid aggregate induced the dissociation of the follicular cells that surround the oocyte, thus leading to a failure in the electro-chemical communication between these cells. This was also evidenced by the suppression of the oscillatory Ca(2+)-dependent ATP-currents, which require proper coupling between oocytes and the follicular cell layer. These observations, made using the X. laevis oocytes as a versatile experimental model, may help to understand the effects of amyloid aggregate on cellular communication.

  20. RNS60, a charge-stabilized nanostructure saline alters Xenopus Laevis oocyte biophysical membrane properties by enhancing mitochondrial ATP production

    PubMed Central

    Choi, Soonwook; Yu, Eunah; Kim, Duk-Soo; Sugimori, Mutsuyuki; Llinás, Rodolfo R

    2015-01-01

    We have examined the effects of RNS60, a 0.9% saline containing charge-stabilized oxygen nanobubble-based structures. RNS60 is generated by subjecting normal saline to Taylor–Couette–Poiseuille (TCP) flow under elevated oxygen pressure. This study, implemented in Xenopus laevis oocytes, addresses both the electrophysiological membrane properties and parallel biological processes in the cytoplasm. Intracellular recordings from defolliculated X. laevis oocytes were implemented in: (1) air oxygenated standard Ringer's solution, (2) RNS60-based Ringer's solution, (3) RNS10.3 (TCP-modified saline without excess oxygen)-based Ringer's, and (4) ONS60 (saline containing high pressure oxygen without TCP modification)-based Ringer's. RNS60-based Ringer's solution induced membrane hyperpolarization from the resting membrane potential. This effect was prevented by: (1) ouabain (a blocker of the sodium/potassium ATPase), (2) rotenone (a mitochondrial electron transfer chain inhibitor preventing usable ATP synthesis), and (3) oligomycin A (an inhibitor of ATP synthase) indicating that RNS60 effects intracellular ATP levels. Increased intracellular ATP levels following RNS60 treatment were directly demonstrated using luciferin/luciferase photon emission. These results indicate that RNS60 alters intrinsic the electrophysiological properties of the X. laevis oocyte membrane by increasing mitochondrial-based ATP synthesis. Ultrastructural analysis of the oocyte cytoplasm demonstrated increased mitochondrial length in the presence of RNS60-based Ringer's solution. It is concluded that the biological properties of RNS60 relate to its ability to optimize ATP synthesis. PMID:25742953

  1. Asymmetries in Cell Division, Cell Size, and Furrowing in the Xenopus laevis Embryo.

    PubMed

    Tassan, Jean-Pierre; Wühr, Martin; Hatte, Guillaume; Kubiak, Jacek

    2017-01-01

    Asymmetric cell divisions produce two daughter cells with distinct fate. During embryogenesis, this mechanism is fundamental to build tissues and organs because it generates cell diversity. In adults, it remains crucial to maintain stem cells. The enthusiasm for asymmetric cell division is not only motivated by the beauty of the mechanism and the fundamental questions it raises, but has also very pragmatic reasons. Indeed, misregulation of asymmetric cell divisions is believed to have dramatic consequences potentially leading to pathogenesis such as cancers. In diverse model organisms, asymmetric cell divisions result in two daughter cells, which differ not only by their fate but also in size. This is the case for the early Xenopus laevis embryo, in which the two first embryonic divisions are perpendicular to each other and generate two pairs of blastomeres, which usually differ in size: one pair of blastomeres is smaller than the other. Small blastomeres will produce embryonic dorsal structures, whereas the larger pair will evolve into ventral structures. Here, we present a speculative model on the origin of the asymmetry of this cell division in the Xenopus embryo. We also discuss the apparently coincident asymmetric distribution of cell fate determinants and cell-size asymmetry of the 4-cell stage embryo. Finally, we discuss the asymmetric furrowing during epithelial cell cytokinesis occurring later during Xenopus laevis embryo development.

  2. Entire mesodermal mantle behaves as Spemann's organizer in dorsoanterior enhanced Xenopus laevis embryos

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kao, K.R.; Elinson, R.P.

    1988-05-01

    The body plan of Xenopus laevis can be respecified by briefly exposing early cleavage stage embryos to lithium. Such embryos develop exaggerated dorsoanterior structures such as a radial eye and cement gland. In this paper, we demonstrate that the enhanced dorsoanterior phenotype results from an overcommitment of mesoderm to dorsoanterior mesoderm. Histological and immunohistochemical observations reveal that the embryos have a greatly enlarged notochord with very little muscle tissue. In addition, they develop a radial, beating heart, suggesting that lithium also specifies anterior mesoderm and pharyngeal endoderm. Randomly oriented diametrically opposed marginal zone grafts from lithium-treated embryos, when transplanted intomore » ultraviolet (uv)-irradiated axis-deficient hosts, rescue dorsal axial structures. These transplantation experiments demonstrate that the entire marginal zone of the early gastrula consists of presumptive dorsal mesoderm. Vital dye marking experiments also indicate that the entire marginal zone maps to the prominent proboscis that is composed of chordamesoderm and represents the long axis of the embryo. These results suggest that lithium respecifies the mesoderm of Xenopus laevis embryos so that it differentiates into the Spemann organizer. We suggest that the origin of the dorsoanterior enhanced phenotypes generated by lithium and the dorsoanterior deficient phenotypes generated by uv irradiation are due to relative quantities of organizer. Our evidence demonstrates the existence of a continuum of body plan phenotypes based on this premise.« less

  3. Distribution of Single-Wall Carbon Nanotubes in the Xenopus laevis Embryo after Microinjection

    PubMed Central

    Holt, Brian D.; Shawky, Joseph H.; Dahl, Kris Noel; Davidson, Lance A.; Islam, Mohammad F.

    2016-01-01

    Single-wall carbon nanotubes (SWCNTs) are advanced materials with the potential for a myriad of diverse applications, including biological technologies and largescale usage with the potential for environmental impacts. SWCNTs have been exposed to developing organisms to determine their effects on embryogenesis, and results have been inconsistent arising, in part, from differing material quality, dispersion status, material size, impurity from catalysts, and stability. For this study, we utilized highly purified SWCNT samples with short, uniform lengths (145 ± 17 nm) well dispersed in solution. To test high exposure doses, we microinjected > 500 μg mL-1 SWCNT concentrations into the well-established embryogenesis model, Xenopus laevis, and determined embryo compatibility and sub-cellular localization during development. SWCNTs localized within cellular progeny of the microinjected cells, but heterogeneously distributed throughout the target-injected tissue. Co-registering unique Raman spectral intensity of SWCNTs with images of fluorescently labelled sub-cellular compartments demonstrated that even at the regions of highest SWCNT concentration, there were no gross alterations to sub-cellular microstructures, including filamentous actin, endoplasmic reticulum and vesicles. Furthermore, SWCNTs did not aggregate or localize to the perinuclear sub-cellular region. Combined, these results suggest that purified and dispersed SWCNTs are not toxic to X. laevis animal cap ectoderm and may be suitable candidate materials for biological applications. PMID:26510384

  4. Expression of cardiac sarcolemmal Na sup + -Ca sup 2+ exchange activity in Xenopus laevis oocytes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Longoni, S.; Coady, M.J.; Ikeda, T.

    1988-12-01

    Injection of Xenopus laevis oocytes with rabbit heart poly(A){sup +}RNA results in expression of Na{sup +} inside (Na{sub i}{sup +})-dependent Ca{sup 2+} uptake activity. The activity was measured by first loading the oocytes with Na{sup +} using nystatin and then incubating the oocytes in K{sup +} or Na{sup +} medium containing {sup 45}Ca. The expressed Na{sup +} gradient-dependent Ca{sup 2+} uptake was five to eight times that observed with water-injected oocytes or with poly(A){sup +}RNA-injected oocytes for which the Na{sup +} load step had been omitted. Induced activity was related to the amount of RNA injected and was insensitive tomore » nifedipine. Fractionation of the poly(A){sup +}RNA on a sucrose gradient determined that the active message had a size range between 3 and 8 kb. The properties of the Na{sup +} gradient-dependent Ca{sup 2+} uptake indicated that Na{sup +}-Ca{sup 2+} exchange activity had been expressed in X. laevis oocytes. The result may be useful for cloning and identifying the molecular component responsible for Na{sup +}-Ca{sup 2+} exchange.« less

  5. DNA polymerase gamma from Xenopus laevis. I. The identification of a high molecular weight catalytic subunit by a novel DNA polymerase photolabeling procedure

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Insdorf, N.F.; Bogenhagen, D.F.

    1989-12-25

    DNA polymerase gamma has been purified over 10,000-fold from mitochondria of Xenopus laevis ovaries. We have developed a novel technique which specifically photolabels DNA polymerases. This procedure, the DNA polymerase trap, was used to identify a catalytic subunit of 140,000 Da from X. laevis DNA polymerase gamma. Additional catalytically active polypeptides of 100,000 and 55,000 Da were identified in the highly purified enzyme. These appear to be products of degradation of the 140,000-Da subunit. The DNA polymerase trap, which does not require large amounts of enzyme or renaturation from sodium dodecyl sulfate, is an alternative to the classic activity gel.

  6. Carotenoids of two freshwater amphipod species (Gammarus pulex and G. roeseli) and their common acanthocephalan parasite Polymorphus minutus.

    PubMed

    Gaillard, Maria; Juillet, Cédrik; Cézilly, Frank; Perrot-Minnot, Marie-Jeanne

    2004-09-01

    Carotenoid compositions of two freshwater Gammarus species (Crustacea: Amphipoda) and of their common acanthocephalan parasite Polymorphus minutus were characterized. The effect of carotenoid uptake by the parasite was addressed by comparing the carotenoid content of uninfected and infected female hosts. Using high-pressure liquid chromatography (HPLC), co-chromatography of reference pigments and electron ionization mass spectrometry of collected HPLC fractions (EI-MS), several xanthophylls and non-polar compounds were identified. Seven kinds of carotenoids, mainly xanthophylls, were identified in gammarids. Astaxanthin was predominant, amounting to 40 wt.% of total carotenoid in both uninfected G. pulex and G. roeseli. By contrast, we found only non-polar compounds with a predominance of esterified forms of astaxanthin in P. minutus larvae. No significant effect of infection on carotenoid content was evidenced in G. pulex and G. roeseli females. Our study highlights the use of a Matrix Solid Phase Dispersion as an efficient extraction method of both xanthophylls and non-polar pigments in small samples, including lipid-rich ones as P. minutus parasite. We discuss on the presumptive pathway leading to the formation of free astaxanthin in gammarids via hydroxy compounds, and on the accumulation of esters of astaxanthin in parasites.

  7. Intra-population variation in behavior modification by the acanthocephalan Acanthocephalus dirus: are differences mediated by host condition?

    PubMed

    Caddigan, Sara C; Barkauskas, Rima T; Sparkes, Timothy C

    2014-11-01

    The acanthocephalan parasite Acanthocephalus dirus infects the freshwater isopod Caecidotea intermedius as an intermediate host before completing its life cycle in a fish. Male C. intermedius infected by A. dirus parasites are less likely to engage in mating behavior than uninfected males but there is a significant intra-population variation in the occurrence of this behavioral change. Previous studies on uninfected isopods have shown that glycogen content is a predictor of male mating behavior and we examined whether the intra-population variation in the mating behavior of infected male C. intermedius could be explained by this relationship. A field-based behavioral experiment was used to quantify intra-population variation in male mating behavior, which showed that 50% of infected males were responsive to females and 50% were not responsive. Biochemical analysis of responsive and non-responsive males revealed that glycogen content was a predictor of the mating behavior for uninfected males but was not a predictor of mating behavior for infected males. For infected males, parasite intensity was a predictor of mating behavior. Males that contained more A. dirus parasites were less likely to undergo modification of mating behavior. We propose that the intra-population variation in the mating behavior of infected C. intermedius identified in nature was not mediated by host condition.

  8. Hybridization between the African clawed frogs Xenopus laevis and Xenopus muelleri (Pipidae) increases the multiplicity of antimicrobial peptides in skin secretions of female offspring.

    PubMed

    Mechkarska, Milena; Meetani, Mohammed; Michalak, Pawel; Vaksman, Zalman; Takada, Koji; Conlon, J Michael

    2012-09-01

    Peptidomic analysis was used to compare the distribution of host-defense peptides in norepinephrine-stimulated skin secretions from laboratory-generated female F1 hybrids of the common clawed frog Xenopus laevis (Daudin, 1802) and Mueller's clawed frog Xenopus muelleri (Peters, 1844) with the corresponding distribution in skin secretions from the parent species. A total of 18 peptides were identified in secretions from the hybrid frogs. Eleven peptides (magainin-1, magainin-2, CPF-1, CPF-3, CPF-4, CPF-5, CPF-6, CPF-7, XPF-1, XPF-2, and PGLa) were identified in secretions of both the hybrids and X. laevis. Four peptides (magainin-M1, XPF-M1, CPF-M1, and tigerinin-M1) were previously found in skin secretions of X. muelleri but magainin-M2 and CPF-M2 from X. muelleri were not detected. Three previously undescribed peptides (magainin-LM1, PGLa-LM1, and CPF-LM1) were purified from the secretions of the hybrid frogs that were not detected in secretions from either X. laevis or X. muelleri. Magainin-LM1 differs from magainin-2 from X. laevis by a single amino acid substitution (Gly(13)→Ala) but PGLa-LM1 and CPF-LM1 differ appreciably in structure from orthologs in the parent species. CPF-LM1 shows potent, broad-spectrum antimicrobial activity and is hemolytic. The data indicate that hybridization increases the multiplicity of skin host-defense peptides in skin secretions. As the female F1 hybrids are fertile, hybridization may represent an adaptive strategy among Xenopus species to increase protection against pathogenic microorganisms in the environment. Copyright © 2012 Elsevier Inc. All rights reserved.

  9. Distribution of two species of sea snakes, Aipysurus laevis and Emydocephalus annulatus, in the southern Great Barrier Reef: metapopulation dynamics, marine protected areas and conservation

    NASA Astrophysics Data System (ADS)

    Lukoschek, V.; Heatwole, H.; Grech, A.; Burns, G.; Marsh, H.

    2007-06-01

    Aipysurus laevis and Emydocephalus annulatus typically occur in spatially discrete populations, characteristic of metapopulations; however, little is known about the factors influencing the spatial and temporal stability of populations or whether specific conservation strategies, such as networks of marine protected areas, will ensure the persistence of species. Classification tree analyses of 35 years of distribution data (90 reefs, surveyed 1-11 times) in the southern Great Barrier Reef (GBR) revealed that longitude was a major factor determining the status of A. laevis on reefs (present = 38, absent = 38 and changed = 14). Reef exposure and reef area were also important; however, these factors did not specifically account for the population fluctuations and the recent local extinctions of A. laevis in this region. There were no relationships between the status of E. annulatus (present = 16, absent = 68 and changed = 6) and spatial or physical variables. Moreover, prior protection status of reefs did not account for the distribution of either species. Biotic factors, such as habitat and prey availability and the distribution of predators, which may account for the observed patterns of distribution, are discussed. The potential for inter-population exchange among sea snake populations is poorly understood, as is the degree of protection that will be afforded to sea snakes by the recently implemented network of No-take areas in the GBR. Data from this study provide a baseline for evaluating the responses of A. laevis and E. annulatus populations to changes in biotic factors and the degree of protection afforded on reefs within an ecosystem network of No-take marine protected areas in the southern GBR.

  10. Prominin-1 Localizes to the Open Rims of Outer Segment Lamellae in Xenopus laevis Rod and Cone Photoreceptors

    PubMed Central

    Han, Zhou; Anderson, David W.

    2012-01-01

    Purpose. Prominin-1 expresses in rod and cone photoreceptors. Mutations in the prominin-1 gene cause retinal degeneration in humans. In this study, the authors investigated the expression and subcellular localization of xlProminin-1 protein, the Xenopus laevis ortholog of prominin-1, in rod and cone photoreceptors of this frog. Methods. Antibodies specific for xlProminin-1 were generated. Immunoblotting was used to study the expression and posttranslational processing of xlProminin-1 protein. Immunocytochemical light and electron microscopy and transgenesis were used to study the subcellular distribution of xlProminin-1. Results. xlProminin-1 is expressed and is subject to posttranslational proteolytic processing in the retina, brain, and kidney. xlProminin-1 is differently expressed and localized in outer segments of rod and cone photoreceptors of X. laevis. Antibodies specific for the N or C termini of xlProminin-1 labeled the open rims of lamellae of cone outer segments (COS) and the open lamellae at the base of rod outer segments (ROS). By contrast, anti–peripherin-2/rds antibody, Xper5A11, labeled the closed rims of cone lamellae adjacent to the ciliary axoneme and the rims of the closed ROS disks. The extent of labeling of the basal ROS by anti–xlProminin-1 antibodies varied with the light cycle in this frog. The entire ROS was also faintly labeled by both antibodies, a result that contrasts with the current notion that prominin-1 localizes only to the basal ROS. Conclusions. These findings suggest that xlProminin-1 may serve as an anti–fusogenic factor in the regulation of disk morphogenesis and may help to maintain the open lamellar structure of basal ROS and COS disks in X. laevis photoreceptors. PMID:22076989

  11. Rotation in Xenopus laevis embryos during the second cell cycle.

    PubMed

    Starodubov, Sergey M; Golychenkov, Vladimir A

    2009-01-01

    Using time-lapse video recording and comparing successive digital images, we found that 38% of Xenopus laevis embryos (n=118) exhibited rotation during the second cell cycle. This rotation, which we term the second rotation, started approximately during the appearance of the first cleavage furrow and proceeded clockwise or counterclockwise around the vertical axis. Rotations lasted for 5-30 minutes, i.e. up to the beginning of the third cell cycle. The mean rotation angle was 36.4 degrees, with a maximum rotation of 77 degrees. No mortality was observed among the embryos exhibiting rotation. The second rotation was observed to be similar to the well-known fertilization rotation which takes place during the first cell cycle. The possible nature and significance of the second rotation are discussed.

  12. Inverse Effects on Growth and Development Rates by Means of Endocrine Disruptors in African Clawed Frog Tadpoles ("Xenopus Laevis")

    ERIC Educational Resources Information Center

    Hackney, Zachary Carl

    2007-01-01

    Previous work on fish, frogs, and salamanders, showed the ability for estrogen (EE2) and anthropogenic endocrine disruptors to skew sex ratios and cause hermaphrodism. This study addressed the effects of estrogens on growth and development rates of African clawed frog tadpoles ("Xenopus laevis") during their gender determination stages. The…

  13. Hyperinnervation improves Xenopus laevis limb regeneration.

    PubMed

    Mitogawa, Kazumasa; Makanae, Aki; Satoh, Akira

    2018-01-15

    Xenopus laevis (an anuran amphibian) shows limb regeneration ability between that of urodele amphibians and that of amniotes. Xenopus frogs can initiate limb regeneration but fail to form patterned limbs. Regenerated limbs mainly consist of cone-shaped cartilage without any joints or branches. These pattern defects are thought to be caused by loss of proper expressions of patterning-related genes. This study shows that hyperinnervation surgery resulted in the induction of a branching regenerate. The hyperinnervated blastema allows the identification and functional analysis of the molecules controlling this patterning of limb regeneration. This paper focuses on the nerve affects to improve Xenopus limb patterning ability during regeneration. The nerve molecules, which regulate limb patterning, were also investigated. Blastemas grown in a hyperinnervated forelimb upregulate limb patterning-related genes (shh, lmx1b, and hoxa13). Nerves projecting their axons to limbs express some growth factors (bmp7, fgf2, fgf8, and shh). Inputs of these factors to a blastema upregulated some limb patterning-related genes and resulted in changes in the cartilage patterns in the regenerates. These results indicate that additional nerve factors enhance Xenopus limb patterning-related gene expressions and limb regeneration ability, and that bmp, fgf, and shh are candidate nerve substitute factors. Copyright © 2017 The Authors. Published by Elsevier Inc. All rights reserved.

  14. Establishment of polarities in the oocyte of Xenopus laevis: the provisional axial symmetry of the full-grown oocyte of Xenopus laevis.

    PubMed

    Ubbels, G A

    1997-04-01

    We aimed at understanding of formation and function of the "Nieuwkoop Centre" in embryonic pattern formation. Discussed are data on genesis of cytoplasmic localizations in ovarian oocytes, transient modifications of cytoskeletal structures creating cytoplasmic asymmetries in fertilized eggs, the axis determining "vegetal cortical rotation" and fate of distinct cells, as shown by injection of specific molecular markers into particular blastomeres at specific times. Egg rotation and centrifugation suggested that sperm that gravity cooperate in symmetrization of the axially symmetrical anuran egg. After fertilization in space or in a fast rotating clinostate, axis formation and embryonic development were normal although the blastocoel was transiently abnormal. Normal tadpoles came back on Earth after ovulation, fertilization and culture in space. They metamorphosed normally and got healthy Earth-born F1 offspring. We conclude that neither sperm nor gravity are required for determination of the bilateral symmetry in the embryo of Xenopus laevis. In normal development sperm and gravity, either alone or in collaboration, may overrule an initial bilaterality inherent to, the full-grown oocyte, residing in some still unidentified component(s)/or mechanisms.

  15. Confocal Imaging of Early Heart Development in Xenopus laevis

    PubMed Central

    Kolker, Sandra J.; Tajchman, Urszula; Weeks, Daniel L.

    2013-01-01

    Xenopus laevis provides a number of advantages for studies on cardiovascular development. The embryos are fairly large, easy to obtain, and can develop at ambient temperature in simple buffer solutions. Although classic descriptions of heart development exist, the ability to use whole mount immunohistochemical methods and confocal microscopy may enhance the ability to understand both normal and experimentally perturbed cardiovascular development. We have started to examine the early stages of cardiac development in Xenopus, seeking to identify antibodies and fixatives that allow easy examination of the developing heart. We have used monoclonal antibodies (mAbs) raised against bovine cardiac troponin T and chicken tropomyosin to visualize cardiac muscle, a goat antibody recognizing bovine type VI collagen to stain the lining of vessels, and the JB3 mAb raised against chicken fibrillin which allows the visualization of a variety of cardiovascular tissues during early development. Results from embryonic stages 24–46 are presented. PMID:10644411

  16. Full-grown oocytes from Xenopus laevis resume growth when placed in culture

    PubMed Central

    Wallace, Robin A.; Misulovin, Ziva; Etkin, Laurence D.

    1981-01-01

    When most full-grown, follicle cell-invested oocytes from Xenopus laevis are placed in an appropriate culture medium, they resume growth and remain physiologically healthy for at least 2-3 weeks. Rates of growth by full-grown oocytes in vitro generally approximate and can even exceed the most rapid growth rate achieved by vitellogenic oocytes in vivo. Resumption of oocyte growth can be correlated with the loss of investing follicle cells, which under normal conditions appear to interfere with vitellogenin and nutrient access to the oocyte. The final size reached by the oocyte within the ovary is thus not an intrinsic property of the oocyte but is extrinsically imposed by the somatic environment. Images PMID:16593019

  17. Characterization of Pax3 and Sox10 transgenic Xenopus laevis embryos as tools to study neural crest development.

    PubMed

    Alkobtawi, Mansour; Ray, Heather; Barriga, Elias H; Moreno, Mauricio; Kerney, Ryan; Monsoro-Burq, Anne-Helene; Saint-Jeannet, Jean-Pierre; Mayor, Roberto

    2018-03-06

    The neural crest is a multipotent population of cells that originates a variety of cell types. Many animal models are used to study neural crest induction, migration and differentiation, with amphibians and birds being the most widely used systems. A major technological advance to study neural crest development in mouse, chick and zebrafish has been the generation of transgenic animals in which neural crest specific enhancers/promoters drive the expression of either fluorescent proteins for use as lineage tracers, or modified genes for use in functional studies. Unfortunately, no such transgenic animals currently exist for the amphibians Xenopus laevis and tropicalis, key model systems for studying neural crest development. Here we describe the generation and characterization of two transgenic Xenopus laevis lines, Pax3-GFP and Sox10-GFP, in which GFP is expressed in the pre-migratory and migratory neural crest, respectively. We show that Pax3-GFP could be a powerful tool to study neural crest induction, whereas Sox10-GFP could be used in the study of neural crest migration in living embryos. Copyright © 2018 The Authors. Published by Elsevier Inc. All rights reserved.

  18. Lectins and substitution for helper function in anti-hapten responses in Xenopus laevis.

    PubMed

    Clothier, R H; James, H S; Ruben, L N; Balls, M

    1984-08-01

    Substitution by lectins for the carrier-priming requirement in thymus-dependent, antigen-binding responses in Xenopus laevis has been examined. Concanavalin A (Con A) was found to substitute for carrier priming in control, early-thymectomized and adult-thymectomized animals, but not in animals given a single, high dose of N-methyl-N-nitrosourea, which has a permanent effect on certain thymus-dependent functions in this species. Lipopolysaccharide and other lectins, such as peanut agglutinin and wheat germ agglutinin, were unable to substitute for carrier priming. These effects of Con A are discussed in terms of substitution via amplifier T cells or a helper T cell subset.

  19. Endocrine effects of 2,2{prime},4,4{prime}-tetrachlorobiphenyl in the African clawed frog (Xenopus laevis)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Diana, S.; Hansen, L.; Foley, G.

    1995-12-31

    Ortho-substituted polychlorinated biphenyls are known to exhibit estrogenic activity and, in some cases, to enhance excretion of tetraiodothyronine (T4), resulting in hypothyroxinemia in mammals. Since thyroxine activity is essential for amphibian metamorphosis, and amphibian sex determination can be altered or reversed by exposure to exogenous estrogens or androgens, the effects of exposure of larvae of the African clawed frog (Xenopus laevis) to 2,2{prime},4,4{prime}-tetrachlorobiphenyl (CB 47) were investigated. Eggs and larvae of X. laevis were exposed to nominal concentrations of CB 47 of 0.05 or 0.25 ppm (1 ppm was found to result in 100% mortality) throughout the period of larvalmore » development, and effects on rates of metamorphosis and body growth and on gonad morphology were determined. Stage of metamorphosis, body length and body weight did not differ between treatment and control groups, following exposure to these sub-lethal concentrations, at any time during larval development. Effects of exposure on gonad morphology will be discussed. The failure of CB 47 to delay or prevent metamorphosis under these conditions may be due to poor responsiveness of hepatic UDP-glucuronyl transferases to induction, or novel systems of thyroxine and/or PCB transport, metabolism and excretion in larval amphibians.« less

  20. Extinction of an introduced warm-climate alien species, Xenopus laevis, by extreme weather events.

    PubMed

    Tinsley, Richard C; Stott, Lucy C; Viney, Mark E; Mable, Barbara K; Tinsley, Matthew C

    Invasive, non-native species represent a major threat to biodiversity worldwide. The African amphibian Xenopus laevis is widely regarded as an invasive species and a threat to local faunas. Populations originating at the Western Cape, South Africa, have been introduced on four continents, mostly in areas with a similar Mediterranean climate. Some introduced populations are also established in cooler environments where persistence for many decades suggests a capacity for long-term adaptation. In these cases, recent climate warming might enhance invasion ability, favouring range expansion, population growth and negative effects on native faunas. In the cool temperate UK, populations have been established for about 50 years in Wales and for an unknown period, probably >20 years, in England (Lincolnshire). Our field studies over 30 and 10 years, respectively, show that in favourable conditions there may be good recruitment, fast individual growth rates and large body size; maximum longevity exceeds 23 years. Nevertheless, areas of distribution remained limited, with numbers <500 in each population. In 2010, only a single individual was captured at each locality and further searching failed to record any others in repeated sampling up to 2014. We conclude that both populations are now extinct. The winters of 2009-2010 and 2010-2011 experienced extreme cold and drought (December 2010 was the coldest in 120 years and the third driest in 100 years). The extinction of X. laevis in these areas indicates that even relatively long-established alien species remain vulnerable to rare extreme weather conditions.

  1. Expression analysis of some genes regulated by retinoic acid in controls and triadimefon-exposed embryos: is the amphibian Xenopus laevis a suitable model for gene-based comparative teratology?

    PubMed

    Di Renzo, Francesca; Rossi, Federica; Bacchetta, Renato; Prati, Mariangela; Giavini, Erminio; Menegola, Elena

    2011-06-01

    The use of nonmammal models in teratological studies is a matter of debate and seems to be justified if the embryotoxic mechanism involves conserved processes. Published data on mammals and Xenopus laevis suggest that azoles are teratogenic by altering the endogenous concentration of retinoic acid (RA). The expression of some genes (Shh, Ptch-1, Gsc, and Msx2) controlled by retinoic acid is downregulated in rat embryos exposed at the phylotypic stage to the triazole triadimefon (FON). In order to propose X. laevis as a model for gene-based comparative teratology, this work evaluates the expression of Shh, Ptch-1, Gsc, and Msx2 in FON-exposed X. laevis embryos. Embryos, exposed to a high concentration level (500 µM) of FON from stage 13 till 17, were examined at stages 17, 27, and 47. Stage 17 and 27 embryos were processed to perform quantitative RT-PCR. The developmental rate was never affected by FON at any considered stage. FON-exposed stage 47 larvae showed the typical craniofacial malformations. A significant downregulation of Gsc was observed in FON-exposed stage 17 embryos. Shh, Ptch-1, Msx2 showed a high fluctuation of expression both in control and in FON-exposed samples both at stages 17 and 27. The downregulation of Gsc mimics the effects of FON on rat embryos, showing for this gene a common effect of FON in the two vertebrate classes. The high fluctuation observed in the gene expression of the other genes, however, suggests that X. laevis at this stage has limited utility for gene-based comparative teratology. © 2011 Wiley-Liss, Inc.

  2. Overland movement in African clawed frogs (Xenopus laevis): empirical dispersal data from within their native range.

    PubMed

    De Villiers, F André; Measey, John

    2017-01-01

    Dispersal forms are an important component of the ecology of many animals, and reach particular importance for predicting ranges of invasive species. African clawed frogs ( Xenopus laevis ) move overland between water bodies, but all empirical studies are from invasive populations with none from their native southern Africa. Here we report on incidents of overland movement found through a capture-recapture study carried out over a three year period in Overstrand, South Africa. The maximum distance moved was 2.4 km with most of the 91 animals, representing 5% of the population, moving ∼150 m. We found no differences in distances moved by males and females, despite the former being smaller. Fewer males moved overland, but this was no different from the sex bias found in the population. In laboratory performance trials, we found that males outperformed females, in both distance moved and time to exhaustion, when corrected for size. Overland movement occurred throughout the year, but reached peaks in spring and early summer when temporary water bodies were drying. Despite permanent impoundments being located within the study area, we found no evidence for migrations of animals between temporary and permanent water bodies. Our study provides the first dispersal kernel for X. laevis and suggests that it is similar to many non-pipid anurans with respect to dispersal.

  3. Modeling of DNA local parameters predicts encrypted architectural motifs in Xenopus laevis ribosomal gene promoter

    PubMed Central

    Roux-Rouquie, Magali; Marilley, Monique

    2000-01-01

    We have modeled local DNA sequence parameters to search for DNA architectural motifs involved in transcription regulation and promotion within the Xenopus laevis ribosomal gene promoter and the intergenic spacer (IGS) sequences. The IGS was found to be shaped into distinct topological domains. First, intrinsic bends split the IGS into domains of common but different helical features. Local parameters at inter-domain junctions exhibit a high variability with respect to intrinsic curvature, bendability and thermal stability. Secondly, the repeated sequence blocks of the IGS exhibit right-handed supercoiled structures which could be related to their enhancer properties. Thirdly, the gene promoter presents both inherent curvature and minor groove narrowing which may be viewed as motifs of a structural code for protein recognition and binding. Such pre-existing deformations could simply be remodeled during the binding of the transcription complex. Alternatively, these deformations could pre-shape the promoter in such a way that further remodeling is facilitated. Mutations shown to abolish promoter curvature as well as intrinsic minor groove narrowing, in a variant which maintained full transcriptional activity, bring circumstantial evidence for structurally-preorganized motifs in relation to transcription regulation and promotion. Using well documented X.laevis rDNA regulatory sequences we showed that computer modeling may be of invaluable assistance in assessing encrypted architectural motifs. The evidence of these DNA topological motifs with respect to the concept of structural code is discussed. PMID:10982860

  4. Modeling of DNA local parameters predicts encrypted architectural motifs in Xenopus laevis ribosomal gene promoter.

    PubMed

    Roux-Rouquie, M; Marilley, M

    2000-09-15

    We have modeled local DNA sequence parameters to search for DNA architectural motifs involved in transcription regulation and promotion within the Xenopus laevis ribosomal gene promoter and the intergenic spacer (IGS) sequences. The IGS was found to be shaped into distinct topological domains. First, intrinsic bends split the IGS into domains of common but different helical features. Local parameters at inter-domain junctions exhibit a high variability with respect to intrinsic curvature, bendability and thermal stability. Secondly, the repeated sequence blocks of the IGS exhibit right-handed supercoiled structures which could be related to their enhancer properties. Thirdly, the gene promoter presents both inherent curvature and minor groove narrowing which may be viewed as motifs of a structural code for protein recognition and binding. Such pre-existing deformations could simply be remodeled during the binding of the transcription complex. Alternatively, these deformations could pre-shape the promoter in such a way that further remodeling is facilitated. Mutations shown to abolish promoter curvature as well as intrinsic minor groove narrowing, in a variant which maintained full transcriptional activity, bring circumstantial evidence for structurally-preorganized motifs in relation to transcription regulation and promotion. Using well documented X. laevis rDNA regulatory sequences we showed that computer modeling may be of invaluable assistance in assessing encrypted architectural motifs. The evidence of these DNA topological motifs with respect to the concept of structural code is discussed.

  5. Studies of Xenopus laevis mitochondrial DNA: D-loop mapping and characterization of DNA-binding proteins

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cairns, S.S.

    1987-01-01

    In X. laevis oocytes, mitochondrial DNA accumulates to 10/sup 5/ times the somatic cell complement, and is characterized by a high frequency of a triple-stranded displacement hoop structure at the origin of replication. To map the termini of the single strands, it was necessary to correct the nucleotide sequence of the D-loop region. The revised sequence of 2458 nucleotides contains 54 discrepancies in comparison to a previously published sequence. Radiolabeling of the nascent strands of the D-loop structure either at the 5' end or at the 3' end identifies a major species with a length of 1670 nucleotides. Cleavage ofmore » the 5' labeled strands reveals two families of ends located near several matches to an element, designated CSB-1, that is conserved in this location in several vertebrate genomes. Cleavage of 3' labeled strands produced one fragment. The unique 3' end maps to about 15 nucleotides preceding the tRNA/sup Pro/ gene. A search for proteins which may bind to mtDNA in this region to regulate nucleic acid synthesis has identified three activities in lysates of X. laevis mitochondria. The DNA-binding proteins were assayed by monitoring their ability to retard the migration of labeled double- or single-stranded DNA fragments in polyacrylamide gels. The DNA binding preference was determined by competition with an excess of either ds- or ssDNA.« less

  6. The colloidal thyroxine (T4) ring as a novel biomarker of perchlorate exposure in the African clawed frog Xenopus laevis

    USGS Publications Warehouse

    Hu, F.; Sharma, Bibek; Mukhi, S.; Patino, R.; Carr, J.A.

    2006-01-01

    The purpose of this study was to determine if changes in colloidal thyroxine (T4) immunoreactivity can be used as a biomarker of perchlorate exposure in amphibian thyroid tissue. Larval African clawed frogs (Xenopus laevis) were exposed to 0, 1, 8, 93, and 1131 ??g perchlorate/l for 38 and 69 days to cover the normal period of larval development and metamorphosis. The results of this study confirmed the presence of an immunoreactive colloidal T4 ring in thyroid follicles of X. laevis and demonstrated that the intensity of this ring is reduced in a concentration-dependent manner by perchlorate exposure. The smallest effective concentration of perchlorate capable of significantly reducing colloidal T4 ring intensity was 8 ??g perchlorate/l. The intensity of the immunoreactive colloidal T4 ring is a more sensitive biomarker of perchlorate exposure than changes in hind limb length, forelimb emergence, tail resorption, thyrocyte hypertrophy, or colloid depletion. We conclude that the colloidal T4 ring can be used as a sensitive biomarker of perchlorate-induced thyroid disruption in amphibians. ?? Copyright 2006 Oxford University Press.

  7. Boric Acid Is Reproductively Toxic to Adult Xenopus laevis, but Not Endocrine Active.

    PubMed

    Fort, Douglas J; Fort, Troy D; Mathis, Michael B; Ball, R Wayne

    2016-11-01

    The potential reproductive and endocrine toxicity of boric acid (BA) in the African clawed frog, Xenopus laevis, was evaluated using a 30-day exposure of adult frogs. Adult female and male frogs established as breeders were exposed to a culture water control and 4 target (nominal) test concentrations [5.0, 7.5, 10.0, and 15 mg boron (B)/L, equivalent to 28.5, 42.8, 57.0, and 85.5 mg BA/L] using flow-through diluter exposure system. The primary endpoints measured were adult survival, growth (weight and snout-vent length [SVL]), necropsy data, reproductive fecundity, and development of progeny (F1) from the exposed frogs. Necropsy endpoints included gonad weight, gonado-somatic index (GSI), ovary profile (oocyte normalcy and stage distribution), sperm count, and dysmorphology. Endocrine endpoints included plasma estradiol (E2), testosterone (T), dihydrotestosteone (DHT), gonadal CYP 19 (aromatase), and gonadal 5α-reductase (5-AR). BA exposure to adult female X. laevis increased the proportion of immature oocytes (< stage II) in the ovaries of females, reduced sperm counts and increased sperm cell dysmorphology frequency in male frogs exposed to 15 mg B/L. No effects on the other general, developmental (F1), or endocrine endpoints were observed. Based on the results of the present study, the no observed adverse effects concentration (NOAEC) for the reproductive endpoints was 10 mg B/L; and 15 mg B/L for reproductive fecundity, F1 embryo larval development, and endocrine function. These results confirmed that although BA is capable of inducing reproductive toxicity at high concentrations, it is not an endocrine disrupting agent. © The Author 2016. Published by Oxford University Press on behalf of the Society of Toxicology. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  8. Sex chromosome differentiation and the W- and Z-specific loci in Xenopus laevis.

    PubMed

    Mawaribuchi, Shuuji; Takahashi, Shuji; Wada, Mikako; Uno, Yoshinobu; Matsuda, Yoichi; Kondo, Mariko; Fukui, Akimasa; Takamatsu, Nobuhiko; Taira, Masanori; Ito, Michihiko

    2017-06-15

    Genetic sex-determining systems in vertebrates include two basic types of heterogamety; XX (female)/XY (male) and ZZ (male)/ZW (female) types. The African clawed frog Xenopus laevis has a ZZ/ZW-type sex-determining system. In this species, we previously identified a W-specific sex (female)-determining gene dmw, and specified W and Z chromosomes, which could be morphologically indistinguishable (homomorphic). In addition to dmw, we most recently discovered two genes, named scanw and ccdc69w, and one gene, named capn5z in the W- and Z-specific regions, respectively. In this study, we revealed the detail structures of the W/Z-specific loci and genes. Sequence analysis indicated that there is almost no sequence similarity between 278kb W-specific and 83kb Z-specific sequences on chromosome 2Lq32-33, where both the transposable elements are abundant. Synteny and phylogenic analyses indicated that all the W/Z-specific genes might have emerged independently. Expression analysis demonstrated that scanw and ccdc69w or capn5z are expressed in early differentiating ZW gonads or testes, thereby suggesting possible roles in female or male development, respectively. Importantly, the sex-determining gene (SDG) dmw might have been generated after allotetraploidization, thereby indicating the construction of the new sex-determining system by dmw after species hybridization. Furthermore, by direct genotyping, we confirmed that diploid WW embryos developed into normal female frogs, which indicate that the Z-specific region is not essential for female development. Overall, these findings indicate that sex chromosome differentiation has started, although no heteromorphic sex chromosomes are evident yet, in X. laevis. Homologous recombination suppression might have promoted the accumulation of mutations and transposable elements, and enlarged the W/Z-specific regions, thereby resulting in differentiation of the W/Z chromosomes. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Expressed ryanodine receptor can substitute for the inositol 1,4,5-trisphosphate receptor in Xenopus laevis oocytes during progesterone-induced maturation.

    PubMed

    Kobrinsky, E; Ondrias, K; Marks, A R

    1995-12-01

    Two structurally related forms of intracellular calcium release channels that can mediate the release of intracellular calcium have been identified: the ryanodine receptors (RyR) and the inositol 1,4,5-trisphosphate receptors (IP3R). Each channel responds to distinct pathways for activation. The IP3R is activated by IP3 and the RyR is thought to be activated by calcium or by another second messenger cADP ribose. It has been proposed that each type of channel subserves a specialized pool of intracellular calcium, and it is not understood why some cell types require more than one form of intracellular calcium release channel. The present study was designed to examine whether the RyR can substitute for the IP3R during oocyte maturation. IP3R expression was inhibited in Xenopus laevis oocytes using antisense oligonucleotides. These oocytes, with reduced levels of IP3R, demonstrated a marked delay in the time course of progesterone-induced maturation. The cloned skeletal muscle RyR1 was then expressed in X. laevis oocytes that were deficient in IP3R. Functional studies showed that the properties of the cloned RyR1, expressed in oocytes, were comparable to those of the native RyR1. X. laevis oocytes deficient in IP3R, but expressing RyR1, were able to undergo progesterone-induced maturation with a time course comparable to that seen in wild-type oocytes when caffeine was used to activate RyR and induce intracellular calcium release. These studies show that RyR1 can substitute for the IP3R as the intracellular calcium release channel required for Xenopus oocyte maturation and that intracellular calcium release is important for controlling the rate of progesterone-induced maturation.

  10. Planar induction of anteroposterior pattern in the developing central nervous system of Xenopus laevis

    NASA Technical Reports Server (NTRS)

    Doniach, T.; Phillips, C. R.; Gerhart, J. C.

    1992-01-01

    It has long been thought that anteroposterior (A-P) pattern in the vertebrate central nervous system is induced in the embryo's dorsal ectoderm exclusively by signals passing vertically from underlying, patterned dorsal mesoderm. Explants from early gastrulae of the frog Xenopus laevis were prepared in which vertical contact between dorsal ectoderm and mesoderm was prevented but planar contact was maintained. In these, four position-specific neural markers (engrailed-2, Krox-20, XlHbox 1, and XlHbox 6) were expressed in the ectoderm in the same A-P order as in the embryo. Thus, planar signals alone, following a path available in the normal embryo, can induce A-P neural pattern.

  11. Regeneration of Xenopus laevis spinal cord requires Sox2/3 expressing cells

    PubMed Central

    Muñoz, Rosana; Edwards-Faret, Gabriela; Moreno, Mauricio; Zuñiga, Nikole; Cline, Hollis; Larraín, Juan

    2016-01-01

    Spinal cord regeneration is very inefficient in humans, causing paraplegia and quadriplegia. Studying model organisms that can regenerate the spinal cord in response to injury could be useful for understanding the cellular and molecular mechanisms that explain why this process fails in humans. Here, we use Xenopus laevis as a model organism to study spinal cord repair. Histological and functional analyses showed that larvae at pre-metamorphic stages restore anatomical continuity of the spinal cord and recover swimming after complete spinal cord transection. These regenerative capabilities decrease with onset of metamorphosis. The ability to study regenerative and non-regenerative stages in Xenopus laevis makes it a unique model system to study regeneration. We studied the response of Sox2/3 expressing cells to spinal cord injury and their function in the regenerative process. We found that cells expressing Sox2 and/or Sox3 are present in the ventricular zone of regenerative animals and decrease in non-regenerative froglets. Bromodeoxyuridine (BrdU) experiments and in vivo time-lapse imaging studies using green fluorescent protein (GFP) expression driven by the Sox3 promoter showed a rapid, transient and massive proliferation of Sox2/3+ cells in response to injury in the regenerative stages. The in vivo imaging also demonstrated that Sox2/3+ neural progenitor cells generate neurons in response to injury. In contrast, these cells showed a delayed and very limited response in non-regenerative froglets. Sox2 knockdown and overexpression of a dominant negative form of Sox2 disrupts locomotor and anatomical-histological recovery. We also found that neurogenesis markers increase in response to injury in regenerative but not in non-regenerative animals. We conclude that Sox2 is necessary for spinal cord regeneration and suggest a model whereby spinal cord injury activates proliferation of Sox2/3 expressing cells and their differentiation into neurons, a mechanism that is

  12. Nondestructive Imaging of Internal Structures of Frog (Xenopus laevis) Embryos by Shadow-Projection X-Ray Microtomography

    NASA Astrophysics Data System (ADS)

    Aoki, Sadao; Yoneda, Ikuo; Nagai, Takeharu; Ueno, Naoto; Murakami, Kazuo

    1994-04-01

    Nondestructive high-resolution imaging of frog ( Xenopus laevis) embryos has been developed by X-ray microtomography. Shadow-projection X-ray microtomography with a brilliant fine focus laboratory X-ray source could image fine structures of Xenopus embryos which were embedded in paraffin wax. The imaging system enabled us to not only distinguish endoderm from ectoderm at the gastrula stage, but also to obtain a cross-section view of the tail bud embryo showing muscle, notochord and neural tube without staining. Furthermore, the distribution of myosin was also imaged in combination with whole-mount immunohistochemistry.

  13. Xenopus laevis - A success story in biological research in Space

    NASA Astrophysics Data System (ADS)

    Horn, E.

    A feature of sensory, neuronal and motor systems is the existence of a critical period during their development. Environmental modifications, in particular stimulus depri-vation, during this period of life affects development in a long-term manner. For gravity sensory systems, space flights offer the only opportunity for deprivation conditions. Studies in the amphibian Xenopus laevis presented the most complete picture. The presentation demonstrates the importance of Xenopus laevis as an ex-perimental model animal in the past and even for future research in Space. Studies are presented which range from fertilization in Space and anatomical studies during early development under weightlessness up to post-flight studies on the anatomy of the peripheral sense organ, the spinal motor activity and behavior. Gravity depriva-tion induces anatomical as well as behavioral and neurophysiological modifications, which are normalized either during flight (thickening of the blastocoel roof) or after reentry in 1g-conditions (swimming and reflex behavior, spinal motor activity). The physiological changes can be explained by mechanisms of physiological adaptation. However, the studies also revealed stages which were insensitive to gravity depriva-tion; they point to the existence of a critical period. Observations on morphological mal-formations are described which are reversible after termination of microgravity and which are linked to a depression of vestibular reflex behavior. They might be caused by a competition between dorsalization and ventralization inducing growth factors. This observation offers the possibility for a genetic approach in finding ba-sics for microgravity effects on the development of Xenopus, and in a general frame, on the development of vertebrates including men. At the present stage of research, it remains open whether adaptive processes during exposure to altered gravity or the existence of a critical period in vestibular development are responsible for

  14. Relative information content of polymorphic microsatellites and mitochondrial DNA for inferring dispersal and population genetic structure in the olive sea snake, Aipysurus laevis.

    PubMed

    Lukoschek, V; Waycott, M; Keogh, J S

    2008-07-01

    Polymorphic microsatellites are widely considered more powerful for resolving population structure than mitochondrial DNA (mtDNA) markers, particularly for recently diverged lineages or geographically proximate populations. Weaker population subdivision for biparentally inherited nuclear markers than maternally inherited mtDNA may signal male-biased dispersal but can also be attributed to marker-specific evolutionary characteristics and sampling properties. We discriminated between these competing explanations with a population genetic study on olive sea snakes, Aipysurus laevis. A previous mtDNA study revealed strong regional population structure for A. laevis around northern Australia, where Pleistocene sea-level fluctuations have influenced the genetic signatures of shallow-water marine species. Divergences among phylogroups dated to the Late Pleistocene, suggesting recent range expansions by previously isolated matrilines. Fine-scale population structure within regions was, however, poorly resolved for mtDNA. In order to improve estimates of fine-scale genetic divergence and to compare population structure between nuclear and mtDNA, 354 olive sea snakes (previously sequenced for mtDNA) were genotyped for five microsatellite loci. F statistics and Bayesian multilocus genotype clustering analyses found similar regional population structure as mtDNA and, after standardizing microsatellite F statistics for high heterozygosities, regional divergence estimates were quantitatively congruent between marker classes. Over small spatial scales, however, microsatellites recovered almost no genetic structure and standardized F statistics were orders of magnitude smaller than for mtDNA. Three tests for male-biased dispersal were not significant, suggesting that recent demographic expansions to the typically large population sizes of A. laevis have prevented microsatellites from reaching mutation-drift equilibrium and local populations may still be diverging.

  15. Morphometric and genetic analysis of Arcella intermedia and Arcella intermedia laevis (Amoebozoa, Arcellinida) illuminate phenotypic plasticity in microbial eukaryotes.

    PubMed

    Porfírio-Sousa, Alfredo L; Ribeiro, Giulia M; Lahr, Daniel J G

    2017-04-01

    Testate amoebae are eukaryotic microorganisms characterized by the presence of an external shell (test). The shell morphology is used as a diagnostic character, but discordance between morphological and molecular data has been demonstrated in groups of arcellinids (Amoebozoa), one of the principal groups of testate amoebae. Morphology of the test is supposed to differentiate genera and species and it is applied in ecological, monitoring and paleontological studies. However, if phenotype does not reflect genotype, conclusions in these types of studies become severely impaired. The objective of this work is to evaluate the morphometrical and morphological variation of the closely related and morphologically similar taxa Arcella intermedia laevis Tsyganov and Mazei, 2006 and Arcella intermedia (Deflandre 1928) Tsyganov and Mazei, 2006 in nature and in cultured individuals and see how these are correlated with molecular data. Our results demonstrate that phenotypic plasticity in Arcella intermedia make morphological distinctions impossible in both taxa. Arcella intermedia and Arcella intermedia laevis are molecularly identical for SSU rDNA and a mitochondrial molecular marker (NAD9/7). We conclude that morphological techniques alone cannot identify phenotypic plasticity from natural populations. More work is clearly needed to better understand the morphological, morphometric and molecular variability in these organisms. Copyright © 2016 Elsevier GmbH. All rights reserved.

  16. Anticariogenic activity of the active fraction from Isertia laevis against S. mutans and S. sobrinus: comparison of two extraction methods.

    PubMed

    Téllez, Nohemí; Téllez, Mayra; Perdomo, Margarita; Alvarado, Andrea; Gamboa, Fredy

    2010-01-01

    Dental caries is considered a multi-factorial, infectious, chronic, localized, post-eruptive, transmissible disease that leads to the destruction of dental hard tissue. The recognition of Streptococcus mutans as the major bacterial species involved in dental caries has led to the implementation of prevention and control measures for eliminating or reducing it in oral cavity. The main goal of research on medicinal plants is the search for substances or compounds with antimicrobial activity. The aim of this study was to evaluate the antimicrobial activity of fractions obtained by two methods from Isertia laevis against S. mutans and S. sobrinus. The plant material was collected in Medina (Colombia), at an elevation of 550 meters above sea level. From the ethanol extract of leaves of I. laevis, fractions were obtained by two methods: extraction by column vacuum chromatography (CVC) and extraction by continuous liquid/liquid partitioning (CLLP). The evaluation of the antimicrobial activity of fractions against S. mutans and S. sobrinus was performed by well diffusion and bioautography assays. From the CVC technique, only the methanol and methanol-dichloromethane fractions showed activity against S. mutans and S. sobrinus, with a minimum inhibitory concentration of 2 mg/well. From the CLLP technique, only the dichloromethane fraction showed activity against both microorganisms, with a minimum inhibitory concentration of 1 mg/well. Compounds C1 and C2 were isolated from the three active fractions, and showed a minimum inhibitory concentration of 0.4 mg/well for S. mutans and S. sobrinus, with zones of inhibition measuring 6.5 and 6.2 mm, respectively. 1) the three active fractions of I. laevis showed activity against S. mutans and S. sobrinus, 2) compounds C1 and C2 were presen equally in the three active fractions showing activity against the two bacteria, 3) compounds C1 and C2 may be triterpenoid and/or steroidal saponin structures, and 4) the two extraction methods

  17. Atomic force microscopy on plasma membranes from Xenopus laevis oocytes containing human aquaporin 4.

    PubMed

    Orsini, Francesco; Santacroce, Massimo; Cremona, Andrea; Gosvami, Nitya N; Lascialfari, Alessandro; Hoogenboom, Bart W

    2014-11-01

    Atomic force microscopy (AFM) is a unique tool for imaging membrane proteins in near-native environment (embedded in a membrane and in buffer solution) at ~1 nm spatial resolution. It has been most successful on membrane proteins reconstituted in 2D crystals and on some specialized and densely packed native membranes. Here, we report on AFM imaging of purified plasma membranes from Xenopus laevis oocytes, a commonly used system for the heterologous expression of membrane proteins. Isoform M23 of human aquaporin 4 (AQP4-M23) was expressed in the X. laevis oocytes following their injection with AQP4-M23 cRNA. AQP4-M23 expression and incorporation in the plasma membrane were confirmed by the changes in oocyte volume in response to applied osmotic gradients. Oocyte plasma membranes were then purified by ultracentrifugation on a discontinuous sucrose gradient, and the presence of AQP4-M23 proteins in the purified membranes was established by Western blotting analysis. Compared with membranes without over-expressed AQP4-M23, the membranes from AQP4-M23 cRNA injected oocytes showed clusters of structures with lateral size of about 10 nm in the AFM topography images, with a tendency to a fourfold symmetry as may be expected for higher-order arrays of AQP4-M23. In addition, but only infrequently, AQP4-M23 tetramers could be resolved in 2D arrays on top of the plasma membrane, in good quantitative agreement with transmission electron microscopy analysis and the current model of AQP4. Our results show the potential and the difficulties of AFM studies on cloned membrane proteins in native eukaryotic membranes. Copyright © 2014 John Wiley & Sons, Ltd.

  18. Exposure to butachlor causes thyroid endocrine disruption and promotion of metamorphosis in Xenopus laevis.

    PubMed

    Li, Shuying; Li, Meng; Wang, Qiangwei; Gui, Wenjun; Zhu, Guonian

    2016-06-01

    Butachlor is extensively applied in rice paddy ecosystem in china, and has been widespread contaminant in the aquatic environment. Here, Xenopus laevis was used for the evaluation of teratogenesis developmental toxicity, and disruption of thyroid system when exposure to different concentrations of butachlor by window phase exposure. Acute toxicity investigation shown that 96 h-LC50 value of butachlor was 1.424 mg L(-1) and 0.962 mg L(-1) for tadpoles (starting from stages 46/47) and embryos (starting from stages 8/9), respectively. Exposure to butachlor caused malformation, including abnormal eye, pericardial edema, enlarged proctodaeum and bent tail. Window phase exposure test indicated that butachlor significantly promote the contents of whole-body thyroid hormones (THs, T3 and T4) at higher levels, indicating thyroid endocrine disruption. At 7 days, exposure to butachlor up-regulated the mRNA expression of genes involved in THs synthesis and metabolism (tshα, tg, tpo and dio1) and THs receptors (trα and trβ). At 14 days, up-regulation of the mRNA expression of genes related to THs synthesis and metabolism (tshα, tshβ, tg, tpo, dio1, dio2 and ttr) and THs receptors (trβ) were also observed after the exposure to butachlor. At 21 days, butachlor up-regulated the mRNA expression of tshα, tg, tpo genes and down-regulated the mRNA expression of tshβ, tg, dio1, ttr and trα genes. These results showed that butachlor could change the mRNA expression of genes involved in the HPT axis and increase whole-body thyroid hormones levels of X. laevis tadpoles in a dose- and time-dependent manner, causing thyroid endocrine disruption and developmental toxicity. Copyright © 2016 Elsevier Ltd. All rights reserved.

  19. Single olfactory organ associated with prosencephalic malformation and cyclopia in a Xenopus laevis tadpole.

    PubMed

    Magrassi, L; Graziadei, P P

    1987-06-02

    A cyclops Xenopus laevis tadpole with a single olfactory organ is described. At a stage comparable to 48, the telencephalon was severely atrophic and only the region where the olfactory fibres terminated appeared to have the cytoarchitecture of the olfactory bulb. In this animal the central nervous system (CNS) appeared normally developed only posterior to the preoptic area. The hypothesis of a diencephalic origin of the region where the olfactory fibres terminated is discussed in the light of our previous results of olfactory placode transplantation. By analogy between this case and other malformations (cyclopia, holoprosencephaly) in higher vertebrates and humans, the need is emphasized for a more precise anatomical description of the olfactory input in related malformations.

  20. Urocortins of the South African clawed frog, Xenopus laevis: conservation of structure and function in tetrapod evolution.

    PubMed

    Boorse, Graham C; Crespi, Erica J; Dautzenberg, Frank M; Denver, Robert J

    2005-11-01

    Several corticotropin-releasing factor (CRF) family genes have been identified in vertebrates. Mammals have four paralogous genes that encode CRF or the urocortins 1, 2, and 3. In teleost fishes, a CRF, urotensin I (a fish ortholog of mammalian urocortin 1) and urocortin 3 have been identified, suggesting that at least three of the four mammalian lineages arose in a common ancestor of modern bony fishes and tetrapods. Here we report the isolation of genes orthologous to mammalian urocortin 1 and urocortin 3 from the South African clawed frog, Xenopus laevis. We characterize the pharmacology of the frog peptides and show that X. laevis urocortin 1 binds to and activates the frog CRF1 and CRF2 receptors at picomolar concentrations. Similar to mammals, frog urocortin 3 is selective for the CRF2 receptor. Only frog urocortin 1 binds to the CRF-binding protein, although with significantly lower affinity than frog CRF. Both urocortin genes are expressed in brain, pituitary, heart, and kidney of juvenile frogs; urocortin 1 is also expressed in skin. We also identified novel urocortin sequences in the genomes of pufferfish, zebrafish, chicken, and dog. Phylogenetic analysis supports the view that four paralogous lineages of CRF-like peptides arose before the divergence of the actinopterygian and sarcopterygian fishes. Our findings show that the functional relationships among CRF ligands and binding proteins, and their anorexigenic actions mediated by the CRF2 receptor, arose early in vertebrate evolution.

  1. Nucleotide sequence of the L1 ribosomal protein gene of Xenopus laevis: remarkable sequence homology among introns.

    PubMed Central

    Loreni, F; Ruberti, I; Bozzoni, I; Pierandrei-Amaldi, P; Amaldi, F

    1985-01-01

    Ribosomal protein L1 is encoded by two genes in Xenopus laevis. The comparison of two cDNA sequences shows that the two L1 gene copies (L1a and L1b) have diverged in many silent sites and very few substitution sites; moreover a small duplication occurred at the very end of the coding region of the L1b gene which thus codes for a product five amino acids longer than that coded by L1a. Quantitatively the divergence between the two L1 genes confirms that a whole genome duplication took place in Xenopus laevis approximately 30 million years ago. A genomic fragment containing one of the two L1 gene copies (L1a), with its nine introns and flanking regions, has been completely sequenced. The 5' end of this gene has been mapped within a 20-pyridimine stretch as already found for other vertebrate ribosomal protein genes. Four of the nine introns have a 60-nucleotide sequence with 80% homology; within this region some boxes, one of which is 16 nucleotides long, are 100% homologous among the four introns. This feature of L1a gene introns is interesting since we have previously shown that the activity of this gene is regulated at a post-transcriptional level and it involves the block of the normal splicing of some intron sequences. Images Fig. 3. Fig. 5. PMID:3841512

  2. Polystyrene nanoparticles affect Xenopus laevis development

    NASA Astrophysics Data System (ADS)

    Tussellino, Margherita; Ronca, Raffaele; Formiggini, Fabio; Marco, Nadia De; Fusco, Sabato; Netti, Paolo Antonio; Carotenuto, Rosa

    2015-02-01

    Exposing living organisms to nanoparticulates is potentially hazardous, in particular when it takes place during embryogenesis. In this investigation, we have studied the effects of 50-nm-uncoated polystyrene nanoparticles (PSNPs) as a model to investigate the suitability of their possible future employments. We have used the standardized Frog Embryo Teratogenesis Assay- Xenopus test during the early stages of larval development of Xenopus laevis, and we have employed either contact exposure or microinjections. We found that the embryos mortality rate is dose dependent and that the survived embryos showed high percentage of malformations. They display disorders in pigmentation distribution, malformations of the head, gut and tail, edema in the anterior ventral region, and a shorter body length compared with sibling untreated embryos. Moreover, these embryos grow more slowly than the untreated embryos. Expressions of the mesoderm markers, bra (T-box Brachyury gene), myod1 (myogenic differentiation1), and of neural crest marker sox9 (sex SRY (determining region Y-box 9) transcription factor sox9), are modified. Confocal microscopy showed that the nanoparticles are localized in the cytoplasm, in the nucleus, and in the periphery of the digestive gut cells. Our data suggest that PSNPs are toxic and show a potential teratogenic effect for Xenopus larvae. We hypothesize that these effects may be due either to the amount of NPs that penetrate into the cells and/or to the "corona" effect caused by the interaction of PSNPs with cytoplasm components. The three endpoints of our study, i.e., mortality, malformations, and growth inhibition, suggest that the tests we used may be a powerful and flexible bioassay in evaluating pollutants in aquatic embryos.

  3. In vivo Assessment and Potential Diagnosis of Xenobiotics that Perturb the Thyroid Pathway: Proteomic Analysis of Xenopus laevis Brain Tissue following Exposure to Model T4 Inhibitors

    EPA Science Inventory

    As part of a multi-endpoint systems approach to develop comprehensive methods for assessing endocrine stressors in vertebrates, differential protein profiling was used to investigate expression profiles in the brain of an amphibian model (Xenopus laevis) following in vivo exposur...

  4. Potential ecotoxic effects of polychlorinated biphenyls on Xenopus laevis.

    PubMed

    Qin, Zhan-Fen; Zhou, Jing-Ming; Cong, Lin; Xu, Xiao-Bai

    2005-10-01

    We examined potential ecotoxic effects of polychlorinated biphenyl (PCB)3, PCB5, Aroclor 1254, and Aroclor 1242 on Xenopus laevis. Tadpoles were exposed to PCBs from stage 46/47 (system of Nieuwkoop and Faber) to the completion of metamorphosis. We demonstrated, to our knowledge for the first time, forelimb malformations caused by PCBs (malformation rate, > 70%). The malformed forelimbs were fixed in the adduction-backward rotation position and could not move. Therefore, malformed male frogs were destined to have no offspring, because they could not grasp the females with their forelimbs to mate. Alcian blue-alizarin red double-staining indicated that the forelimb malformation resulted from the shoulder abnormality. Compared with the normal shoulder joint, the proximal humerus with the humerus inter-rotated 90 degrees in the abnormal shoulder joint. Moreover, testes from more than a third of male frogs with exposed to PCBs exhibited feminization to different degrees at gross morphology and histology, with fewer or abnormal spermatogonia and oocytes. Gonadal abnormalities would lead directly to reproductive dysfunction and population decline. These results suggest that PCBs have potentially ecotoxic effects on amphibian populations. We infer that PCBs could play roles in amphibian malformations and population declines, at least at sites that are polluted heavily with PCBs.

  5. Evaluation and Refinement of Euthanasia Methods for Xenopus laevis

    PubMed Central

    Torreilles, Stéphanie L; McClure, Diane E; Green, Sherril L

    2009-01-01

    The most common method of euthanasia for Xenopus species is by immersion in tricaine methane sulfonate solution (MS222). A wide range of doses of MS222 (0.5 to 5 g/L) have been recommended, but few reports describe dose–response testing, the time to loss of consciousness, or the reliability of euthanasia. The objective of this study is to evaluate the efficacy of immersing individual and groups of frogs in MS222 at concentrations ranging from 1 to 5 g/L for euthanasia and of 3 less-common methods: intracoelomic injection of MS222, intracoelomic injection of sodium pentobarbital with phenytoin, and ventral cutaneous application of benzocaine gel. Our results indicate that immersion for at least 1 h in a 5-g/L buffered solution of MS222, intracoelomic injection of 1100 mg/kg sodium pentobarbital with sodium phenytoin (equivalent to 0.3 mL solution per frog), or ventral cutaneous application of 182 mg/kg benzocaine (equivalent to a 2 cm × 1 mm of 20% benzocaine gel) is necessary to euthanize adult X. laevis and ensure complete cessation of the heartbeat without recovery. These doses are considerably higher than those previously recommended for this species. PMID:19807972

  6. Computational study of antimalarial pyrazole alkaloids from Newbouldia laevis.

    PubMed

    Mammino, Liliana; Bilonda, Mireille K

    2014-11-01

    Six pyrazole alkaloids of natural origin (isolated from Newbouldia laevis in DR Congo) that exhibit antimalarial activity-namely withasomnine, newbouldine, and their para-hydroxy and -methoxy derivatives-were investigated theoretically. The nitro derivatives of withasomnine and para-hydroxywithasomnine, which show enhanced antimalarial activity, were also studied in this manner. A thorough conformational study was performed in vacuo and in three solvents (chloroform, acetonitrile, and water) at different levels of theory (HF, DFT/B3LYP, and MP2) using different basis sets. Adducts with explicit water molecules were calculated at the HF level. Due to the rigidity of the pyrazole system and the benzene ring, the only factor that influences the energies of withasomnine and newbouldine is the relative orientation of the two ring systems; two orientations are equally preferred. The para-hydroxy and -methoxy derivatives show a preference for a planar orientation of the OH and OC bonds. The main stabilizing influence on the nitro derivative of para-hydroxywithasomnine is the intramolecular hydrogen bond between the two consecutive functional groups. The calculated adducts show the preferred arrangements of water molecules in the vicinity of the N atoms of the pyrazole system and, for the derivatives, also in the vicinity of the substituents on the benzene ring.

  7. Nucleosome Translational Position, Not Histone Acetylation, Determines TFIIIA Binding to Nucleosomal Xenopus laevis 5S rRNA Genes

    PubMed Central

    Howe, LeAnn; Ausió, Juan

    1998-01-01

    We sought to study the binding constraints placed on the nine-zinc-finger protein transcription factor IIIA (TFIIIA) by a histone octamer. To this end, five overlapping fragments of the Xenopus laevis oocyte and somatic 5S rRNA genes were reconstituted into nucleosomes, and it was subsequently shown that nucleosome translational positioning is a major determinant of the binding of TFIIIA to the 5S rRNA genes. Furthermore, it was found that histone acetylation cannot override the TFIIIA binding constraints imposed by unfavorable translational positions. PMID:9488430

  8. Mitochondrial DNA diversity in the acanthocephalan Prosthenorchis elegans in Colombia based on cytochrome c oxidase I (COI) gene sequence.

    PubMed

    Falla, Ana Carolina; Brieva, Claudia; Bloor, Paul

    2015-12-01

    Prosthenorchis elegans is a member of the Phylum Acanthocephala and is an important parasite affecting New World Primates in the wild in South America and in captivity around the world. It is of significant management concern due to its pathogenicity and mode of transmission through intermediate hosts. Current diagnosis of P. elegans is based on the detection of eggs by coprological examination. However, this technique lacks both specificity and sensitivity, since eggs of most members of the genus are morphologically indistinguishable and shed intermittently, making differential diagnosis difficult, and coprological examinations are often negative in animals severely infected at death. We examined sequence variation in 633 bp of mitochondrial DNA (mtDNA) cytochrome c oxidase I (COI) sequence in 37 isolates of P. elegans from New World monkeys (Saguinus leucopus and Cebus albifrons) in Colombia held in rescue centers and from the wild. Intraspecific divergence ranged from 0.0 to 1.6% and was comparable with corresponding values within other species of acanthocephalans. Furthermore, comparisons of patterns of sequence divergence within the Acanthocephala suggest that Prosthenorchis represents a separate genus within the Oligacanthorhynchida. Six distinct haplotypes were identified within P. elegans which grouped into one of two well-supported mtDNA haplogroups. No association between haplogroup/haplotype, holding facility and species was found. This information will help pave the way to the development of molecular-based diagnostic tools for the detection of P. elegans as well as furthering research into the life cycle, intermediate hosts and epidemiological aspects of the species.

  9. Transmembrane voltage potential controls embryonic eye patterning in Xenopus laevis

    PubMed Central

    Pai, Vaibhav P.; Aw, Sherry; Shomrat, Tal; Lemire, Joan M.; Levin, Michael

    2012-01-01

    Uncovering the molecular mechanisms of eye development is crucial for understanding the embryonic morphogenesis of complex structures, as well as for the establishment of novel biomedical approaches to address birth defects and injuries of the visual system. Here, we characterize change in transmembrane voltage potential (Vmem) as a novel biophysical signal for eye induction in Xenopus laevis. During normal embryogenesis, a striking hyperpolarization demarcates a specific cluster of cells in the anterior neural field. Depolarizing the dorsal lineages in which these cells reside results in malformed eyes. Manipulating Vmem of non-eye cells induces well-formed ectopic eyes that are morphologically and histologically similar to endogenous eyes. Remarkably, such ectopic eyes can be induced far outside the anterior neural field. A Ca2+ channel-dependent pathway transduces the Vmem signal and regulates patterning of eye field transcription factors. These data reveal a new, instructive role for membrane voltage during embryogenesis and demonstrate that Vmem is a crucial upstream signal in eye development. Learning to control bioelectric initiators of organogenesis offers significant insight into birth defects that affect the eye and might have significant implications for regenerative approaches to ocular diseases. PMID:22159581

  10. Identification of metalloprotease/disintegrins in Xenopus laevis testis with a potential role in fertilization.

    PubMed

    Shilling, F M; Krätzschmar, J; Cai, H; Weskamp, G; Gayko, U; Leibow, J; Myles, D G; Nuccitelli, R; Blobel, C P

    1997-06-15

    Proteins containing a membrane-anchored metalloprotease domain, a disintegrin domain, and a cysteine-rich region (MDC proteins) are thought to play an important role in mammalian fertilization, as well as in somatic cell-cell interactions. We have identified PCR sequence tags encoding the disintegrin domain of five distinct MDC proteins from Xenopus laevis testis cDNA. Four of these sequence tags (xMDC9, xMDC11.1, xMDC11.2, and xMDC13) showed strong similarity to known mammalian MDC proteins, whereas the fifth (xMDC16) apparently represents a novel family member. Northern blot analysis revealed that the mRNA for xMDC16 was only expressed in testis, and not in heart, muscle, liver, ovaries, or eggs, whereas the mRNAs corresponding to the four other PCR products were expressed in testis and in some or all somatic tissues tested. The xMDC16 protein sequence, as predicted from the full-length cDNA, contains a metalloprotease domain with the active-site sequence HEXXH, a disintegrin domain, a cysteine-rich region, an EGF repeat, a transmembrane domain, and a short cytoplasmic tail. To study a potential role for these xMDC proteins in fertilization, peptides corresponding to the predicted integrin-binding domain of each protein were tested for their ability to inhibit X. laevis fertilization. Cyclic and linear xMDC16 peptides inhibited fertilization in a concentration-dependent manner, whereas xMDC16 peptides that were scrambled or had certain amino acid replacements in the predicted integrin-binding domain did not affect fertilization. Cyclic and linear xMDC9 peptides and linear xMDC13 peptides also inhibited fertilization similarly to xMDC16 peptides, whereas peptides corresponding to the predicted integrin-binding site of xMDC11.1 and xMDC11.2 did not. These results are discussed in the context of a model in which multiple MDC protein-receptor interactions are necessary for fertilization to occur.

  11. Antioxidant metabolism in Xenopus laevis embryos is affected by stratospheric balloon flight.

    PubMed

    Rizzo, Angela M; Rossi, Federica; Zava, Stefania; Montorfano, Gigliola; Adorni, Laura; Cotronei, Vittorio; Zanini, Alba; Berra, Bruno

    2007-07-01

    To test the effects of low levels of radiation from space on living organisms, we flew Xenopus laevis embryos at different stages of development on a stratospheric balloon (BI.R.BA mission). After recovery, different parameters were analyzed to assess the effects of flight, with particular regard to oxidative stress damage. Because of failed temperature control during flight, the flight shielded embryos (FC) could not be used for biochemical or morphological comparisons. In contrast, the incubation conditions (i.e. temperature, containers, volumes) for the flight embryos (F) were parallel to those for the ground controls. Mortality data show that younger embryos (16 h) flown on the balloon (F) are more sensitive to radiation exposure than older ones (40 h and 6 days). Exposure during flight lowered the antioxidant potential in all embryos, particularly older ones. These preliminary data demonstrate that flight on a stratospheric balloon might affect antioxidant metabolism, though it is not yet possible to correlate these results with low radiation exposure during flight.

  12. Functional joint regeneration is achieved using reintegration mechanism in Xenopus laevis

    PubMed Central

    Yamada, Shigehito

    2016-01-01

    Abstract A functional joint requires integration of multiple tissues: the apposing skeletal elements should form an interlocking structure, and muscles should insert into skeletal tissues via tendons across the joint. Whereas newts can regenerate functional joints after amputation, Xenopus laevis regenerates a cartilaginous rod without joints, a “spike.” Previously we reported that the reintegration mechanism between the remaining and regenerated tissues has a significant effect on regenerating joint morphogenesis during elbow joint regeneration in newt. Based on this insight into the importance of reintegration, we amputated frogs’ limbs at the elbow joint and found that frogs could regenerate a functional elbow joint between the remaining tissues and regenerated spike. During regeneration, the regenerating cartilage was partially connected to the remaining articular cartilage to reform the interlocking structure of the elbow joint at the proximal end of the spike. Furthermore, the muscles of the remaining part inserted into the regenerated spike cartilage via tendons. This study might open up an avenue for analyzing molecular and cellular mechanisms of joint regeneration using Xenopus. PMID:27499877

  13. A transgenic reporter under control of an es1 promoter/enhancer marks wound epidermis and apical epithelial cap during tail regeneration in Xenopus laevis tadpole.

    PubMed

    Sato, Kentaro; Umesono, Yoshihiko; Mochii, Makoto

    2018-01-15

    Rapid wound healing and subsequent formation of the apical epithelial cap (AEC) are believed to be required for successful appendage regeneration in amphibians. Despite the significant role of AEC in limb regeneration, its role in tail regeneration and the mechanisms that regulate the wound healing and AEC formation are not well understood. We previously identified Xenopus laevis es1, which is preferentially expressed in wounded regions, including the AEC after tail regeneration. In this study we established and characterized transgenic Xenopus laevis lines harboring the enhanced green fluorescent protein (EGFP) gene under control of an es1 gene regulatory sequence (es1:egfp). The EGFP reporter expression was clearly seen in several regions of the embryo and then declined to an undetectable level in larvae, recapitulating the endogenous es1 expression. After amputation of the tadpole tail, EGFP expression was re-activated at the edge of the stump epidermis and then increased in the wound epidermis (WE) covering the amputation surface. As the stump started to regenerate, the EGFP expression became restricted to the most distal epidermal region, including the AEC. EGFP was preferentially expressed in the basal or deep cells but not in the superficial cells of the WE and AEC. We performed a small-scale pharmacological screening for chemicals that affected the expression of EGFP in the stump epidermis after tail amputation. The EGFP expression was attenuated by treatment with an inhibitor for ERK, TGF-β or reactive oxygen species (ROS) signaling. These treatments also impaired wound closure of the amputation surface, suggesting that the three signaling activities are required for es1 expression in the WE and successful wound healing after tail amputation. These findings showed that es1:egfp Xenopus laevis should be a useful tool to analyze molecular mechanisms regulating wound healing and appendage regeneration. Copyright © 2017 Elsevier Inc. All rights reserved.

  14. Characterization of X-OCRL, a Xenopus laevis homologue of OCRL-1, the Lowe oculocerebrorenal syndrome candidate gene

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Reilly, D.S.; Nussbaum, R.L.

    1994-09-01

    The Lowe oculocerebrorenal syndrome (OCRL) is an X-linked disease characterized by congenital cataract, mental retardation, and renal tubular dysfunction. A candidate cDNA, OCRL-1, was identified by positional cloning and mutations in OCRL-1 have been detected in patients with Lowe syndrome. The OCRL-1 nucleotide sequence encodes a predicted protein of 968 amino acids and shares 51% amino acid identity with a human inositol polyphosphate-5-phosphatase. This suggests that the underlying defect in OCRL may be due to a defect in inositol phosphate metabolism. The isolation of OCRL-1 provides the opportunity to investigate its function through the use of animal model systems. Wemore » have isolated a partial cDNA clone encoding an OCRL-1 homologue, X-OCRL, from the South African clawed frog, Xenopus laevis. We used a portion of the human cDNA to screen a Xenopus laevis embryo cDNA library and isolated four positive clones. One clone, 42-5A, is a 650 bp insert with over 75% amino acid identity to the corresponding region of the human OCRL-1 sequence. 42-5A detects messenger RNA in adult Xenopus brain, stomach, small intestine, skin, muscle, lung, blood, and oviduct. X-OCRL messenger RNA is first detected during late gastrula and continues to be expressed throughout Xenopus development. In situ hybridization studies are underway to identify the cellular localization of X-OCRL expression in Xenopus embryos and adult tissues. We are especially interested in characterizing X-OCRL expression during formation of the amphibian lens since congenital cataracts are a constant feature of the human disease.« less

  15. Psf2 plays important roles in normal eye development in Xenopus laevis

    PubMed Central

    Walter, Brian E.; Perry, Kimberly J.; Fukui, Lisa; Malloch, Erica L.; Wever, Jason

    2008-01-01

    Purpose Psf2 (partner of Sld5 2) represents a member of the GINS (go, ichi, ni, san) heterotetramer [1] and functions in DNA replication as a “sliding clamp.” Previous in situ hybridization analyses revealed that Psf2 is expressed during embryonic development in a tissue-specific manner, including the optic cup (retina) and the lens [2]. This article provides an analysis of Psf2 function during eye development in Xenopus laevis. Methods A morpholino targeted to Psf2 mRNA was designed to knockdown Psf2 translation and was injected into specific embryonic cells during early cleavage stages in the frog, Xenopus laevis. Injected embryos were assayed for specific defects in morphology, cell proliferation, and apoptosis. Synthetic Psf2 RNA was also co-injected with the morpholino to rescue morpholino-mediated developmental defects. It is well known that reciprocal inductive interactions control the development of the optic cup and lens. Therefore, control- and morpholino-injected embryos were used for reciprocal transplantation experiments to distinguish the intrinsic role of Psf2 in the development of the optic cup (retina) versus the lens. Results Morpholino-mediated knockdown of Psf2 expression resulted in dosage-dependent phenotypes, which included microphthalmia, incomplete closure of the ventral retinal fissure, and retinal and lens dysgenesis. Defects were also observed in other embryonic tissues that normally express Psf2 including the pharyngeal arches and the otic vesicle, although other tissues that express Psf2 were not found to be grossly defective. Eye defects could be rescued by co-injection of synthetic Psf2 RNA. Examination of cell proliferation via an antibody against phospho-histone H3 S10P revealed no significant differences in the retina and lens following Psf2 knockdown. However, there was a significant increase in the level of apoptosis in retinal as well as forebrain tissues, as revealed by TUNEL (terminal deoxynucleotide transferase dUTP nick

  16. ( sup 125 I)Bolton-Hunter neuropeptide-Y-binding sites on folliculo-stellate cells of the pars intermedia of Xenopus laevis: A combined autoradiographic and immunocytochemical study

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    De Rijk, E.P.; Cruijsen, P.M.; Jenks, B.G.

    1991-02-01

    It has previously been established that neuropeptide-Y (NPY) is a potent inhibitor of alpha MSH release from the pars intermedia of the amphibian Xenopus laevis. The location of binding sites for NPY in the pars intermedia of the pituitary has now been studied with light microscopic autoradiography, using a dispersed cell labeling method with the specific NPY receptor ligand ({sup 125}I)Bolton-Hunter NPY. The majority of radioactive labeling was associated with folliculo-stellate cells; the percentage of labeling as well as the mean number of grains were approximately 5 times higher for folliculo-stellate cells than for melanotropes. An excess of nonlabeled NPYmore » drastically reduced radiolabeling of folliculo-stellate cells, but had no effect on the degree of labeling of melanotropes. These results show that folliculo-stellate cells of X. laevis possess specific binding sites for NPY and indicate that NPY exerts its inhibitory action on the release of alpha MSH in an indirect fashion, by acting on the folliculo-stellate cells.« less

  17. Calcium dependent current recordings in Xenopus laevis oocytes in microgravity

    NASA Astrophysics Data System (ADS)

    Wuest, Simon L.; Roesch, Christian; Ille, Fabian; Egli, Marcel

    2017-12-01

    Mechanical unloading by microgravity (or weightlessness) conditions triggers profound adaptation processes at the cellular and organ levels. Among other mechanisms, mechanosensitive ion channels are thought to play a key role in allowing cells to transduce mechanical forces. Previous experiments performed under microgravity have shown that gravity affects the gating properties of ion channels. Here, a method is described to record a calcium-dependent current in native Xenopus laevis oocytes under microgravity conditions during a parabolic flight. A 3-voltage-step protocol was applied to provoke a calcium-dependent current. This current increased with extracellular calcium concentration and could be reduced by applying extracellular gadolinium. The custom-made ;OoClamp; hardware was validated by comparing the results of the 3-voltage-step protocol to results obtained with a well-established two-electrode voltage clamp (TEVC). In the context of the 2nd Swiss Parabolic Flight Campaign, we tested the OoClamp and the method. The setup and experiment protocol worked well in parabolic flight. A tendency that the calcium-dependent current was smaller under microgravity than under 1 g condition could be observed. However, a conclusive statement was not possible due to the small size of the data base that could be gathered.

  18. Changes in hepatic levels of tyrosine aminotransferase messenger RNA during induction by hydrocortisone. [Xenopus laevis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nickol, J.M.; Lee, K.L.; Kenney, F.T.

    Messenger RNA specific for tyrosine aminotransferase was quantitated by microinjection into oocytes of Xenopus laevis. The heterologously translated enzyme was identified by specific immunoprecipitation and found to be identical with authentic aminotransferase by several criteria. The level of functional message present in rat liver increases during hydrocortisone induction, and this increase is directly proportional to the increased rate of synthesis of the enzyme. Kinetic analysis of the changes in tyrosine aminotransferase mRNA levels during induction and withdrawal indicates that the steroid does not affect the stability of the message, which has a half-life of approximately 1.2 h. Hydrocortisone, therefore, actsmore » to increase the rate of synthesis of the specific messenger by stimulating either its transcription or processing to functional mRNA.« less

  19. Dynein-Based Accumulation of Membranes Regulates Nuclear Expansion in Xenopus laevis Egg Extracts.

    PubMed

    Hara, Yuki; Merten, Christoph A

    2015-06-08

    Nuclear size changes dynamically during development and has long been observed to correlate with the space surrounding the nucleus, as well as with the volume of the cell. Here we combine an in vitro cell-free system of Xenopus laevis egg extract with microfluidic devices to systematically analyze the effect of spatial constraints. The speed of nuclear expansion depended on the available space surrounding the nucleus up to a threshold volume in the nanoliter range, herein referred to as the nuclear domain. Under spatial constraints smaller than this nuclear domain, the size of microtubule-occupied space surrounding the nucleus turned out to be limiting for the accumulation of membranes around the nucleus via the motor protein dynein, therefore determining the speed of nuclear expansion. This mechanism explains how spatial information surrounding the nucleus, such as the positioning of the nucleus inside the cell, can control nuclear expansion. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Adrenocorticotropin receptors: Functional expression from rat adrenal mRNA in Xenopus laevis oocytes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Mertz, L.M.; Catt, K.J.

    1991-10-01

    The adrenocorticotropin (ACTH) receptor, which binds corticotropin and stimulates adenylate cyclase and steroidogenesis in adrenocortical cells, was expressed in Xenopus laevis oocytes microinjected with rat adrenal poly(A){sup +} RNA. Expression of the ACTH receptor in individual stage 5 and 6 oocytes was monitored by radioimmunoassay of ligand-stimulated cAMP production. Injection of 5-40 ng of adrenal mRNA caused dose-dependent increases in ACTH-responsive cAMP production. Size fractionation of rat adrenal poly(A){sup +}RNA by sucrose density-gradient centrifugation revealed that mRNA encoding the ACTH receptor was present in the 1.1-to 2.0-kilobase fraction. These data indicate that ACTH receptors can be expressed from adrenal mRNAmore » in Xenopus oocytes and are fully functional in terms of ligand specificity and signal generation. The extracellular cAMP response to ACTH is a sensitive and convenient index of receptor expression. This system should permit more complete characterization and expression cloning of the ACTH receptor.« less

  1. Repair and recombination of X-irradiated plasmids in Xenopus laevis oocytes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sweigert, S.E.; Carroll, D.

    1990-11-01

    Plasmid DNA substrates were X-irradiated and injected into the nuclei of Xenopus laevis oocytes. After incubation for 20 h, DNA was recovered from the oocytes and analyzed simultaneously for repair and for intermolecular homologous recombination by electrophoresis and bacterial transformation. Oocyte-mediated repair of DNA strand breaks was observed with both methods. Using a repair-deficient mutant Escherichia coli strain and its repair-proficient parent as hosts for the transformation assay, we also demonstrated that oocytes repaired oxidative-type DNA base damage induced by X-rays. X-irradiation of a circular DNA stimulated its potential to recombine with a homologous linear partner. Recombination products were detectedmore » directly by Southern blot hybridization and as bacterial transformant clones expressing two antibiotic resistance markers originally carried separately on the two substrates. The increase in recombination was dependent on X-ray dose. There is some suggestion that lesions other than double-strand breaks contribute to the stimulation of oocyte-mediated homologous recombination. In summary, oocytes have considerable capacity to repair X-ray-induced damage, and some X-ray lesions stimulate homologous recombination in these cells.« less

  2. The synthetic gestagen levonorgestrel directly affects gene expression in thyroid and pituitary glands of Xenopus laevis tadpoles.

    PubMed

    Lorenz, Claudia; Opitz, Robert; Trubiroha, Achim; Lutz, Ilka; Zikova, Andrea; Kloas, Werner

    2016-08-01

    The synthetic gestagen levonorgestrel (LNG) was previously shown to perturb thyroid hormone-dependent metamorphosis in Xenopus laevis. However, so far the mechanisms underlying the anti-metamorphic effects of LNG remained unknown. Therefore, a series of in vivo and ex vivo experiments was performed to identify potential target sites of LNG action along the pituitary-thyroid axis of X. laevis tadpoles. Prometamorphic tadpoles were treated in vivo with LNG (0.01-10nM) for 72h and brain-pituitary and thyroid tissue was analyzed for marker gene expression. While no treatment-related changes were observed in brain-pituitary tissue, LNG treatment readily affected thyroidal gene expression in tadpoles including decreased slc5a5 and iyd mRNA expression and a strong induction of dio2 and dio3 expression. When using an ex vivo organ explant culture approach, direct effects of LNG on both pituitary and thyroid gland gene expression were detecTable Specifically, treatment of pituitary explants with 10nM LNG strongly stimulated dio2 expression and concurrently suppressed tshb expression. In thyroid glands, ex vivo LNG treatment induced dio2 and dio3 mRNA expression in a thyrotropin-independent manner. When thyroid explants were cultured in thyrotropin-containing media, LNG caused similar gene expression changes as seen after 72h in vivo treatment including a very strong repression of thyrotropin-induced slc5a5 expression. Concerning the anti-thyroidal activity of LNG as seen under in vivo conditions, our ex vivo data provide clear evidence that LNG directly affects expression of genes important for thyroidal iodide handling as well as genes involved in negative feedback regulation of pituitary tshb expression. Copyright © 2016 Elsevier B.V. All rights reserved.

  3. The Expression of TALEN before Fertilization Provides a Rapid Knock-Out Phenotype in Xenopus laevis Founder Embryos.

    PubMed

    Miyamoto, Kei; Suzuki, Ken-Ichi T; Suzuki, Miyuki; Sakane, Yuto; Sakuma, Tetsushi; Herberg, Sarah; Simeone, Angela; Simpson, David; Jullien, Jerome; Yamamoto, Takashi; Gurdon, J B

    2015-01-01

    Recent advances in genome editing using programmable nucleases have revolutionized gene targeting in various organisms. Successful gene knock-out has been shown in Xenopus, a widely used model organism, although a system enabling less mosaic knock-out in founder embryos (F0) needs to be explored in order to judge phenotypes in the F0 generation. Here, we injected modified highly active transcription activator-like effector nuclease (TALEN) mRNA to oocytes at the germinal vesicle (GV) stage, followed by in vitro maturation and intracytoplasmic sperm injection, to achieve a full knock-out in F0 embryos. Unlike conventional injection methods to fertilized embryos, the injection of TALEN mRNA into GV oocytes allows expression of nucleases before fertilization, enabling them to work from an earlier stage. Using this procedure, most of developed embryos showed full knock-out phenotypes of the pigmentation gene tyrosinase and/or embryonic lethal gene pax6 in the founder generation. In addition, our method permitted a large 1 kb deletion. Thus, we describe nearly complete gene knock-out phenotypes in Xenopus laevis F0 embryos. The presented method will help to accelerate the production of knock-out frogs since we can bypass an extra generation of about 1 year in Xenopus laevis. Meantime, our method provides a unique opportunity to rapidly test the developmental effects of disrupting those genes that do not permit growth to an adult able to reproduce. In addition, the protocol shown here is considerably less invasive than the previously used host transfer since our protocol does not require surgery. The experimental scheme presented is potentially applicable to other organisms such as mammals and fish to resolve common issues of mosaicism in founders.

  4. Label-free real-time imaging of myelination in the Xenopus laevis tadpole by in vivo stimulated Raman scattering microscopy

    NASA Astrophysics Data System (ADS)

    Hu, Chun-Rui; Zhang, Delong; Slipchenko, Mikhail N.; Cheng, Ji-Xin; Hu, Bing

    2014-08-01

    The myelin sheath plays an important role as the axon in the functioning of the neural system, and myelin degradation is a hallmark pathology of multiple sclerosis and spinal cord injury. Electron microscopy, fluorescent microscopy, and magnetic resonance imaging are three major techniques used for myelin visualization. However, microscopic observation of myelin in living organisms remains a challenge. Using a newly developed stimulated Raman scattering microscopy approach, we report noninvasive, label-free, real-time in vivo imaging of myelination by a single-Schwann cell, maturation of a single node of Ranvier, and myelin degradation in the transparent body of the Xenopus laevis tadpole.

  5. Autocatalytic microtubule nucleation determines the size and mass of Xenopus laevis egg extract spindles

    PubMed Central

    Decker, Franziska; Oriola, David; Dalton, Benjamin

    2018-01-01

    Regulation of size and growth is a fundamental problem in biology. A prominent example is the formation of the mitotic spindle, where protein concentration gradients around chromosomes are thought to regulate spindle growth by controlling microtubule nucleation. Previous evidence suggests that microtubules nucleate throughout the spindle structure. However, the mechanisms underlying microtubule nucleation and its spatial regulation are still unclear. Here, we developed an assay based on laser ablation to directly probe microtubule nucleation events in Xenopus laevis egg extracts. Combining this method with theory and quantitative microscopy, we show that the size of a spindle is controlled by autocatalytic growth of microtubules, driven by microtubule-stimulated microtubule nucleation. The autocatalytic activity of this nucleation system is spatially regulated by the limiting amounts of active microtubule nucleators, which decrease with distance from the chromosomes. This mechanism provides an upper limit to spindle size even when resources are not limiting. PMID:29323637

  6. Plasticity of lung development in the amphibian, Xenopus laevis

    PubMed Central

    Rose, Christopher S.; James, Brandon

    2013-01-01

    Summary Contrary to previous studies, we found that Xenopus laevis tadpoles raised in normoxic water without access to air can routinely complete metamorphosis with lungs that are either severely stunted and uninflated or absent altogether. This is the first demonstration that lung development in a tetrapod can be inhibited by environmental factors and that a tetrapod that relies significantly on lung respiration under unstressed conditions can be raised to forego this function without adverse effects. This study compared lung development in untreated, air-deprived (AD) and air-restored (AR) tadpoles and frogs using whole mounts, histology, BrdU labeling of cell division and antibody staining of smooth muscle actin. We also examined the relationship of swimming and breathing behaviors to lung recovery in AR animals. Inhibition and recovery of lung development occurred at the stage of lung inflation. Lung recovery in AR tadpoles occurred at a predictable and rapid rate and correlated with changes in swimming and breathing behavior. It thus presents a new experimental model for investigating the role of mechanical forces in lung development. Lung recovery in AR frogs was unpredictable and did not correlate with behavioral changes. Its low frequency of occurrence could be attributed to developmental, physical and behavioral changes, the effects of which increase with size and age. Plasticity of lung inflation at tadpole stages and loss of plasticity at postmetamorphic stages offer new insights into the role of developmental plasticity in amphibian lung loss and life history evolution. PMID:24337117

  7. Plasticity of lung development in the amphibian, Xenopus laevis.

    PubMed

    Rose, Christopher S; James, Brandon

    2013-12-15

    Contrary to previous studies, we found that Xenopus laevis tadpoles raised in normoxic water without access to air can routinely complete metamorphosis with lungs that are either severely stunted and uninflated or absent altogether. This is the first demonstration that lung development in a tetrapod can be inhibited by environmental factors and that a tetrapod that relies significantly on lung respiration under unstressed conditions can be raised to forego this function without adverse effects. This study compared lung development in untreated, air-deprived (AD) and air-restored (AR) tadpoles and frogs using whole mounts, histology, BrdU labeling of cell division and antibody staining of smooth muscle actin. We also examined the relationship of swimming and breathing behaviors to lung recovery in AR animals. Inhibition and recovery of lung development occurred at the stage of lung inflation. Lung recovery in AR tadpoles occurred at a predictable and rapid rate and correlated with changes in swimming and breathing behavior. It thus presents a new experimental model for investigating the role of mechanical forces in lung development. Lung recovery in AR frogs was unpredictable and did not correlate with behavioral changes. Its low frequency of occurrence could be attributed to developmental, physical and behavioral changes, the effects of which increase with size and age. Plasticity of lung inflation at tadpole stages and loss of plasticity at postmetamorphic stages offer new insights into the role of developmental plasticity in amphibian lung loss and life history evolution.

  8. Further Characterization of an Interleukin-2-1Ike Cytokine Produced by Xenopus Laevis T Lymphocytes

    PubMed Central

    Haynes, Laura

    1993-01-01

    A T-cell growth factor (TCGF) is produced by antigen- or mitogen-stimulated T lymphocytes from the South African clawed frog Xenopus laevis. This study further defines the physical and biological properties of this cytokine and demonstrates that TCGF is biochemically similar to mammalian interleukin-2 (IL-2). Biologically active TCGF eluted from SDS-PAGE displays a Mr of 16 kD and lectin-affinity chromatography indicates that the three-dimensionmal configuration of carbohydrates on TCGF and human IL-2 is similar. Secretion of TCGF is detectable 1 day after stimulation of splenocytes with the T-cell mitogen phytohemagglutinin (PHA) and peaks following 2 to 3 days of stimulation. Finally, despite the biological and physical similarities between Xenopus TCGF and mammalian IL-2, anti-human IL-2 monoclonal antibodies do not recognize Xenopus TCGF. PMID:8281036

  9. Exposure to 3,3',5-triiodothyronine affects histone and RNA polymerase II modifications, but not DNA methylation status, in the regulatory region of the Xenopus laevis thyroid hormone receptor βΑ gene.

    PubMed

    Kasai, Kentaro; Nishiyama, Norihito; Izumi, Yushi; Otsuka, Shunsuke; Ishihara, Akinori; Yamauchi, Kiyoshi

    2015-11-06

    Thyroid hormones (THs) play a critical role in amphibian metamorphosis, during which the TH receptor (TR) gene, thrb, is upregulated in a tissue-specific manner. The Xenopus laevis thrb gene has 3 TH response elements (TREs) in the 5' flanking regulatory region and 1 TRE in the exon b region, around which CpG sites are highly distributed. To clarify whether exposure to 3,3',5-triiodothyronine (T3) affects histone and RNA polymerase II (RNAPII) modifications and the level of DNA methylation in the 5' regulatory region, we conducted reverse transcription-quantitative polymerase chain reaction, bisulfite sequencing and chromatin immunoprecipitation assay using X. laevis cultured cells and premetamorphic tadpoles treated with or without 2 nM T3. Exposure to T3 increased the amount of the thrb transcript, in parallel with enhanced histone H4 acetylation and RNAPII recruitment, and probably phosphorylation of RNAPII at serine 5, in the 5' regulatory and exon b regions. However, the 5' regulatory region remained hypermethylated even with exposure to T3, and there was no significant difference in the methylation status between DNAs from T3-untreated and -treated cultured cells or tadpole tissues. Our results demonstrate that exposure to T3 induced euchromatin-associated epigenetic marks by enhancing histone acetylation and RNAPII recruitment, but not by decreasing the level of DNA methylation, in the 5' regulatory region of the X. laevis thrb gene. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Environmental estrogens alter early development in Xenopus laevis.

    PubMed

    Bevan, Cassandra L; Porter, Donna M; Prasad, Anita; Howard, Marthe J; Henderson, Leslie P

    2003-04-01

    A growing number of environmental toxicants found in pesticides, herbicides, and industrial solvents are believed to have deleterious effects on development by disrupting hormone-sensitive processes. We exposed Xenopus laevis embryos at early gastrula to the commonly encountered environmental estrogens nonylphenol, octylphenol, and methoxychlor, the antiandrogen, p,p-DDE, or the synthetic androgen, 17 alpha-methyltestosterone at concentrations ranging from 10 nM to 10 microM and examined them at tailbud stages (approximately 48 hr of treatment). Exposure to the three environmental estrogens, as well as to the natural estrogen 17 beta-estradiol, increased mortality, induced morphologic deformations, increased apoptosis, and altered the deposition and differentiation of neural crest-derived melanocytes in tailbud stage embryos. Although neural crest-derived melanocytes were markedly altered in embryos treated with estrogenic toxicants, expression of the early neural crest maker Xslug, a factor that regulates both the induction and subsequent migration of neural crest cells, was not affected, suggesting that the disruption induced by these compounds with respect to melanocyte development may occur at later stages of their differentiation. Co-incubation of embryos with the pure antiestrogen ICI 182,780 blocked the ability of nonylphenol to induce abnormalities in body shape and in melanocyte differentiation but did not block the effects of methoxychlor. Our data indicate not only that acute exposure to these environmental estrogens induces deleterious effects on early vertebrate development but also that different environmental estrogens may alter the fate of a specific cell type via different mechanisms. Finally, our data suggest that the differentiation of neural crest-derived melanocytes may be particularly sensitive to the disruptive actions of these ubiquitous chemical contaminants.

  11. Maitotoxin Is a Potential Selective Activator of the Endogenous Transient Receptor Potential Canonical Type 1 Channel in Xenopus laevis Oocytes

    PubMed Central

    Flores, Pedro L.; Rodríguez, Emma; Zapata, Estrella; Carbó, Roxana; Farías, José María; Martínez, Martín

    2017-01-01

    Maitotoxin (MTX) is the most potent marine toxin known to date. It is responsible for a particular human intoxication syndrome called ciguatera fish poisoning (CFP). Several reports indicate that MTX is an activator of non-selective cation channels (NSCC) in different cell types. The molecular identity of these channels is still an unresolved topic, and it has been proposed that the transient receptor potential (TRP) channels are involved in this effect. In Xenopus laevis oocytes, MTX at picomolar (pM) concentrations induces the activation of NSCC with functional and pharmacological properties that resemble the activity of TRP channels. The purpose of this study was to characterize the molecular identity of the TRP channel involved in the MTX response, using the small interference RNA (siRNA) approach and the two-electrode voltage-clamp technique (TEVC). The injection of a specifically designed siRNA to silence the transient receptor potential canonical type 1 (TRPC1) protein expression abolished the MTX response. MTX had no effect on oocytes, even at doses 20-fold higher compared to cells without injection. Total mRNA and protein levels of TRPC1 were notably diminished. The TRPC4 siRNA did not change the MTX effect, even though it was important to note that the protein level was reduced by the silencing of TRPC4. Our results suggest that MTX could be a selective activator of TRPC1 channels in X. laevis oocytes and a useful pharmacological tool for further studies on these TRP channels. PMID:28672825

  12. Maitotoxin Is a Potential Selective Activator of the Endogenous Transient Receptor Potential Canonical Type 1 Channel in Xenopus laevis Oocytes.

    PubMed

    Flores, Pedro L; Rodríguez, Emma; Zapata, Estrella; Carbó, Roxana; Farías, José María; Martínez, Martín

    2017-06-25

    Maitotoxin (MTX) is the most potent marine toxin known to date. It is responsible for a particular human intoxication syndrome called ciguatera fish poisoning (CFP). Several reports indicate that MTX is an activator of non-selective cation channels (NSCC) in different cell types. The molecular identity of these channels is still an unresolved topic, and it has been proposed that the transient receptor potential (TRP) channels are involved in this effect. In Xenopus laevis oocytes, MTX at picomolar (pM) concentrations induces the activation of NSCC with functional and pharmacological properties that resemble the activity of TRP channels. The purpose of this study was to characterize the molecular identity of the TRP channel involved in the MTX response, using the small interference RNA (siRNA) approach and the two-electrode voltage-clamp technique (TEVC). The injection of a specifically designed siRNA to silence the transient receptor potential canonical type 1 (TRPC1) protein expression abolished the MTX response. MTX had no effect on oocytes, even at doses 20-fold higher compared to cells without injection. Total mRNA and protein levels of TRPC1 were notably diminished. The TRPC4 siRNA did not change the MTX effect, even though it was important to note that the protein level was reduced by the silencing of TRPC4. Our results suggest that MTX could be a selective activator of TRPC1 channels in X. laevis oocytes and a useful pharmacological tool for further studies on these TRP channels.

  13. Designation of the anterior/posterior axis in pregastrula Xenopus laevis.

    PubMed

    Lane, M C; Sheets, M D

    2000-09-01

    A new fate map for mesodermal tissues in Xenopus laevis predicted that the prime meridian, which runs from the animal pole to the vegetal pole through the center of Spemann's organizer, is the embryo's anterior midline, not its dorsal midline (M. C. Lane and W. C. Smith, 1999, Development 126, 423-434). In this report, we demonstrate by lineage labeling that the column 1 blastomeres at st. 6, which populate the prime meridian, give rise to the anterior end of the embryo. In addition, we surgically isolate and culture tissue centered on this meridian from early gastrulae. This tissue forms a patterned head with morphologically distinct ventral and dorsal structures. In situ hybridization and immunostaining reveal that the cultured heads contain the anterior tissues of all three germ layers, correctly patterned. Regardless of how we dissect early gastrulae along meridians running from the animal to the vegetal pole, both the formation of head structures and the expression of anterior marker genes always segregate with the prime meridian passing through Spemann's organizer. The prime meridian also gives rise to dorsal, axial mesoderm, but not uniquely, as specification tests show that dorsal mesoderm arises in fragments of the embryo which exclude the prime meridian. These results support the hypothesis that the midline that bisects Spemann's organizer is the embryo's anterior midline. Copyright 2000 Academic Press.

  14. Autocatalytic microtubule nucleation determines the size and mass of Xenopus laevis egg extract spindles.

    PubMed

    Decker, Franziska; Oriola, David; Dalton, Benjamin; Brugués, Jan

    2018-01-11

    Regulation of size and growth is a fundamental problem in biology. A prominent example is the formation of the mitotic spindle, where protein concentration gradients around chromosomes are thought to regulate spindle growth by controlling microtubule nucleation. Previous evidence suggests that microtubules nucleate throughout the spindle structure. However, the mechanisms underlying microtubule nucleation and its spatial regulation are still unclear. Here, we developed an assay based on laser ablation to directly probe microtubule nucleation events in Xenopus laevis egg extracts. Combining this method with theory and quantitative microscopy, we show that the size of a spindle is controlled by autocatalytic growth of microtubules, driven by microtubule-stimulated microtubule nucleation. The autocatalytic activity of this nucleation system is spatially regulated by the limiting amounts of active microtubule nucleators, which decrease with distance from the chromosomes. This mechanism provides an upper limit to spindle size even when resources are not limiting. © 2018, Decker et al.

  15. Xenopus laevis ribosomal protein genes: isolation of recombinant cDNA clones and study of the genomic organization.

    PubMed Central

    Bozzoni, I; Beccari, E; Luo, Z X; Amaldi, F

    1981-01-01

    Poly-A+ mRNA from Xenopus laevis oocytes, partially enriched for r-protein coding capacity has been used as starting material for preparing a cDNA bank in plasmid pBR322. The clones containing sequences specific for r-proteins have been selected by translation of the complementary mRNAs. Clones for six different r-proteins have been identified and utilized as probes for studying their genomic organization. Two gene copies per haploid genome were found for r-proteins L1, L14, S19, and four-five for protein S1, S8 and L32. Moreover a population polymorphism has been observed for the genomic regions containing sequences for r-protein S1, S8 and L14. Images PMID:6112733

  16. Biochemical effects of chlorpyrifos on two developmental stages of Xenopus laevis.

    PubMed

    Richards, Sean M; Kendall, Ron J

    2002-09-01

    Abstract-The effects of a 96-h static exposure to chlorpyrifos were examined in two developmental stages of larval Xenopus laevis (premetamorph and metamorph). Measures of effect included mortality, deformity, cholinesterase (ChE) activity, and DNA and protein concentration. All parameters indicated that metamorphs were more sensitive than were premetamorphs. For larvae exposed as premetamorphs, the median lethal concentration and median effective concentration were 14.6 mg/L and 1.71 mg/L; for those exposed as metamorphs, values were 0.56 mg/L and 0.24 mg/L, respectively. Cholinesterase activity was the most sensitive biochemical parameter. Exposure to chlorpyrifos at 0.01 mg/L caused significant decreases in the ChE activity of metamorphs; 0.1 mg/L significantly decreased premetamorph ChE activity. Metamorph DNA was significantly decreased at 0.1 mg/L; premetamorph DNA was not reduced until exposure to 1.0 mg/L. Whole-body protein was the least sensitive biochemical measure of effect. Premetamorphs did not experience a reduction in protein concentrations. Metamorph protein concentration was significantly decreased at 1.0 mg/L. Based on current surface water data, the most sensitive effect would not have a high probability (< or = 4.2%) of occurring in the environment.

  17. The Effect of Plasma Exposure on Tail Regeneration of Tadpoles Xenopus Laevis

    NASA Astrophysics Data System (ADS)

    June, Joyce; Rivie, Adonis; Ezuduemoih, Raphael; Menon, Jaishri; Martus, Kevin

    2014-03-01

    Wound healing requires a balanced combination of nutrients and growth factors for healing and tissue regeneration. The effect of plasma exposure on tail regeneration of tadpoles, Xenopus laevis is investigated. The exposure of the wound to the helium plasma immediately followed the amputation of 40% of the tail. Amputation of the tail initiates regeneration of spinal cord, muscle, notochord, skin and connective tissues. By 24 h, the wound was covered by wound epithelium and blastema was formed by day 5. There was increased angiogenesis in plasma exposed tail regenerate compared to the control following 5 d post amputation. Observed was an increase in NO production in the regenerate of plasma exposed tadpoles was derived from increased activity of nNOS and iNOS. Western blot analysis for vascular endothelial growth factor showed stronger bands for the protein in amputated tadpoles of both the groups. Analysis of the composition and characteristics of the plasma using optical emission spectroscopy indicates excited state species consisting of N2, N2+,and OH is present in the plasma. This study was supported, in part, by the NSF Grant 1040108.

  18. Neurotransmitter signaling pathways required for normal development in Xenopus laevis embryos: a pharmacological survey screen

    PubMed Central

    Sullivan, Kelly G.; Levin, Michael

    2016-01-01

    Neurotransmitters are not only involved in brain function but are also important signaling molecules for many diverse cell types. Neurotransmitters are widely conserved, from evolutionarily ancient organisms lacking nervous systems through man. Here, we report results from a loss- and gain-of-function survey, using pharmacologic modulators of several neurotransmitter pathways to examine possible roles in normal embryogenesis. Applying reagents targeting the glutamatergic, adrenergic, and dopaminergic pathways to embryos of Xenopus laevis from gastrulation to organogenesis stages, we observed and quantified numerous malformations including craniofacial defects, hyperpigmentation, muscle mispatterning, and miscoiling of the gut. These data implicate several key neurotransmitters in new embryonic patterning roles, reveal novel earlier stages for processes involved in eye development, suggest new targets for subsequent molecular-genetic investigation, and highlight the necessity for in-depth toxicology studies of psychoactive compounds to which human embryos might be exposed during pregnancy. PMID:27060969

  19. Inositol 1,4,5-trisphosphate-induced calcium release in the organelle layers of the stratified, intact egg of Xenopus laevis

    PubMed Central

    1990-01-01

    Using double-barreled, Ca2(+)-sensitive microelectrodes, we have examined the characteristics of the Ca2+ release by inositol 1,4,5- trisphosphate (Ins(1,4,5)P3) in the various layers of Xenopus laevis eggs in which the organelles had been stratified by centrifugation. Centrifugation of living eggs stratifies the organelles yet retains them in the normal cytoplasmic milieu. The local increase in intracellular free Ca2+ in each layer was directly measured under physiological conditions using theta-tubing, double-barreled, Ca2(+)- sensitive microelectrodes in which one barrel was filled with the Ca2+ sensor and the other was filled with Ins(1,4,5)P3 for microinjection. The two tips of these electrodes were very close to each other (3 microns apart) enabling us to measure the kinetics of both the highly localized intracellular Ca2+ release and its subsequent removal in response to Ins(1,4,5)P3 injection. Upon Ins(1,4,5)P3 injection, the ER- enriched layer exhibited the largest release of Ca2+ in a dosage- dependent manner, whereas the other layers, mitochondria, lipid, and yolk, released 10-fold less Ca2+ in a dosage-independent manner. The removal of released Ca2+ took place within approximately 1 min. The sensitivity to Ins(1,4,5)P3 and the time course of intracellular Ca2+ release in the unstratified (unactivated) egg is nearly identical to that observed in the ER layer of the stratified egg. Our data suggest that the ER is the major organelle of the Ins(1,4,5)P3-sensitive Ca2+ store in the egg of Xenopus laevis. PMID:2324195

  20. Fertilization and development of eggs of the South African clawed toad, Xenopus laevis, on sounding rockets in space

    NASA Astrophysics Data System (ADS)

    Ubbels, Geertje A.; Berendsen, Willem; Kerkvliet, Sonja; Narraway, Jenny

    Egg rotation and centrifugation experiments strongly suggest a role for gravity in the determination of the spatial structure of amphibian embryos. Decisive experiments can only be made in Space. Eggs of Xenopus laevis, the South African clawed toad, were the first vertebrate eggs which were successfully fertilized on Sounding Rockets in Space. Unfixed, newly fertilized eggs survived reentry, and a reasonable number showed a seemingly normal gastrulation but died between gastrulation and neurulation. Only a few reached the larval stage, but these developed abnormally. In the future, we inted to test whether this abnormal morphogenesis is due to reentry perturbations, or due to a real microgravity effect, through perturbation of the reinitiation of meiosis and other processes, or started by later sperm penetration.

  1. Fertilization and development of eggs of the South African clawed toad, Xenopus laevis, on sounding rockets in space.

    PubMed

    Ubbels, G A; Berendsen, W; Kerkvliet, S; Narraway, J

    1992-01-01

    Egg rotation and centrifugation experiments strongly suggest a role for gravity in the determination of the spatial structure of amphibian embryos. Decisive experiments can only be made in Space. Eggs of Xenopus laevis, the South African clawed toad, were the first vertebrate eggs which were successfully fertilized on Sounding Rockets in Space. Unfixed, newly fertilized eggs survived reentry, and a reasonable number showed a seemingly normal gastrulation but died between gastrulation and neurulation. Only a few reached the larval stage, but these developed abnormally. In the future, we intend to test whether this abnormal morphogenesis is due to reentry perturbations, or due to a real microgravity effect, through perturbation of the reinitiation of meiosis and other processes, or started by later sperm penetration.

  2. The G-protein-coupled receptor, GPR84, is important for eye development in Xenopus laevis.

    PubMed

    Perry, Kimberly J; Johnson, Verity R; Malloch, Erica L; Fukui, Lisa; Wever, Jason; Thomas, Alvin G; Hamilton, Paul W; Henry, Jonathan J

    2010-11-01

    G-protein-coupled receptors (GPCRs) represent diverse, multifamily groups of cell signaling receptors involved in many cellular processes. We identified Xenopus laevis GPR84 as a member of the A18 subfamily of GPCRs. During development, GPR84 is detected in the embryonic lens placode, differentiating lens fiber cells, retina, and cornea. Anti-sense morpholino oligonucleotide-mediated knockdown and RNA rescue experiments demonstrate GPR84's importance in lens, cornea, and retinal development. Examination of cell proliferation using an antibody against histone H3 S10P reveals significant increases in the lens and retina following GPR84 knockdown. Additionally, there was also an increase in apoptosis in the retina and lens, as revealed by TUNEL assay. Reciprocal transplantation of the presumptive lens ectoderm between uninjected controls and morpholino-injected embryos demonstrates that GPR84 is necessary in the retina for proper development of the retina, as well as other eye tissues including the lens and cornea. © 2010 Wiley-Liss, Inc.

  3. The G-protein-coupled receptor, GPR84, is important for eye development in Xenopus laevis

    PubMed Central

    Perry, Kimberly J.; Johnson, Verity R.; Malloch, Erica L.; Fukui, Lisa; Wever, Jason; Thomas, Alvin G.; Hamilton, Paul W.; Henry, Jonathan J.

    2010-01-01

    G-protein-coupled receptors (GPCRs) represent diverse, multifamily groups of cell signaling receptors involved in many cellular processes. We identified Xenopus laevis GPR84 as a member of the A18 subfamily of GPCRs. During development, GPR84 is detected in the embryonic lens placode, differentiating lens fiber cells, retina and cornea. Anti-sense morpholino oligonucleotide-mediated knockdown and RNA rescue experiments demonstrate GPR84’s importance in lens, cornea and retinal development. Examination of cell proliferation using an antibody against histone H3 S10P reveals significant increases in the lens and retina following GPR84 knockdown. Additionally, there was also an increase in apoptosis in the retina and lens, as revealed by TUNEL assay. Reciprocal transplantation of the presumptive lens ectoderm between uninjected controls and morpholino injected embryos demonstrates that GPR84 is necessary in the retina for proper development of the retina, as well as other eye tissues including the lens and cornea. PMID:20925114

  4. Skeletal advance and arrest in giant non-metamorphosing African clawed frog tadpoles (Xenopus laevis: Daudin)

    PubMed Central

    Kerney, Ryan; Wassersug, Richard; Hall, Brian K

    2010-01-01

    This study examines the skeletons of giant non-metamorphosing (GNM) Xenopus laevis tadpoles, which arrest their development indefinitely before metamorphosis, and grow to excessively large sizes in the absence of detectable thyroid glands. Cartilage growth is isometric; however, chondrocyte size is smaller in GNM tadpoles than in controls. Most cartilages stain weakly with alcian blue, and several cartilages are calcified (unlike controls). However, cartilages subjacent to periosteum-derived bone retain strong affinities for alcian blue, indicating a role for periosteum-derived bone in the retention of glycosaminoglycans during protracted larval growth. Bone formation in the head, limb, and axial skeletons is advanced in comparison with stage-matched controls, but arrests at various mid-metamorphic states. Both dermal and periosteum-derived bones grow to disproportionately large sizes in comparison to controls. Additionally, mature monocuspid teeth form in several GNM tadpoles. Advances in skeletal development are attributable to the old ages and large sizes of these tadpoles, and reveal unexpected developmental potentials of the pre-metamorphic skeleton. PMID:20402828

  5. Budgett's frog (Lepidobatrachus laevis): A new amphibian embryo for developmental biology.

    PubMed

    Amin, Nirav M; Womble, Mandy; Ledon-Rettig, Cristina; Hull, Margaret; Dickinson, Amanda; Nascone-Yoder, Nanette

    2015-09-15

    The large size and rapid development of amphibian embryos has facilitated ground-breaking discoveries in developmental biology. Here, we describe the embryogenesis of the Budgett's frog (Lepidobatrachus laevis), an unusual species with eggs that are over twice the diameter of laboratory Xenopus, and embryos that can tolerate higher temperatures to develop into a tadpole four times more rapidly. In addition to detailing their early development, we demonstrate that, like Xenopus, these embryos are amenable to explant culture assays and can express exogenous transcripts in a tissue-specific manner. Moreover, the steep developmental trajectory and large scale of Lepidobatrachus make it exceptionally well-suited for morphogenesis research. For example, the developing organs of the Budgett's frog are massive compared to those of most model species, and are composed of larger individual cells, thereby affording increased subcellular resolution of early vertebrate organogenesis. Furthermore, we found that complete limb regeneration, which typically requires months to achieve in most vertebrate models, occurs in a matter of days in the Budgett's tadpole, which substantially accelerates the pace of experimentation. Thus, the unusual combination of the greater size and speed of the Budgett's frog model provides inimitable advantages for developmental studies-and a novel inroad to address the mechanisms of spatiotemporal scaling during evolution. Copyright © 2015 Elsevier Inc. All rights reserved.

  6. Budgett’s frog (Lepidobatrachus laevis): a new amphibian embryo for developmental biology

    PubMed Central

    Amin, Nirav M.; Womble, Mandy; Ledon-Rettig, Cris; Hull, Margaret; Dickinson, Amanda; Nascone-Yoder, Nanette

    2015-01-01

    The large size and rapid development of amphibian embryos has facilitated ground-breaking discoveries in developmental biology. Here, we describe the embryogenesis of the Budgett’s frog (Lepidobatrachus laevis), an unusual species with eggs that are over twice the diameter of laboratory Xenopus, and embryos that can tolerate higher temperatures to develop into a tadpole four times more rapidly. In addition to detailing their early development, we demonstrate that, like Xenopus, these embryos are amenable to explant culture assays and can express exogenous transcripts in a tissue-specific manner. Moreover, the steep developmental trajectory and large scale of Lepidobatrachus make it exceptionally well-suited for morphogenesis research. For example, the developing organs of the Budgett’s frog are massive compared to those of most model species, and are composed of larger individual cells, thereby affording increased subcellular resolution of early vertebrate organogenesis. Furthermore, we found that complete limb regeneration, which typically requires months to achieve in most vertebrate models, occurs in a matter of days in the Budgett’s tadpole, which substantially accelerates the pace of experimentation. Thus, the unusual combination of the greater size and speed of the Budgett’s frog model provides inimitable advantages for developmental studies—and a novel inroad to address the mechanisms of spatiotemporal scaling during evolution. PMID:26169245

  7. Noggin4 is a long-range inhibitor of Wnt8 signalling that regulates head development in Xenopus laevis.

    PubMed

    Eroshkin, Fedor M; Nesterenko, Alexey M; Borodulin, Alexander V; Martynova, Natalia Yu; Ermakova, Galina V; Gyoeva, Fatima K; Orlov, Eugeny E; Belogurov, Alexey A; Lukyanov, Konstantin A; Bayramov, Andrey V; Zaraisky, Andrey G

    2016-03-14

    Noggin4 is a Noggin family secreted protein whose molecular and physiological functions remain unknown. In this study, we demonstrate that in contrast to other Noggins, Xenopus laevis Noggin4 cannot antagonise BMP signalling; instead, it specifically binds to Wnt8 and inhibits the Wnt/β -catenin pathway. Live imaging demonstrated that Noggin4 diffusivity in embryonic tissues significantly exceeded that of other Noggins. Using the Fluorescence Recovery After Photobleaching (FRAP) assay and mathematical modelling, we directly estimated the affinity of Noggin4 for Wnt8 in living embryos and determined that Noggin4 fine-tune the Wnt8 posterior-to-anterior gradient. Our results suggest a role for Noggin4 as a unique, freely diffusing, long-range inhibitor of canonical Wnt signalling, thus explaining its ability to promote head development.

  8. Microneedle-based analysis of the micromechanics of the metaphase spindle assembled in Xenopus laevis egg extracts

    PubMed Central

    Shimamoto, Yuta; Kapoor, Tarun M.

    2014-01-01

    SUMMARY To explain how micron-sized cellular structures generate and respond to forces we need to characterize their micromechanical properties. Here we provide a protocol to build and use a dual force-calibrated microneedle-based set-up to quantitatively analyze the micromechanics of a metaphase spindle assembled in Xenopus laevis egg extracts. This cell-free extract system allows for controlled biochemical perturbations of spindle components. We describe how the microneedles are prepared and how they can be used to apply and measure forces. A multi-mode imaging system allows tracking of microtubules, chromosomes and needle tips. This set-up can be used to analyze the viscoelastic properties of the spindle on time-scales ranging from minutes to sub-seconds. A typical experiment, along with data analysis, is also detailed. We anticipate that our protocol can be readily extended to analyze the micromechanics of other cellular structures assembled in cell-free extracts. The entire procedure can take 3-4 days. PMID:22538847

  9. Lectin from embryos and oocytes of Xenopus laevis. Purification and properties.

    PubMed

    Roberson, M M; Barondes, S H

    1982-07-10

    Soluble extracts of Xenopus laevis blastula stage embryos, oocytes, and adult liver contain lectin activities detected by agglutination of trypsinized, glutaraldehyde-fixed rabbit erythrocytes. Lectin from the embryos and oocytes was purified by affinity chromatography on a column derivatized with melibiose. Trace contaminants were removed either by preparative isoelectric focusing or by gel filtration. Based on its behavior on Sepharose 6B the purified oocyte lectin has an apparent molecular weight of approximately 480,000. On sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions there were two major bands with molecular weight ranges of about 43,000 and 45,000, with diffuse trails. Since the purified lectin contains about 20% saccharides by weight and since both bands are glycosylated, diffuseness might be due to variable glycosylation. Heterogeneity was indicated by isoelectric focusing in polyacrylamide gels, which showed four protein bands with isoelectric points ranging from 4.4 to 4.9. Lectins from both embryos and oocytes comprised about 1 to 2% of the total soluble protein and could not be distinguished by sodium dodecyl sulfate polyacrylamide gel electrophoresis. However, the specific hemagglutination activity of the purified oocyte lectin was, on the average, 7-fold higher. Levels in crude extracts of liver were 3 orders of magnitude lower than those from oocytes. The hemagglutination activities of the lectins from embryos, oocytes, and adult liver required Ca2+ and were blocked by similar concentrations of both alpha- and beta-galactosides.

  10. The Pharmacokinetics of Enrofloxacin in Adult African Clawed Frogs (Xenopus laevis)

    PubMed Central

    Howard, Antwain M; Papich, Mark G; Felt, Stephen A; Long, Charles T; McKeon, Gabriel P; Bond, Emmitt S; Torreilles, Stéphanie L; Luong, Richard H; Green, Sherril L

    2010-01-01

    Pharmacokinetics of enrofloxacin, a fluoroquinolone antibiotic, was determined in adult female Xenopus laevis after single-dose administration (10 mg/kg) by intramuscular or subcutaneous injection. Frogs were evaluated at various time points until 8 h after injection. Plasma was analyzed for antibiotic concentration levels by HPLC. We computed pharmacokinetic parameters by using noncompartmental analysis of the pooled concentrations (naive pooled samples). After intramuscular administration of enrofloxacin, the half-life was 5.32 h, concentration maximum was 10.85 µg/mL, distribution volume was 841.96 mL/kg, and area under the time–concentration curve was 57.59 µg×h/mL; after subcutaneous administration these parameters were 4.08 h, 9.76 µg/mL, 915.85 mL/kg, and 47.42 µg×h/mL, respectively. According to plasma pharmacokinetics, Xenopus seem to metabolize enrofloxacin in a manner similar to mammals: low levels of the enrofloxacin metabolite, ciprofloxacin, were detected in the frogs’ habitat water and plasma. At necropsy, there were no gross or histologic signs of toxicity after single-dose administration; toxicity was not evaluated for repeated dosing. The plasma concentrations reached levels considered effective against common aquatic pathogens and suggest that a single, once-daily dose would be a reasonable regimen to consider when treating sick frogs. The treatment of sick frogs should be based on specific microbiologic identification of the pathogen and on antibiotic susceptibility testing. PMID:21205443

  11. Development of the Larval Amphibian Growth and Development Assay: Effects of benzophenone-2 exposure in Xenopus laevis from embryo to juvenile.

    PubMed

    Haselman, Jonathan T; Sakurai, Maki; Watanabe, Naoko; Goto, Yasushi; Onishi, Yuta; Ito, Yuki; Onoda, Yu; Kosian, Patricia A; Korte, Joseph J; Johnson, Rodney D; Iguchi, Taisen; Degitz, Sigmund J

    2016-12-01

    The Larval Amphibian Growth and Development Assay (LAGDA) is a globally harmonized chemical testing guideline developed by the U.S. Environmental Protection Agency in collaboration with Japan's Ministry of Environment to support risk assessment. The assay is employed as a higher tiered approach to evaluate effects of chronic chemical exposure throughout multiple life stages in a model amphibian species, Xenopus laevis. To evaluate the utility of the initial LAGDA design, the assay was performed using a mixed mode of action endocrine disrupting chemical, benzophenone-2 (BP-2). X. laevis embryos were exposed in flow-through conditions to 0, 1.5, 3.0 or 6.0 mg l -1 BP-2 until 2 months post-metamorphosis. Overt toxicity was evident throughout the exposure period in the 6.0 mg l -1 treatment due to elevated mortality rates and observed liver and kidney pathologies. Concentration-dependent increases in severity of thyroid follicular cell hypertrophy and hyperplasia occurred in larval tadpoles indicating BP-2-induced impacts on the thyroid axis. Additionally, gonads were impacted in all treatments with some genetic males showing both testis and ovary tissues (1.5 mg l -1 ) and 100% of the genetic males in the 3.0 and 6.0 mg l -1 treatments experiencing complete male-to-female sex reversal. Concentration-dependent vitellogenin induction occurred in both genders with associated accumulations of protein in the livers, kidneys and gonads, which was likely vitellogenin and other estrogen-responsive yolk proteins. This is the first study that demonstrates the endocrine effects of this mixed mode of action chemical in an amphibian species and demonstrates the utility of the LAGDA design for supporting chemical risk assessment. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  12. The Nedd4 binding protein 3 is required for anterior neural development in Xenopus laevis.

    PubMed

    Kiem, Lena-Maria; Dietmann, Petra; Linnemann, Alexander; Schmeisser, Michael J; Kühl, Susanne J

    2017-03-01

    The Fezzin family member Nedd4-binding protein 3 (N4BP3) is known to regulate axonal and dendritic branching. Here, we show that n4bp3 is expressed in the neural tissue of the early Xenopus laevis embryo including the eye, the brain and neural crest cells. Knockdown of N4bp3 in the Xenopus anterior neural tissue results in severe developmental impairment of the eye, the brain and neural crest derived cranial cartilage structures. Moreover, we demonstrate that N4bp3 depletion leads to a significant reduction of both eye and brain specific marker genes and reduced neural crest cell migration. Finally, we demonstrate an impact of N4bp3 deficiency on cell apoptosis and proliferation. Our studies indicate that N4bp3 is required for early anterior neural development of vertebrates. This is in line with a study implicating that genetic disruption of N4BP3 in humans might be related to neurodevelopmental disease. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Survey of O-GlcNAc level variations in Xenopus laevis from oogenesis to early development.

    PubMed

    Dehennaut, Vanessa; Lefebvre, Tony; Leroy, Yves; Vilain, Jean-Pierre; Michalski, Jean-Claude; Bodart, Jean-François

    2009-04-01

    Little is known about the impact of O-linked-N-acetylglucosaminylation (O-GlcNAc) in gametes production and developmental processes. Here we investigated changes in O-GlcNAc, UDP-GlcNAc and O-GlcNAc transferase (OGT) levels in Xenopus laevis from oogenesis to embryo hatching. We showed that in comparison to stage VI, stages I-V oocytes expressed higher levels of O-GlcNAc correlating changes in OGT expression, but not in UDP-GlcNAc pools. Upon progesterone stimulation, an O-GlcNAc level burst occurred during meiotic resumption long before MPF and Mos-Erk2 pathways activations. Finally, we observed high levels of O-GlcNAc, UDP-GlcNAc and OGT during segmentation that decreased concomitantly at the onset of gastrulation. Nevertheless, no correlation between the glycosylation, the nucleotide-sugar and the glycosyltransferase was observed after neurulation. Our results show that O-GlcNAc is regulated throughout oogenesis and development within a complex pattern and suggest that dysfunctions in the dynamics of this glycosylation could lead to developmental abnormalities.

  14. A ribosomal orphon sequence from Xenopus laevis flanked by novel low copy number repetitive elements.

    PubMed

    Guimond, A; Moss, T

    1999-02-01

    We have used a differential cloning approach to isolate ribosomal/non-ribosomal frontier sequences from Xenopus laevis. A ribosomal intergenic spacer sequence (IGS) was cloned and shown not to be physically linked with the ribosomal locus. This ribosomal orphon contained the IGS sequences found immediately downstream of the 28S gene and included an array of enhancer repetitions and a non-functional spacer promoter. The orphon sequence was flanked by a member of the novel 'Frt' low copy repetitive element family. Three individual Frt repeats were sequenced and all members of this family were shown to lie clustered at two chromosomal sites, one of which contained the ribosomal orphon. One of the Frt elements contained an insertion of 297 bp that showed extensive homology to sequences within at least three other Xenopus genes. Each homology region was flanked by members of the T2 family of short interspersed repetitive elements, (SINEs), and by its target insertion sequence, suggesting multiple translocation events. The data are discussed in terms of the evolution of the ribosomal gene locus.

  15. In vitro antioxidant capacity and free radical scavenging evaluation of active metabolite constituents of Newbouldia laevis ethanolic leaf extract.

    PubMed

    Habu, Josiah Bitrus; Ibeh, Bartholomew Okechukwu

    2015-03-14

    The aim of the present study was to evaluate the in vitro antioxidant and free radical scavenging capacity of bioactive metabolites present in Newbouldia laevis leaf extract. Chromatographic and spectrophotometric methods were used in the study and modified where necessary in the study. Bioactivity of the extract was determined at 10 μg/ml, 50 μg/ml, 100 μg/ml, 200 μg/ml and 400 μg/ml concentrations expressed in % inhibition. The yield of the ethanolic leaf extract of N.laevis was 30.3 g (9.93%). Evaluation of bioactive metabolic constituents gave high levels of ascorbic acid (515.53 ± 12 IU/100 g [25.7 mg/100 g]), vitamin E (26.46 ± 1.08 IU/100 g), saponins (6.2 ± 0.10), alkaloids (2.20 ± 0.03), cardiac glycosides(1.48 ± 0.22), amino acids and steroids (8.01 ± 0.04) measured in mg/100 g dry weight; moderate levels of vitamin A (188.28 ± 6.19 IU/100 g), tannins (0.09 ± 0.30), terpenoids (3.42 ± 0.67); low level of flavonoids (1.01 ± 0.34 mg/100 g) and absence of cyanogenic glycosides, carboxylic acids and aldehydes/ketones. The extracts percentage inhibition of DPPH, hydroxyl radical (OH.), superoxide anion (O2 .-), iron chelating, nitric oxide radical (NO), peroxynitrite (ONOO-), singlet oxygen (1O2), hypochlorous acid (HOCl), lipid peroxidation (LPO) and FRAP showed a concentration-dependent antioxidant activity with no significant difference with the controls. Though, IC50 of the extract showed significant difference only in singlet oxygen (1O2) and iron chelating activity when compared with the controls. The extract is a potential source of antioxidants/free radical scavengers having important metabolites which maybe linked to its ethno-medicinal use.

  16. Effects of dietary exposure of polycyclic musk HHCB on the metamorphosis of Xenopus laevis.

    PubMed

    Pablos, María Victoria; Jiménez, María Ángeles; San Segundo, Laura; Martini, Federica; Beltrán, Eulalia; Fernández, Carlos

    2016-06-01

    The compound 1,3,4,6,7,8-hexahydro-4,6,6,7,8,8-hexamethylcyclopenta-[γ]-2-benzopyrane (HHCB; galaxolide, Chemical Abstracts Service number 1222-05-5) is a synthetic musk used extensively as a fragrance in many consumer products and classified as an emerging pollutant. The ecotoxicological information available for HHCB addresses exposure via water, but this compound is frequently adsorbed into particulate matter. The goal of the present study was to assess the effects of dietary exposure to several environmentally relevant HHCB concentrations adsorbed in food during Xenopus laevis metamorphosis. The authors sought to determine if such exposure to this synthetic musk resulted in histological changes in the thyroid gland in conjunction with changes in development (staging, timing to metamorphosis), body weight, and length. Developmental acceleration on day 14, together with hypertrophy of the thyroid follicular epithelium in tadpoles, suggested a possible agonistic effect of HHCB, which would have been compensated after metamorphosis by regulatory mechanisms to maintain homeostasis. Further research into the potential thyroid-related mechanisms of action of HHCB should be conducted. Environ Toxicol Chem 2016;35:1428-1435. © 2015 SETAC. © 2015 SETAC.

  17. Effect of chronic copper and pentachlorophenol exposure to early life stages of Xenopus laevis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fort, D.J.; Stover, E.L.

    1995-12-31

    An evaluation of the effects of low-level copper and pentachlorophenol exposure on various early life stages of the South African clawed frog, Xenopus laevis was performed using stage-specific and long-term continuous exposures. Stage-specific exposure experiments were conducted such that separate subsets of embryos and larvae from the same clutch were exposed to two toxicants, copper and pentachlorophenol, from 0 d to 4 d (standard Frog Embryo Teratagenesis Assay Xenopus [FETAX]), 4 d to 8 d, 8 d to 12 d, and 12 d to 16 d. Results from two separate concentration-response experiments indicated that sensitivity to either toxicant increased inmore » each successive time period. Continuous exposure studies conducted for 60 to 75 days indicated that copper, but not pentachlorophenol induced reduction deficiency malformations of the hind limb at concentrations as low as 0.05 mg/L. Pentachlorophenol concentrations as low as 0.5/{micro}g/L inhibited tail resorption. However, copper did not adversely affect the process of tail resorption. These results indicated that studies evaluating longer-term developmental processes are important in ecological hazard evaluation.« less

  18. Male-Male Clasping May Be Part of an Alternative Reproductive Tactic in Xenopus laevis

    PubMed Central

    Rhodes, Heather J.; Stevenson, Rachel J.; Ego, Courtney L.

    2014-01-01

    Male Xenopus laevis frogs have been observed to clasp other males in a sustained, amplectant position, the purpose of which is unknown. We examined three possible hypotheses for this counter-intuitive behavior: 1) clasping males fail to discriminate the sex of the frogs they clasp; 2) male-male clasping is an aggressive or dominant behavior; or 3) that males clasp other males to gain proximity to breeding events and possibly engage in sperm competition. Our data, gathered through a series of behavioral experiments in the laboratory, refute the first two hypotheses. We found that males did not clasp indiscriminately, but showed a sex preference, with most males preferentially clasping a female, but a proportion preferentially clasping another male. Males that clasped another male when there was no female present were less likely to “win” reproductive access in a male-male-female triad, indicating that they did not establish dominance through clasping. However, those males did gain proximity to oviposition by continued male-male clasping in the presence of the female. Thus, our findings are consistent with, but cannot confirm, the third hypothesis of male-male clasping as an alternative reproductive tactic. PMID:24849114

  19. A Tunable Silk Hydrogel Device for Studying Limb Regeneration in Adult Xenopus Laevis

    PubMed Central

    Golding, Anne; Levin, Michael; Kaplan, David L.

    2016-01-01

    In certain amphibian models limb regeneration can be promoted or inhibited by the local wound bed environment. This research introduces a device that can be utilized as an experimental tool to characterize the conditions that promotes limb regeneration in the adult frog (Xenopus laevis) model. In particular, this device was designed to manipulate the local wound environment via a hydrogel insert. Initial characterization of the hydrogel insert revealed that this interaction had a significant influence on mechanical forces to the animal, due to the contraction of the hydrogel. The material and mechanical properties of the hydrogel insert were a factor in the device design in relation to the comfort of the animal and the ability to effectively manipulate the amputation site. The tunable features of the hydrogel were important in determining the pro-regenerative effects in limb regeneration, which was measured by cartilage spike formation and quantified by micro-computed tomography. The hydrogel insert was a factor in the observed morphological outcomes following amputation. Future work will focus on characterizing and optimizing the device’s observed capability to manipulate biological pathways that are essential for limb regeneration. However, the present work provides a framework for the role of a hydrogel in the device and a path forward for more systematic studies. PMID:27257960

  20. Coding rate and duration of vocalizations of the frog, Xenopus laevis.

    PubMed

    Zornik, Erik; Yamaguchi, Ayako

    2012-08-29

    Vocalizations involve complex rhythmic motor patterns, but the underlying temporal coding mechanisms in the nervous system are poorly understood. Using a recently developed whole-brain preparation from which "fictive" vocalizations are readily elicited in vitro, we investigated the cellular basis of temporal complexity of African clawed frogs (Xenopus laevis). Male advertisement calls contain two alternating components--fast trills (∼300 ms) and slow trills (∼700 ms) that contain clicks repeated at ∼60 and ∼30 Hz, respectively. We found that males can alter the duration of fast trills without changing click rates. This finding led us to hypothesize that call rate and duration are regulated by independent mechanisms. We tested this by obtaining whole-cell patch-clamp recordings in the "fictively" calling isolated brain. We discovered a single type of premotor neuron with activity patterns correlated with both the rate and duration of fast trills. These "fast-trill neurons" (FTNs) exhibited long-lasting depolarizations (LLDs) correlated with each fast trill and action potentials that were phase-locked with motor output-neural correlates of call duration and rate, respectively. When depolarized without central pattern generator activation, FTNs produced subthreshold oscillations and action potentials at fast-trill rates, indicating FTN resonance properties are tuned to, and may dictate, the fast-trill rhythm. NMDA receptor (NMDAR) blockade eliminated LLDs in FTNs, and NMDAR activation in synaptically isolated FTNs induced repetitive LLDs. These results suggest FTNs contain an NMDAR-dependent mechanism that may regulate fast-trill duration. We conclude that a single premotor neuron population employs distinct mechanisms to regulate call rate and duration.

  1. New and already known acanthocephalans from amphibians and reptiles in Vietnam, with keys to species of Pseudoacanthocephalus Petrochenko, 1956 (Echinorhynchidae) and Sphaerechinorhynchus Johnston and Deland, 1929 (Plagiorhynchidae).

    PubMed

    Amin, Omar M; Ha, Ngyuen Van; Heckmann, Richard A

    2008-02-01

    Adults of 2 new species in 2 orders of acanthocephalans obtained from the intestines of terrestrial amphibians and reptiles collected between 1998 and 2004 in Vietnam are described here. Pseudoacanthocephalus nguyenthileae n. sp. (Palaeacnthocephala: Echinorhynchidae) was collected from 5 species of terrestrial amphibians: (1) the common Sunda toad Bufo melanostictus Schneider (Bufonidae); (2) Paa verucospinosa (Bourret); (3) Gunther's Amoy frog Rana guentheri Boulenger; (4) Taipei frog R. taipehensis Denburgh (Ranidae), and (5) the Burmese whipping frog Polypedates mutus (Smith) (Racophoridae); as well as from the Chinese cobra Naja atra Cantor (Reptilia: Elapidae) and house gecko Hemidactylus frenatus Dumeril and Bibron (Reptilia: Gekkonidae). Sphaerechinorhynchus maximesospinus n. sp. (Plagiorhynchidae: Sphaerechinorhynchinae) was isolated from a king cobra Ophiophagus hannah (cantor) (Reptilia: Elapidae). Cystacanths of Porrorchis houdemeri (Joyeux and Baer, 1935) Schmidt and Kuntz, 1967 (Plagiorhynchidae: Porrorchinae) obtained from the mesenteries of banded krait Bungarus fasciatus (Schneider) (Reptilia: Elapidae), a paratenic host, are reported for the first time. Keys to the species of Pseudoacanthocephalus and Sphaerechinorhynchus are included. Characteristic features distinguishing the new species from related taxa include: P. nguyenthileae has 15-19 (usually 16-18) proboscis hook rows, each with 5-6 hooks that progressively increase in length and size posteriorly. The largest, intermediate, and smallest proboscis hooks of S. maximesospinus are the middle, anterior, and posterior hooks, respectively; the proboscis and neck are enclosed in a membrane. Morphometric characteristics of P. nguyenthileae show host-related variability.

  2. An adhesome comprising laminin, dystroglycan and myosin IIA is required during notochord development in Xenopus laevis.

    PubMed

    Buisson, Nicolas; Sirour, Cathy; Moreau, Nicole; Denker, Elsa; Le Bouffant, Ronan; Goullancourt, Aline; Darribère, Thierry; Bello, Valérie

    2014-12-01

    Dystroglycan (Dg) is a transmembrane receptor for laminin that must be expressed at the right time and place in order to be involved in notochord morphogenesis. The function of Dg was examined in Xenopus laevis embryos by knockdown of Dg and overexpression and replacement of the endogenous Dg with a mutated form of the protein. This analysis revealed that Dg is required for correct laminin assembly, for cell polarization during mediolateral intercalation and for proper differentiation of vacuoles. Using mutations in the cytoplasmic domain, we identified two sites that are involved in cell polarization and are required for mediolateral cell intercalation, and a site that is required for vacuolation. Furthermore, using a proteomic analysis, the cytoskeletal non-muscle myosin IIA has been identified for the first time as a molecular link between the Dg-cytoplasmic domain and cortical actin. The data allowed us to identify the adhesome laminin-Dg-myosin IIA as being required to maintain the cortical actin cytoskeleton network during vacuolation, which is crucial to maintain the shape of notochordal cells. © 2014. Published by The Company of Biologists Ltd.

  3. Histone H1 is essential for mitotic chromosome architecture and segregation in Xenopus laevis egg extracts

    PubMed Central

    Maresca, Thomas J.; Freedman, Benjamin S.; Heald, Rebecca

    2005-01-01

    During cell division, condensation and resolution of chromosome arms and the assembly of a functional kinetochore at the centromere of each sister chromatid are essential steps for accurate segregation of the genome by the mitotic spindle, yet the contribution of individual chromatin proteins to these processes is poorly understood. We have investigated the role of embryonic linker histone H1 during mitosis in Xenopus laevis egg extracts. Immunodepletion of histone H1 caused the assembly of aberrant elongated chromosomes that extended off the metaphase plate and outside the perimeter of the spindle. Although functional kinetochores assembled, aligned, and exhibited poleward movement, long and tangled chromosome arms could not be segregated in anaphase. Histone H1 depletion did not significantly affect the recruitment of known structural or functional chromosomal components such as condensins or chromokinesins, suggesting that the loss of H1 affects chromosome architecture directly. Thus, our results indicate that linker histone H1 plays an important role in the structure and function of vertebrate chromosomes in mitosis. PMID:15967810

  4. Developmental changes in head movement kinematics during swimming in Xenopus laevis tadpoles.

    PubMed

    Hänzi, Sara; Straka, Hans

    2017-01-15

    During the post-embryonic developmental growth of animals, a number of physiological parameters such as locomotor performance, dynamics and behavioural repertoire are adjusted to match the requirements determined by changes in body size, proportions and shape. Moreover, changes in movement parameters also cause changes in the dynamics of self-generated sensory stimuli, to which motion-detecting sensory systems have to adapt. Here, we examined head movements and swimming kinematics of Xenopus laevis tadpoles with a body length of 10-45 mm (developmental stage 46-54) and compared these parameters with fictive swimming, recorded as ventral root activity in semi-intact in vitro preparations. Head movement kinematics was extracted from high-speed video recordings of freely swimming tadpoles. Analysis of these locomotor episodes indicated that the swimming frequency decreased with development, along with the angular velocity and acceleration of the head, which represent self-generated vestibular stimuli. In contrast, neither head oscillation amplitude nor forward velocity changed with development despite the ∼3-fold increase in body size. The comparison between free and fictive locomotor dynamics revealed very similar swimming frequencies for similarly sized animals, including a comparable developmental decrease of the swimming frequency. Body morphology and the motor output rhythm of the spinal central pattern generator therefore develop concurrently. This study thus describes development-specific naturalistic head motion profiles, which form the basis for more natural stimuli in future studies probing the vestibular system. © 2017. Published by The Company of Biologists Ltd.

  5. Comparative studies of the endonucleases from two related Xenopus laevis retrotransposons, Tx1L and Tx2L: target site specificity and evolutionary implications.

    PubMed

    Christensen, S; Pont-Kingdon, G; Carroll, D

    2000-01-01

    In the genome of the South African frog, Xenopus laevis, there are two complex families of transposable elements, Tx1 and Tx2, that have identical overall structures, but distinct sequences. In each family there are approximately 1500 copies of an apparent DNA-based element (Tx1D and Tx2D). Roughly 10% of these elements in each family are interrupted by a non-LTR retrotransposon (Tx1L and Tx2L). Each retrotransposon is flanked by a 23-bp target duplication of a specific D element sequence. In earlier work, we showed that the endonuclease domain (Tx1L EN) located in the second open reading frame (ORF2) of Tx1L encodes a protein that makes a single-strand cut precisely at the expected site within its target sequence, supporting the idea that Tx1L is a site-specific retrotransposon. In this study, we express the endonuclease domain of Tx2L (Tx2L EN) and compare the target preferences of the two enzymes. Each endonuclease shows some preference for its cognate target, on the order of 5-fold over the non-cognate target. The observed discrimination is not sufficient, however, to explain the observation that no cross-occupancy is observed - that is, L elements of one family have never been found within D elements of the other family. Possible sources of additional specificity are discussed. We also compare two hypotheses regarding the genome duplication event that led to the contemporary pseudotetraploid character of Xenopus laevis in light of the Tx1L and Tx2L data.

  6. Actions of Hydrogen Sulfide on Sodium Transport Processes across Native Distal Lung Epithelia (Xenopus laevis)

    PubMed Central

    Erb, Alexandra; Althaus, Mike

    2014-01-01

    Hydrogen sulfide (H2S) is well known as a highly toxic environmental chemical threat. Prolonged exposure to H2S can lead to the formation of pulmonary edema. However, the mechanisms of how H2S facilitates edema formation are poorly understood. Since edema formation can be enhanced by an impaired clearance of electrolytes and, consequently, fluid across the alveolar epithelium, it was questioned whether H2S may interfere with transepithelial electrolyte absorption. Electrolyte absorption was electrophysiologically measured across native distal lung preparations (Xenopus laevis) in Ussing chambers. The exposure of lung epithelia to H2S decreased net transepithelial electrolyte absorption. This was due to an impairment of amiloride-sensitive sodium transport. H2S inhibited the activity of the Na+/K+-ATPase as well as lidocaine-sensitive potassium channels located in the basolateral membrane of the epithelium. Inhibition of these transport molecules diminishes the electrochemical gradient which is necessary for transepithelial sodium absorption. Since sodium absorption osmotically facilitates alveolar fluid clearance, interference of H2S with the epithelial transport machinery provides a mechanism which enhances edema formation in H2S-exposed lungs. PMID:24960042

  7. Neurally Derived Tissues in Xenopus laevis Embryos Exhibit a Consistent Bioelectrical Left-Right Asymmetry

    PubMed Central

    Pai, Vaibhav P.; Vandenberg, Laura N.; Blackiston, Douglas; Levin, Michael

    2012-01-01

    Consistent left-right asymmetry in organ morphogenesis is a fascinating aspect of bilaterian development. Although embryonic patterning of asymmetric viscera, heart, and brain is beginning to be understood, less is known about possible subtle asymmetries present in anatomically identical paired structures. We investigated two important developmental events: physiological controls of eye development and specification of neural crest derivatives, in Xenopus laevis embryos. We found that the striking hyperpolarization of transmembrane potential (V mem) demarcating eye induction usually occurs in the right eye field first. This asymmetry is randomized by perturbing visceral left-right patterning, suggesting that eye asymmetry is linked to mechanisms establishing primary laterality. Bilateral misexpression of a depolarizing channel mRNA affects primarily the right eye, revealing an additional functional asymmetry in the control of eye patterning by V mem. The ATP-sensitive K+ channel subunit transcript, SUR1, is asymmetrically expressed in the eye primordia, thus being a good candidate for the observed physiological asymmetries. Such subtle asymmetries are not only seen in the eye: consistent asymmetry was also observed in the migration of differentiated melanocytes on the left and right sides. These data suggest that even anatomically symmetrical structures may possess subtle but consistent laterality and interact with other developmental left-right patterning pathways. PMID:23346115

  8. Neurally Derived Tissues in Xenopus laevis Embryos Exhibit a Consistent Bioelectrical Left-Right Asymmetry.

    PubMed

    Pai, Vaibhav P; Vandenberg, Laura N; Blackiston, Douglas; Levin, Michael

    2012-01-01

    Consistent left-right asymmetry in organ morphogenesis is a fascinating aspect of bilaterian development. Although embryonic patterning of asymmetric viscera, heart, and brain is beginning to be understood, less is known about possible subtle asymmetries present in anatomically identical paired structures. We investigated two important developmental events: physiological controls of eye development and specification of neural crest derivatives, in Xenopus laevis embryos. We found that the striking hyperpolarization of transmembrane potential (V(mem)) demarcating eye induction usually occurs in the right eye field first. This asymmetry is randomized by perturbing visceral left-right patterning, suggesting that eye asymmetry is linked to mechanisms establishing primary laterality. Bilateral misexpression of a depolarizing channel mRNA affects primarily the right eye, revealing an additional functional asymmetry in the control of eye patterning by V(mem). The ATP-sensitive K(+) channel subunit transcript, SUR1, is asymmetrically expressed in the eye primordia, thus being a good candidate for the observed physiological asymmetries. Such subtle asymmetries are not only seen in the eye: consistent asymmetry was also observed in the migration of differentiated melanocytes on the left and right sides. These data suggest that even anatomically symmetrical structures may possess subtle but consistent laterality and interact with other developmental left-right patterning pathways.

  9. Temperature-independent energy expenditure in early development of the African clawed frog Xenopus laevis.

    PubMed

    Nagano, Yatsuhisa; Ode, Koji L

    2014-08-01

    The thermal dissipation of activated eggs and embryos undergoing development from cleavage to the tailbud stage of the African clawed frog Xenopus laevis was measured as a function of incubation time at temperatures ranging from T = 288.2 K to 295.2 K, using a high-precision isothermal calorimeter. A23187-mediated activation of mature eggs induced stable periodic thermal oscillations lasting for 8-34 h. The frequency agreed well with the cell cycle frequency of initial cleavages at the identical temperature. In the developing embryo, energy metabolism switches from embryonic to adult features during gastrulation. The thermal dissipation after gastrulation fit well with a single modified Avrami equation, which has been used for modeling crystal-growth. Both the oscillation frequency of the activated egg and the growth rate of the embryo strongly depend on temperature with the same apparent activation energy of approximately 87 kJ mole(-1). This result suggests that early development proceeds as a single biological time, attributable to a single metabolic rate. A temperature-independent growth curve was derived by scaling the thermogram to the biological time, indicating that the amount of energy expenditure during each developmental stage is constant over the optimal temperature range.

  10. The Sperm-surface glycoprotein, SGP, is necessary for fertilization in the frog, Xenopus laevis.

    PubMed

    Nagai, Keita; Ishida, Takuya; Hashimoto, Takafumi; Harada, Yuichirou; Ueno, Shuichi; Ueda, Yasushi; Kubo, Hideo; Iwao, Yasuhiro

    2009-06-01

    To identify a molecule involved in sperm-egg plasma membrane binding at fertilization, a monoclonal antibody against a sperm-surface glycoprotein (SGP) was obtained by immunizing mice with a sperm membrane fraction of the frog, Xenopus laevis, followed by screening of the culture supernatants based on their inhibitory activity against fertilization. The fertilization of both jellied and denuded eggs was effectively inhibited by pretreatment of sperm with intact anti-SGP antibody as well as its Fab fragment, indicating that the antibody recognizes a molecule on the sperm's surface that is necessary for fertilization. On Western blots, the anti-SGP antibody recognized large molecules, with molecular masses of 65-150 kDa and minor smaller molecules with masses of 20-28 kDa in the sperm membrane vesicles. SGP was distributed over nearly the entire surface of the sperm, probably as an integral membrane protein in close association with microfilaments. More membrane vesicles containing SGP bound to the surface were found in the animal hemisphere compared with the vegetal hemisphere in unfertilized eggs, but the vesicle-binding was not observed in fertilized eggs. These results indicate that SGP mediates sperm-egg membrane binding and is responsible for the establishment of fertilization in Xenopus.

  11. Acanthocephaliasis

    USGS Publications Warehouse

    Cole, Rebecca A.

    1999-01-01

    The phylum Acanthocephala contains parasitic worms referred to as thorny-headed worms because both the larval and adult parasites have a retractable proboscis or a tubular structure at the head, which has sharp, recurved hooks or spines. Much like the cestodes or tapeworms, they lack digestive tracts and absorb nutrients from the bird’s intestinal canal. This may weaken the bird and may make it more susceptible to other diseases and to predation.Adult acanthocephalans are found in a variety of bird species and in other vertebrates. More than 50 species of acanthocephalans have been reported in waterfowl, but reevaluations of acanthocephalan taxonomy are resulting in revised speciation. Nevertheless, numerous species within the phylum are found in birds.

  12. Transforming Growth Factor Beta (TGFβ) Is Produced by and Influences the Proliferative Response of Xenopus laevis Lymphocytes

    PubMed Central

    Haynes, Laura

    1993-01-01

    Both TGF/β2 and 5 have been described in the South African clawed frog Xenopus laevis and have been cloned from the tadpole-derived fibroblast cell line, XTC. Because TGFβ has such a profound inhibitory effect on the mammalian immune system, this study was performed to determine whether TGFβ: (a) has any in vitro effects on the growth of Xenopus lymphoblasts, and (b) is produced by mitogen-activated Xenopus lymphocytes. Following stimulation with mitogen or alloantigen, T lymphocytes from Xenopus secrete a T-cell growth factor (TCGF) that is functionally homologous to mammalian interleukin-2 (IL-2). Both recombinant human TGFβ1 and Xenopus TGFβ5 inhibit TCGF-induced proliferation of Xenopus splenic blasts and this inhibition can be reversed with anti-pan TGFβ antiserum. The Xenopus mitogen-induced saturated ammonium sulfate precipitated TCGF-containing supernatant (SAS TCGF SN) also contains latent TGFβ as assayed on mink lung fibroblasts and Xenopus splenic blasts, and experiments utilizing anti-TGFβ antiserum showed that only TGFβ5 is present in this supernatant. PMID:8281035

  13. Effect of low-level copper and pentachlorophenol exposure on various early life stages of Xenopus laevis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Fort, D.J.; Stover, E.L.

    1996-12-31

    An evaluation of the effects of low-level copper and pentachlorophenol exposure on various early life stages of the South African clawed frog, Xenopus laevis, was performed using stage-specific and long-term continuous exposures. Stage-specific exposure experiments were conducted such that separate subsets of embryos and larvae from the same clutch were exposed to two toxicants, copper and pentachlorphenol, from 0 d to 4 d (standard Frog Embryo Teratogenesis Assay--Xenopus [FETAX]), 4 d to 8 d, 8 d to 12 d, and 12 d to 16 d. Results from two separate concentration-response experiments indicated that sensitivity to either toxicant increased in eachmore » successive time period. Longer-term exposure studies conducted for 60 to 75 days indicated that copper, but not pentachlorophenol induced reduction deficiency malformations of the hind limb at concentrations as low as 0.05 mg/L. Pentachlorophenol concentrations as low as 0.5 {micro}g/L inhibited tail resorption. However, copper did not adversely affect the process of tail resorption. These results indicated that studies evaluating longer-term developmental processes are important in ecological hazard evaluation.« less

  14. Gonadal development of larval male Xenopus laevis exposed to atrazine in outdoor microcosms

    USGS Publications Warehouse

    Jooste, A.M.; Du Preez, L.H.; Carr, J.A.; Giesy, J.P.; Gross, T.S.; Kendall, R.J.; Smith, E.E.; Van Der Kraak, G. L.; Solomon, K.R.

    2005-01-01

    The potential effects of atrazine on gonadal development in metamorphs and subadults of the African clawed frog (Xenopus laevis) were studied under conditions of natural photoperiod and temperatures in outdoor microcosms from August 2002 to June 2003 in South Africa. Triplicate 1100 L microcosms for each nominal concentration of 0.0, 1, 10, and 25 ??g of atrazine/L were used. Measured atrazine concentrations varied <25% throughout the study, and no atrazine was detected in the control microcosms. Tadpoles developed well at all concentrations. On the basis of histological examination of testes of recently metamorphosed stage 66 frogs, 57% of the individuals in the reference group exhibited testicular oocytes as compared with 57, 59, and 39% of the 1, 10, and 25 ??g/L atrazine groups, respectively. The average prevalence of testicular oocytes for all of the treatments including the controls was 54% in a single testis, while, in 35% of individuals, testicular oocytes were observed in both testes. The number of testicular oocytes per individual ranged from 0 to 58 with means of 9.5, 9.8, 8.5, and 11.1 for the 0.0, 1, 10, and 25 ??g of atrazine/L groups, respectively. Ten months after metamorphosis, another subset of juveniles was examined, and the maximum number of testicular oocytes observed was five in one animal. The presence of testicular oocytes was not related to exposure to atrazine and may be a natural phenomenon during ontogeny. ?? 2005 American Chemical Society.

  15. Changes in Oscillatory Dynamics in the Cell Cycle of Early Xenopus laevis Embryos

    PubMed Central

    Tsai, Tony Y.-C.; Theriot, Julie A.; Ferrell, James E.

    2014-01-01

    During the early development of Xenopus laevis embryos, the first mitotic cell cycle is long (∼85 min) and the subsequent 11 cycles are short (∼30 min) and clock-like. Here we address the question of how the Cdk1 cell cycle oscillator changes between these two modes of operation. We found that the change can be attributed to an alteration in the balance between Wee1/Myt1 and Cdc25. The change in balance converts a circuit that acts like a positive-plus-negative feedback oscillator, with spikes of Cdk1 activation, to one that acts like a negative-feedback-only oscillator, with a shorter period and smoothly varying Cdk1 activity. Shortening the first cycle, by treating embryos with the Wee1A/Myt1 inhibitor PD0166285, resulted in a dramatic reduction in embryo viability, and restoring the length of the first cycle in inhibitor-treated embryos with low doses of cycloheximide partially rescued viability. Computations with an experimentally parameterized mathematical model show that modest changes in the Wee1/Cdc25 ratio can account for the observed qualitative changes in the cell cycle. The high ratio in the first cycle allows the period to be long and tunable, and decreasing the ratio in the subsequent cycles allows the oscillator to run at a maximal speed. Thus, the embryo rewires its feedback regulation to meet two different developmental requirements during early development. PMID:24523664

  16. A new sawshark, Pristiophorus laevis, from the Eocene of Antarctica with comments on Pristiophorus lanceolatus.

    PubMed

    Engelbrecht, Andrea; Mörs, Thomas; Reguero, Marcelo A; Kriwet, Jürgen

    2017-01-01

    The highly fossiliferous Eocene deposits of the Antarctic Peninsula are among the most productive sites for fossil remains in the Southern Hemisphere and offer rare insights into high-latitude faunas during the Palaeogene. Chondrichthyans, which are represented by abundant isolated remains, seemingly dominate the marine assemblages. Eocene Antarctic sawsharks have only been known from few isolated rostral spines up to now, that were assigned to Pristiophorus lanceolatus . Here, we present the first oral teeth of a sawshark from the Eocene of Seymour Island and a re-evaluation of previously described Pristiophorus remains from Gondwana consisting exclusively of rostral spines. The holotype of Pristiophorus lanceolatus represents a single, abraded and insufficiently illustrated spine from the Oligocene of New Zealand. All other Cenozoic rostral spines assigned to this species are morphologically very indistinct and closely resemble those of living taxa. Consequently, we regard this species as dubious and introduce a new species, Pristiophorus laevis , based on oral teeth. The combination of dental characteristics of the new species makes it unique compared to all other described species based on oral teeth. Rostral spines from the Eocene of Seymour Island are assigned to this new species whereas those from other Cenozoic Gondwana localities remain ambiguous.

  17. CELL SEGREGATION, MIXING, AND TISSUE PATTERN IN THE SPINAL CORD OF THE XENOPUS LAEVIS NEURULA

    PubMed Central

    Davidson, Lance A.; Keller, Raymond E.

    2014-01-01

    Background During Xenopus laevis neurulation, neural ectodermal cells of the spinal cord are patterned at the same time that they intercalate mediolaterally and radially, moving within and between two cell layers. Curious if these rearrangements disrupt early cell identities, we lineage-traced cells in each layer from neural plate stages to the closed neural tube, and used in situ hybridization to assay gene expression in the moving cells. Results Our biotin- and fluorescent labeling of deep and superficial cells reveals that mediolateral intercalation does not disrupt cell cohorts, in other words it is conservative. However, outside the midline notoplate, later radial intercalation does displace superficial cells dorsoventrally, radically disrupting cell cohorts. The tube roof is composed almost exclusively of superficial cells, including some displaced from ventral positions; gene expression in these displaced cells must now be surveyed further. Superficial cells also flank the tube’s floor, which is, itself, almost exclusively composed of deep cells. Conclusions Our data provide: 1) a fate map of superficial- and deep-cell positions within the Xenopus neural tube, 2) the paths taken to these positions, and 3) preliminary evidence of re-patterning in cells carried out of one environment and into another, during neural morphogenesis. PMID:23813905

  18. A new sawshark, Pristiophorus laevis, from the Eocene of Antarctica with comments on Pristiophorus lanceolatus

    PubMed Central

    Engelbrecht, Andrea; Mörs, Thomas; Reguero, Marcelo A.; Kriwet, Jürgen

    2017-01-01

    The highly fossiliferous Eocene deposits of the Antarctic Peninsula are among the most productive sites for fossil remains in the Southern Hemisphere and offer rare insights into high-latitude faunas during the Palaeogene. Chondrichthyans, which are represented by abundant isolated remains, seemingly dominate the marine assemblages. Eocene Antarctic sawsharks have only been known from few isolated rostral spines up to now, that were assigned to Pristiophorus lanceolatus. Here, we present the first oral teeth of a sawshark from the Eocene of Seymour Island and a re-evaluation of previously described Pristiophorus remains from Gondwana consisting exclusively of rostral spines. The holotype of Pristiophorus lanceolatus represents a single, abraded and insufficiently illustrated spine from the Oligocene of New Zealand. All other Cenozoic rostral spines assigned to this species are morphologically very indistinct and closely resemble those of living taxa. Consequently, we regard this species as dubious and introduce a new species, Pristiophorus laevis, based on oral teeth. The combination of dental characteristics of the new species makes it unique compared to all other described species based on oral teeth. Rostral spines from the Eocene of Seymour Island are assigned to this new species whereas those from other Cenozoic Gondwana localities remain ambiguous. PMID:28579693

  19. Embryonic wound healing by apical contraction and ingression in Xenopus laevis.

    PubMed

    Davidson, Lance A; Ezin, Akouavi M; Keller, Ray

    2002-11-01

    We have characterized excisional wounds in the animal cap of early embryos of the frog Xenopus laevis and found that these wounds close accompanied by three distinct processes: (1) the assembly of an actin purse-string in the epithelial cells at the wound margin, (2) contraction and ingression of exposed deep cells, and (3) protrusive activity of epithelial cells at the margin. Microsurgical manipulation allowing fine control over the area and depth of the wound combined with videomicroscopy and confocal analysis enabled us to describe the kinematics and challenge the mechanics of the closing wound. Full closure typically occurs only when the deep, mesenchymal cell-layer of the ectoderm is left intact; in contrast, when deep cells are removed along with the superficial, epithelial cell-layer of the ectoderm, wounds do not close. Actin localizes to the superficial epithelial cell-layer at the wound margin immediately after wounding and forms a contiguous "purse-string" in those cells within 15 min. However, manipulation and closure kinematics of shaped wounds and microsurgical cuts made through the purse-string rule out a major force-generating role for the purse-string. Further analysis of the cell behaviors within the wound show that deep, mesenchymal cells contract their apical surfaces and ingress from the exposed surface. High resolution time-lapse sequences of cells at the leading edge of the wound show that these cells undergo protrusive activity only during the final phases of wound closure as the ectoderm reseals. We propose that assembly of the actin purse-string works to organize and maintain the epithelial sheet at the wound margin, that contraction and ingression of deep cells pulls the wound margins together, and that protrusive activity of epithelial cells at the wound margin reseals the ectoderm and re-establishes tissue integrity during wound healing in the Xenopus embryonic ectoderm. Copyright 2002 Wiley-Liss, Inc.

  20. Expression of XNOA 36 in the mitochondrial cloud of Xenopus laevis oocytes.

    PubMed

    Vaccaro, M C; Wilding, M; Dale, B; Campanella, C; Carotenuto, R

    2012-08-01

    In Xenopus laevis oocytes a mitochondrial cloud (MC) is found between the nucleus and the plasma membrane at stages I-II of oogenesis. The MC contains RNAs that are transported to the future vegetal pole at stage II of oogenesis. In particular, germinal plasm mRNAs are found in the Message Transport Organiser (METRO) region, the MC region opposite to the nucleus. At stages II-III, a second pathway transports Vg1 and VegT mRNAs to the area where the MC content merges with the vegetal cortex. Microtubules become polarized at the sites of migration of Vg1 and VegT mRNAs through an unknown signalling mechanism. In early meiotic stages, the centrioles are almost completely lost with their remnants being dispersed into the cytoplasm and the MC, which may contain a MTOC to be used in the later localization pathway of the mRNAs. In mammals, XNOA 36 encodes a member of a highly conserved protein family and localises to the nucleolus or in the centromeres. In the Xenopus late stage I oocyte, XNOA 36 mRNA is transiently segregated in one half of the oocyte, anchored by a cytoskeletal network that contains spectrin. Here we found that XNOA 36 transcript also localises to the nucleoli and in the METRO region. XNOA 36 protein immunolocalization, using an antibody employed for the library immunoscreening that depicted XNOA 36 expression colonies, labels the migrating MC, the cytoplasm of stage I oocytes and in particular the vegetal cortex facing the MC. The possible role of XNOA 36 in mRNA anchoring to the vegetal cortex or in participating in early microtubule reorganization is discussed.

  1. An integrated field-effect microdevice for monitoring membrane transport in Xenopus laevis oocytes via lateral proton diffusion.

    PubMed

    Schaffhauser, Daniel Felix; Patti, Monica; Goda, Tatsuro; Miyahara, Yuji; Forster, Ian Cameron; Dittrich, Petra Stephanie

    2012-01-01

    An integrated microdevice for measuring proton-dependent membrane activity at the surface of Xenopus laevis oocytes is presented. By establishing a stable contact between the oocyte vitelline membrane and an ion-sensitive field-effect (ISFET) sensor inside a microperfusion channel, changes in surface pH that are hypothesized to result from facilitated proton lateral diffusion along the membrane were detected. The solute diffusion barrier created between the sensor and the active membrane area allowed detection of surface proton concentration free from interference of solutes in bulk solution. The proposed sensor mechanism was verified by heterologously expressing membrane transport proteins and recording changes in surface pH during application of the specific substrates. Experiments conducted on two families of phosphate-sodium cotransporters (SLC20 & SLC34) demonstrated that it is possible to detect phosphate transport for both electrogenic and electroneutral isoforms and distinguish between transport of different phosphate species. Furthermore, the transport activity of the proton/amino acid cotransporter PAT1 assayed using conventional whole cell electrophysiology correlated well with changes in surface pH, confirming the ability of the system to detect activity proportional to expression level.

  2. Anisakid nematodes (Nematoda: Anisakidae) from the marine fishes Plectropomus laevis Lacépède (Serranidae) and Sphyraena qenie Klunzinger (Sphyraenidae) off New Caledonia, including two new species of Hysterothylacium Ward & Magath, 1917.

    PubMed

    Moravec, František; Justine, Jean-Lou

    2015-11-01

    Based on light and scanning electron microscopical studies, two new species of Hysterothylacium Ward & Magath, 1917 (Nematoda: Anisakidae) are described from the digestive tract of perciform fishes off New Caledonia: H. alatum n. sp. from Plectropomus laevis (Lacépède) (Serranidae) and H. sphyraenae n. sp. from Sphyraena qenie Klunzinger (Sphyraenidae). The former species (H. alatum) is mainly characterised by its large body (male 42.05 mm, gravid females 51.18-87.38 mm long), the shape of the dorsal lip, conspicuously broad cervical alae, a short caecum and a long ventricular appendix, the length of the spicules (925 µm), the number (25 pairs) and distribution of the genital papillae and the tail tip bearing numerous minute cuticular protuberances. The other species (H. sphyraenae) is mainly characterised by the presence of narrow lateral alae, a short caecum and a long ventricular appendix, the length (762-830 µm) and shape of the spicules, the number (37-38 pairs) and arrangement of the genital papillae, and by the tail tip which lacks any distinct cuticular projections visible under the light microscope. In addition, and unidentifiable at the species level, conspicuously large (45.71-66.10 mm long) larvae of Contracaecum Railliet & Henry, 1912, were found in the body cavity of P. laevis, which serves as a paratenic host for this parasite.

  3. Ovarian hyperstimulation syndrome in gonadotropin-treated laboratory South African clawed frogs (Xenopus laevis).

    PubMed

    Green, Sherril L; Parker, John; Davis, Corrine; Bouley, Donna M

    2007-05-01

    Ovarian hyperstimulation syndrome (OHS) is a rare but sometimes fatal iatrogenic complication of ovarian stimulation associated with the administration of exogenous gonadotropins to women undergoing treatment for infertility. Laboratory Xenopus spp are commonly treated with human chorionic gonadotropin (hCG) to stimulate ovulation and optimize the number of oocytes harvested for use in biomedical research. Here we report cases of OHS in 2 gonadotropin-treated laboratory Xenopus laevis. After receiving hCG, the frogs developed severe subcutaneous accumulation of fluid, coelomic distention, and whole-body edema and were unable to dive, although they continued to eat and swim. At postmortem examination, extensive subcutaneous edema was present; ascites and massive numbers of free-floating eggs were found in the coelomic cavity and in aberrant locations: around the heart-sac and adhered to the liver capsule. Whole-body edema, gross enlargement of the ovaries, ascites, and abdominal distention are findings comparable to those observed in women with OHS. The pathophysiology of OHS is thought to be related to hormonally induced disturbances of vasoactive mediators, one of which may be vascular endothelial growth factor secreted by theca and granulosa cells. We know of no other report describing OHSlike symptoms in gonadotropin-treated frogs, and the cases described here are 2 of the 3 we have observed at our respective institutions over the last 6 y. According to these results, OHS appears to be rare in gonadotropin-treated laboratory Xenopus. However, the condition should be included in the differential diagnosis for the bloated frog.

  4. Expression of the adhesion G protein-coupled receptor A2 (adgra2) during Xenopus laevis development.

    PubMed

    Seigfried, Franziska A; Dietmann, Petra; Kühl, Michael; Kühl, Susanne J

    2018-06-01

    The adhesion G protein-coupled receptor A2 (Adgra2) is a seven transmembrane receptor that has been described to be a regulator for angiogenesis in mice. Furthermore, the zebrafish ouchless mutant is unable to develop dorsal root ganglia through a disrupted trafficking of Adgra2. Besides RNA sequencing data, nothing is reported about Adgra2 in the south African crawled frog Xenopus laevis. In this study, we investigated for the first time the spatio-temporal expression of adgra2 during early Xenopus embryogenesis in detail. In silico approaches showed that the genomic adgra2 region as well as the Adgra2 protein sequence is highly conserved among different species including Xenopus. RT-PCR experiments confirmed that embryonic adgra2 expression is primarily detected at the beginning of neurulation and is then present throughout the whole Xenopus embryogenesis until stage 42. Whole mount in situ hybridization approaches visualized adgra2 expression in many tissues during Xenopus embryogenesis such as the cardiovascular system including the heart, the migrating neural crest cells and the developing eye including the periocular mesenchyme. Our results indicate a role of Adgra2 for embryogenesis and are a good starting point for further functional studies during early vertebrate development. Copyright © 2018 Elsevier B.V. All rights reserved.

  5. The Midblastula Transition Defines the Onset of Y RNA-Dependent DNA Replication in Xenopus laevis

    PubMed Central

    Collart, Clara; Christov, Christo P.; Smith, James C.; Krude, Torsten

    2011-01-01

    Noncoding Y RNAs are essential for the initiation of chromosomal DNA replication in mammalian cell extracts, but their role in this process during early vertebrate development is unknown. Here, we use antisense morpholino nucleotides (MOs) to investigate Y RNA function in Xenopus laevis and zebrafish embryos. We show that embryos in which Y RNA function is inhibited by MOs develop normally until the midblastula transition (MBT) but then fail to replicate their DNA and die before gastrulation. Consistent with this observation, Y RNA function is not required for DNA replication in Xenopus egg extracts but is required for replication in a post-MBT cell line. Y RNAs do not bind chromatin in karyomeres before MBT, but they associate with interphase nuclei after MBT in an origin recognition complex (ORC)-dependent manner. Y RNA-specific MOs inhibit the association of Y RNAs with ORC, Cdt1, and HMGA1a proteins, suggesting that these molecular associations are essential for Y RNA function in DNA replication. The MBT is thus a transition point between Y RNA-independent and Y RNA-dependent control of vertebrate DNA replication. Our data suggest that in vertebrates Y RNAs function as a developmentally regulated layer of control over the evolutionarily conserved eukaryotic DNA replication machinery. PMID:21791613

  6. Altered dopamine levels induced by the parasite Profilicollis antarcticus on its intermediate host, the crab Hemigrapsus crenulatus.

    PubMed

    Rojas, José Miguel; Ojeda, F Patricio

    2005-01-01

    A serotonergic pathway is apparently involved in parasite-host interactions. Previous studies conducted in our laboratory showed increased rates in oxygen consumption and alterations in body posture in the crab Hemigrapsus crenulatus parasitized by the acanthocephalan, Profilicollis antarcticus. Such changes may be related to the functions described for biogenic amines in crustaceans. During the infective stage the acanthocephalans live freely in the hemocelomic cavity, suggesting that the possible alteration induced by biogenic amines may be related to their neurohormonal function in crustaceans. To test whether the presence of P. antarcticus produced neurohormonal changes in its intermediate host, H. crenulatus, we analyzed serotonin and dopamine levels in the host using HPLC with electrochemical detection. Two groups of 11 female crabs were studied; one group was artificially inoculated with two cystacanths while the other was used as the control. Our results show a dramatic increase in hemolymph dopamine, but not serotonin in H. crenulatus parasitized by the acanthocephalan P. antarcticus. Our results, along with those reported by Maynard (1996), suggest a parasite-specific strategy involved in the behavior alteration caused by the acanthocephalans on their intermediate host. The use of a biogenic amine as a mechanism of interaction by the parasites gives them an endless number of alternative potential actions on their intermediate hosts.

  7. Prepatterning and patterning of the thalamus along embryonic development of Xenopus laevis

    PubMed Central

    Bandín, Sandra; Morona, Ruth; González, Agustín

    2015-01-01

    Previous developmental studies of the thalamus (alar part of the diencephalic prosomere p2) have defined the molecular basis for the acquisition of the thalamic competence (preparttening), the subsequent formation of the secondary organizer in the zona limitans intrathalamica, and the early specification of two anteroposterior domains (rostral and caudal progenitor domains) in response to inducing activities and that are shared in birds and mammals. In the present study we have analyzed the embryonic development of the thalamus in the anuran Xenopus laevis to determine conserved or specific features in the amphibian diencephalon. From early embryonic stages to the beginning of the larval period, the expression patterns of 22 markers were analyzed by means of combined In situ hybridization (ISH) and immunohistochemical techniques. The early genoarchitecture observed in the diencephalon allowed us to discern the boundaries of the thalamus with the prethalamus, pretectum, and epithalamus. Common molecular features were observed in the thalamic prepatterning among vertebrates in which Wnt3a, Fez, Pax6 and Xiro1 expression were of particular importance in Xenopus. The formation of the zona limitans intrathalamica was observed, as in other vertebrates, by the progressive expression of Shh. The largely conserved expressions of Nkx2.2 in the rostral thalamic domain vs. Gbx2 and Ngn2 (among others) in the caudal domain strongly suggest the role of Shh as morphogen in the amphibian thalamus. All these data showed that the molecular characteristics observed during preparttening and patterning in the thalamus of the anuran Xenopus (anamniote) share many features with those described during thalamic development in amniotes (common patterns in tetrapods) but also with zebrafish, strengthening the idea of a basic organization of this diencephalic region across vertebrates. PMID:26321920

  8. Growth and fatbody cycles in feral populations of the African clawed frog, Xenopus laevis (Pipidae), in California with comments on reproduction

    USGS Publications Warehouse

    McCoid, M.J.; Fritts, T.H.

    1989-01-01

    Feral populations of the African clawed frog (Xenopus laevis) exist in several areas of southern California. By following the first cohort of progeny produced by African clawed frogs at a recently colonized site, data on the growth rates and age at first maturity were obtained in field conditions. Females reached maturity at an earlier age than males, grew faster than males, and attained body lengths up to 25% larger than males. Larger females were capable of producing larger numbers of eggs than small females and, therefore, had greater reproductive potential. The relatively stable ambient temperatures of southern California contributed to the possibility of reproduction of clawed frogs during all but the coolest periods of the year. Cycles detected in the mass of fatbodies suggested that nutrients were mobilized from fat prior to and during ovulation. The amount of fat in females varied widely, but fat in males tended to accumulate as the males grew during the study period.

  9. A glyphosate micro-emulsion formulation displays teratogenicity in Xenopus laevis.

    PubMed

    Bonfanti, Patrizia; Saibene, M; Bacchetta, R; Mantecca, P; Colombo, A

    2018-02-01

    Glyphosate is the active ingredient in broad-spectrum herbicide formulations used in agriculture, domestic area and aquatic weed control worldwide. Its market is growing steadily concurrently with the cultivation of glyphosate-tolerant transgenic crops and emergence of weeds less sensitive to glyphosate. Ephemeral and lentic waters near to agricultural lands, representing favorite habitats for amphibian reproduction and early life-stage development, may thus be contaminated by glyphosate based herbicides (GBHs) residues. Previous studies on larval anuran species highlighted increased mortality and growth effects after exposure to different GBHs in comparison to glyphosate itself, mainly because of the surfactants such as polyethoxylated tallow amine present in the formulations. Nevertheless, these conclusions are not completely fulfilled when the early development, characterized by primary organogenesis events, is considered. In this study, we compare the embryotoxicity of Roundup ® Power 2.0, a new GBH formulation currently authorized in Italy, with that of technical grade glyphosate using the Frog Embryo Teratogenesis Assay-Xenopus (FETAX). Our results evidenced that glyphosate was not embryolethal and only at the highest concentration (50 mg a.e./L) caused edemas. Conversely, Roundup ® Power 2.0 exhibited a 96 h LC50 of 24.78 mg a.e./L and a 96 h EC50 of 7.8 mg a.e./L. A Teratogenic Index of 3.4 was derived, pointing out the high teratogenic potential of the Roundup ® Power 2.0. Specific concentration-dependent abnormal phenotypes, such as craniofacial alterations, microphthalmia, narrow eyes and forebrain regionalization defects were evidenced by gross malformation screening and histopathological analysis. These phenotypes are coherent with those evidenced in Xenopus laevis embryos injected with glyphosate, allowing us to hypothesize that the teratogenicity observed for Roundup ® Power 2.0 may be related to the improved efficacy in delivering

  10. Cytoskeleton and gravity at work in the establishment of dorso-ventral polarity in the egg of Xenopus laevis

    NASA Astrophysics Data System (ADS)

    Ubbels, Geertje A.; Brom, Tim G.

    The establishment of polarities during early embryogenesis is essential for normal development. Amphibian eggs are appropriate models for studies on embryonic pattern formation. The animal-vegetal axis of the axially symmetrical amphibian egg originates during oogenesis and foreshadows the main body axis of the embryo. The dorso-ventral polarity is epigenetically established before first cleavage. Recent experiments strongly suggest that in the monospermic eggs of the anuran Xenopus laevis both the cytoskeleton and gravity act in the determination of the dorso-ventral polarity. In order to test the role of gravity in this process, eggs will be fertilized under microgravity conditions during the SL-D1 flight in 1985. In a fully automatic experiment container eggs will be kept under well-defined conditions and artificially fertilized as soon as microgravity is reached; eggs and embryos at different stages will then be fixed for later examination. Back on earth the material will be analysed and we will know whether fertilization under microgravity conditions is possible. If so, the relation of the dorso-ventral axis to the former sperm entry point will be determined on the whole embryos; in addition eggs and embryos will be analysed cytologically.

  11. Patterned femtosecond-laser ablation of Xenopus laevis melanocytes for studies of cell migration, wound repair, and developmental processes

    PubMed Central

    Mondia, Jessica P.; Adams, Dany S.; Orendorff, Ryan D.; Levin, Michael; Omenetto, Fiorenzo G.

    2011-01-01

    Ultrafast (femtosecond) lasers have become an important tool to investigate biological phenomena because of their ability to effect highly localized tissue removal in surgical applications. Here we describe programmable, microscale, femtosecond-laser ablation of melanocytes found on Xenopus laevis tadpoles, a technique that is applicable to biological studies in development, regeneration, and cancer research. We illustrate laser marking of individual melanocytes, and the drawing of patterns on melanocyte clusters to help track their migration and/or regeneration. We also demonstrate that this system can upgrade scratch tests, a technique used widely with cultured cells to study cell migration and wound healing, to the more realistic in vivo realm, by clearing a region of melanocytes and monitoring their return over time. In addition, we show how melanocyte ablation can be used for loss-of-function experiments by damaging neighboring tissue, using the example of abnormal tail regeneration following localized spinal cord damage. Since the size, shape, and depth of melanocytes vary as a function of tadpole age and melanocyte location (head or tail), an ablation threshold chart is given. Mechanisms of laser ablation are also discussed. PMID:21833375

  12. Patterned femtosecond-laser ablation of Xenopus laevis melanocytes for studies of cell migration, wound repair, and developmental processes.

    PubMed

    Mondia, Jessica P; Adams, Dany S; Orendorff, Ryan D; Levin, Michael; Omenetto, Fiorenzo G

    2011-08-01

    Ultrafast (femtosecond) lasers have become an important tool to investigate biological phenomena because of their ability to effect highly localized tissue removal in surgical applications. Here we describe programmable, microscale, femtosecond-laser ablation of melanocytes found on Xenopus laevis tadpoles, a technique that is applicable to biological studies in development, regeneration, and cancer research. We illustrate laser marking of individual melanocytes, and the drawing of patterns on melanocyte clusters to help track their migration and/or regeneration. We also demonstrate that this system can upgrade scratch tests, a technique used widely with cultured cells to study cell migration and wound healing, to the more realistic in vivo realm, by clearing a region of melanocytes and monitoring their return over time. In addition, we show how melanocyte ablation can be used for loss-of-function experiments by damaging neighboring tissue, using the example of abnormal tail regeneration following localized spinal cord damage. Since the size, shape, and depth of melanocytes vary as a function of tadpole age and melanocyte location (head or tail), an ablation threshold chart is given. Mechanisms of laser ablation are also discussed.

  13. EYA1 mutations associated with the branchio-oto-renal syndrome result in defective otic development in Xenopus laevis

    PubMed Central

    Li, Youe; Manaligod, Jose M.; Weeks, Daniel L.

    2009-01-01

    Background information. The BOR (branchio-oto-renal) syndrome is a dominant disorder most commonly caused by mutations in the EYA1 (Eyes Absent 1) gene. Symptoms commonly include deafness and renal anomalies. Results. We have used the embryos of the frog Xenopus laevis as an animal model for early ear development to examine the effects of different EYA1 mutations. Four eya1 mRNAs encoding proteins correlated with congenital anomalies in human were injected into early stage embryos. We show that the expression of mutations associated with BOR, even in the presence of normal levels of endogenous eya1 mRNA, leads to morphologically abnormal ear development as measured by overall otic vesicle size, establishment of sensory tissue and otic innervation. The molecular consequences of mutant eya1 expression were assessed by QPCR (quantitative PCR) analysis and in situ hybridization. Embryos expressing mutant eya1 showed altered levels of multiple genes (six1, dach, neuroD, ngnr-1 and nt3) important for normal ear development. Conclusions. These studies lend support to the hypothesis that dominant-negative effects of EYA1 mutations may have a role in the pathogenesis of BOR. PMID:19951260

  14. Vestibular lesion-induced developmental plasticity in spinal locomotor networks during Xenopus laevis metamorphosis.

    PubMed

    Beyeler, Anna; Rao, Guillaume; Ladepeche, Laurent; Jacques, André; Simmers, John; Le Ray, Didier

    2013-01-01

    During frog metamorphosis, the vestibular sensory system remains unchanged, while spinal motor networks undergo a massive restructuring associated with the transition from the larval to adult biomechanical system. We investigated in Xenopus laevis the impact of a pre- (tadpole stage) or post-metamorphosis (juvenile stage) unilateral labyrinthectomy (UL) on young adult swimming performance and underlying spinal locomotor circuitry. The acute disruptive effects on locomotion were similar in both tadpoles and juvenile frogs. However, animals that had metamorphosed with a preceding UL expressed restored swimming behavior at the juvenile stage, whereas animals lesioned after metamorphosis never recovered. Whilst kinematic and electrophysiological analyses of the propulsive system showed no significant differences in either juvenile group, a 3D biomechanical simulation suggested that an asymmetry in the dynamic control of posture during swimming could account for the behavioral restoration observed in animals that had been labyrinthectomized before metamorphosis. This hypothesis was subsequently supported by in vivo electromyography during free swimming and in vitro recordings from isolated brainstem/spinal cord preparations. Specifically, animals lesioned prior to metamorphosis at the larval stage exhibited an asymmetrical propulsion/posture coupling as a post-metamorphic young adult. This developmental alteration was accompanied by an ipsilesional decrease in propriospinal coordination that is normally established in strict left-right symmetry during metamorphosis in order to synchronize dorsal trunk muscle contractions with bilateral hindlimb extensions in the swimming adult. Our data thus suggest that a disequilibrium in descending vestibulospinal information during Xenopus metamorphosis leads to an altered assembly of adult spinal locomotor circuitry. This in turn enables an adaptive compensation for the dynamic postural asymmetry induced by the vestibular imbalance

  15. Downregulation of surface sodium pumps by endocytosis during meiotic maturation of Xenopus laevis oocytes

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Schmalzing, G.; Eckard, P.; Kroener, S.P.

    1990-01-01

    During meiotic maturation, plasma membranes of Xenopus laevis oocytes completely lose the capacity to transport Na and K and to bind ouabain. To explore whether the downregulation might be due to an internalization of the sodium pump molecules, the intracellular binding of ouabain was determined. Selective permeabilization of the plasma membrane of mature oocytes (eggs) by digitonin almost failed to disclose ouabain binding sites. However, when the eggs were additionally treated with 0.02% sodium dodecyl sulfate (SDS) to permeabilize inner membranes, all sodium pumps present before maturation were recovered. Phosphorylation by (gamma-32P)ATP combined with SDS-polyacrylamide gel electrophoresis (PAGE) and autoradiographymore » showed that sodium pumps were greatly reduced in isolated plasma membranes of eggs. According to sucrose gradient fractionation, maturation induced a shift of sodium pumps from the plasma membrane fraction to membranes of lower buoyant density with a protein composition different from that of the plasma membrane. Endocytosed sodium pumps identified on the sucrose gradient from (3H)ouabain bound to the cell surface before maturation could be phosphorylated with inorganic (32P)phosphate. The findings suggest that downregulation of sodium pumps during maturation is brought about by translocation of surface sodium pumps to an intracellular compartment, presumably endosomes. This contrasts the mechanism of downregulation of Na-dependent cotransport systems, the activities of which are reduced as a consequence of a maturation-induced depolarization of the membrane without a removal of the corresponding transporter from the plasma membrane.« less

  16. Williams Syndrome Transcription Factor is critical for neural crest cell function in Xenopus laevis

    PubMed Central

    Barnett, Chris; Yazgan, Oya; Kuo, Hui-Ching; Malakar, Sreepurna; Thomas, Trevor; Fitzgerald, Amanda; Harbour, Billy; Henry, Jonathan J.; Krebs, Jocelyn E.

    2012-01-01

    Williams Syndrome Transcription Factor (WSTF) is one of ~25 haplodeficient genes in patients with the complex developmental disorder Williams Syndrome (WS). WS results in visual/spatial processing defects, cognitive impairment, unique behavioral phenotypes, characteristic “elfin” facial features, low muscle tone and heart defects. WSTF exists in several chromatin remodeling complexes and has roles in transcription, replication, and repair. Chromatin remodeling is essential during embryogenesis, but WSTF’s role in vertebrate development is poorly characterized. To investigate the developmental role of WSTF, we knocked down WSTF in Xenopus laevis embryos using a morpholino that targets WSTF mRNA. BMP4 shows markedly increased and spatially aberrant expression in WSTF-deficient embryos, while SHH, MRF4, PAX2, EPHA4 and SOX2 expression are severely reduced, coupled with defects in a number of developing embryonic structures and organs. WSTF-deficient embryos display defects in anterior neural development. Induction of the neural crest, measured by expression of the neural crest-specific genes SNAIL and SLUG, is unaffected by WSTF depletion. However, at subsequent stages WSTF knockdown results in a severe defect in neural crest migration and/or maintenance. Consistent with a maintenance defect, WSTF knockdowns display a specific pattern of increased apoptosis at the tailbud stage in regions corresponding to the path of cranial neural crest migration. Our work is the first to describe a role for WSTF in proper neural crest function, and suggests that neural crest defects resulting from WSTF haploinsufficiency may be a major contributor to the pathoembryology of WS. PMID:22691402

  17. Do Nanoparticle Physico-Chemical Properties and Developmental Exposure Window Influence Nano ZnO Embryotoxicity in Xenopus laevis?

    PubMed

    Bonfanti, Patrizia; Moschini, Elisa; Saibene, Melissa; Bacchetta, Renato; Rettighieri, Leonardo; Calabri, Lorenzo; Colombo, Anita; Mantecca, Paride

    2015-07-28

    The growing global production of zinc oxide nanoparticles (ZnONPs) suggests a realistic increase in the environmental exposure to such a nanomaterial, making the knowledge of its biological reactivity and its safe-by-design synthesis mandatory. In this study, the embryotoxicity of ZnONPs (1-100 mg/L) specifically synthesized for industrial purposes with different sizes, shapes (round, rod) and surface coatings (PEG, PVP) was tested using the frog embryo teratogenesis assay-Xenopus (FETAX) to identify potential target tissues and the most sensitive developmental stages. The ZnONPs did not cause embryolethality, but induced a high incidence of malformations, in particular misfolded gut and abdominal edema. Smaller, round NPs were more effective than the bigger, rod ones, and PEGylation determined a reduction in embryotoxicity. Ingestion appeared to be the most relevant exposure route. Only the embryos exposed from the stomodeum opening showed anatomical and histological lesions to the intestine, mainly referable to a swelling of paracellular spaces among enterocytes. In conclusion, ZnONPs differing in shape and surface coating displayed similar toxicity in X. laevis embryos and shared the same target organ. Nevertheless, we cannot exclude that the physico-chemical characteristics may influence the severity of such effects. Further research efforts are mandatory to ensure the synthesis of safer nano-ZnO-containing products.

  18. Differential recognition of ACE inhibitors in Xenopus laevis oocytes expressing rat PEPT1 and PEPT2.

    PubMed

    Zhu, T; Chen, X Z; Steel, A; Hediger, M A; Smith, D E

    2000-05-01

    To examine the mechanism of inhibition of glycylsarcosine (GlySar) transport by quinapril and enalapril, and whether or not angiotensin converting enzyme (ACE) inhibitors are transported by PEPT2 as well as by PEPT1. Xenopus laevis oocytes were cRNA-injected with rat PEPT1 or PEPT2 and the transport kinetics of radiolabeled GlySar were studied in the absence and presence of quinapril and enalapril. The two-microelectrode voltage-clamp technique was also performed to probe the electrogenic uptake of captopril, quinapril and enalapril. Kinetic analyses demonstrated that quinapril inhibited the uptake of GlySar in a noncompetitive manner in Xenopus oocytes injected with PEPT1 or PEPT2 (Ki = 0.8 or 0.4 mM, respectively). In contrast, a competitive interaction was observed between GlySar and enalapril (Ki = 10.8 mM for PEPT1 or 4.3 mM for PEPT2). Most significantly, captopril and enalapril, but not quinapril, induced inwardly-directed currents in both PEPT1- and PEPT2-expressed oocytes. These results are unique in providing direct evidence for the substrate recognition and transport of some ACE inhibitors by the high- and low-affinity oligopeptide transporters. Our findings point to differences between PEPT1 and PEPT2 in their affinity to, rather than in their specificity for, ACE inhibitors.

  19. A role for FoxN3 in the development of cranial cartilages and muscles in Xenopus laevis (Amphibia: Anura: Pipidae) with special emphasis on the novel rostral cartilages

    PubMed Central

    Schmidt, Jennifer; Schuff, Maximilian; Olsson, Lennart

    2011-01-01

    The origin of morphological novelties is a controversial topic in evolutionary developmental biology. The heads of anuran larvae have several unique structures, including the supra- and infrarostral cartilages, the specialised structure of the gill basket (used for filtration), and novel cranial muscle arrangements. FoxN3, a member of the forkhead/winged helix family of transcription factors, has been implicated as important for normal craniofacial development in the pipid anuran Xenopus laevis. We have investigated the effects of functional knockdown of FoxN3 (using antisense oligonucleotide morpholino) on the development of the larval head skeleton and the associated cranial muscles in X. laevis. Our data complement earlier studies and provide a more complete account of the requirement of FoxN3 in chondrocranium development. In addition, we analyse the effects of FoxN3 knockdown on cranial muscle development. We show that FoxN3 knockdown primarily affects the novel skeletal structures unique to anuran larvae, i.e. the rostralia or the fine structure of the gill apparatus. The articulation between the infrarostral and Meckel's cartilage is malformed and the filigreed processes of the gill basket do not develop. Because these features do not develop after FoxN3 knockdown, the head morphology resembles that in the less specialised larvae of salamanders. Furthermore, the development of all cartilages derived from the neural crest is delayed and cranial muscle fibre development incomplete. The cartilage precursors initially condense in their proper position but later differentiate incompletely; several visceral arch muscles start to differentiate at their origin but fail to extend toward their insertion. Our findings indicate that FoxN3 is essential for the development of novel cartilages such as the infrarostral and other cranial tissues derived from the neural crest and, indirectly, also for muscle morphogenesis. PMID:21050205

  20. Concentrations of 17 elements in the zebra mussel (Dreissena polymorpha), in different tissues of perch (Perca fluviatilis), and in perch intestinal parasites (Acanthocephalus lucii) from the subalpine lake Mondsee, Austria

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sures, B.; Steiner, W.; Rydlo, M.

    1999-11-01

    Concentrations of the elements Al, Ag, Ba, ca, Cd, Co, Cr, cu, Fe, Ga, Mg, Mn, Ni, Pb, Sr, Tl, and Zn were analyzed by inductively coupled plasma mass spectrometry in the acanthocephalan Acanthocephalus lucii (Mueller); in its host, Perca fluviatilis (L.), and in the soft tissue of the zebra mussel, Dreissena polymorpha (Pallas). All animals were collected from the same sampling site in a subalpine lake, Mondsee, in Austria. Most of the elements were found at significantly higher concentrations in the acanthocephalan than in different tissues (muscle, liver, and intestinal wall) of its perch host. Only Co was concentratedmore » in the liver of perch to a level that was significantly higher than that found in the parasite. Most of the analyzed elements were also present at significantly higher concentrations in A. lucii than in D. polymorpha. Barium and Cr were the only elements recorded at higher concentrations in the mussel compared with the acanthocephalan. Thus, when comparing the accumulation of elements, the acanthocephalans appear to be even more suitable than the zebra mussels in terms of their use in the detection of metal contamination within aquatic biotopes. Spearman correlation analysis revealed that the concentrations of several elements within the parasites decreased with increasing infrapopulation. Furthermore, the levels of some elements in the perch liver were negatively correlated with the weight of A. lucii in the intestine. Thus, it emerged that not only is there competition for elements between acanthocephalans inside the gut but there is also competition for these elements between the host and the parasites. The elevated element concentrations demonstrated here in the parasitic worm A. lucii provide support for further investigations of these common helminthes and of their accumulation properties.« less

  1. Sclerocollum saudii Al-Jahdali, 2010 (Acanthocephala: Cavisomidae) as a sentinel for heavy-metal pollution in the Red Sea.

    PubMed

    Hassanine, R M El-S; Al-Hasawi, Z M; Hariri, M S; Touliabah, H El-S

    2018-02-07

    Currently, fish helminth parasites, especially cestodes and acanthocephalans, are regarded as sentinel organisms to elucidate metal pollution in aquatic ecosystems. Here, 34 specimens of the fish Siganus rivulatus were collected in the Red Sea, from a seriously polluted, small lagoon named Sharm-Elmaya Bay, at Sharm El-Sheikh, South Sinai, Egypt; 22 (64.7%) were infected by Sclerocollum saudii (Acanthocephala: Cavisomidae). Thus, 22 natural infrapopulations (26-245 individuals) of this parasite were collected from infected fish. Samples of water and sediments from the bay, samples of muscle, intestine and liver from each fish, and samples from the parasite were taken for analysis of heavy metals (cadmium (Cd) and lead (Pb)). Both Cd and Pb concentrations in sediments were higher than those in water. The concentration of these metals were significantly higher in tissues (intestine, liver and muscle) of non-infected fish than those in infected fish, with Pb concentrations consistently higher than those of Cd, and both were drastically decreased in the order: liver > intestine > muscle. Metal concentrations in this acanthocephalan were much higher than those in its fish host. There were strong negative relationships between metal concentrations in tissues (intestine, liver and muscle) of infected fish and infrapopulation size, and between metal concentrations in the acanthocephalan and its infrapopulation size. These relationships strongly suggest competition for these metals between the fish host and its acanthocephalan parasite, and intraspecific competition among acanthocephalan individuals for available metals in the fish intestine. Bioconcentration factors were relatively high, since the mean Cd concentration in S. saudii was 239, 68 and 329 times higher than those in intestine, liver and muscle tissues, respectively, of its fish host. Also, mean Pb concentration was 55, 13 and 289 times higher than those in these tissues, respectively. The host-parasite system

  2. Growth and fat-body cycles in feral populations of the African clawed frog, Xenopus laevis (Pipidae), in California with comments on reproduction

    USGS Publications Warehouse

    McCoid, Michael J.; Fritts, Thomas H.

    1989-01-01

    Feral populations of the African clawed frog (Xenopus laevis) exist in several areas of southern California. By following the first cohort of progeny produced by African clawed frogs at a recently colonized site, data on the growth rates and age at first maturity were obtained in field conditions. Females reached maturity at an earlier age than males, grew faster than males, and attained body lengths up to 25% larger than males. Larger females were capable of producing larger numbers of eggs than small females and, therefore, had greater reproductive potential. The relatively stable ambient temperatures of southern California contributed to the possibility of reproduction of clawed frogs during all but the coolest periods of the year. Cycles detected in the mass of fatbodies suggested that nutrients were mobilized from fat prior to and during ovulation. The amount of fat in females varies widely, but fat in males tended to accumulate as the males grew during the study period.

  3. The B-subdomain of the Xenopus laevis XFIN KRAB-AB domain is responsible for its weaker transcriptional repressor activity compared to human ZNF10/Kox1.

    PubMed

    Born, Nadine; Thiesen, Hans-Jürgen; Lorenz, Peter

    2014-01-01

    The Krüppel-associated box (KRAB) domain interacts with the nuclear hub protein TRIM28 to initiate or mediate chromatin-dependent processes like transcriptional repression, imprinting or suppression of endogenous retroviruses. The prototype KRAB domain initially identified in ZNF10/KOX1 encompasses two subdomains A and B that are found in hundreds of zinc finger transcription factors studied in human and murine genomes. Here we demonstrate for the first time transcriptional repressor activity of an amphibian KRAB domain. After sequence correction, the updated KRAB-AB domain of zinc finger protein XFIN from the frog Xenopus laevis was found to confer transcriptional repression in reporter assays in Xenopus laevis A6 kidney cells as well as in human HeLa, but not in the minnow Pimephales promelas fish cell line EPC. Binding of the XFIN KRAB-AB domain to human TRIM28 was demonstrated in a classical co-immunoprecipitation approach and visualized in a single-cell compartmentalization assay. XFIN-AB displayed reduced potency in repression as well as lower strength of interaction with TRIM28 compared to ZNF10 KRAB-AB. KRAB-B subdomain swapping between the two KRAB domains indicated that it was mainly the KRAB-B subdomain of XFIN that was responsible for its lower capacity in repression and binding to human TRIM28. In EPC fish cells, ZNF10 and XFIN KRAB repressor activity could be partially restored to low levels by adding exogenous human TRIM28. In contrast to XFIN, we did not find any transcriptional repression activity for the KRAB-like domain of human PRDM9 in HeLa cells. PRDM9 is thought to harbor an evolutionary older domain related to KRAB whose homologs even occur in invertebrates. Our results support the notion that functional bona fide KRAB domains which confer transcriptional repression and interact with TRIM28 most likely co-evolved together with TRIM28 at the beginning of tetrapode evolution.

  4. Essential roles of LEM-domain protein MAN1 during organogenesis in Xenopus laevis and overlapping functions of emerin.

    PubMed

    Reil, Michael; Dabauvalle, Marie-Christine

    2013-01-01

    Mutations in nuclear envelope proteins are linked to an increasing number of human diseases, called envelopathies. Mutations in the inner nuclear membrane protein emerin lead to X-linked Emery-Dreifuss muscular dystrophy, characterized by muscle weakness or wasting. Conversely, mutations in nuclear envelope protein MAN1 are linked to bone and skin disorders. Both proteins share a highly conserved domain, called LEM-domain. LEM proteins are known to interact with Barrier-to-autointegration factor and several transcription factors. Most envelopathies are tissue-specific, but knowledge on the physiological roles of related LEM proteins is still unclear. For this reason, we investigated the roles of MAN1 and emerin during Xenopus laevis organogenesis. Morpholino-mediated knockdown of MAN1 revealed that MAN1 is essential for the formation of eye, skeletal and cardiac muscle tissues. The MAN1 knockdown could be compensated by ectopic expression of emerin, leading to a proper organ development. Further investigations revealed that MAN1 is involved in regulation of genes essential for organ development and tissue homeostasis. Thereby our work supports that LEM proteins might be involved in signalling essential for organ development during early embryogenesis and suggests that loss of MAN1 may cause muscle and retina specific diseases. Copyright © 2013 Elsevier GmbH. All rights reserved.

  5. C8orf46 homolog encodes a novel protein Vexin that is required for neurogenesis in Xenopus laevis.

    PubMed

    Moore, Kathryn B; Logan, Mary A; Aldiri, Issam; Roberts, Jacqueline M; Steele, Michael; Vetter, Monica L

    2018-05-01

    Neural basic helix-loop helix (bHLH) transcription factors promote progenitor cell differentiation by activation of downstream target genes that coordinate neuronal differentiation. Here we characterize a neural bHLH target gene in Xenopus laevis, vexin (vxn; previously sbt1), that is homologous to human c8orf46 and is conserved across vertebrate species. C8orf46 has been implicated in cancer progression, but its function is unknown. Vxn is transiently expressed in differentiating progenitors in the developing central nervous system (CNS), and is required for neurogenesis in the neural plate and retina. Its function is conserved, since overexpression of either Xenopus or mouse vxn expands primary neurogenesis and promotes early retinal cell differentiation in cooperation with neural bHLH factors. Vxn protein is localized to the cell membrane and the nucleus, but functions in the nucleus to promote neural differentiation. Vxn inhibits cell proliferation, and works with the cyclin-dependent kinase inhibitor p27Xic1 (cdkn1b) to enhance neurogenesis and increase levels of the proneural protein Neurog2. We propose that vxn provides a key link between neural bHLH activity and execution of the neurogenic program. Copyright © 2018 Elsevier Inc. All rights reserved.

  6. Transdifferentiation from cornea to lens in Xenopus laevis depends on BMP signalling and involves upregulation of Wnt signalling

    PubMed Central

    2011-01-01

    Background Surgical removal of the lens from larval Xenopus laevis results in a rapid transdifferention of central corneal cells to form a new lens. The trigger for this process is understood to be an induction event arising from the unprecedented exposure of the cornea to the vitreous humour that occurs following lens removal. The molecular identity of this trigger is unknown. Results Here, we have used a functional transgenic approach to show that BMP signalling is required for lens regeneration and a microarray approach to identify genes that are upregulated specifically during this process. Analysis of the array data strongly implicates Wnt signalling and the Pitx family of transcription factors in the process of cornea to lens transdifferentiation. Our analysis also captured several genes associated with congenital cataract in humans. Pluripotency genes, in contrast, were not upregulated, supporting the idea that corneal cells transdifferentiate without returning to a stem cell state. Several genes from the array were expressed in the forming lens during embryogenesis. One of these, Nipsnap1, is a known direct target of BMP signalling. Conclusions Our results strongly implicate the developmental Wnt and BMP signalling pathways in the process of cornea to lens transdifferentiation (CLT) in Xenopus, and suggest direct transdifferentiation between these two anterior eye tissues. PMID:21896182

  7. Caging, but not air deprivation, slows tadpole growth and development in the amphibian Xenopus laevis.

    PubMed

    Rose, Christopher S

    2014-08-01

    Xenopus laevis tadpoles raised in submerged cages in normoxic water develop more slowly than tadpoles raised with access to air. This study distinguishes between the effects of being caged and being deprived access to air on development and growth. Tadpoles were raised in high and low density control tanks and in cages in the same tank that were either completely submerged or with the top exposed to air. Experiments were repeated with the cages in different positions relative to the air stones and with and without the water flow from air stones supplemented with a pump. Whereas caging tadpoles has a large effect on their development and growth, additionally depriving them of air has a small effect and this effect can be removed by optimizing water flow through the cage. The effect of caging, though significant in this study, is small compared to the variation in growth and developmental rates that is commonly encountered within and among controls in lab studies. Caging effects can also be diminished by optimizing rearing conditions and/or having exceptionally vigorous tadpoles. The effects of air deprivation and caging thus pose less of a problem for experimenting on air-deprived (AD) and air-restored Xenopus tadpoles than their inherent variability in growth and developmental rates and their susceptibility to growth and developmental arrest. Further, the effect of air deprivation in this air-breathing amphibian does not pose a conflict with evolutionary hypotheses for lung loss involving lengthening of the larval period and delay in the onset of air breathing. © 2014 Wiley Periodicals, Inc.

  8. Do Nanoparticle Physico-Chemical Properties and Developmental Exposure Window Influence Nano ZnO Embryotoxicity in Xenopus laevis?

    PubMed Central

    Bonfanti, Patrizia; Moschini, Elisa; Saibene, Melissa; Bacchetta, Renato; Rettighieri, Leonardo; Calabri, Lorenzo; Colombo, Anita; Mantecca, Paride

    2015-01-01

    The growing global production of zinc oxide nanoparticles (ZnONPs) suggests a realistic increase in the environmental exposure to such a nanomaterial, making the knowledge of its biological reactivity and its safe-by-design synthesis mandatory. In this study, the embryotoxicity of ZnONPs (1–100 mg/L) specifically synthesized for industrial purposes with different sizes, shapes (round, rod) and surface coatings (PEG, PVP) was tested using the frog embryo teratogenesis assay-Xenopus (FETAX) to identify potential target tissues and the most sensitive developmental stages. The ZnONPs did not cause embryolethality, but induced a high incidence of malformations, in particular misfolded gut and abdominal edema. Smaller, round NPs were more effective than the bigger, rod ones, and PEGylation determined a reduction in embryotoxicity. Ingestion appeared to be the most relevant exposure route. Only the embryos exposed from the stomodeum opening showed anatomical and histological lesions to the intestine, mainly referable to a swelling of paracellular spaces among enterocytes. In conclusion, ZnONPs differing in shape and surface coating displayed similar toxicity in X. laevis embryos and shared the same target organ. Nevertheless, we cannot exclude that the physico-chemical characteristics may influence the severity of such effects. Further research efforts are mandatory to ensure the synthesis of safer nano-ZnO-containing products. PMID:26225989

  9. Embryotoxic effects of environmental chemicals: tests with the South African clawed toad (Xenopus laevis)

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dumpert, K.

    1987-06-01

    In the course of the investigations reported below, it was shown that p-chloroaniline has a lethal effect on the embryos of Xenopus laevis at a concentration of 100 ppm and is development inhibiting (teratogenic) at concentrations of 1 and 10 ppm, respectively. In the case of aniline, a significant development-inhibiting effect was observed at a concentration as low as 1 ppm. A toxic effect was caused by concentrations between 30 and 40 ppm during embryogenesis and by concentrations above 40 ppm during larval development. A very conspicuous finding was an inhibiting effect of 20 to 40 ppm aniline on pigmentationmore » during embryogenesis and of a concentration as low as 1 ppm on the body size of the young toads. In the case of potassium dichromate, it was possible to barely detect a weak development-inhibiting effect during embryogenesis but no development-retarding effect during larval development. Toxic effects of potassium dichromate occurred during embryogenesis at concentrations of 5 and 7.5 ppm and during the larval development at concentrations above 10 ppm. Sodium dodecylbenzenesulfonic acid at a concentration of 50 ppm was found to have such a strong embryolethal effect that 80% of the eggs showed no cell division at all and the remaining 20% developed to only the bicellular stage. A teratogenic effect of this substance was not observed. Phenol, too, was found to be toxic at a concentration of 50 ppm; in contrast to sodium dodecylbenzenesulfonic acid, however, it did not show any lethal effect on the embryos but it did on the tadpoles, mainly in the first stages of larval development. Lower concentrations of phenol (5 and 10 ppm) had a nonsignificant inhibiting effect on the growth of the larvae. A teratogenic effect of phenol was not detected.« less

  10. Expression of the mammalian calcium signaling response to Trypanosoma cruzi in Xenopus laevis oocytes.

    PubMed

    Leite, M F; Moyer, M S; Andrews, N W

    1998-04-01

    Infective stages of the protozoan parasite Trypanosoma cruzi contain a soluble factor that induces elevation in the intracellular free Ca2+ concentration ([Ca2+]i) of mammalian cells. The process is pertussis toxin (PTx)-sensitive, and involves phospholipase C (PLC) activation, inositol 1,4,5-trisphosphate (IP3) formation and Ca2+ release from intracellular stores (Tardieux I, et al. J Exp Med 1994;179:1017-1022; Rodriguez A, et al. J Cell Biol 1995;129:1263-1273). We now report that a molecule exposed on the surface of the target cells is required to trigger the signaling cascade, and that a response with identical characteristics can be induced in Xenopus laevis oocytes injected with mRNA from normal rat kidney (NRK) fibroblasts. Xenopus oocytes do not show an endogenous response to the trypomastigote Ca2+ signaling factor, but a vigorous response in the form of a propagating Ca2+ wave is expressed after injection of NRK cell mRNA. As previously demonstrated for mammalian cells, the response is inhibited when injected oocytes are pretreated with PTx, implicating Galphai or Galphao trimeric G-proteins, and with thapsigargin, which depletes intracellular Ca2+ stores. Moreover, the [Ca2+]i transients triggered by the T. cruzi soluble factor in mRNA-injected oocytes are blocked by the same inhibitors of the parasite oligopeptidase B that abolish the [Ca2+]i response in NRK cells (Burleigh B, Andrews NW. J Biol Chem 1995;270:5172-5180; Burleigh BA et al. J Cell Biol 1997;136:609-620). The NRK mRNA fraction that induces expression of the [Ca2+]i response to the T. cruzi signaling factor contains messages from 1.5 to 2.0 kb, a size range consistent with the family of seven-transmembrane G-protein-coupled receptors.

  11. Protein 4.1 and its interaction with other cytoskeletal proteins in Xenopus laevis oogenesis.

    PubMed

    Carotenuto, Rosa; Petrucci, Tamara C; Correas, Isabel; Vaccaro, Maria C; De Marco, Nadia; Dale, Brian; Wilding, Martin

    2009-06-01

    In human red blood cells, protein 4.1 (4.1R) is an 80-kDa polypeptide that stabilizes the spectrin-actin network and anchors it to the plasma membrane. In non-erythroid cells there is a great variety of 4.1R isoforms, mainly generated by alternative pre-mRNA splicing, which localize at various intracellular sites, including the nucleus. We studied protein 4.1R distribution in relation to beta-spectrin, actin and cytokeratin during Xenopus oogenesis. Immunoprecipitation experiments indicate that at least two isoforms of protein 4.1R are present in Xenopus laevis oocytes: a 56-kDa form in the cytoplasm and a 37-kDa form in the germinal vesicle (GV). Antibodies to beta-spectrin reveal two bands of 239 and 100 kDa in the cytoplasm. Coimmunoprecipitation experiments indicate that both the 37- and 56-kDa isoforms of protein 4.1R associate with the 100-kDa isoform of beta-spectrin. Moreover, the 56-kDa form coimmunoprecipitates with a cytokeratin of the same molecular weight. Confocal immunolocalization shows that protein 4.1R distribution is in the peripheral cytoplasm, in the mitochondrial cloud (MC) and in the GV of previtellogenic oocytes. In the cytoplasm of vitellogenic oocytes, a loose network of fibers stained by the anti-protein 4.1R antibody spreads across the cytoplasm. beta-Spectrin has a similar distribution. Protein 4.1R was found to colocalize with actin in the cortex of oocytes in the form of fluorescent dots. Double immunolocalization of protein 4.1R and cytokeratin depicts two separate networks that overlap throughout the whole cytoplasm. Protein 4.1R filaments partially colocalize with cytokeratin in both the animal and vegetal hemispheres. We hypothesize that protein 4.1R could function as a linker protein between cytokeratin and the actin-based cytoskeleton.

  12. Activation of Src and release of intracellular calcium by phosphatidic acid during Xenopus laevis fertilization

    PubMed Central

    Bates, Ryan C.; Fees, Colby P.; Holland, William L.; Winger, Courtney C.; Batbayar, Khulan; Ancar, Rachel; Bergren, Todd; Petcoff, Douglas; Stith, Bradley J.

    2014-01-01

    We report a new step in the fertilization in Xenopus laevis which has been found to involve activation of Src tyrosine kinase to stimulate phospholipase C-γ (PLC- γ) which increases inositol 1,4,5-trisphosphate (IP3) to release intracellular calcium ([Ca]i). Molecular species analysis and mass measurements suggested that sperm activate phospholipase D (PLD) to elevate phosphatidic acid (PA). We now report that PA mass increased 2.7 fold by 1 minute after insemination and inhibition of PA production by two methods inhibited activation of Src and PLCγ, increased [Ca]i and other fertilization events. As compared to 14 other lipids, PA strongly bound Xenopus Src but not PLCγ. Addition of synthetic PA activated egg Src (an action requiring intact lipid rafts) and PLCγ as well as doubling the amount of PLCγ in rafts. In the absence of elevated [Ca]i, PA addition elevated IP3 mass to levels equivalent to that induced by sperm (but twice that achieved by calcium ionophore). Finally, PA induced [Ca]i release that was blocked by an IP3 receptor inhibitor. As only PLD1b message was detected, and Western blotting did not detect PLD2, we suggest that sperm activate PLD1b to elevate PA which then binds to and activates Src leading to PLCγ stimulation, IP3 elevation and [Ca]i release. Due to these and other studies, PA may also play a role in membrane fusion events such as sperm-egg fusion, cortical granule exocytosis, the elevation of phosphatidylinositol 4,5-bisphosphate and the large, late increase in sn 1,2-diacylglycerol in fertilization. PMID:24269904

  13. Activation of Src and release of intracellular calcium by phosphatidic acid during Xenopus laevis fertilization.

    PubMed

    Bates, Ryan C; Fees, Colby P; Holland, William L; Winger, Courtney C; Batbayar, Khulan; Ancar, Rachel; Bergren, Todd; Petcoff, Douglas; Stith, Bradley J

    2014-02-01

    We report a new step in the fertilization in Xenopus laevis which has been found to involve activation of Src tyrosine kinase to stimulate phospholipase C-γ (PLC-γ) which increases inositol 1,4,5-trisphosphate (IP3) to release intracellular calcium ([Ca](i)). Molecular species analysis and mass measurements suggested that sperm activate phospholipase D (PLD) to elevate phosphatidic acid (PA). We now report that PA mass increased 2.7 fold by 1 min after insemination and inhibition of PA production by two methods inhibited activation of Src and PLCγ, increased [Ca](i) and other fertilization events. As compared to 14 other lipids, PA specifically bound Xenopus Src but not PLCγ. Addition of synthetic PA activated egg Src (an action requiring intact lipid rafts) and PLCγ as well as doubling the amount of PLCγ in rafts. In the absence of elevated [Ca](i), PA addition elevated IP3 mass to levels equivalent to that induced by sperm (but twice that achieved by calcium ionophore). Finally, PA induced [Ca](i) release that was blocked by an IP3 receptor inhibitor. As only PLD1b message was detected, and Western blotting did not detect PLD2, we suggest that sperm activate PLD1b to elevate PA which then binds to and activates Src leading to PLCγ stimulation, IP3 elevation and [Ca](i) release. Due to these and other studies, PA may also play a role in membrane fusion events such as sperm-egg fusion, cortical granule exocytosis, the elevation of phosphatidylinositol 4,5-bisphosphate and the large, late increase in sn 1,2-diacylglycerol in fertilization. © 2013 The Authors. Published by Elsevier Inc. All rights reserved.

  14. Spatial distribution of the capacity to initiate a secondary embryo in the 32-cell embryo of Xenopus laevis.

    PubMed

    Kageura, H

    1990-12-01

    To examine the spatial distribution of dorsal determinants in the early embryos of Xenopus laevis, individual cells from the 32-cell embryo were transplanted into the same tier of the ventral side of a synchronous recipient. Their abilities to initiate a secondary embryo were measured by the incidence of secondary embryos and by the length of the secondary axis relative to the primary embryo. The ability was found to be localized in all cells (A1, B1, C1, and D1) of the dorsal most column and in the vegetal cells (C2 and D2) of the dorsolateral column. Transplanted C1 (subequatorial) cells caused the highest incidence of a secondary embryo and the average relative length of the secondary embryo was also greatest. Effectiveness decreased in the order: D1, B1, D2, C2, and A1. When these results were compared with Dale and Slack's fate map of the 32-cell embryo, it was concluded that the distribution of dorsal determinants is unique and does not coincide with the prospective regions for any tissues, though it is somewhat similar to the prospective region of dorsal endoderm or notochord. From these results it seems that dorsal determinants do not determine a particular tissue in an embryo but rather the "dorsal" region of an embryo.

  15. Transcription factor COUP-TFII is indispensable for venous and lymphatic development in zebrafish and Xenopus laevis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Aranguren, Xabier L., E-mail: xabier.lopezaranguren@med.kuleuven.be; Beerens, Manu, E-mail: manu.beerens@med.kuleuven.be; Vandevelde, Wouter, E-mail: woutervandevelde@gmail.com

    Highlights: {yields} COUP-TFII deficiency in zebrafish affects arterio-venous EC specification. {yields} COUP-TFII is indispensable for lymphatic development in zebrafish. {yields} COUP-TFII knockdown in Xenopus disrupts lymphatic EC differentiation and migration. {yields} COUP-TFII's role in EC fate decisions is evolutionary conserved. -- Abstract: Transcription factors play a central role in cell fate determination. Gene targeting in mice revealed that Chicken Ovalbumin Upstream Promoter-Transcription Factor II (COUP-TFII, also known as Nuclear Receptor 2F2 or NR2F2) induces a venous phenotype in endothelial cells (ECs). More recently, NR2F2 was shown to be required for initiating the expression of Prox1, responsible for lymphatic commitment ofmore » venous ECs. Small animal models like zebrafish embryos and Xenopus laevis tadpoles have been very useful to elucidate mechanisms of (lymph) vascular development. Therefore, the role of NR2F2 in (lymph) vascular development was studied by eliminating its expression in these models. Like in mice, absence of NR2F2 in zebrafish resulted in distinct vascular defects including loss of venous marker expression, major trunk vessel fusion and vascular leakage. Both in zebrafish and Xenopus the development of the main lymphatic structures was severely hampered. NR2F2 knockdown significantly decreased prox1 expression in zebrafish ECs and the same manipulation affected lymphatic (L)EC commitment, migration and function in Xenopus tadpoles. Therefore, the role of NR2F2 in EC fate determination is evolutionary conserved.« less

  16. Pitx2c attenuation results in cardiac defects and abnormalities of intestinal orientation in developing Xenopus laevis.

    PubMed

    Dagle, John M; Sabel, Jaime L; Littig, Jennifer L; Sutherland, Lillian B; Kolker, Sandra J; Weeks, Daniel L

    2003-10-15

    The experimental manipulation of early embryologic events, resulting in the misexpression of the homeobox transcription factor pitx2, is associated with subsequent defects of laterality in a number of vertebrate systems. To clarify the role of one pitx2 isoform, pitx2c, in determining the left-right axis of amphibian embryos, we examined the heart and gut morphology of Xenopus laevis embryos after attenuating pitx2c mRNA levels using chemically modified antisense oligonucleotides. We demonstrate that the partial depletion of pitx2c mRNA in these embryos results in alteration of both cardiac morphology and intestinal coiling. The most common cardiac abnormality seen was a failure of rightward migration of the outflow tract, while the most common intestinal laterality phenotype seen was a full reversal in the direction of coiling, each present in 23% of embryos injected with the pitx2c antisense oligonucleotide. An abnormality in either the heart or gut further predisposed to a malformation in the other. In addition, a number of other cardiac anomalies were observed after pitx2c mRNA attenuation, including abnormalities of atrial septation, extracellular matrix restriction, relative atrial-ventricular chamber positioning, and restriction of ventricular development. Many of these findings correlate with cardiac defects previously reported in pitx2 null and hypomorphic mice, but can now be assigned specifically to attenuation of the pitx2c isoform in Xenopus.

  17. Differential muscle regulatory factor gene expression between larval and adult myogenesis in the frog Xenopus laevis: adult myogenic cell-specific myf5 upregulation and its relation to the notochord suppression of adult muscle differentiation.

    PubMed

    Yamane, Hitomi; Nishikawa, Akio

    2013-08-01

    During Xenopus laevis metamorphosis, larval-to-adult muscle conversion depends on the differential responses of adult and larval myogenic cells to thyroid hormone. Essential differences in cell growth, differentiation, and hormone-dependent life-or-death fate have been reported between cultured larval (tail) and adult (hindlimb) myogenic cells. A previous study revealed that tail notochord cells suppress terminal differentiation in adult (but not larval) myogenic cells. However, little is known about the differences in expression patterns of myogenic regulatory factors (MRF) and the satellite cell marker Pax7 between adult and larval myogenic cells. In the present study, we compared mRNA expression of these factors between the two types. At first, reverse transcription polymerase chain reaction analysis of hindlimb buds showed sequential upregulation of myf5, myogenin, myod, and mrf4 during stages 50-54, when limb buds elongate and muscles begin to form. By contrast, in the tail, there was no such increase during the same period. Secondary, these results were duplicated in vitro: adult myogenic cells upregulated myf5, myod, and pax7 in the early culture period, followed by myogenin upregulation and myotube differentiation, while larval myogenic cells did not upregulate these genes and precociously started myotube differentiation. Thirdly, myf5 upregulation and early-phase proliferation in adult myogenic cells were potently inhibited by the presence of notochord cells, suggesting that notochord cells suppress adult myogenesis through inhibiting the transition from Myf5(-) stem cells to Myf5(+) committed myoblasts. All of the data presented here suggest that myf5 upregulation can be a good criterion for the activation of adult myogenesis during X. laevis metamorphosis.

  18. The cloning and characterization of a localized maternal transcript in Xenopus laevis whose zygotic counterpart is detected in the CNS.

    PubMed

    Reddy, B A; Kloc, M; Etkin, L D

    1992-12-01

    We have cloned a cDNA (xlan4) from a Xenopus laevis oocyte cDNA library whose cognate mRNA is localized in the animal pole region of full grown oocytes. The cDNA can be translated in vitro to produce a predicted size protein of 35 kDa and, is also expressed in E. coli as a fusion protein. The conceptual protein encoded by the xlan4 cDNA is 17.5% proline rich and possesses several PEST sequences found in proteins with short half-lives. The xlan4 mRNA is 2.6 kb and during early development its titer decreases until the neurula stage after which it begins to reaccumulate. Northern blots on dissected embryos and in situ hybridization revealed that the zygotic expression is limited to the dorsal axial structures consisting primarily of the CNS. UV irradiation of the vegetal pole region immediately following fertilization that produces ventralized embryos results in a loss of zygotic xlan4 expression. In the adult, xlan4 mRNA is limited primarily to the brain. The presence of this mRNA in animal pole region which contributes to the future neural cell lineages suggests that this gene product may function either in the specification of neural cell types or in a neural specific function.

  19. Polyunsaturated fatty acids are potent openers of human M-channels expressed in Xenopus laevis oocytes.

    PubMed

    Liin, S I; Karlsson, U; Bentzen, B H; Schmitt, N; Elinder, F

    2016-09-01

    Polyunsaturated fatty acids have been reported to reduce neuronal excitability, in part by promoting inactivation of voltage-gated sodium and calcium channels. Effects on neuronal potassium channels are less explored and experimental data ambiguous. The aim of this study was to investigate anti-excitable effects of polyunsaturated fatty acids on the neuronal M-channel, important for setting the resting membrane potential in hippocampal and dorsal root ganglion neurones. Effects of fatty acids and fatty acid analogues on mouse dorsal root ganglion neurones and on the human KV 7.2/3 channel expressed in Xenopus laevis oocytes were studied using electrophysiology. Extracellular application of physiologically relevant concentrations of the polyunsaturated fatty acid docosahexaenoic acid hyperpolarized the resting membrane potential (-2.4 mV by 30 μm) and increased the threshold current to evoke action potentials in dorsal root ganglion neurones. The polyunsaturated fatty acids docosahexaenoic acid, α-linolenic acid and eicosapentaenoic acid facilitated opening of the human M-channel, comprised of the heteromeric human KV 7.2/3 channel expressed in Xenopus oocytes, by shifting the conductance-vs.-voltage curve towards more negative voltages (by -7.4 to -11.3 mV by 70 μm). Uncharged docosahexaenoic acid methyl ester and monounsaturated oleic acid did not facilitate opening of the human KV 7.2/3 channel. These findings suggest that circulating polyunsaturated fatty acids, with a minimum requirement of multiple double bonds and a charged carboxyl group, dampen excitability by opening neuronal M-channels. Collectively, our data bring light to the molecular targets of polyunsaturated fatty acids and thus a possible mechanism by which polyunsaturated fatty acids reduce neuronal excitability. © 2016 Scandinavian Physiological Society. Published by John Wiley & Sons Ltd.

  20. Developmental disorders in embryos of the frog Xenopus laevis induced by chloroacetanilide herbicides and their degradation products.

    PubMed

    Osano, Odipo; Admiraal, Wim; Otieno, Dismas

    2002-02-01

    Pesticides are known to transform in the environment, but so far the study of their effects in the environment has concentrated on the parent compounds, thereby neglecting the effects of the degradation products. The embryotoxic, developmental, and teratogenic effects of chloroacetanilide herbicides and their environmentally stable aniline degradation products were investigated in this study in view of the massive application of alachlor and metolachlor. Embryos at midblastula to early gastrula stages of a locally abundant African clawed frog Xenopus laevis were used as test organisms. The embryos were exposed to the test chemicals for 96 h in each experiment. Alachlor is more embryotoxic (the concentration causing 50% embryo lethality, 96-h LC50 = 23 microM [6.1 mg/L]) and teratogenic (teratogenic index [TI] = 1.7) than metolachlor (96-h LC50 = 48 microM [13.6 mg/L], TI = 0.2). The degradation products of alachlor and metolachlor, respectively, 2,6-diethylaniline (96-h LC50 = 13 microM [19.4 mg/L], TI = 2.1) and 2-ethyl-6-methyaniline (96-h LC50 = 509 microM [68.8 mg/L], TI = 2.7), are less embryotoxic but more teratogenic than their parent compounds. The most common teratogenic effects observed were edema for alachlor as opposed to axial flexures and eye abnormalities for 2,6-diethylaniline and 2-ethyl-6-methylaniline. Metolachlor is found to be an example of a nonteratogenic herbicide that upon degradation loses toxicity but gains teratogenicity, and both the herbicides, metolachlor and alachlor, are potential sources of teratogenic transformation products.

  1. NSF- and SNARE-mediated membrane fusion is required for nuclear envelope formation and completion of nuclear pore complex assembly in Xenopus laevis egg extracts.

    PubMed

    Baur, Tina; Ramadan, Kristijan; Schlundt, Andreas; Kartenbeck, Jürgen; Meyer, Hemmo H

    2007-08-15

    Despite the progress in understanding nuclear envelope (NE) reformation after mitosis, it has remained unclear what drives the required membrane fusion and how exactly this is coordinated with nuclear pore complex (NPC) assembly. Here, we show that, like other intracellular fusion reactions, NE fusion in Xenopus laevis egg extracts is mediated by SNARE proteins that require activation by NSF. Antibodies against Xenopus NSF, depletion of NSF or the dominant-negative NSF(E329Q) variant specifically inhibited NE formation. Staging experiments further revealed that NSF was required until sealing of the envelope was completed. Moreover, excess exogenous alpha-SNAP that blocks SNARE function prevented membrane fusion and caused accumulation of non-flattened vesicles on the chromatin surface. Under these conditions, the nucleoporins Nup107 and gp210 were fully recruited, whereas assembly of FxFG-repeat-containing nucleoporins was blocked. Together, we define NSF- and SNARE-mediated membrane fusion events as essential steps during NE formation downstream of Nup107 recruitment, and upstream of membrane flattening and completion of NPC assembly.

  2. Changes in Acetyl CoA Levels during the Early Embryonic Development of Xenopus laevis

    PubMed Central

    Tsuchiya, Yugo; Pham, Uyen; Hu, Wanzhou; Ohnuma, Shin-ichi; Gout, Ivan

    2014-01-01

    Coenzyme A (CoA) is a ubiquitous and fundamental intracellular cofactor. CoA acts as a carrier of metabolically important carboxylic acids in the form of CoA thioesters and is an obligatory component of a multitude of catabolic and anabolic reactions. Acetyl CoA is a CoA thioester derived from catabolism of all major carbon fuels. This metabolite is at a metabolic crossroads, either being further metabolised as an energy source or used as a building block for biosynthesis of lipids and cholesterol. In addition, acetyl CoA serves as the acetyl donor in protein acetylation reactions, linking metabolism to protein post-translational modifications. Recent studies in yeast and cultured mammalian cells have suggested that the intracellular level of acetyl CoA may play a role in the regulation of cell growth, proliferation and apoptosis, by affecting protein acetylation reactions. Yet, how the levels of this metabolite change in vivo during the development of a vertebrate is not known. We measured levels of acetyl CoA, free CoA and total short chain CoA esters during the early embryonic development of Xenopus laevis using HPLC. Acetyl CoA and total short chain CoA esters start to increase around midblastula transition (MBT) and continue to increase through stages of gastrulation, neurulation and early organogenesis. Pre-MBT embryos contain more free CoA relative to acetyl CoA but there is a shift in the ratio of acetyl CoA to CoA after MBT, suggesting a metabolic transition that results in net accumulation of acetyl CoA. At the whole-embryo level, there is an apparent correlation between the levels of acetyl CoA and levels of acetylation of a number of proteins including histones H3 and H2B. This suggests the level of acetyl CoA may be a factor, which determines the degree of acetylation of these proteins, hence may play a role in the regulation of embryogenesis. PMID:24831956

  3. Isolation and characterization of Xenopus laevis homologs of the mouse inv gene and functional analysis of the conserved calmodulin binding sites.

    PubMed

    Yasuhiko, Yukuto; Shiokawa, Koichiro; Mochizuki, Toshio; Asashima, Makoto; Yokoyama, Takahiko

    2006-04-01

    The homozygous inv (inversion of embryonic turning) mouse mutant shows situs inversus and polycystic kidney disease, both of which result from the lack of the inv gene. Previously, we suggested that inv may be important for the left-right axis formation, not only in mice but also in Xenopus, and that calmodulin regulates this inv protein function. Here, we isolated and characterized two Xenopus laevis homologs (Xinv-1 and Xinv-2) of the mouse inv gene, and performed functional analysis of the conserved IQ motifs that interact with calmodulin. Xinv-1 expresses early in development in the same manner as mouse inv does. Unexpectedly, a full-length Xenopus inv mRNA did not randomize cardiac orientation when injected into Xenopus embryos, which is different from mouse inv mRNA. Contrary to mouse inv mRNA, Xenopus inv mRNA with mutated IQ randomized cardiac orientation. The present study indicates that calmodulin binding sites (IQ motifs) are crucial in controlling the biological activity of both mouse and Xenopus inv proteins. Although mouse and Xenopus inv genes have a quite similar structure, the interaction with calmodulin and IQ motifs of Xenopus inv and mouse inv proteins may regulate their function in different ways.

  4. Controlling the Messenger: Regulated Translation of Maternal mRNAs in Xenopus laevis Development

    PubMed Central

    Fox, Catherine A.; Dowdle, Megan E.; Blaser, Susanne Imboden; Chung, Andy; Park, Sookhee

    2017-01-01

    The selective translation of maternal mRNAs encoding cell-fate determinants drives the earliest decisions of embryogenesis that establish the vertebrate body plan. This chapter will discuss studies in Xenopus laevis that provide insights into mechanisms underlying this translational control. Xenopus has been a powerful model organism for many discoveries relevant to the translational control of maternal mRNAs because of the large size of its oocytes and eggs that allow for microinjection of molecules and the relative ease of manipulating the oocyte to egg transition (maturation) and fertilization in culture. Consequently, many key studies have focused on the expression of maternal mRNAs during the oocyte to egg transition (the meiotic cell cycle) and the rapid cell divisions immediately following fertilization. This research has made seminal contributions to our understanding of translational regulatory mechanisms, but while some of the mRNAs under consideration at these stages encode cell-fate determinants, many encode cell cycle regulatory proteins that drive these early cell cycles. In contrast, while maternal mRNAs encoding key developmental (i.e., cell-fate) regulators that function after the first cleavage stages may exploit aspects of these foundational mechanisms, studies reveal that these mRNAs must also rely on distinct and, as of yet, incompletely understood mechanisms. These findings are logical because the functions of such developmental regulatory proteins have requirements distinct from cell cycle regulators, including becoming relevant only after fertilization and then only in specific cells of the embryo. Indeed, key maternal cell-fate determinants must be made available in exquisitely precise amounts (usually low), only at specific times and in specific cells during embryogenesis. To provide an appreciation for the regulation of maternal cell-fate determinant expression, an overview of the maternal phase of Xenopus embryogenesis will be presented

  5. Analysis of thyroid hormone receptor {beta}A mRNA expression in Xenopus laevis tadpoles as a means to detect agonism and antagonism of thyroid hormone action

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Opitz, Robert; Lutz, Ilka; Nguyen, Ngoc-Ha

    2006-04-01

    Amphibian metamorphosis represents a unique biological model to study thyroid hormone (TH) action in vivo. In this study, we examined the utility of thyroid hormone receptors {alpha} (TR{alpha}) and {beta}A (TR{beta}A) mRNA expression patterns in Xenopus laevis tadpoles as molecular markers indicating modulation of TH action. During spontaneous metamorphosis, only moderate changes were evident for TR{alpha} gene expression whereas a marked up-regulation of TR{beta}A mRNA occurred in hind limbs (prometamorphosis), head (late prometamorphosis), and tail tissue (metamorphic climax). Treatment of premetamorphic tadpoles with 1 nM 3,5,3'-triiodothyronine (T3) caused a rapid induction of TR{beta}A mRNA in head and tail tissue withinmore » 6 to 12 h which was maintained for at least 72 h after initiation of T3 treatment. Developmental stage had a strong influence on the responsiveness of tadpole tissues to induce TR{beta}A mRNA during 24 h treatment with thyroxine (0, 1, 5, 10 nM T4) or T3 (0, 1, 5, 10 nM). Premetamorphic tadpoles were highly sensitive in their response to T4 and T3 treatments, whereas sensitivity to TH was decreased in early prometamorphic tadpoles and strongly diminished in late prometamorphic tadpoles. To examine the utility of TR{beta}A gene expression analysis for detection of agonistic and antagonistic effects on T3 action, mRNA expression was assessed in premetamorphic tadpoles after 48 h of treatment with the synthetic agonist GC-1 (0, 10, 50, 250 nM), the synthetic antagonist NH-3 (0, 40, 200, 1000 nM), and binary combinations of NH-3 (0, 40, 200, 1000 nM) and T3 (1 nM). All tested concentrations of GC-1 as well as the highest concentration of NH-3 caused an up-regulation of TR{beta}A expression. Co-treatment with NH-3 and T3 revealed strong antagonistic effects by NH-3 on T3-induced TR{beta}A mRNA up-regulation. Results of this study suggest that TR{beta}A mRNA expression analysis could serve as a sensitive molecular testing approach to study

  6. Pattern of calbindin-D28k and calretinin immunoreactivity in the brain of Xenopus laevis during embryonic and larval development.

    PubMed

    Morona, Ruth; González, Agustín

    2013-01-01

    The present study represents a detailed spatiotemporal analysis of the localization of calbindin-D28k (CB) and calretinin (CR) immunoreactive structures in the brain of Xenopus laevis throughout development, conducted with the aim to correlate the onset of the immunoreactivity with the development of compartmentalization of distinct subdivisions recently identified in the brain of adult amphibians and primarily highlighted when analyzed within a segmental paradigm. CR and CB are expressed early in the brain and showed a progressively increasing expression throughout development, although transient expression in some neuronal subpopulations was also noted. Common and distinct characteristics in Xenopus, as compared with reported features during development in the brain of mammals, were observed. The development of specific regions in the forebrain such as the olfactory bulbs, the components of the basal ganglia and the amygdaloid complex, the alar and basal hypothalamic regions, and the distinct diencephalic neuromeres could be analyzed on the basis of the distinct expression of CB and CR in subregions. Similarly, the compartments of the mesencephalon and the main rhombencephalic regions, including the cerebellum, were differently highlighted by their specific content in CB and CR throughout development. Our results show the usefulness of the analysis of the distribution of these proteins as a tool in neuroanatomy to interpret developmental aspects of many brain regions. Copyright © 2012 Wiley Periodicals, Inc.

  7. Development of selective blockers for Ca2+-activated Cl- channel using Xenopus laevis oocytes with an improved drug screening strategy

    PubMed Central

    Oh, Soo-Jin; Park, Jung Hwan; Han, Sungyu; Lee, Jae Kyun; Roh, Eun Joo; Lee, C Justin

    2008-01-01

    Background Ca2+-activated Cl- channels (CaCCs) participate in many important physiological processes. However, the lack of effective and selective blockers has hindered the study of these channels, mostly due to the lack of good assay system. Here, we have developed a reliable drug screening method for better blockers of CaCCs, using the endogeneous CaCCs in Xenopus laevis oocytes and two-electrode voltage-clamp (TEVC) technique. Results Oocytes were prepared with a treatment of Ca2+ ionophore, which was followed by a treatment of thapsigargin which depletes Ca2+ stores to eliminate any contribution of Ca2+ release. TEVC was performed with micropipette containing chelerythrine to prevent PKC dependent run-up or run-down. Under these conditions, Ca2+-activated Cl- currents induced by bath application of Ca2+ to oocytes showed stable peak amplitude when repetitively activated, allowing us to test several concentrations of a test compound from one oocyte. Inhibitory activities of commercially available blockers and synthesized anthranilic acid derivatives were tested using this method. As a result, newly synthesized N-(4-trifluoromethylphenyl)anthranilic acid with trifluoromethyl group (-CF3) at para position on the benzene ring showed the lowest IC50. Conclusion Our results provide an optimal drug screening strategy suitable for high throughput screening, and propose N-(4-trifluoromethylphenyl)anthranilic acid as an improved CaCC blocker. PMID:18959787

  8. The RNA-binding protein xCIRP2 is involved in apoptotic tail regression during metamorphosis in Xenopus laevis tadpoles.

    PubMed

    Eto, Ko; Iwama, Tomoyuki; Tajima, Tatsuya; Abe, Shin-ichi

    2012-10-01

    Frog metamorphosis induced by thyroid hormone (TH) involves not only cell proliferation and differentiation in reconstituted organs such as limbs, but also apoptotic cell death in degenerated organs such as tails. However, the molecular mechanisms directing the TH-dependent cell fate determination remain unclear. We have previously identified from newts an RNA-binding protein (nRBP) acting as the regulator governing survival and death in germ cells during spermatogenesis. To investigate the molecular events leading the tail resorption during metamorphosis, we analyzed the expression, the functional role in apoptosis, and the regulation of xCIRP2, a frog homolog of nRBP, in tails of Xenopus laevis tadpoles. At the prometamorphic stage, xCIRP2 protein is expressed in fibroblast, epidermal, nerve, and muscular cells and localized in their cytoplasm. When spontaneous metamorphosis progressed, the level of xCIRP2 mRNA remained unchanged but the amount of the protein decreased. In organ cultures of tails at the prometamorphic stage, xCIRP2 protein decreased before their lengths shortened during TH-dependent metamorphosis. The inhibition of calpain or proteasome attenuated the TH-induced decrease of xCIRP2 protein in tails, impairing their regression. These results suggest that xCIRP2 protein is downregulated through calpain- and proteasome-mediated proteolysis in response to TH at the onset of metamorphosis, inducing apoptosis in tails and thereby degenerating them. Copyright © 2012 Elsevier Inc. All rights reserved.

  9. Induction of mortality and malformation in Xenopus laevis embryos by water sources associated with field frog deformities.

    PubMed

    Burkhart, J G; Helgen, J C; Fort, D J; Gallagher, K; Bowers, D; Propst, T L; Gernes, M; Magner, J; Shelby, M D; Lucier, G

    1998-12-01

    Water samples from several ponds in Minnesota were evaluated for their capacity to induce malformations in embryos of Xenopus laevis. The FETAX assay was used to assess the occurrence of malformations following a 96-hr period of exposure to water samples. These studies were conducted following reports of high incidences of malformation in natural populations of frogs in Minnesota wetlands. The purpose of these studies was to determine if a biologically active agent(s) was present in the waters and could be detected using the FETAX assay. Water samples from ponds with high incidences of frog malformations (affected sites), along with water samples from ponds with unaffected frog populations (reference sites), were studied. Initial experiments clearly showed that water from affected sites induced mortality and malformation in Xenopus embryos, while water from reference sites had little or no effect. Induction of malformation was dose dependent and highly reproducible, both with stored samples and with samples taken at different times throughout the summer. The biological activity of the samples was reduced or eliminated when samples were passed through activated carbon. Limited evidence from these samples indicates that the causal factor(s) is not an infectious organism nor are ion concentrations or metals responsible for the effects observed. Results do indicate that the water matrix has a significant effect on the severity of toxicity. Based on the FETAX results and the occurrence of frog malformations observed in the field, these studies suggest that water in the affected sites contains one or more unknown agents that induce developmental abnormalities in Xenopus. These same factors may contribute to the increased incidence of malformation in native species.

  10. E-cigarette aerosol exposure can cause craniofacial defects in Xenopus laevis embryos and mammalian neural crest cells

    PubMed Central

    Kennedy, Allyson E.; Kandalam, Suraj; Olivares-Navarrete, Rene

    2017-01-01

    Since electronic cigarette (ECIG) introduction to American markets in 2007, vaping has surged in popularity. Many, including women of reproductive age, also believe that ECIG use is safer than traditional tobacco cigarettes and is not hazardous when pregnant. However, there are few studies investigating the effects of ECIG exposure on the developing embryo and nothing is known about potential effects on craniofacial development. Therefore, we have tested the effects of several aerosolized e-cigarette liquids (e-cigAM) in an in vivo craniofacial model, Xenopus laevis, as well as a mammalian neural crest cell line. Results demonstrate that e-cigAM exposure during embryonic development induces a variety of defects, including median facial clefts and midface hypoplasia in two of e-cigAMs tested e-cigAMs. Detailed quantitative analyses of the facial morphology revealed that nicotine is not the main factor in inducing craniofacial defects, but can exacerbate the effects of the other e-liquid components. Additionally, while two different e-cigAMs can have very similar consequences on facial appearances, there are subtle differences that could be due to the differences in e-cigAM components. Further assessment of embryos exposed to these particular e-cigAMs revealed cranial cartilage and muscle defects and a reduction in the blood supply to the face. Finally, the expression of markers for vascular and cartilage differentiation was reduced in a mammalian neural crest cell line corroborating the in vivo effects. Our work is the first to show that ECIG use could pose a potential hazard to the developing embryo and cause craniofacial birth defects. This emphasizes the need for more testing and regulation of this new popular product. PMID:28957438

  11. E-cigarette aerosol exposure can cause craniofacial defects in Xenopus laevis embryos and mammalian neural crest cells.

    PubMed

    Kennedy, Allyson E; Kandalam, Suraj; Olivares-Navarrete, Rene; Dickinson, Amanda J G

    2017-01-01

    Since electronic cigarette (ECIG) introduction to American markets in 2007, vaping has surged in popularity. Many, including women of reproductive age, also believe that ECIG use is safer than traditional tobacco cigarettes and is not hazardous when pregnant. However, there are few studies investigating the effects of ECIG exposure on the developing embryo and nothing is known about potential effects on craniofacial development. Therefore, we have tested the effects of several aerosolized e-cigarette liquids (e-cigAM) in an in vivo craniofacial model, Xenopus laevis, as well as a mammalian neural crest cell line. Results demonstrate that e-cigAM exposure during embryonic development induces a variety of defects, including median facial clefts and midface hypoplasia in two of e-cigAMs tested e-cigAMs. Detailed quantitative analyses of the facial morphology revealed that nicotine is not the main factor in inducing craniofacial defects, but can exacerbate the effects of the other e-liquid components. Additionally, while two different e-cigAMs can have very similar consequences on facial appearances, there are subtle differences that could be due to the differences in e-cigAM components. Further assessment of embryos exposed to these particular e-cigAMs revealed cranial cartilage and muscle defects and a reduction in the blood supply to the face. Finally, the expression of markers for vascular and cartilage differentiation was reduced in a mammalian neural crest cell line corroborating the in vivo effects. Our work is the first to show that ECIG use could pose a potential hazard to the developing embryo and cause craniofacial birth defects. This emphasizes the need for more testing and regulation of this new popular product.

  12. In vivo time-lapse imaging of cell proliferation and differentiation in the optic tectum of Xenopus laevis tadpoles

    PubMed Central

    Bestman, Jennifer E.; Lee-Osbourne, Jane; Cline, Hollis T.

    2012-01-01

    We analyzed the function of neural progenitors in the developing CNS of Xenopus laevis tadpoles using in vivo time-lapse confocal microscopy to collect images through the tectum at intervals of 2 to 24 hours over 3 days. Neural progenitor cells were labeled with fluorescent protein reporters based on expression of endogenous Sox2 transcription factor. With this construct, we identified Sox2-expressing cells as radial glia and as a component of the progenitor pool of cells in the developing tectum that gives rise to neurons and other radial glia. Lineage analysis of individual radial glia and their progeny demonstrated that less than 10% of radial glia undergo symmetric divisions resulting in two radial glia, while the majority of radial glia divide asymmetrically to generate neurons and radial glia. Time-lapse imaging revealed the direct differentiation of radial glia into neurons. Although radial glia may guide axons as they navigate to superficial tectum, we find no evidence that radial glia function as a scaffold for neuronal migration at early stages of tectal development. Over three days, the number of labeled cells increased 20%, as the fraction of radial glia dropped and the proportion of neuronal progeny increased to approximately 60% of the labeled cells. Tadpoles provided with short-term visual enhancement generated significantly more neurons, with a corresponding decrease in cell proliferation. Together these results demonstrate that radial glial cells are neural progenitors in the developing optic tectum and reveal that visual experience increases the proportion of neurons generated in an intact animal. PMID:22113462

  13. Ca2+-recruitment in tachykinin-induced contractions of gut smooth muscle from African clawed frog, Xenopus laevis and rainbow trout, Oncorhynchus mykiss.

    PubMed

    Johansson, Agot; Holmgren, Susanne

    2003-04-01

    Changes in intracellular Ca(2+) concentration control many essential cellular functions like the contraction of smooth muscle cells. The aim of this study was to investigate if the tachykinin substance P (SP) engages external Ca(2+)-sources, internal Ca(2+)-sources, or both in the contraction of the gastrointestinal smooth muscle of rainbow trout (Oncorhynchus mykiss) and the African clawed frog (Xenopus laevis). Strip preparations made of either longitudinal smooth muscle of proximal intestine or circular smooth muscle of cardiac stomach were mounted in organ baths and the tension was recorded via force transducers. Ca(2+)-free Ringer's solution containing the Ca(2+) chelating agent EGTA (2mM) abolished all spontaneous contractions. Exposure to SP in Ca(2+)-free solution decreased the response. Preparations were also treated with the Ca(2+)-ATPase inhibitor thapsigargin (10 microM) during 30 min. Thapsigargin reduced the effect of SP on intestinal longitudinal smooth muscle in rainbow trout and on stomach circular smooth muscle in the African clawed frog and to a less extent in the intestinal longitudinal smooth muscle. The results show that external Ca(2+) is of great importance, but is not the only source of Ca(2+) recruitment in SP-activation of gastrointestinal smooth muscle in rainbow trout and the African clawed frog.

  14. Intestinal parasite Acanthocephalus lucii (Acanthocephala) from European perch (Perca fluviatilis) as a bioindicator for lead pollution in the stream "Jevanský potok" near Prague, Czech Republic.

    PubMed

    Jankovská, Ivana; Miholová, Daniela; Petrtýl, Miloslav; Romočuský, Stěpán; Kalous, Lukáš; Vadlejch, Jaroslav; Cadková, Zuzana; Langrová, Iva

    2011-03-01

    Lead concentrations in the tissues of perch and its parasites were determined as mg/kg dw. Lead was found at higher concentrations in the acanthocephalans (11.56) than in different tissues (liver, gonads and muscle with skin and bone) of perch. With respect to fish tissues, the highest concentrations of lead were present in the liver (1.24), followed by the gonads (0.57) whereas the lowest concentrations were in the muscle with skin and bone (0.21). The bioconcentration factors for lead indicated that parasites accumulate metals to a higher degree than fish tissues--lead concentrations in acanthocephalans were 9.32, 19.27 and 55.05 higher than in liver, gonads and muscles of host, respectively.

  15. Are Fish and Standardized FETAX Assays Protective Enough for Amphibians? A Case Study on Xenopus laevis Larvae Assay with Biologically Active Substances Present in Livestock Wastes

    PubMed Central

    Martini, Federica; Tarazona, José V.; Pablos, M. Victoria

    2012-01-01

    Biologically active substances could reach the aquatic compartment when livestock wastes are considered for recycling. Recently, the standardized FETAX assay has been questioned, and some researchers have considered that the risk assessment performed on fish could not be protective enough to cover amphibians. In the present study a Xenopus laevis acute assay was developed in order to compare the sensitivity of larvae relative to fish or FETAX assays; veterinary medicines (ivermectin, oxytetracycline, tetracycline, sulfamethoxazole, and trimethoprim) and essential metals (zinc, copper, manganese, and selenium) that may be found in livestock wastes were used for the larvae exposure. Lethal (LC50) and sublethal effects were estimated. Available data in both, fish and FETAX studies, were in general more protective than values found out in the current study, but not in all cases. Moreover, the presence of nonlethal effects, caused by ivermectin, zinc, and copper, suggested that several physiological mechanisms could be affected. Thus, this kind of effects should be deeply investigated. The results obtained in the present study could expand the information about micropollutants from livestock wastes on amphibians. PMID:22629159

  16. Induction of mortality and malformation in Xenopus laevis embryos by water sources associated with field frog deformities.

    PubMed Central

    Burkhart, J G; Helgen, J C; Fort, D J; Gallagher, K; Bowers, D; Propst, T L; Gernes, M; Magner, J; Shelby, M D; Lucier, G

    1998-01-01

    Water samples from several ponds in Minnesota were evaluated for their capacity to induce malformations in embryos of Xenopus laevis. The FETAX assay was used to assess the occurrence of malformations following a 96-hr period of exposure to water samples. These studies were conducted following reports of high incidences of malformation in natural populations of frogs in Minnesota wetlands. The purpose of these studies was to determine if a biologically active agent(s) was present in the waters and could be detected using the FETAX assay. Water samples from ponds with high incidences of frog malformations (affected sites), along with water samples from ponds with unaffected frog populations (reference sites), were studied. Initial experiments clearly showed that water from affected sites induced mortality and malformation in Xenopus embryos, while water from reference sites had little or no effect. Induction of malformation was dose dependent and highly reproducible, both with stored samples and with samples taken at different times throughout the summer. The biological activity of the samples was reduced or eliminated when samples were passed through activated carbon. Limited evidence from these samples indicates that the causal factor(s) is not an infectious organism nor are ion concentrations or metals responsible for the effects observed. Results do indicate that the water matrix has a significant effect on the severity of toxicity. Based on the FETAX results and the occurrence of frog malformations observed in the field, these studies suggest that water in the affected sites contains one or more unknown agents that induce developmental abnormalities in Xenopus. These same factors may contribute to the increased incidence of malformation in native species. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 PMID:9831545

  17. Insulin-like growth factor 1 regulation of proliferation and differentiation of Xenopus laevis myogenic cells in vitro.

    PubMed

    Miyata, Sairi; Yada, Tomotaka; Ishikawa, Natsuko; Taheruzzaman, Kazi; Hara, Ryohei; Matsuzaki, Takashi; Nishikawa, Akio

    2017-03-01

    To understand the mechanism of muscle remodeling during Xenopus laevis metamorphosis, we examined the in vitro effect of insulin-like growth factor 1 (IGF-1) on growth and differentiation of three different-fate myogenic cell populations: tadpole tail, tadpole dorsal, and young adult leg muscle. IGF-1 promoted growth and differentiation of both tail and leg myogenic cells only under conditions where these cells could proliferate. Inhibition of cell proliferation by DNA synthesis inhibitor cytosine arabinoside completely canceled the IGF-1's cell differentiation promotion, suggesting the possibility that IGF-1's differentiation-promotion effect is an indirect effect via IGF-1's cell proliferation promotion. IGF-1 promoted differentiation dose dependently with maximum effect at 100-500 ng/ml. RT-PCR analysis revealed the upregulation (11-fold) of ifg1 mRNA expression in developing limbs, suggesting that IGF-1 plays a role in promoting muscle differentiation during limb development. The combined effect of triiodo-L-thyronine (T 3 ) and IGF-1 was also examined. In adult leg cells, IGF-1 promoted growth and differentiation irrespective of the presence of T 3 . In larval tail cells, cell count was 76% lower in the presence of T 3 , and IGF-1 did not promote proliferation and differentiation in T 3 -containing medium. In larval dorsal cells, cell count was also lower in the presence of T 3 , but IGF-1 enhanced proliferation and differentiation in T 3 -containing medium. This result is likely due to the presence among dorsal cells of both adult and larval types (1:1). Thus, IGF-1 affects only adult-type myogenic cells in the presence of T 3 and helps accelerate dorsal muscle remodeling during metamorphosis.

  18. Different forms of soluble cytoplasmic mRNA binding proteins and particles in Xenopus laevis oocytes and embryos

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Murray, M.T.; Krohne, G.; Franke, W.W.

    1991-01-01

    To gain insight into the mechanisms involved in the formation of maternally stored mRNPs during Xenopus laevis development, we searched for soluble cytoplasmic proteins of the oocyte that are able to selectively bind mRNAs, using as substrate radiolabeled mRNA. In vitro mRNP assembly in solution was followed by UV-cross-linking and RNase digestion, resulting in covalent tagging of polypeptides by nucleotide transfer. Five polypeptides of approximately 54, 56 60, 70, and 100 kD (p54, p56, p60, p70, and p100) have been found to selectively bind mRNA and assemble into mRNPs. These polypeptides, which correspond to previously described native mRNP components, occurmore » in three different particle classes of approximately 4.5S, approximately 6S, and approximately 15S, as also determined by their reactions with antibodies against p54 and p56. Whereas the approximately 4.5S class contains p42, p60, and p70, probably each in the form of individual molecules or small complexes, the approximately 6S particles appears to consist only of p54 and p56, which occur in a near-stoichiometric ratio suggestive of a heterodimer complex. The approximately 15S particles contain, in addition to p54 and p56, p60 and p100 and this is the single occurring form of RNA-binding p100. We have also observed changes in the in vitro mRNA binding properties of these polypeptides during oogenesis and early embryonic development, in relation to their phosphorylation state and to the activity of an approximately 15S particle-associated protein kinase, suggesting that these proteins are involved in the developmental translational regulation of maternal mRNAs.« less

  19. Bio-insecticidal activities of halophytic plant extracts against Tribolium confusum (Coleoptera, Tenebrionidae).

    PubMed

    Saïdana, D; Ben Halima-Kamel, M; Ben Tiba, B; Haouas, D; Mahjoub, M A; Mighri, Z; Helal, A N

    2005-01-01

    Salt marsh plants were tested for their insecticidal activities against larvae of the confused flour beetle Tribolium confusum (Tenebrionidae). 16 aerial part extracts were obtained using organic solvents of increasing polarity and tested for their anti-feedant and toxicity effects. Responses varied with plant material and extract type. Ethyl acetate extracts of F. laevis, S. echioides and. T. boveana and petroleum ether extract of F. laevis presented, anti-feedant property. However, S. fructicosa seemed to be attractive to the tested flour beetle. Mortalities of 97, 87, 97 and 80% were observed by using respectively ethyl acetate extracts of F. laevis, S. echioides and T. boveana and petroleum ether extract of F. laevis, when applied at a dose of 1%, mixed with the insect diet. This preliminary study showed that F. laevis, S. echioides and T. boveana presented potential bio-insecticidal activity with ethyl acetate extracts, similar result was found with petroleum ether extract of F. laevis. More complementary studies are needed for the use of these extracts to control T. confusum.

  20. Captive breeding of the barndoor skate (Dipturus laevis) at the Montreal Biodome, with comparison notes on two other captive-bred skate species.

    PubMed

    Parent, Serge; Pépin, Serge; Genet, Jean-Pierre; Misserey, Laurent; Rojas, Salvador

    2008-03-01

    In 1997, the Montreal Biodome obtained five barndoor skates (Dipturus laevis) from the waters off Boston, Massachusetts. Six years later, those specimens began reproducing, and the first egg case was collected in November 2003. Since then, 73 hatchlings have been born and raised. Egg cases were observed year round, and annual fecundity was measured for the first time: one female laid 69 eggs in 2005, 85 in 2006 and 115 in 2007. Egg incubation was longer than believed previously, ranging from 342 to 494 days. Hatching occurred throughout the year. Hatchlings averaged 193 mm total length and 128 mm disk width and weighed 32 g. They were fed krill and diced fish. All but one survived the first month. A photo identification system was useful in recognizing two groups of 10 specimens during their first year, and transponders could be inserted in the wing muscles of 1-year-old skates. Total lengths at birth and at age 2 were similar to the data reported from the wild, suggesting a similar growth pattern in captivity. The reproduction characteristics of the barndoor skate were compared with those of two other skate species currently bred at the Montreal Biodome, the winter skate (Leucoraja ocellata) and the thorny skate (Amblyraja radiata). Zoo Biol 27:145-153, 2008. (c) 2008 Wiley-Liss, Inc.

  1. The progestin norethisterone affects thyroid hormone-dependent metamorphosis of Xenopus laevis tadpoles at environmentally relevant concentrations.

    PubMed

    Lorenz, Claudia; Krüger, Angela; Schöning, Viola; Lutz, Ilka

    2018-04-15

    Previously, levonorgestrel (LNG) has been shown to be an endocrine disruptor of the amphibian thyroid system. In the present study, we investigated whether anti-thyroidal effects are a common property of progestins other than LNG. Premetamorphic Xenopus laevis tadpoles were exposed to norethisterone (NET) and dienogest DIE (each at 0.1-10nM) and LNG (10nM) until completion of metamorphosis. LNG and NET at all concentrations caused a significant developmental retardation whereas DIE did not impair time to metamorphosis. In LNG and 10nM NET exposed animals, tsh mRNA levels increased considerably later than the developmental delay occurred and thyroid histopathology showed no signs of TSH-hyperstimulation. Instead, thyroid glands from these treatments appeared inactive in producing thyroid hormones. Thyroidal transcript levels of dio2 and dio3 were increased by treatments with LNG and NET at 1nM and 10nM, whereas iyd mRNA was reduced by LNG and 10nM NET. Expression of slc5α5 was not changed by any treatment. Effects of DIE differed from those induced by LNG and NET. No developmental delay was measurable; however, tshβ and dio2 mRNAs were increased in pituitary glands of tadpoles exposed to 1.0nM and 10nM DIE. Thyroid histopathology displayed no abnormalities and thyroidal mRNA expression of the genes analyzed (slc5α5, iyd, dio2, dio3) was not changed by DIE. Overall, our results provide evidence that the anti-thyroidal effects already known from LNG are also present in another progestin, namely NET, even at environmentally relevant concentrations. In conclusion we suggest that progestins do not only pose an environmental risk in terms of their impact on reproductive success of aquatic vertebrates, but also with respect to their anti-thyroidal properties affecting amphibian metamorphosis. Copyright © 2017 Elsevier Inc. All rights reserved.

  2. cis- and trans-acting elements of the estrogen-regulated vitellogenin gene B1 of Xenopus laevis.

    PubMed

    Wahli, W; Martinez, E; Corthésy, B; Cardinaux, J R

    1989-01-01

    Vitellogenin genes are expressed under strict estrogen control in the liver of female oviparous vertebrates. Gene transfer experiments using estrogen-responsive cells have shown that the 13 bp perfect palindromic element GGTCACTGTGACC found upstream of the Xenopus laevis vitellogenin gene A2 promoter mediates hormonal stimulation and thus, was called the estrogen-responsive element (ERE). In the Xenopus vitellogenin genes B1 and B2 there are two closely adjacent EREs with one or more base substitutions when compared to the consensus ERE GGTCANNNTGACC. On their own, these degenerated elements have only a low or no regulatory capacity at all but act together synergistically to form an estrogen-responsive unit (ERU) with the same strength as the perfect palindromic 13 bp element. Analysis of estrogen receptor binding to the gene B1 ERU revealed a cooperative interaction of receptor dimers to the two adjacent imperfect EREs which most likely explains the synergistic stimulation observed in vivo. Furthermore, a promoter activator element located between positions --113 and --42 of the gene B1 and functional in the human MCF-7 and the Xenopus B3.2 cells has been identified and shown to be involved in the high level of induced transcription activity when the ERE is placed at a distance from the promoter. Finally, a hormone-controlled in vitro transcription system derived from Xenopus liver nuclear extracts was exploited to characterize two additional novel cis-acting elements within the vitellogenin gene B1 promoter. One of them, a negative regulatory element (NRE), is responsible for repression of promoter activity in the absence of hormone. The second is related to the NF-I binding site and is required, together with the ERE, to mediate hormonal induction. Moreover, we detected three trans-acting activities in Xenopus liver nuclear extracts that interact with these regions and demonstrated that they participate in the regulation of the expression of the vitellogenin

  3. Genes encoding Xenopus laevis Ig L chains: Implications for the evolution of [kappa] and [lambda] chains

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Zezza, D.J.; Stewart, S.E.; Steiner, L.A.

    1992-12-15

    Xenopus laevis Ig contain two distinct types of L chains, designated [rho] or L1 and [sigma] or L2. The authors have analyzed Xenopus genomic DNA by Southern blotting with cDNA probes specific for L1 V and C regions. Many fragments hybridized to the V probe, but only one or two fragments hybridized to the C probe. Corresponding C, J, and V gene segments were identified on clones isolated from a genomic library prepared from the same DNA. One clone contains a C gene segment separated from a J gene segment by an intron of 3.4 kb. The J and Cmore » gene segments are nearly identical in sequence to cDNA clones analyzed previously. The C segment is somewhat more similar and the J segment considerably more similar in sequence to the corresponding segments of mammalian [kappa] chains than to those of mammalian [lambda] chains. Upstream of the J segment is a typical recombination signal sequence with a spacer of 23 bp, as in J[kappa]. A second clone from the library contains four V gene segments, separated by 2.1 to 3.6 kb. Two of these, V1 and V3, have the expected structural and regulatory features of V genes, and are very similar in sequence to each other and to mammalian V[kappa]. A third gene segment, V2, resembles V1 and V3 in its coding region and nearby 5[prime]-flanking region, but diverges in sequence 5[prime] to position [minus]95 with loss of the octamer promoter element. The fourth V-like segment is similar to the others at the 3[prime]-end, but upstream of codon 64 bears no resemblance in sequence to any Ig V region. All four V segments have typical recombination signal sequences with 12-bp spacers at their 3[prime]-ends, as in V[kappa]. Taken together, the data suggest that Xenopus L1 L chain genes are members of the [kappa] gene family. 80 refs., 9 figs.« less

  4. Optomotor behaviour in Xenopus laevis tadpoles as a measure of the effect of gravity on visual and vestibular neural integration

    NASA Technical Reports Server (NTRS)

    Pronych, S. P.; Souza, K. A.; Neff, A. W.; Wassersug, R. J.

    1996-01-01

    The ability of aquatic vertebrates to maintain their position requires integration of visual and vestibular sensory information. To understand better how aquatic animals integrate such information, we measured the optomotor behaviour of Xenopus laevis tadpoles raised in growth chambers in microgravity (< 10(-3)g), normal gravity (1 g), hypergravity (3 g) and on a slowly rotating clinostat (simulated microgravity). The goal of this research was to determine how development in an altered gravitational force field affects the visual- and vestibular-dependent behaviour of tadpoles. This research represents the first time that the optomotor behaviour of an organism raised from fertilization in microgravity has been tested. Significant differences were observed in the optomotor behaviour among the four gravity treatments. When first exposed to normal gravity, the microgravity-raised tadpoles exhibited the strongest (or most positive) optomotor behaviour, while the 3 g centrifuge tadpoles showed no optomotor response. Some abnormal behaviours (such as erratic swimming, lying motionless and abnormal swimming posture) were observed in the tadpoles raised in altered gravity on the initial day of testing. One day later, the tadpoles raised in hypergravity did not differ significantly in their optomotor behaviour from control tadpoles raised in normal gravity. However, tadpoles raised in microgravity still displayed an exaggerated optomotor response. One week after the tadpoles had been introduced to normal gravity, there was no longer a significant difference in optomotor behaviour among the different gravity treatments. This convergence of optomotor behaviour by tadpoles from the different treatment reflects the acclimation of their vestibular systems to normal gravity.

  5. Unusual development of light-reflecting pigment cells in intact and regenerating tail in the periodic albino mutant of Xenopus laevis.

    PubMed

    Fukuzawa, Toshihiko

    2010-10-01

    Unusual light-reflecting pigment cells, "white pigment cells", specifically appear in the periodic albino mutant (a(p) /a(p)) of Xenopus laevis and localize in the same place where melanophores normally differentiate in the wild-type. The mechanism responsible for the development of unusual pigment cells is unclear. In this study, white pigment cells in the periodic albino were compared with melanophores in the wild-type, using a cell culture system and a tail-regenerating system. Observations of both intact and cultured cells demonstrate that white pigment cells are unique in (1) showing characteristics of melanophore precursors at various stages of development, (2) accumulating reflecting platelets characteristic of iridophores, and (3) exhibiting pigment dispersion in response to α-melanocyte stimulating hormone (α-MSH) in the same way that melanophores do. When a tadpole tail is amputated, a functionally competent new tail is regenerated. White pigment cells appear in the mutant regenerating tail, whereas melanophores differentiate in the wild-type regenerating tail. White pigment cells in the mutant regenerating tail are essentially similar to melanophores in the wild-type regenerating tail with respect to their localization, number, and response to α-MSH. In addition to white pigment cells, iridophores which are never present in the intact tadpole tail appear specifically in the somites near the amputation level in the mutant regenerating tail. Iridophores are distinct from white pigment cells in size, shape, blue light-induced fluorescence, and response to α-MSH. These findings strongly suggest that white pigment cells in the mutant arise from melanophore precursors and accumulate reflecting platelets characteristic of iridophores.

  6. Unusual development of light-reflecting pigment cells in intact and regenerating tail in the periodic albino mutant of Xenopus laevis

    PubMed Central

    2010-01-01

    Unusual light-reflecting pigment cells, “white pigment cells”, specifically appear in the periodic albino mutant (ap/ap) of Xenopus laevis and localize in the same place where melanophores normally differentiate in the wild-type. The mechanism responsible for the development of unusual pigment cells is unclear. In this study, white pigment cells in the periodic albino were compared with melanophores in the wild-type, using a cell culture system and a tail-regenerating system. Observations of both intact and cultured cells demonstrate that white pigment cells are unique in (1) showing characteristics of melanophore precursors at various stages of development, (2) accumulating reflecting platelets characteristic of iridophores, and (3) exhibiting pigment dispersion in response to α-melanocyte stimulating hormone (α-MSH) in the same way that melanophores do. When a tadpole tail is amputated, a functionally competent new tail is regenerated. White pigment cells appear in the mutant regenerating tail, whereas melanophores differentiate in the wild-type regenerating tail. White pigment cells in the mutant regenerating tail are essentially similar to melanophores in the wild-type regenerating tail with respect to their localization, number, and response to α-MSH. In addition to white pigment cells, iridophores which are never present in the intact tadpole tail appear specifically in the somites near the amputation level in the mutant regenerating tail. Iridophores are distinct from white pigment cells in size, shape, blue light-induced fluorescence, and response to α-MSH. These findings strongly suggest that white pigment cells in the mutant arise from melanophore precursors and accumulate reflecting platelets characteristic of iridophores. PMID:20859642

  7. Transcriptome Data Reveal Syndermatan Relationships and Suggest the Evolution of Endoparasitism in Acanthocephala via an Epizoic Stage

    PubMed Central

    Rieger, Benjamin; Rosenkranz, David; Witek, Alexander; Welch, David B. Mark; Ebersberger, Ingo; Hankeln, Thomas

    2014-01-01

    The taxon Syndermata comprises the biologically interesting wheel animals (“Rotifera”: Bdelloidea + Monogononta + Seisonidea) and thorny-headed worms (Acanthocephala), and is central for testing superordinate phylogenetic hypotheses (Platyzoa, Gnathifera) in the metazoan tree of life. Recent analyses of syndermatan phylogeny suggested paraphyly of Eurotatoria (free-living bdelloids and monogononts) with respect to endoparasitic acanthocephalans. Data of epizoic seisonids, however, were absent, which may have affected the branching order within the syndermatan clade. Moreover, the position of Seisonidea within Syndermata should help in understanding the evolution of acanthocephalan endoparasitism. Here, we report the first phylogenomic analysis that includes all four higher-ranked groups of Syndermata. The analyzed data sets comprise new transcriptome data for Seison spec. (Seisonidea), Brachionus manjavacas (Monogononta), Adineta vaga (Bdelloidea), and Paratenuisentis ambiguus (Acanthocephala). Maximum likelihood and Bayesian trees for a total of 19 metazoan species were reconstructed from up to 410 functionally diverse proteins. The results unanimously place Monogononta basally within Syndermata, and Bdelloidea appear as the sister group to a clade comprising epizoic Seisonidea and endoparasitic Acanthocephala. Our results support monophyly of Syndermata, Hemirotifera (Bdelloidea + Seisonidea + Acanthocephala), and Pararotatoria (Seisonidea + Acanthocephala), rejecting monophyly of traditional Rotifera and Eurotatoria. This serves as an indication that early acanthocephalans lived epizoically or as ectoparasites on arthropods, before their complex lifecycle with arthropod intermediate and vertebrate definite hosts evolved. PMID:24520404

  8. Transcriptome data reveal Syndermatan relationships and suggest the evolution of endoparasitism in Acanthocephala via an epizoic stage.

    PubMed

    Wey-Fabrizius, Alexandra R; Herlyn, Holger; Rieger, Benjamin; Rosenkranz, David; Witek, Alexander; Welch, David B Mark; Ebersberger, Ingo; Hankeln, Thomas

    2014-01-01

    The taxon Syndermata comprises the biologically interesting wheel animals ("Rotifera": Bdelloidea + Monogononta + Seisonidea) and thorny-headed worms (Acanthocephala), and is central for testing superordinate phylogenetic hypotheses (Platyzoa, Gnathifera) in the metazoan tree of life. Recent analyses of syndermatan phylogeny suggested paraphyly of Eurotatoria (free-living bdelloids and monogononts) with respect to endoparasitic acanthocephalans. Data of epizoic seisonids, however, were absent, which may have affected the branching order within the syndermatan clade. Moreover, the position of Seisonidea within Syndermata should help in understanding the evolution of acanthocephalan endoparasitism. Here, we report the first phylogenomic analysis that includes all four higher-ranked groups of Syndermata. The analyzed data sets comprise new transcriptome data for Seison spec. (Seisonidea), Brachionus manjavacas (Monogononta), Adineta vaga (Bdelloidea), and Paratenuisentis ambiguus (Acanthocephala). Maximum likelihood and Bayesian trees for a total of 19 metazoan species were reconstructed from up to 410 functionally diverse proteins. The results unanimously place Monogononta basally within Syndermata, and Bdelloidea appear as the sister group to a clade comprising epizoic Seisonidea and endoparasitic Acanthocephala. Our results support monophyly of Syndermata, Hemirotifera (Bdelloidea + Seisonidea + Acanthocephala), and Pararotatoria (Seisonidea + Acanthocephala), rejecting monophyly of traditional Rotifera and Eurotatoria. This serves as an indication that early acanthocephalans lived epizoically or as ectoparasites on arthropods, before their complex lifecycle with arthropod intermediate and vertebrate definite hosts evolved.

  9. Assessing host-parasite specificity through coprological analysis: a case study with species of Corynosoma (Acanthocephala: Polymorphidae) from marine mammals.

    PubMed

    Aznar, F J; Hernández-Orts, J; Suárez, A A; García-Varela, M; Raga, J A; Cappozzo, H L

    2012-06-01

    In this paper we report an investigation of the utility of coprological analysis as an alternative technique to study parasite specificity whenever host sampling is problematic; acanthocephalans from marine mammals were used as a model. A total of 252 scats from the South American sea lion, Otaria flavescens, and rectal faeces from 43 franciscanas, Pontoporia blainvillei, from Buenos Aires Province, were examined for acanthocephalans. Specimens of two species, i.e. Corynosoma australe and C. cetaceum, were collected from both host species. In sea lions, 78 out of 145 (37.9%) females of C. australe were gravid and the sex ratio was strongly female-biased. However, none of the 168 females of C. cetaceum collected was gravid and the sex ratio was not female-biased. Conversely, in franciscanas, 14 out of 17 (82.4%) females of C. cetaceum were gravid, but none of 139 females of C. australe was, and the sex ratio of C. cetaceum, but not that of C. australe, was female-biased. In putative non-hosts, the size of worms was similar to that from specimens collected from prey. Results suggest that both acanthocephalans contact sea lions and franciscanas regularly. However, C. australe and C. cetaceum cannot apparently reproduce, nor even grow, in franciscanas and sea lions, respectively. Coprological analysis may represent a useful supplementary method to investigate parasite specificity, particularly when host carcasses are difficult to obtain.

  10. Effects of ethanol and anesthetics on type 1 and 5 metabotropic glutamate receptors expressed in Xenopus laevis oocytes.

    PubMed

    Minami, K; Gereau, R W; Minami, M; Heinemann, S F; Harris, R A

    1998-01-01

    Previous studies have demonstrated that ethanol and volatile anesthetics inhibit the function of some metabotropic (G protein-coupled) receptors, including the 5-hydroxytryptamine2 and muscarinic cholinergic receptors. The metabotropic glutamate receptors (mGluRs) show little sequence homology with most other metabotropic receptors and are important modulators of synaptic transmission in the mammalian central nervous system. It was of interest to determine drug actions on these receptors, and we investigated the effects of ethanol, halothane, the anesthetic compound F3 (1-chloro-1,2,2-trifluorocyclobutane), and the nonanesthetics F6 (1,2-dichlorohexafluorocyclobutane) and F8 (2,3-chlorooctafluorobutane) on the function of mGluR1 and mGluR5 expressed in Xenopus laevis oocytes. Halothane, F3, and ethanol inhibited mGluR5-induced Ca(2+)-dependent Cl- currents, yet pharmacologically relevant concentrations of these compounds had little effect on the glutamate-induced currents in the oocytes expressing mGluR1. F6 had inhibitory effects on both receptors, and F8 did not affect either mGluR1 or mGluR5 function. The protein kinase C (PKC) inhibitor GF109203X enhanced the glutamate-induced current, and the PKC activator phorbol-12-myristate-13-acetate inhibited this current in the oocytes expressing mGluR5, but these compounds had little effect on mGluR1 function. GF109203X abolished the inhibitory effects of halothane, F3, and ethanol on mGluR5s. Conversely, the phosphatase inhibitor calyculin A prolonged the action of halothane and ethanol. Furthermore, mutation of a PKC consensus site (Ser890) of mGluR5 abolished the inhibitory effects of halothane, F3, and ethanol. These results suggest that ethanol and volatile anesthetics inhibit mGluR5 because they promote PKC-mediated phosphorylation.

  11. The vestibuloocular reflex of tadpoles (Xenopus laevis) after knock-down of the isthmus-related transcription factor XTcf-4.

    PubMed

    Horn, Eberhard R; El-Yamany, Nabil A; Gradl, Dietmar

    2013-02-15

    Development of the amphibian vestibular organ is regulated by molecular and neuronal mechanisms and by environmental input. The molecular component includes inductive signals derived from neural tissue of the hindbrain and from the surrounding mesoderm. The integrity of hindbrain patterning, on the other hand, depends on instructive signals from the isthmus organizer of the midbrain, including the transcription factor XTcf-4. If the development of the vestibular system depends on the integrity of the isthmus as the organizing centre, suppression of isthmus maintenance should modify vestibular morphology and function. We tested this hypothesis by downregulation of the transcription factor XTcf-4. 10 pmol l(-1) XTcf-4-specific antisense morpholino oligonucleotide was injected in one blastomere of two-cell-stage embryos of Xenopus laevis. For reconstitution experiments, 500 pg mRNA of the repressing XTcf-4A isoform or the activating XTcf-4C isoform were co-injected. Overexpression experiments were included using the same isoforms. Otoconia formation and vestibular controlled behaviour such as the roll-induced vestibuloocular reflex (rVOR) and swimming were recorded two weeks later. In 50% of tadpoles, downregulation of XTcf-4 induced (1) a depression of otoconia formation accompanied by a reduction of the rVOR, (2) abnormal tail development and (3) loop swimming behaviour. (4) All effects were rescued by co-injection of XTcf-4C but not, or only partially, by XTcf-4A. (5) Overexpression of XTcf-4A caused similar morphological and rVOR modifications as XTcf-4 depletion, while overexpression of XTcf-4C had no effect. Because XTcf-4C has been described as an essential factor for isthmus development, we postulate that the isthmus is strongly involved in vestibular development.

  12. Effects of cyanobacterial biomass and purified microcystins on malformations in Xenopus laevis: teratogenesis assay (FETAX).

    PubMed

    Dvoráková, Dagmar; Dvoráková, Katerina; Bláha, Ludek; Marsálek, Blahoslav; Knotková, Zora

    2002-12-01

    Xenopus laevis (African clawed frog) embryos in a 96-h teratogenesis assay (FETAX) were exposed to 0-250 microg/L and 500 microg/L of purified microcystin-LR (MCYST-LR) for the estimation of lethality, as well as to equivalent concentrations of biomass containing MCYST-LR (natural water bloom dominated by Microcystis aeruginosa) and biomass without MCYST-LR (bloom dominated by Microcystis wesenbergii). The highest tested concentrations of purified MCYST-LR caused up to 30% lethality after a 96-h exposure, corresponding to a LC(25) of 380 microg/L. Cyanobacterial biomass containing MCYST-LR caused significant lethality up to 50% at the highest tested concentrations (300 mg/L, i.e., 250 microg/L of MCYST-LR). The estimated 96-h LC(25) values varied from 125 mg/L (biomass containing MCYST-LR) up to 232 mg/L (biomass without MCYST-LR). A statistically significant increase in the number of malformed embryos was observed after exposure to cyanobacterial samples. Purified MCYST-LR at and above 25 microg/L significantly increased the number of malformations, with 53% of surviving embryos malformed in the highest tested concentration, 250 microg/L (EC(25) = 27 microg/L). Exposure to the highest concentration of MCYST-LR containing biomass resulted in more than 60% of the embryos being malformed and an EC(25) of 52 mg/L (i.e., 43 microg of MCYST-LR/L). Cyanobacterial biomass with no natural microcystin also induced substantial malformations-about 50% aberrant embryos at the highest concentration, 300 mg/L (EC(25) = 75 mg/L). External additions of purified MCYST-LR to the biomass that was originally without microcystins resulted in a slight additional increase in the rate of malformations (80% at the highest concentration, 300 mg of biomass plus 250 microg of MCYST-LR per liter). A comparison of lethality and effects on malformations (teratogenic index, TI = LC(25)/EC(25)) showed that all samples had significant teratogenic potential in the FETAX assay (TI(MCYST-LR) = 14; TI

  13. Efficacy of Tricaine Methanesulfonate (MS-222) as an Anesthetic Agent for Blocking Sensory-Motor Responses in Xenopus laevis Tadpoles

    PubMed Central

    Ramlochansingh, Carlana; Branoner, Francisco; Chagnaud, Boris P.; Straka, Hans

    2014-01-01

    Anesthetics are drugs that reversibly relieve pain, decrease body movements and suppress neuronal activity. Most drugs only cover one of these effects; for instance, analgesics relieve pain but fail to block primary fiber responses to noxious stimuli. Alternately, paralytic drugs block synaptic transmission at neuromuscular junctions, thereby effectively paralyzing skeletal muscles. Thus, both analgesics and paralytics each accomplish one effect, but fail to singularly account for all three. Tricaine methanesulfonate (MS-222) is structurally similar to benzocaine, a typical anesthetic for anamniote vertebrates, but contains a sulfate moiety rendering this drug more hydrophilic. MS-222 is used as anesthetic in poikilothermic animals such as fish and amphibians. However, it is often argued that MS-222 is only a hypnotic drug and its ability to block neural activity has been questioned. This prompted us to evaluate the potency and dynamics of MS-222-induced effects on neuronal firing of sensory and motor nerves alongside a defined motor behavior in semi-intact in vitro preparations of Xenopus laevis tadpoles. Electrophysiological recordings of extraocular motor discharge and both spontaneous and evoked mechanosensory nerve activity were measured before, during and after administration of MS-222, then compared to benzocaine and a known paralytic, pancuronium. Both MS-222 and benzocaine, but not pancuronium caused a dose-dependent, reversible blockade of extraocular motor and sensory nerve activity. These results indicate that MS-222 as benzocaine blocks the activity of both sensory and motor nerves compatible with the mechanistic action of effective anesthetics, indicating that both caine-derivates are effective as single-drug anesthetics for surgical interventions in anamniotes. PMID:24984086

  14. Expression, Purification, and Structural Insights for the Human Uric Acid Transporter, GLUT9, Using the Xenopus laevis Oocytes System

    PubMed Central

    Clémençon, Benjamin; Lüscher, Benjamin P.; Fine, Michael; Baumann, Marc U.; Surbek, Daniel V.; Bonny, Olivier; Hediger, Matthias A.

    2014-01-01

    The urate transporter, GLUT9, is responsible for the basolateral transport of urate in the proximal tubule of human kidneys and in the placenta, playing a central role in uric acid homeostasis. GLUT9 shares the least homology with other members of the glucose transporter family, especially with the glucose transporting members GLUT1-4 and is the only member of the GLUT family to transport urate. The recently published high-resolution structure of XylE, a bacterial D-xylose transporting homologue, yields new insights into the structural foundation of this GLUT family of proteins. While this represents a huge milestone, it is unclear if human GLUT9 can benefit from this advancement through subsequent structural based targeting and mutagenesis. Little progress has been made toward understanding the mechanism of GLUT9 since its discovery in 2000. Before work can begin on resolving the mechanisms of urate transport we must determine methods to express, purify and analyze hGLUT9 using a model system adept in expressing human membrane proteins. Here, we describe the surface expression, purification and isolation of monomeric protein, and functional analysis of recombinant hGLUT9 using the Xenopus laevis oocyte system. In addition, we generated a new homology-based high-resolution model of hGLUT9 from the XylE crystal structure and utilized our purified protein to generate a low-resolution single particle reconstruction. Interestingly, we demonstrate that the functional protein extracted from the Xenopus system fits well with the homology-based model allowing us to generate the predicted urate-binding pocket and pave a path for subsequent mutagenesis and structure-function studies. PMID:25286413

  15. Effects of endocrine-disrupting contaminants on amphibian oogenesis: methoxychlor inhibits progesterone-induced maturation of Xenopus laevis oocytes in vitro.

    PubMed Central

    Pickford, D B; Morris, I D

    1999-01-01

    There is currently little evidence of pollution-induced endocrine dysfunction in amphibia, in spite of widespread concern over global declines in this ecologically diverse group. Data regarding the potential effects of endocrine-disrupting contaminants (EDCs) on reproductive function in amphibia are particularly lacking. We hypothesized that estrogenic EDCs may disrupt progesterone-induced oocyte maturation in the adult amphibian ovary, and tested this with an in vitro germinal vesicle breakdown assay using defolliculated oocytes from the African clawed frog, Xenopus laevis. While a variety of natural and synthetic estrogens and xenoestrogens were inactive in this system, the proestrogenic pesticide methoxychlor was a surprisingly potent inhibitor of progesterone-induced oocyte maturation (median inhibitive concentration, 72 nM). This inhibitory activity was specific to methoxychlor, rather than to its estrogenic contaminants or metabolites, and was not antagonized by the estrogen receptor antagonist ICI 182,780, suggesting that this activity is not estrogenic per se. The inhibitory activity of methoxychlor was dose dependent, reversible, and early acting. However, washout was unable to reverse the effect of short methoxychlor exposure, and methoxychlor did not competitively displace [3H]progesterone from a specific binding site in the oocyte plasma membrane. Therefore, methoxychlor may exert its action not directly at the site of progesterone action, but downstream on early events in maturational signaling, although the precise mechanism of action is unclear. The activity of methoxychlor in this system indicates that xenobiotics may exert endocrine-disrupting effects through interference with progestin-regulated processes and through mechanisms other than receptor antagonism. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Figure 9 PMID:10090707

  16. Thyroid Hormone‐Induced Activation of Notch Signaling is Required for Adult Intestinal Stem Cell Development During Xenopus Laevis Metamorphosis

    PubMed Central

    Fujimoto, Kenta; Kajita, Mitsuko; Fu, Liezhen; Shi, Yun‐Bo; Ishizuya‐Oka, Atsuko

    2016-01-01

    Abstract In Xenopus laevis intestine during metamorphosis, the larval epithelial cells are removed by apoptosis, and the adult epithelial stem (AE) cells appear concomitantly. They proliferate and differentiate to form the adult epithelium (Ep). Thyroid hormone (TH) is well established to trigger this remodeling by regulating the expression of various genes including Notch receptor. To study the role of Notch signaling, we have analyzed the expression of its components, including the ligands (DLL and Jag), receptor (Notch), and targets (Hairy), in the metamorphosing intestine by real‐time reverse transcription‐polymerase chain reaction and in situ hybridization or immunohistochemistry. We show that they are up‐regulated during both natural and TH‐induced metamorphosis in a tissue‐specific manner. Particularly, Hairy1 is specifically expressed in the AE cells. Moreover, up‐regulation of Hairy1 and Hairy2b by TH was prevented by treating tadpoles with a γ‐secretase inhibitor (GSI), which inhibits Notch signaling. More importantly, TH‐induced up‐regulation of LGR5, an adult intestinal stem cell marker, was suppressed by GSI treatment. Our results suggest that Notch signaling plays a role in stem cell development by regulating the expression of Hairy genes during intestinal remodeling. Furthermore, we show with organ culture experiments that prolonged exposure of tadpole intestine to TH plus GSI leads to hyperplasia of secretory cells and reduction of absorptive cells. Our findings here thus provide evidence for evolutionarily conserved role of Notch signaling in intestinal cell fate determination but more importantly reveal, for the first time, an important role of Notch pathway in the formation of adult intestinal stem cells during vertebrate development. Stem Cells 2017;35:1028–1039 PMID:27870267

  17. Blastomeres show differential fate changes in 8-cell Xenopus laevis embryos that are rotated 90 degrees before first cleavage

    NASA Technical Reports Server (NTRS)

    Huang, S.; Johnson, K. E.; Wang, H. Z.

    1998-01-01

    To study the mechanisms of dorsal axis specification, the alteration in dorsal cell fate of cleavage stage blastomeres in axis-respecified Xenopus laevis embryos was investigated. Fertilized eggs were rotated 90 degrees with the sperm entry point up or down with respect to the gravitational field. At the 8-cell stage, blastomeres were injected with the lineage tracers, Texas Red- or FITC-Dextran Amines. The distribution of the labeled progeny was mapped at the tail-bud stages (stages 35-38) and compared with the fate map of an 8-cell embryo raised in a normal orientation. As in the normal embryos, each blastomere in the rotated embryos has a characteristic and predictable cell fate. After 90 degrees rotation the blastomeres in the 8-cell stage embryo roughly switched their position by 90 degrees, but the fate of the blastomeres did not simply show a 90 degrees switch appropriate for their new location. Four types of fate change were observed: (i) the normal fate of the blastomere is conserved with little change; (ii) the normal fate is completely changed and a new fate is adopted according to the blastomere's new position: (iii) the normal fate is completely changed, but the new fate is not appropriate for its new position; and (4) the blastomere partially changed its fate and the new fate is a combination of its original fate and a fate appropriate to its new location. According to the changed fates, the blastomeres that adopt dorsal fates were identified in rotated embryos. This identification of dorsal blastomeres provides basic important information for further study of dorsal signaling in Xenopus embryos.

  18. Gastrointestinal helminths of Gentoo penguins (Pygoscelis papua) from Stranger Point, 25 de Mayo/King George Island, Antarctica.

    PubMed

    Diaz, Julia Inés; Fusaro, Bruno; Longarzo, Lucrecia; Coria, Néstor Rubén; Vidal, Virginia; Jerez, Silvia; Ortiz, Juana; Barbosa, Andrés

    2013-05-01

    The aim of this work is to contribute to the knowledge of gastrointestinal parasites of the Gentoo penguin (Pygoscelis papua) from 25 de Mayo/King George Island (South Shetlands, Antarctica). Gastrointestinal tracts of 37 fresh dead individuals (21 chicks, 10 juveniles, and 6 adults) were collected from December 2006 to February 2012 and examined for macroparasites. Four adult parasite species were found: one Cestoda species (Parorchites zederi), two Nematoda species (Stegophorus macronectes and Tetrameres wetzeli), and one Acanthocephalan (Corynosoma shackletoni). Two species of immature acanthocephalans, Corynosoma hamanni and Corynosoma bullosum, were found in a single host. This is the first record of Tetrameres wetzeli in Gentoo penguins. The low parasite richness observed could be related to the stenophagic and pelagic diet of this host species which feeds almost exclusively on krill.

  19. Purification, physicochemical characterization, and immunohistochemical localization of a major 11.7 S glycoprotein from the jelly coats of the anuran Lepidobatrachus laevis.

    PubMed

    Carroll, E J; Wei, S H; Nagel, G M

    1991-02-01

    Embryos of the frog Lepidobatrachus laevis are encased by a fertilization envelope and two jelly layers, termed J1 (innermost) and J2 (outermost). From preparations of total jelly solubilized from cleavage-stage embryos by a solution of alkaline beta-mercaptoethanol we have purified one jelly coat glycoprotein to homogeneity via FPLC gel permeation chromatography on Superose 6H. The purified glycoprotein was 94% protein and 6% carbohydrate, had an s0(20),w of 11.7 S, with a molecular weight of 245,000 measured by sedimentation equilibrium and 263,000 by gel permeation chromatography. SDS-PAGE revealed that the glycoprotein is composed of a single subunit near 29,700 molecular weight; thus we propose that eight of these subunits comprise the native molecule. Amino acid analysis of the glycoprotein indicated a high content of Glx + Asx (32.4 mole%), a low content of basic amino acids (Arg + Lys = 12.2 mole%), and a single cysteine residue per subunit. The N-terminal amino acid was threonine and the sequence of the first twenty amino acids was determined. Monospecific antisera to the glycoprotein were prepared in rabbits and were used to immunohistochemically localize the glycoprotein throughout the matrix of both jelly layers. Antiserum against the glycoprotein had virtually no effect on the fertilizability of jellied eggs in vitro; thus we hypothesize that the glycoprotein fulfills a structural role in both jelly layers.

  20. Differences in mobility at the range edge of an expanding invasive population of Xenopus laevis in the west of France.

    PubMed

    Louppe, Vivien; Courant, Julien; Herrel, Anthony

    2017-01-15

    Theoretical models predict that spatial sorting at the range edge of expanding populations should favor individuals with increased mobility relative to individuals at the center of the range. Despite the fact that empirical evidence for the evolution of locomotor performance at the range edge is rare, data on cane toads support this model. However, whether this can be generalized to other species remains largely unknown. Here, we provide data on locomotor stamina and limb morphology in individuals from two sites: one from the center and one from the periphery of an expanding population of the clawed frog Xenopus laevis in France where it was introduced about 30 years ago. Additionally, we provide data on the morphology of frogs from two additional sites to test whether the observed differences can be generalized across the range of this species in France. Given the known sexual size dimorphism in this species, we also test for differences between the sexes in locomotor performance and morphology. Our results show significant sexual dimorphism in stamina and morphology, with males having longer legs and greater stamina than females. Moreover, in accordance with the predictions from theoretical models, individuals from the range edge had a greater stamina. This difference in locomotor performance is likely to be driven by the significantly longer limb segments observed in animals in both sites sampled in different areas along the range edge. Our data have implications for conservation because spatial sorting on the range edge may lead to an accelerated increase in the spread of this invasive species in France. © 2017. Published by The Company of Biologists Ltd.

  1. Evidence that the rabbit proton-peptide co-transporter PepT1 is a multimer when expressed in Xenopus laevis oocytes.

    PubMed

    Panitsas, Konstantinos-E; Boyd, C A R; Meredith, David

    2006-04-01

    To test whether the rabbit proton-coupled peptide transporter PepT1 is a multimer, we have employed a combination of transport assays, luminometry and site-directed mutagenesis. A functional epitope-tagged PepT1 construct (PepT1-FLAG) was co-expressed in Xenopus laevis oocytes with a non-functional but normally trafficked mutant form of the same transporter (W294F-PepT1). The amount of PepT1-FLAG cRNA injected into the oocytes was kept constant, while the amount of W294F-PepT1 cRNA was increased over the mole fraction range of 0 to 1. The uptake of [(3)H]-D: -Phe-L: -Gln into the oocytes was measured at pH(out) 5.5, and the surface expression of PepT1-FLAG was quantified by luminometry. As the mole fraction of injected W294F-PepT1 increased, the uptake of D: -Phe-L: -Gln decreased. This occurred despite the surface expression of PepT1-FLAG remaining constant, and so we can conclude that PepT1 must be a multimer. Assuming that PepT1 acts as a homomultimer, the best fit for the modelling suggests that PepT1 could be a tetramer, with a minimum requirement of two functional subunits in each protein complex. Western blotting also showed the presence of higher-order complexes of PepT1-FLAG in oocyte membranes. It should be noted that we cannot formally exclude the possibility that PepT1 interacts with unidentified Xenopus protein(s). The finding that PepT1 is a multimer has important implications for the molecular modelling of this protein.

  2. Thyroid Hormone-Induced Activation of Notch Signaling is Required for Adult Intestinal Stem Cell Development During Xenopus Laevis Metamorphosis.

    PubMed

    Hasebe, Takashi; Fujimoto, Kenta; Kajita, Mitsuko; Fu, Liezhen; Shi, Yun-Bo; Ishizuya-Oka, Atsuko

    2017-04-01

    In Xenopus laevis intestine during metamorphosis, the larval epithelial cells are removed by apoptosis, and the adult epithelial stem (AE) cells appear concomitantly. They proliferate and differentiate to form the adult epithelium (Ep). Thyroid hormone (TH) is well established to trigger this remodeling by regulating the expression of various genes including Notch receptor. To study the role of Notch signaling, we have analyzed the expression of its components, including the ligands (DLL and Jag), receptor (Notch), and targets (Hairy), in the metamorphosing intestine by real-time reverse transcription-polymerase chain reaction and in situ hybridization or immunohistochemistry. We show that they are up-regulated during both natural and TH-induced metamorphosis in a tissue-specific manner. Particularly, Hairy1 is specifically expressed in the AE cells. Moreover, up-regulation of Hairy1 and Hairy2b by TH was prevented by treating tadpoles with a γ-secretase inhibitor (GSI), which inhibits Notch signaling. More importantly, TH-induced up-regulation of LGR5, an adult intestinal stem cell marker, was suppressed by GSI treatment. Our results suggest that Notch signaling plays a role in stem cell development by regulating the expression of Hairy genes during intestinal remodeling. Furthermore, we show with organ culture experiments that prolonged exposure of tadpole intestine to TH plus GSI leads to hyperplasia of secretory cells and reduction of absorptive cells. Our findings here thus provide evidence for evolutionarily conserved role of Notch signaling in intestinal cell fate determination but more importantly reveal, for the first time, an important role of Notch pathway in the formation of adult intestinal stem cells during vertebrate development. Stem Cells 2017;35:1028-1039. © 2016 The Authors STEM CELLS published by Wiley Periodicals, Inc. on behalf of AlphaMed Press.

  3. Effects of cadmium on growth, metamorphosis and gonadal sex differentiation in tadpoles of the African clawed frog, Xenopus laevis

    USGS Publications Warehouse

    Sharma, Bibek; Patino, R.

    2009-01-01

    Xenopus laevis larvae were exposed to cadmium (Cd) at 0, 1, 8, 85 or 860 ??g L-1 in FETAX medium from 0 to 86 d postfertilization. Premetamorphic tadpoles were sampled on day 31; pre and prometamorphic tadpoles on day 49; and frogs (NF stage 66) between days 50 and 86. Survival, snout-vent length (SVL), tail length, total length, hindlimb length (HLL), initiation of metamorphic climax, size at and completion of metamorphosis, and gonadal condition and sex ratio (assessed histologically) were determined. Survival was unaffected by Cd until day 49, but increased mortality was observed after day 49 at 860 ??g Cd L-1. On day 31, when tadpoles were in early premetamorphosis, inhibitory effects on tadpole growth were observed only at 860 ??g Cd L-1. On day 49, when most tadpoles where in late premetamorphosis/early prometamorphosis, reductions in SVL, HLL and total length were observed at 8 and 860 but not 85 ??g L-1, thus creating a U-shaped size distribution at 0-85 ??g Cd L-1. However, this U-shaped size pattern was not evident in postmetamorphic individuals. In fact, frog size at completion of metamorphosis was slightly smaller at 85 ??g Cd L-1relative to control animals. These observations confirmed a recent report of a Cd concentration-dependent bimodal growth pattern in late-premetamorphic Xenopus tadpoles, but also showed that growth responses to varying Cd concentrations change with development. The fraction of animals initiating or completing metamorphosis during days 50-86 was reduced in a Cd concentration-dependent manner. Testicular histology and population sex ratios were unaffected by Cd suggesting that, unlike mammals, Cd is not strongly estrogenic in Xenopus tadpoles. ?? 2009 Elsevier Ltd.

  4. Effects of cadmium on growth, metamorphosis and gonadal sex differentiation in tadpoles of the African clawed frog, Xenopus laevis

    USGS Publications Warehouse

    Sharma, Bibek; Patino, Reynaldo

    2009-01-01

    Xenopus laevis larvae were exposed to cadmium (Cd) at 0, 1, 8. 85 or 860 mu g L(-1) in FETAX medium from 0 to 86 d postfertilization. Premetamorphic tadpoles were sampled on day 3 1; pre and prometamorphic tadpoles on day 49; and frogs (NF stage 66) between days 50 and 86. Survival, snout-vent length (SVL), tail length, total length, hindlimb length (HLL), initiation of metamorphic climax, size at and completion of metamorphosis, and gonadal condition and sex ratio (assessed histologically) were determined. Survival was unaffected by Cd until day 49, but increased mortality was observed after day 49 at 860 mu g Cd L(-1). On day 31, when tadpoles were in early premetamorphosis, inhibitory effects on tadpole growth were observed only at 860 mu g Cd L(-1). On day 49, when most tadpoles where in late premetamorphosis/early prometamorphosis, reductions in SVL, HLL and total length were observed at 8 and 860 but not 85 mu g L(-1), thus creating a U-shaped size distribution at 0-85 mu g Cd L(-1). However, this U-shaped size pattern was not evident in postmetamorphic individuals. In fact, frog size at completion of metamorphosis was slightly smaller at 85 mu g Cd L(-1) relative to control animals. These observations confirmed a recent report of a Cd concentration-dependent bimodal growth pattern in late-premetamorphic Xenopus tadpoles, but also showed that growth responses to varying Cd concentrations change with development. The fraction of animals initiating or completing metamorphosis during days 50-86 was reduced in a Cd concentration-dependent manner. Testicular histology and population sex ratios were unaffected by Cd suggesting that, unlike mammals, Cd is not strongly estrogenic in Xenopus tadpoles.

  5. Distinct abscisic acid signaling pathways for modulation of guard cell versus mesophyll cell potassium channels revealed by expression studies in Xenopus laevis oocytes

    NASA Technical Reports Server (NTRS)

    Sutton, F.; Paul, S. S.; Wang, X. Q.; Assmann, S. M.; Evans, M. L. (Principal Investigator)

    2000-01-01

    Regulation of guard cell ion transport by abscisic acid (ABA) and in particular ABA inhibition of a guard cell inward K(+) current (I(Kin)) is well documented. However, little is known concerning ABA effects on ion transport in other plant cell types. Here we applied patch clamp techniques to mesophyll cell protoplasts of fava bean (Vicia faba cv Long Pod) plants and demonstrated ABA inhibition of an outward K(+) current (I(Kout)). When mesophyll cell protoplast mRNA (mesophyll mRNA) was expressed in Xenopus laevis oocytes, I(Kout) was generated that displayed similar properties to I(Kout) observed from direct analysis of mesophyll cell protoplasts. I(Kout) expressed by mesophyll mRNA-injected oocytes was inhibited by ABA, indicating that the ABA signal transduction pathway observed in mesophyll cells was preserved in the frog oocytes. Co-injection of oocytes with guard cell protoplast mRNA and cRNA for KAT1, an inward K(+) channel expressed in guard cells, resulted in I(Kin) that was similarly inhibited by ABA. However, oocytes co-injected with mesophyll mRNA and KAT1 cRNA produced I(Kin) that was not inhibited by ABA. These results demonstrate that the mesophyll-encoded signaling mechanism could not substitute for the guard cell pathway. These findings indicate that mesophyll cells and guard cells use distinct and different receptor types and/or signal transduction pathways in ABA regulation of K(+) channels.

  6. An elongated model of the Xenopus laevis transcription factor IIIA-5S ribosomal RNA complex derived from neutron scattering and hydrodynamic measurements.

    PubMed Central

    Timmins, P A; Langowski, J; Brown, R S

    1988-01-01

    The precise molecular composition of the Xenopus laevis TFIIIA-5S ribosomal RNA complex (7S particle) has been established from small angle neutron and dynamic light scattering. The molecular weight of the particle was found to be 95,700 +/- 10,000 and 86,700 +/- 9000 daltons from these two methods respectively. The observed match point of 54.4% D2O obtained from contrast variation experiments indicates a 1:1 molar ratio. It is concluded that only a single molecule of TFIIIA, a zinc-finger protein, and of 5S RNA are present in this complex. At high neutron scattering contrast radius of gyration of 42.3 +/- 2 A was found for the 7S particle. In addition a diffusion coefficient of 4.4 x 10(-11) [m2 s-1] and a sedimentation coefficient of 6.2S were determined. The hydrodynamic radius obtained for the 7S particle is 48 +/- 5 A. A simple elongated cylindrical model with dimensions of 140 A length and 59 A diameter is compatible with the neutron results. A globular model can be excluded by the shallow nature of the neutron scattering curves. It is proposed that the observed difference of 15 A in length between the 7S particle and isolated 5S RNA most likely indicates that part(s) of the protein protrudes from the end(s) of the RNA molecule. There is no biochemical evidence for any gross alteration in 5S RNA conformation upon binding to TFIIIA. PMID:3419928

  7. Urotensin II receptor (UTR) exists in hyaline chondrocytes: a study of peripheral distribution of UTR in the African clawed frog, Xenopus laevis.

    PubMed

    Konno, Norifumi; Fujii, Yuya; Imae, Haruka; Kaiya, Hiroyuki; Mukuda, Takao; Miyazato, Mikiya; Matsuda, Kouhei; Uchiyama, Minoru

    2013-05-01

    Urotensin II (UII) and UII-related peptide (URP) exhibit diverse physiological actions including vasoconstriction, locomotor activity, osmoregulation, and immune response through UII receptor (UTR), which is expressed in the central nervous system and peripheral tissues of fish and mammals. In amphibians, only UII has been identified. As the first step toward elucidating the actions of UII and URP in amphibians, we cloned and characterized URP and UTR from the African clawed frog Xenopus laevis. Functional analysis showed that treatment of UII or URP with Chinese hamster ovary cells transfected with the cloned receptor increased the intracellular calcium concentration in a concentration-dependent manner, whereas the administration of the UTR antagonist urantide inhibited UII- or URP-induced Ca(2+) mobilization. An immunohistochemical study showed that UTR was expressed in the splenocytes and leukocytes isolated from peripheral blood, suggesting that UII and URP are involved in the regulation of the immune system. UTR was also localized in the apical membrane of the distal tubule of the kidney and in the transitional epithelial cells of the urinary bladder. This result supports the view that the UII/URP-UTR system plays an important role in osmoregulation of amphibians. Interestingly, immunopositive labeling for UTR was first detected in the chondrocytes of various hyaline cartilages (the lung septa, interphalangeal joint and sternum). The expression of UTR was also observed in the costal cartilage, tracheal cartilages, and xiphoid process of the rat. These novel findings probably suggest that UII and URP mediate the formation of the cartilaginous matrix. Copyright © 2013 Elsevier Inc. All rights reserved.

  8. Steroid exposure during larval development of Xenopus laevis affects mRNA expression of the reproductive pituitary-gonadal axis in a sex- and stage-dependent manner.

    PubMed

    Urbatzka, Ralph; Lorenz, Claudia; Wiedemann, Caterina; Lutz, Ilka; Kloas, Werner

    2014-03-01

    Steroids are known to influence the reproductive pituitary-gonadal axis in adult amphibians. Here, we studied the effects of hormones on pituitary and gonadal mRNA expression during the development of Xenopus laevis. Tadpoles at NF 58 (prometamorphosis) and at NF 66 (freshly metamorphosed) were exposed for three days to 17β-estradiol (E2), tamoxifen (TAM), testosterone (T), dihydrotestosterone (DHT) at 10(-7)M, and flutamide (FLU) at 10(-6)M. In both genders at NF 58 and 66, T and DHT decreased luteinizing hormone beta (lhβ), but increased follicle stimulating hormone beta (fshβ), while FLU induced lhβ specifically in males. In the testis steroidogenic genes (p450 side chain cleavage enzyme, p450scc; steroid acute regulatory protein, star) at NF 58 showed a similar pattern as for lhβ, while the response at NF 66 was only partially present. In females, TAM induced lhβ at NF 58, while E2 decreased lhβ and increased fshβ at NF 66. In the ovaries, no alterations were observed for the steroidogenic genes. Summarizing, gonadotropic and steroidogenic mRNA expression may indicate control of androgen level during testis differentiation in male tadpoles at NF 58. In females the non-responsiveness of steroidogenic genes could be a sign of gonadal quiescence during pre-pubertal stages. Copyright © 2013 Elsevier Inc. All rights reserved.

  9. Friend of GATA (FOG) Interacts with the Nucleosome Remodeling and Deacetylase Complex (NuRD) to Support Primitive Erythropoiesis in Xenopus laevis

    PubMed Central

    Mimoto, Mizuho S.; Christian, Jan L.

    2012-01-01

    Friend of GATA (FOG) plays many diverse roles in adult and embryonic hematopoiesis, however the mechanisms by which it functions and the roles of potential interaction partners are not completely understood. Previous work has shown that overexpression of FOG in Xenopus laevis causes loss of blood suggesting that in contrast to its role in mammals, FOG might normally function to repress erythropoiesis in this species. Using loss-of-function analysis, we demonstrate that FOG is essential to support primitive red blood cell (RBC) development in Xenopus. Moreover, we show that it is specifically required to prevent excess apoptosis of circulating primitive RBCs and that in the absence of FOG, the pro-apoptotic gene Bim-1 is strongly upregulated. To identify domains of FOG that are essential for blood development and, conversely, to begin to understand the mechanism by which overexpressed FOG represses primitive erythropoiesis, we asked whether FOG mutants that are unable to interact with known co-factors retain their ability to rescue blood formation in FOG morphants and whether they repress erythropoiesis when overexpressed in wild type embryos. We find that interaction of FOG with the Nucleosome Remodeling and Deacetylase complex (NuRD), but not with C-terminal Binding Protein, is essential for normal primitive RBC development. In contrast, overexpression of all mutant and wild type constructs causes a comparable repression of primitive erythropoiesis. Together, our data suggest that a requirement for FOG and its interaction with NuRD during primitive erythropoiesis are conserved in Xenopus and that loss of blood upon FOG overexpression is due to a dominant-interfering effect. PMID:22235346

  10. Nearly 1000 Protein Identifications from 50 ng of Xenopus laevis Zygote Homogenate Using Online Sample Preparation on a Strong Cation Exchange Monolith Based Microreactor Coupled with Capillary Zone Electrophoresis.

    PubMed

    Zhang, Zhenbin; Sun, Liangliang; Zhu, Guijie; Cox, Olivia F; Huber, Paul W; Dovichi, Norman J

    2016-01-05

    A sulfonate-silica hybrid strong cation exchange monolith microreactor was synthesized and coupled to a linear polyacrylamide coated capillary for online sample preparation and capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) bottom-up proteomic analysis. The protein sample was loaded onto the microreactor in an acidic buffer. After online reduction, alkylation, and digestion with trypsin, the digests were eluted with 200 mM ammonium bicarbonate at pH 8.2 for CZE-MS/MS analysis using 1 M acetic acid as the background electrolyte. This combination of basic elution and acidic background electrolytes results in both sample stacking and formation of a dynamic pH junction. 369 protein groups and 1274 peptides were identified from 50 ng of Xenopus laevis zygote homogenate, which is comparable with an offline sample preparation method, but the time required for sample preparation was decreased from over 24 h to less than 40 min. Dramatically improved performance was produced by coupling the reactor to a longer separation capillary (∼100 cm) and a Q Exactive HF mass spectrometer. 975 protein groups and 3749 peptides were identified from 50 ng of Xenopus protein using the online sample preparation method.

  11. Effects of environmentally relevant sub-chronic atrazine concentrations on African clawed frog (Xenopus laevis) survival, growth and male gonad development.

    PubMed

    Rimayi, Cornelius; Odusanya, David; Weiss, Jana M; de Boer, Jacob; Chimuka, Luke; Mbajiorgu, Felix

    2018-06-01

    Sub-chronic toxicity of environmentally relevant atrazine concentrations on exposed tadpoles and adult male African clawed frogs (Xenopus laevis) was evaluated in a quality controlled laboratory for 90 days. The aim of this study was to determine the effects of atrazine on the survival, growth and gonad development of African clawed frogs. After exposure of tadpoles to atrazine concentrations of 0 (control), 0.01, 200 and 500 μg L -1 in water, mortality rates of 0, 0, 3.3 and 70% respectively were recorded for the 90 day exposure period. Morphometry showed significantly reduced tadpole mass in the 500 μg L -1 atrazine exposed tadpoles (p < 0.05). Light microscopy on testes of adult frogs exposed to the same atrazine concentrations using hematoxylin and eosin (H&E) and Van Gieson staining techniques revealed gonadal atrophy, disruption of germ cell lines, seminiferous tubule structure damage and formation of extensive connective tissue around seminiferous tubules of frogs exposed to 200 μg L -1 and 500 μg L -1 atrazine concentrations. Ultrastructural analysis of the cellular organelles using transmission electron microscopy (TEM) revealed significant amounts of damaged mitochondria in testosterone producing Leydig cells as well as Sertoli cells. Biochemical analysis revealed reduced serum testosterone levels in adult frogs at all exposure levels as well as presence of six atrazine metabolites in frog serum and liver. The results indicate that atrazine concentrations greater than the calculated LC50 of 343.7 μg L -1 cause significant mortality in tadpoles, while concentrations ≥200 μg L -1 adversely affect reproductive health of adult frogs and development of tadpoles sub-chronically exposed to atrazine. Copyright © 2018 Elsevier B.V. All rights reserved.

  12. Organization of cytokeratin cytoskeleton and germ plasm in the vegetal cortex of Xenopus laevis oocytes depends on coding and non-coding RNAs: Three-dimensional and ultrastructural analysis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Kloc, Malgorzata; Bilinski, Szczepan; Dougherty, Matthew T.

    2007-05-01

    Recent studies discovered a novel structural role of RNA in maintaining the integrity of the mitotic spindle and cellular cytoskeleton. In Xenopus laevis, non-coding Xlsirts and coding VegT RNAs play a structural role in anchoring localized RNAs, maintaining the organization of the cytokeratin cytoskeleton and germinal granules in the oocyte vegetal cortex and in subsequent development of the germline in the embryo. We studied the ultrastructural effects of antisense oligonucleotide driven ablation of Xlsirts and VegT RNAs on the organization of the cytokeratin, germ plasm and other components of the vegetal cortex. We developed a novel method to immunolabel andmore » visualize cytokeratin at the electron microscopy level, which allowed us to reconstruct the ultrastructural organization of the cytokeratin network relative to the components of the vegetal cortex in Xenopus oocytes. The removal of Xlsirts and VegT RNAs not only disrupts the cytokeratin cytoskeleton but also has a profound transcript-specific effect on the anchoring and distribution of germ plasm islands and their germinal granules and the arrangement of yolk platelets within the vegetal cortex. We suggest that the cytokeratin cytoskeleton plays a role in anchoring of germ plasm islands within the vegetal cortex and germinal granules within the germ plasm islands.« less

  13. Paxillin and embryonic PolyAdenylation Binding Protein (ePABP) engage to regulate androgen-dependent Xenopus laevis oocyte maturation - A model of kinase-dependent regulation of protein expression.

    PubMed

    Miedlich, Susanne U; Taya, Manisha; Young, Melissa Rasar; Hammes, Stephen R

    2017-06-15

    Steroid-triggered Xenopus laevis oocyte maturation is an elegant physiologic model of nongenomic steroid signaling, as it proceeds completely independent of transcription. We previously demonstrated that androgens are the main physiologic stimulator of oocyte maturation in Xenopus oocytes, and that the adaptor protein paxillin plays a crucial role in mediating this process through a positive feedback loop in which paxillin first enhances Mos protein translation, ensued by Erk2 activation and Erk-dependent phosphorylation of paxillin on serine residues. Phosphoserine-paxillin then further augments Mos protein translation and downstream Erk2 activation, resulting in meiotic progression. We hypothesized that paxillin enhances Mos translation by interacting with embryonic PolyAdenylation Binding Protein (ePABP) on polyadenylated Mos mRNA. Knockdown of ePABP phenocopied paxillin knockdown, with reduced Mos protein expression, Erk2 and Cdk1 activation, as well as oocyte maturation. In both Xenopus oocytes and mammalian cells (HEK-293), paxillin and ePABP constitutively interacted. Testosterone (Xenopus) or EGF (HEK-293) augmented ePABP-paxillin binding, as well as ePABP binding to Mos mRNA (Xenopus), in an Erk-dependent fashion. Thus, ePABP and paxillin work together in an Erk-dependent fashion to enhance Mos protein translation and promote oocyte maturation. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. A perchlorate sensitive iodide transporter in frogs

    PubMed Central

    Carr, Deborah L.; Carr, James A.; Willis, Ray E.; Pressley, Thomas A.

    2008-01-01

    Nucleotide sequence comparisons have identified a gene product in the genome database of African clawed frogs (Xenopus laevis) as a probable member of the solute carrier family of membrane transporters. To confirm its identity as a putative iodide transporter, we examined the function of this sequence after heterologous expression in mammalian cells. A green monkey kidney cell line transfected with the Xenopus nucleotide sequence had significantly greater 125I uptake than sham-transfected control cells. The uptake in carrier-transfected cells was significantly inhibited in the presence of perchlorate, a competitive inhibitor of mammalian Na+/iodide symporter. Tissue distributions of the sequence were also consistent with a role in iodide uptake. The mRNA encoding the carrier was found to be expressed in the thyroid gland, stomach, and kidney of tadpoles from X. laevis, as well as the bullfrog Rana catesbeiana. The ovaries of adult X. laevis also were found to express the carrier. Phylogenetic analysis suggested that the putative X. laevis iodide transporter is orthologous to vertebrate Na+-dependent iodide symporters. We conclude that the amphibian sequence encodes a protein that is indeed a functional Na+/iodide symporter in Xenopus laevis, as well as Rana catesbeiana. PMID:18275962

  15. Comparative studies on Pb and Cd levels in parasites of terrestrial and aquatic animals

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Sures, B.; Taraschewski, H.

    1995-12-31

    Several fish parasites (Acanthocephala, Cestoda, Nematoda) and organs of their respective intermediate and final hosts were analyzed for heavy metals by electrothermal atomic absorption spectrometry (ET-AAS). Pb and Cd were also quantified in the liver fluke Fasciola hepatica as well as in different organs of the large intestinal roundworm Ascaris suum. The levels of these heavy metals in the parasites were compared to those of muscle, liver, kidney and intestine of the respective definitive hosts cattle and swine obtained from a slaughter house. Most parasites accumulated significantly higher levels of metals than their final hosts. This was most conspicuous inmore » acanthocephalans which contained up to 3 {times} 10{sup 3} fold more lead than the muscle of their fish hosts and up to 1.1 {times} 10{sup 4} more lead than the water surrounding the fish. In these helminths cadmium was enriched up to 400 fold compared to the muscle of the fish and up to 2.7 {times} 10{sup 4} compared to the water. In contrast to the accumulation capacity of adult acanthocephalans their larvae contained about 30 to 180 times less Pb and Cd. Thus, the predominant accumulation of both metals appears in the adult worms. The cestodes of fish and the liver flukes of cattle accumulated the metals up to 200 fold compared to the muscle of their hosts. The nematodes did not contain higher levels of the metals than their hosts. Thus, parasites, especially acanthocephalans, seem to be sensitive bioindicators of Pb and Cd in their environments.« less

  16. Cultural Resources Investigations of the West Bank Hurricane Protection Project, Jefferson Parish, Louisiana

    DTIC Science & Technology

    1989-09-01

    most common understory plants include thistle (Carduus sp.), violet (Viola sp.), bur- marigold (Bidens laevis), and ferns. Drained cypress-tupelo swamp... marigold (Bidens laevis), violet (Viola sp.), thistle (Carduus sp.), and Southern shield fern (Thelypteris kunthil). Drained marsh makes up the

  17. Over 4,100 protein identifications from a Xenopus laevis fertilized egg digest using reversed-phase chromatographic prefractionation followed by capillary zone electrophoresis - electrospray ionization - tandem mass spectrometry analysis

    PubMed Central

    Yan, Xiaojing; Sun, Liangliang; Zhu, Guijie; Cox, Olivia F.; Dovichi, Norman J.

    2016-01-01

    A tryptic digest generated from Xenopus laevis fertilized embryos was fractionated by reversed phase liquid chromatography. One set of 30 fractions was analyzed by 100-min CZE-ESI-MS/MS separations (50 hr total instrument time), and a second set of 15 fractions was analyzed by 3-hr UPLC-ESI-MS/MS separations (45 hr total instrument time). CZE-MS/MS produced 70% as many protein IDs (4,134 vs. 5,787) and 60% as many peptide IDs (22,535 vs. 36,848) as UPLC-MS/MS with similar instrument time (50 h vs. 45 h) but with 50 times smaller total consumed sample amount (1.5 μg vs. 75 μg). Surprisingly, CZE generated peaks that were 25% more intense than UPLC for peptides that were identified by both techniques, despite the 50-fold lower loading amount; this high sensitivity reflects the efficient ionization produced by the electrokinetically-pumped nanospray interface used in CZE. This report is the first comparison of CZE-MS/MS and UPLC-MS/MS for large-scale eukaryotic proteomic analysis. The numbers of protein and peptide identifications produced by CZE-ESI-MS/MS approach those produced by UPLC-MS/MS, but with nearly two orders of magnitude lower sample amounts. PMID:27723263

  18. The cost of muscle power production: muscle oxygen consumption per unit work increases at low temperatures in Xenopus laevis.

    PubMed

    Seebacher, Frank; Tallis, Jason A; James, Rob S

    2014-06-01

    Metabolic energy (ATP) supply to muscle is essential to support activity and behaviour. It is expected, therefore, that there is strong selection to maximise muscle power output for a given rate of ATP use. However, the viscosity and stiffness of muscle increases with a decrease in temperature, which means that more ATP may be required to achieve a given work output. Here, we tested the hypothesis that ATP use increases at lower temperatures for a given power output in Xenopus laevis. To account for temperature variation at different time scales, we considered the interaction between acclimation for 4 weeks (to 15 or 25°C) and acute exposure to these temperatures. Cold-acclimated frogs had greater sprint speed at 15°C than warm-acclimated animals. However, acclimation temperature did not affect isolated gastrocnemius muscle biomechanics. Isolated muscle produced greater tetanus force, and faster isometric force generation and relaxation, and generated more work loop power at 25°C than at 15°C acute test temperature. Oxygen consumption of isolated muscle at rest did not change with test temperature, but oxygen consumption while muscle was performing work was significantly higher at 15°C than at 25°C, regardless of acclimation conditions. Muscle therefore consumed significantly more oxygen at 15°C for a given work output than at 25°C, and plastic responses did not modify this thermodynamic effect. The metabolic cost of muscle performance and activity therefore increased with a decrease in temperature. To maintain activity across a range of temperature, animals must increase ATP production or face an allocation trade-off at lower temperatures. Our data demonstrate the potential energetic benefits of warming up muscle before activity, which is seen in diverse groups of animals such as bees, which warm flight muscle before take-off, and humans performing warm ups before exercise. © 2014. Published by The Company of Biologists Ltd.

  19. COMPARATIVE TOXICITY OF DIURON ON SURVIVAL AND GROWTH OF PACIFIC TREEFROG, BULLFROG, RED-LEGGED FROG, AND AFRICAN CLAWED FROG EMBRYOS AND TADPOLES

    EPA Science Inventory

    The effects of the herbicide diuron on survival and growth of Pacific treefrog (Pseudacris regilla),bullfrog(Rana catesbeiana), red-legged frog(Rana aurora),and African clawed frog(Xenopus laevis)embryos and tadpoles were determined in static-renewal tests. P.regilla and X.laevis...

  20. Comparison of Etomidate, Benzocaine, and MS222 Anesthesia with and without Subsequent Flunixin Meglumine Analgesia in African Clawed Frogs (Xenopus laevis).

    PubMed

    Smith, Briony D; Vail, Krystal J; Carroll, Gwendolyn L; Taylor, Maggie C; Jeffery, Nicholas D; Vemulapalli, Tracy H; Elliott, James J

    2018-03-01

    Often few alternative anesthetics for exotic species are available, due to the small numbers of these animals used in research. In this study, we evaluated the depth and duration of anesthesia in Xenopus laevis after their immersion in 3 doses of etomidate (15, 22.5, and 30 mg/L) and in 3 doses of benzocaine (0.1%, 0.5%, and 1%) compared with the 'gold standard,' tricaine methanesulfonate (MS222; 2 g/L). We then chose an optimal dose for each alternative anesthetic according to induction time, duration of surgical plane, and time to complete recovery. The optimal etomidate and benzocaine doses (22.5 mg/L and 0.1%, respectively) as well as the MS222 dose were then used to achieve a surgical plane of anesthesia, with the addition of flunixin meglumine (25 or 50 mg/kg) administered in the dorsal lymph sac at the completion of mock oocyte harvest. Efficacy of the analgesic was assessed at 1, 3, 6, and 24 h postoperatively by using acetic acid testing (AAT). Histology of the liver, kidney, and tissues surrounding the dorsal lymph sac was performed at day 3, 14, and 28 in each group of animals. Mild to moderate myocyte degeneration and necrosis were present in tissues surrounding the dorsal lymph sac at both flunixin meglumine doses after etomidate and benzocaine anesthesia. In addition, the 50-mg/kg dose of flunixin meglumine resulted in the death of 5 of the 12 frogs within 24 h, despite an otherwise uneventful anesthetic recovery. In conclusion, benzocaine and etomidate offer alternative anesthetic regimens, according to typical requirements for an anesthetic event. Flunixin meglumine at the 25-mg/kg dose provided analgesic relief at the latest time point during etomidate dosage and at all time points during benzocaine dosage, but further characterization is warranted regarding long-term or repeated analgesic administration.

  1. Adult-type myogenesis of the frog Xenopus laevis specifically suppressed by notochord cells but promoted by spinal cord cells in vitro.

    PubMed

    Yamane, Hitomi; Ihara, Setsunosuke; Kuroda, Masaaki; Nishikawa, Akio

    2011-08-01

    Larval-to-adult myogenic conversion occurs in the dorsal muscle but not in the tail muscle during Xenopus laevis metamorphosis. To know the mechanism for tail-specific suppression of adult myogenesis, response character was compared between adult myogenic cells (Ad-cells) and larval tail myogenic cells (La-cells) to a Sonic hedgehog (Shh) inhibitor, notochord (Nc) cells, and spinal cord (SC) cells in vitro. Cyclopamine, an Shh inhibitor, suppressed the differentiation of cultured Ad (but not La) cells, suggesting the significance of Shh signaling in promoting adult myogenesis. To test the possibility that Shh-producing axial elements (notochord and spinal cord) regulate adult myogenesis, Ad-cells or La-cells were co-cultured with Nc or SC cells. The results showed that differentiation of Ad-cells were strongly inhibited by Nc cells but promoted by SC cells. If Ad-cells were "separately" co-cultured with Nc cells without direct cell-cell interactions, adult differentiation was not inhibited but rather promoted, suggesting that Nc cells have two roles, one is a short-range suppression and another is a long-range promotion for adult myogenesis. Immunohistochemical analysis showed both notochord and spinal cord express the N-terminal Shh fragment throughout metamorphosis. The "spinal cord-promotion" and long-range effect by Nc cells on adult myogenesis is thus involved in Shh signaling, while the signaling concerning the short-range "Nc suppression" will be determined by future studies. Interestingly, these effects, "Nc suppression" and "SC promotion" were not observed for La-cells. Situation where the spinal cord/notochord cross-sectional ratio is quite larger in tadpole trunk than in the tail seems to contribute to trunk-specific promotion and tail-specific suppression of adult myogenesis during Xenopus metamorphosis.

  2. Prevalence of Corynosoma caspicum infection in Gasterosteus aculeatus fish in Caspian Sea, Northern Iran.

    PubMed

    Rahimi-Esboei, Bahman; Najm, Mahdi; Shaker, Morad; Fakhar, Mahdi; Mobedi, Iraj

    2017-09-01

    There is little information about the prevalence of Corynosoma caspicum in fish particularly Gasterosteus aculeatus in Iran and the world. The aim of the present study was to find out the prevalence of acanthocephalan infection in Babolsar district, southern coastal of Caspian Sea, Northern Iran. Between September 2012 and August 2014, a total of 360 G. aculeatus fishes were randomly collected by drift nets from coastal regions in Babolsar and then examined the intestine and body cavity for worm infections. A total of 360 G. aculeatus fishes, 109 (30.3%) were found infected with at least one Corynosoma capsicum , and there was no significant association between genders and the prevalence infection of acanthocephalan. Moreover, there was a significant difference in infected rate between summer (79%, 86/109) and spring (21%, 23/109) (p<0.05). The high occurrence of Corynosoma infection in G. aculeatus indicates the enzootic constancy status of the infection in the southern coastal of Caspian Sea, Northern Iran.

  3. Cytological and Morphological Analyses Reveal Distinct Features of Intestinal Development during Xenopus tropicalis Metamorphosis

    PubMed Central

    Matsuura, Kazuo; Shi, Yun-Bo

    2012-01-01

    Background The formation and/or maturation of adult organs in vertebrates often takes place during postembryonic development, a period around birth in mammals when thyroid hormone (T3) levels are high. The T3-dependent anuran metamorphosis serves as a model to study postembryonic development. Studies on the remodeling of the intestine during Xenopus (X.) laevis metamorphosis have shown that the development of the adult intestine involves de novo formation of adult stem cells in a process controlled by T3. On the other hand, X. tropicalis, highly related to X. laevis, offers a number of advantages for studying developmental mechanisms, especially at genome-wide level, over X. laevis, largely due to its shorter life cycle and sequenced genome. To establish X. tropicalis intestinal metamorphosis as a model for adult organogenesis, we analyzed the morphological and cytological changes in X. tropicalis intestine during metamorphosis. Methodology/Principal Findings We observed that in X. tropicalis, the premetamorphic intestine was made of mainly a monolayer of larval epithelial cells surrounded by little connective tissue except in the single epithelial fold, the typhlosole. During metamorphosis, the larval epithelium degenerates and adult epithelium develops to form a multi-folded structure with elaborate connective tissue and muscles. Interestingly, typhlosole, which is likely critical for adult epithelial development, is present along the entire length of the small intestine in premetamorphic tadpoles, in contrast to X. laevis, where it is present only in the anterior 1/3. T3-treatment induces intestinal remodeling, including the shortening of the intestine and the typhlosole, just like in X. laevis. Conclusions/Significance Our observations indicate that the intestine undergoes similar metamorphic changes in X. laevis and X. tropicalis, making it possible to use the large amount of information available on X. laevis intestinal metamorphosis and the genome sequence

  4. [Antitumor effect of recombinant Xenopus laevis vascular endothelial growth factor (VEGF) as a vaccine combined with adriamycin on EL4 lymphoma in mice].

    PubMed

    Niu, Ting; Liu, Ting; Jia, Yong-Qian; Liu, Ji-Yan; Wu, Yang; Hu, Bing; Tian, Ling; Yang, Li; Kan, Bing; Wei, Yu-Quan

    2005-09-01

    To explore the antitumor effect of immunotherapy with recombinant Xenopus laevis vascular endothelial growth factor (xVEGF) as a vaccine combined with adriamycin on lymphoma model in mice. EL4 lymphoma model was established in C57BL/6 mice. Mice were randomized into four groups: combination therapy, adriamycin alone, xVEGF alone and normal saline (NS) groups, and then were given relevant treatments. The growth of tumor, the survival rate of tumor-bearing mice, and the potential toxicity of regimens above were observed. Anti-VEGF antibody-producing B cells (APBCs) were detected by enzyme-linked immunospot (ELISPOT) assay. In addition, microvessel density (MVD) of tumor was detected by immunohistochemistry, and tumor cell apoptosis was also detected by TUNEL staining. The tumor volumes of mice were significantly smaller in combination group than those in other three groups (P < 0.05). Complete regression of tumor was observed in 3 of 10 mice in combination group. Forty-eight days after inoculation of tumor cells, the survival rate of mice was significantly higher in combination group than in NS group (P < 0.01). The anti-VEGF APBC count in combination group or xVEGF group was significantly higher, compared with that in adriamycin group or NS group (P < 0.01). MVD in tumor tissues was significantly lower in combination group than those in other three groups (P < 0.05). Moreover, tumor cell apoptosis was significantly higher in combination group than those in other three groups (P < 0.05). In this experimental study, the use of xVEGF vaccine and adriamycin as a combination of immunotherapy with chemotherapy has sucessfully produced synergistic antitumor effect on lymphoma in mice.

  5. Impact of maternal n-3 polyunsaturated fatty acid deficiency on dendritic arbor morphology and connectivity of developing Xenopus laevis central neurons in vivo.

    PubMed

    Igarashi, Miki; Santos, Rommel A; Cohen-Cory, Susana

    2015-04-15

    Docosahexaenoic acid (DHA, 22:6n-3) is an essential component of the nervous system, and maternal n-3 polyunsaturated fatty acids (PUFAs) are an important source for brain development. Here, the impact of DHA on developing central neurons was examined using an accessible in vivo model. Xenopus laevis embryos from adult female frogs fed n-3 PUFA-adequate or deficient diets were analyzed every 10 weeks for up to 60 weeks, when frogs were then switched to a fish oil-supplemented diet. Lipid analysis showed that DHA was significantly reduced both in oocytes and tadpoles 40 weeks after deprivation, and brain DHA was reduced by 57% at 60 weeks. In vivo imaging of single optic tectal neurons coexpressing tdTomato and PSD-95-GFP revealed that neurons were morphologically simpler in tadpoles from frogs fed the deficient diet compared with the adequate diet. Tectal neurons had significantly fewer dendrite branches and shorter dendritic arbor over a 48 h imaging period. Postsynaptic cluster number and density were lower in neurons deprived of n-3 PUFA. Moreover, changes in neuronal morphology correlated with a 40% decrease in the levels of BDNF mRNA and mature protein in the brain, but not in TrkB. Importantly, switching to a fish oil-supplemented diet induced a recovery in DHA content in the frog embryos within 20 weeks and diminished the deprivation effects observed on tectal neurons of Stage 45 tadpoles. Consequently, our results indicate that DHA impacts dendrite maturation and synaptic connectivity in the developing brain, and it may be involved in neurotrophic support by BDNF. Copyright © 2015 the authors 0270-6474/15/356079-14$15.00/0.

  6. Exposure of xenopus laevis tadpoles to cadmium reveals concentration-dependent bimodal effects on growth and monotonic effects on development and thyroid gland activity

    USGS Publications Warehouse

    Sharma, Bibek; Patino, R.

    2008-01-01

    Xenopus laevis were exposed to 0-855 ??g cadmium (Cd)/l (measured concentrations) in FETAX medium from fertilization to 47 days postfertilization. Measurements included embryonic survival and, at 47 days, tadpole survival, snout-vent length, tail length, total length, hindlimb length, weight, Nieuwkoop-Faber (NF) stage of development, initiation of metamorphic climax (??? NF 58), and thyroid follicle cell height. Embryonic and larval survival were unaffected by Cd. Relative to control tadpoles, reduced tail and total length were observed at 0.1- 8 and at 855 ??g Cd/l; and reduced snout-vent length, hindlimb length, and weight were observed at 0.1-1 and at 855 ??g Cd/l. Mean stage of development and rate of initiation of climax were unaffected by Cd at 0-84 ??g/l; however, none of the tadpoles exposed to 855 ??g Cd/l progressed beyond mid-premetamorphosis (NF 51). Thyroid glands with fully formed follicles were observed in all tadpoles ??? NF 49 examined. Follicle cell height was unaffected by Cd at 0-84 ??g/l but it was reduced at 855 ??g/l; in the latter, cell height was reduced even when compared with NF 49-51 tadpoles pooled from the 0 to 84 ??g Cd/l groups. In conclusion, (1) Cd affected tadpole growth in a bimodal pattern with the first and second inhibitory modes at concentrations below and above 84 ??g Cd/l, respectively; (2) exposure to high Cd concentrations (855 ??g/l) reduced thyroid activity and arrested tadpole development at mid-premetamorphosis; and (3) unlike its effect on growth, Cd inhibited tadpole development and thyroid function in a seemingly monotonic pattern.

  7. The polarization of the G-protein activated potassium channel GIRK5 to the vegetal pole of Xenopus laevis oocytes is driven by a di-leucine motif.

    PubMed

    Díaz-Bello, Beatriz; Rangel-García, Claudia I; Salvador, Carolina; Carrisoza-Gaytán, Rolando; Escobar, Laura I

    2013-01-01

    The G protein-coupled inwardly-rectifying potassium channels (known as GIRK or Kir3) form functional heterotetramers gated by G-βγ subunits. GIRK channels participate in heart rate modulation and neuronal postsynaptic inhibition in mammals. In Xenopus laevis oocytes, GIRK5 is a functional homomultimer. Previously, we found that phosphorylation of a tyrosine (Y16) at its N-terminus downregulates the surface expression of GIRK5. In this work, we elucidated the subcellular localization and trafficking of GIRK5 in oocytes. Several EGFP-GIRK5 chimeras were produced and an ECFP construct was used to identify the endoplasmic reticulum (ER). Whereas GIRK5-WT was retained in the ER at the animal pole, the phospho-null GIRK5-Y16A was localized to the vegetal pole. Interestingly, a construct with an N-terminal Δ25 deletion produced an even distribution of the channel in the whole oocyte. Through an alanine-scan, we identified an acidic cluster/di-leucine sorting-signal recognition motif between E17 and I22. We quantified the effect of each amino acid residue within this di-leucine motif in determining the distribution of GIRK5 to the animal and vegetal poles. We found that Y16 and I22 contributed to functional expression and were dominant in the polarization of GIRK5. We thus conclude that the N-terminal acidic di-leucine motif of GIRK5 determines its retention and polarized trafficking within Xl oocytes.

  8. The Polarization of the G-Protein Activated Potassium Channel GIRK5 to the Vegetal Pole of Xenopus laevis Oocytes Is Driven by a Di-Leucine Motif

    PubMed Central

    Díaz-Bello, Beatriz; Rangel-García, Claudia I.; Salvador, Carolina; Carrisoza-Gaytán, Rolando; Escobar, Laura I.

    2013-01-01

    The G protein-coupled inwardly-rectifying potassium channels (known as GIRK or Kir3) form functional heterotetramers gated by G-βγ subunits. GIRK channels participate in heart rate modulation and neuronal postsynaptic inhibition in mammals. In Xenopus laevis oocytes, GIRK5 is a functional homomultimer. Previously, we found that phosphorylation of a tyrosine (Y16) at its N-terminus downregulates the surface expression of GIRK5. In this work, we elucidated the subcellular localization and trafficking of GIRK5 in oocytes. Several EGFP-GIRK5 chimeras were produced and an ECFP construct was used to identify the endoplasmic reticulum (ER). Whereas GIRK5-WT was retained in the ER at the animal pole, the phospho-null GIRK5-Y16A was localized to the vegetal pole. Interestingly, a construct with an N-terminal Δ25 deletion produced an even distribution of the channel in the whole oocyte. Through an alanine-scan, we identified an acidic cluster/di-leucine sorting-signal recognition motif between E17 and I22. We quantified the effect of each amino acid residue within this di-leucine motif in determining the distribution of GIRK5 to the animal and vegetal poles. We found that Y16 and I22 contributed to functional expression and were dominant in the polarization of GIRK5. We thus conclude that the N-terminal acidic di-leucine motif of GIRK5 determines its retention and polarized trafficking within Xl oocytes. PMID:23717539

  9. Competition and feeding ecology in two sympatric Xenopus species (Anura: Pipidae)

    PubMed Central

    Vogt, Solveig; de Villiers, F. André; Ihlow, Flora; Rödder, Dennis

    2017-01-01

    The widespread African clawed frog (Xenopus laevis) occurs in sympatry with the IUCN Endangered Cape platanna (Xenopus gilli) throughout its entire range in the south-western Cape, South Africa. In order to investigate aspects of the interspecific competition between populations of X. laevis and X. gilli, an assessment of their niche differentiation was conducted through a comprehensive study on food composition and trophic niche structure at two study sites: the Cape of Good Hope (CoGH) and Kleinmond. A total of 399 stomach contents of X. laevis (n = 183) and X. gilli (n = 216) were obtained together with samples of available prey to determine food preferences using the Electivity index (E*), the Simpson’s index of diversity (1 − D), the Shannon index (H′), and the Pianka index (Ojk). Xenopus gilli diet was more diverse than X. laevis, particularly in Kleimond where the Shannon index was nearly double. Both species were found to consume large amounts of tadpoles belonging to different amphibian species, including congeners, with an overall higher incidence of anurophagy than previously recorded. However, X. laevis also feeds on adult X. gilli, thus representing a direct threat for the latter. While trophic niche overlap was 0.5 for the CoGH, it was almost 1 in Kleinmond, suggesting both species utilise highly congruent trophic niches. Further, subdividing the dataset into three size classes revealed overlap to be higher in small frogs in both study sites. Our study underlines the importance of actively controlling X. laevis at sites with X. gilli in order to limit competition and predation, which is vital for conservation of the south-western Cape endemic. PMID:28439453

  10. The effects of aquatic oxygen concentration, body size and respiratory behaviour on the stamina of obligate aquatic (Bufo americanus) and facultative air-breathing (Xenopus laevis and Rana berlandieri) anuran larvae.

    PubMed

    Wassersug, R J; Feder, M E

    1983-07-01

    Larvae of the anurans Rana berlandieri and Xenopus laevis have lungs and can breathe air as well as irrigate buccal and pharyngeal surfaces for aquatic respiration. Larvae of Bufo americanus lack lungs until just before metamorphosis and are obligately aquatic. We examined the relationship between the locomotor stamina (time to fatigue), aquatic oxygen concentration, body size, and respiratory behaviour of swimming larvae of these species, with the following results: Stamina is size-dependent in all three species. Aquatic hypoxia reduces stamina in larvae of all three species, but most conspicuously in Bufo. Breathing air increases stamina in Rana larvae, especially in large animals and under aquatic hypoxia. In contrast to Rana larvae, Xenopus larvae swimming in normoxic water undergo a reduction in stamina when allowed to breathe air. In hypoxic water, aerial respiration moderates the reduction in stamina seen in Xenopus larvae. Branchial irrigation is associated with increased stamina in Xenopus, and is increased under hypoxia and at high swimming velocities. Respiratory demand, buoyancy and the drag associated with branchial irrigation all affect respiratory behaviour in Xenopus larvae. The great amount of interspecific variation in the relationship between respiratory behaviour and stamina reveals the importance of measuring performance directly when attempting to interpret the functional significance of respiratory structures and behaviour.

  11. In X. laevis embryos high levels of the anti-apoptotic factor p27BBP/eIF6 are stage-dependently found in BrdU and TUNEL-reactive territories.

    PubMed

    De Marco, N; Campanella, C; Carotenuto, R

    2011-05-01

    p27BBP/eIF6 (β4 binding protein/eukaryotic initiation factor 6) is a highly conserved protein necessary for cell life. In adult eIF6 mice, a 50% decrease in the protein levels in all tissues is accompanied by a reduction in cell proliferation only in the liver, fat cells and cultured fibroblasts. During X. laevis embryogenesis expression of p27BBP/eIF6 is abundant in high proliferative territories. However, in Xenopus cell proliferation appears unaffected following p27BBP/eIF6 over-expression or down-regulation. Indeed, p27BBP/eIF6 is an anti-apoptotic factor acting upstream of Bcl2 that reduces endogenous apoptosis. We studied p27BBP/eIF6 protein localization in wild type embryos and compared it to proliferation and apoptosis. At the beginning of embryogenesis, high levels of p27BBP/eIF6, proliferation and apoptosis overlap. In later development stages high proliferation levels are present in the same regions where higher p27BBP/eIF6 expression is observed, while apoptosis does not appear specifically concentrated in the same sites. The higher presence of p27BBP/eIF6 would appear related to an increased need of apoptosis control in the regions where cell death is essential for normal development.

  12. Altered gravity affects ventral root activity during fictive swimming and the static vestibuloocular reflex in young tadpoles (Xenopus laevis).

    PubMed

    Böser, S; Dournon, C; Gualandris-Parisot, L; Horn, E

    2008-03-01

    During early periods of life, modifications of the gravitational environment affect the development of sensory, neuronal and motor systems. The vestibular system exerts significant effects on motor networks that control eye and body posture as well as swimming. The objective of the present study was to study whether altered gravity (AG) affects vestibuloocular and spinal motor systems in a correlated manner. During the French Soyuz taxi flight Andromède to the International Space Station ISS (launch: October 21, 2001; landing: October 31, 2001) Xenopus laevis embryos were exposed for 10 days to microgravity (microg). In addition, a similar experiment with 3g-hypergravity (3g) was performed in the laboratory. At onset of AG, embryos had reached developmental stages 24 to 27. After exposure to AG, each tadpole was tested for its roll-induced vestibuloocular reflex (rVOR) and 3 hours later it was tested for the neuronal activity recorded from the ventral roots (VR) during fictive swimming. During the post-AG recording periods tadpoles had reached developmental stages 45 to 47. It was observed that microgravity affected VR activity during fictive swimming and rVOR. In particular, VR activity changes included a significant decrease of the rostrocaudal delay and a significant increase of episode duration. The rVOR-amplitude was transiently depressed. Hypergravity was less effective on the locomotor pattern; occurring effects on fictive swimming were the opposite of microg effects. As after microgravity, the rVOR was depressed after 3g-exposure. All modifications of the rVOR and VR-activity recovered to normal levels within 4 to 7 days after termination of AG. Significant correlations between the rVOR amplitude and VR activity of respective tadpoles during the recording period have been observed in both tadpoles with or without AG experience. The data are consistent with the assumptions that during this period of life which is characterized by a progressive development

  13. Effects of cadmium, estradiol-17beta and their interaction on gonadal condition and metamorphosis of male and female African clawed frog, Xenopus laevis

    USGS Publications Warehouse

    Sharma, Bibek; Patino, Reynaldo

    2010-01-01

    To assess interaction effects between cadmium (Cd, a putative xenoestrogen) and estradiol-17beta (E(2)) on sex differentiation and metamorphosis, Xenopus laevis were exposed to solvent-control (0.005% ethanol), Cd (10microgL(-1)), E(2) (1microgL(-1)), or Cd and E(2) (Cd+E(2)) in FETAX medium from fertilization to 75d postfertilization. Each treatment was applied to four aquaria, each with 30 fertilized eggs. Mortality was recorded and animals were sampled as they completed metamorphosis (Nieuwkoop and Faber stage 66). Gonadal sex of individuals (including >or= tadpoles NF stage 55 at day 75) was determined gross-morphologically and used to compute sex ratios. Time course and percent completion of metamorphosis, snout-vent length (SVL), hindlimb length (HLL) and weight were analyzed for each gender separately. Survival rates did not differ among treatments. The E(2) and Cd+E(2) treatments significantly skewed sex ratios towards females; however, no sex-ratio differences were observed between the control and Cd treatments or between the E(2) and Cd+E(2) treatments. Time course of metamorphosis was generally delayed and percent completion of metamorphosis was generally reduced in males and females exposed to Cd, E(2) or their combination compared to control animals. In males, but not females, the effect of Cd+E(2) was greater than that of individual chemicals. Weight at completion of metamorphosis was reduced only in females and only by the Cd+E(2) treatment. In conclusion, although Cd at an environmentally relevant concentration did not exhibit direct or indirect feminizing effects in Xenopus tadpoles, the metal and E(2) both had similar inhibitory effects on metamorphosis that were of greater magnitude in males than females.

  14. Effects of cadmium, estradiol-17β and their interaction on gonadal condition and metamorphosis of male and female African clawed frog, Xenopus laevis

    USGS Publications Warehouse

    Sharma, Bibek; Patino, Reynaldo

    2010-01-01

    To assess interaction effects between cadmium (Cd, a putative xenoestrogen) and estradiol-17?? (E2) on sex differentiation and metamorphosis, Xenopus laevis were exposed to solvent-control (0.005% ethanol), Cd (10 ??g L-1), E2 (1 ??g L-1), or Cd and E2 (Cd + E2) in FETAX medium from fertilization to 75 d postfertilization. Each treatment was applied to four aquaria, each with 30 fertilized eggs. Mortality was recorded and animals were sampled as they completed metamorphosis (Nieuwkoop and Faber stage 66). Gonadal sex of individuals (including tadpoles ???NF stage 55 at day 75) was determined gross-morphologically and used to compute sex ratios. Time course and percent completion of metamorphosis, snout-vent length (SVL), hindlimb length (HLL) and weight were analyzed for each gender separately. Survival rates did not differ among treatments. The E2 and Cd + E2 treatments significantly skewed sex ratios towards females; however, no sex-ratio differences were observed between the control and Cd treatments or between the E2 and Cd + E2 treatments. Time course of metamorphosis was generally delayed and percent completion of metamorphosis was generally reduced in males and females exposed to Cd, E2 or their combination compared to control animals. In males, but not females, the effect of Cd + E2 was greater than that of individual chemicals. Weight at completion of metamorphosis was reduced only in females and only by the Cd + E2 treatment. In conclusion, although Cd at an environmentally relevant concentration did not exhibit direct or indirect feminizing effects in Xenopus tadpoles, the metal and E2 both had similar inhibitory effects on metamorphosis that were of greater magnitude in males than females.

  15. Chlorpyrifos exposure affects fgf8, sox9, and bmp4 expression required for cranial neural crest morphogenesis and chondrogenesis in Xenopus laevis embryos.

    PubMed

    Tussellino, Margherita; Ronca, Raffaele; Carotenuto, Rosa; Pallotta, Maria M; Furia, Maria; Capriglione, Teresa

    2016-10-01

    Chlorpyrifos (CPF) is an organophosphate insecticide used primarily to control foliage and soil-borne insect pests on a variety of food and feed crops. In mammals, maternal exposure to CPF has been reported to induce dose-related abnormalities such as slower brain growth and cerebral cortex thinning. In lower vertebrates, for example, fish and amphibians, teratogenic activity of this compound is correlated with several anatomical alterations. Little is known about the effects of CPF on mRNA expression of genes involved in early development of the anatomical structures appearing abnormal in embryos. This study investigated the effects of exposure to different CPF concentrations (10, 15 and 20 mg/L) on Xenopus laevis embryos from stage 4/8 to stage 46. Some of the morphological changes we detected in CPF-exposed embryos included cranial neural crest cell (NCC)-derived structures. For this reason, we analyzed the expression of select genes involved in hindbrain patterning (egr2), cranial neural crest chondrogenesis, and craniofacial development (fgf8, bmp4, sox9, hoxa2 and hoxb2). We found that CPF exposure induced a reduction in transcription of all the genes involved in NCC-dependent chondrogenesis, with largest reductions in fgf8 and sox9; whereas, in hindbrain, we did not find any alterations in egr2 expression. Changes in the expression of fgf8, bmp4, and sox9, which are master regulators of several developmental pathways, have important implications. If these changes are confirmed to belong to a general pattern of alterations in vertebrates prenatally exposed to OP, they might be useful to assess damage during vertebrate embryo development. Environ. Mol. Mutagen. 57:589-604, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  16. Aromatase, steroid-5-alpha-reductase type 1 and type 2 mRNA expression in gonads and in brain of Xenopus laevis during ontogeny.

    PubMed

    Urbatzka, R; Lutz, I; Kloas, W

    2007-01-01

    The key enzymes involved in the production of endogenous sex steroids are steroid-5-alpha-reductase and aromatase converting testosterone (T) into dihydrotestosterone (DHT) and into estradiol (E2), respectively. To gain more insights into the molecular mechanisms of sexual differentiation of amphibians, we determined the mRNA expression of steroid-5-alpha-reductase type1 (Srd5a1), type2 (Srd5a2) and aromatase (Aro) during ontogeny starting from the egg and ending after completion of metamorphosis in Xenopus laevis. Expression of all three enzymes was measured by means of semi-quantitative RT-PCR, determining for the first time Srd5a1 and Srd5a2 mRNA expression in amphibians. mRNA was analyzed in whole body homogenates from stage 12 to 48, while brain and gonads with kidney were studied separately from stage 48 to 66. Different ontogenetic mRNA expression patterns were observed for all genes analyzed, revealing early mRNA expression of Srd5a1 already in the egg at stage 12 whereas Srd5a2 and Aro was detected at stage 39. Sex-specific mRNA expressions of Srd5a2 and of Aro were determined in the gonads with kidney but not in brain. Srd5a2 was two-fold higher expressed in testes than in ovaries while Aro mRNA was ten-fold higher in ovaries. No gender-specific mRNA expression was observed for Srd5a1 in gonads and in brain. The ontogenetic patterns of Aro, Srd5a1 and Srd5a2 suggest that these genes are involved in sexual differentiation of gonads and brain already in early developmental stages. Especially in gonads Srd5a2 seems to be important for physiological regulation of testis development while Aro is associated with the development of ovaries.

  17. Bisphenol A causes malformation of the head region in embryos of Xenopus laevis and decreases the expression of the ESR-1 gene mediated by Notch signaling.

    PubMed

    Imaoka, Susumu; Mori, Tomohiro; Kinoshita, Tsutomu

    2007-02-01

    Bisphenol A (BpA) is widely used in industry and dentistry. Its effects on the embryonic development of Xenopus laevis were investigated. Xenopus embryos at stage 10.5 were treated with BpA. Developmental abnormalities were observed at stage 35; malformation of the head region including eyes and scoliosis. The expression of several markers of embryonic development was investigated by reverse transcription-polymerase chain reaction (RT-PCR). The pan-neural marker SOX-2, the neural stem cell marker nrp-1, the mesodermal marker MyoD, and the endodermal marker sox17alpha, were used. Although the expression of marker genes was not changed by treatment with BpA, that of Pax-6, a key regulator of the morphogenesis of the eyes, was decreased by BpA. Pax-6 is a downstream factor of Notch signaling. So, the expression of a typical Notch-dependent factor, ESR-1, was investigated in the presence of BpA. The expression of ESR-1 was efficiently suppressed by BpA. In whole mount in situ hybridization (WISH), Pax-6 was expressed in the central nervous system and eyes. The expression was lost completely on treatment with BpA. The expression of ESR-1 in the central nervous system and eyes also disappeared with BpA treatment. Injection of the intracellular domain of Notch efficiently recovered ESR-1 expression in the presence of BpA although injection of a ligand for notch, Delta, did not. These results suggest that BpA decreased the expression of ESR-1 by disrupting the Notch signal.

  18. ATP utilization for calcium uptake and force production in skinned muscle fibres of Xenopus laevis.

    PubMed Central

    Stienen, G J; Zaremba, R; Elzinga, G

    1995-01-01

    1. A method has been developed to discriminate between the rate of ATP hydrolysis associated with calcium uptake into the sarcoplasmic reticulum (SR) and force development of the contractile apparatus in mechanically or saponin-skinned skeletal muscle fibres. The rate of ATP hydrolysis was determined in fibres of different types from the iliofibularis muscle of Xenopus laevis by enzymatic coupling of ATP re-synthesis to the oxidation of NADH. 2. The ATPase activity was determined before and after exposure of the preparations for 30 min to a solution containing 0.5% Triton X-100, which effectively abolishes the SR ATPase activity. The fibres were activated in a solution containing 5 mM caffeine to ensure that calcium uptake into the SR was maximal. 3. At saturating Ca2+ concentrations the actomyosin (AM) and SR ATPase activities in fast-twitch fibres, at 4.3 degrees C, amounted to 1.52 +/- 0.07 and 0.58 +/- 0.10 mumol s-1 (g dry wt)-1, respectively (means +/- S.E.M.; n = 25). The SR ATPase activity was 25% of the total ATPase activity. At submaximal calcium concentrations the AM ATPase activity varied in proportion to the isometric force. 4. The calcium sensitivity of the SR ATPase was larger than that of the AM ATPase and its dependence on [Ca2+] was less steep. The AM ATPase activity was half-maximal at a pCa of 6.11 (pCa = -log [Ca2+]) whereas the SR ATPase activity was half-maximal at a pCa of 6.62. 5. In Triton X-100-treated fibres, at different 2,3-butanedione monoxime (BDM) concentrations, the AM ATPase activity and isometric force varied proportionally. The SR ATPase activity determined by extrapolation of the total ATPase activity in mechanically skinned or saponin-treated fibres to zero force, was independent of the BDM concentration in the range studied (0-20 mM). The values obtained for the SR ATPase activity in this way were similar to those obtained with Triton X-100 treatment. 6. The AM ATPase activity in slow-twitch fibres amounted to 0.74 +/- 0

  19. Prevalence of Corynosoma caspicum infection in Gasterosteus aculeatus fish in Caspian Sea, Northern Iran

    PubMed Central

    Rahimi-Esboei, Bahman; Najm, Mahdi; Shaker, Morad; Fakhar, Mahdi; Mobedi, Iraj

    2017-01-01

    Aim: There is little information about the prevalence of Corynosoma caspicum in fish particularly Gasterosteus aculeatus in Iran and the world. The aim of the present study was to find out the prevalence of acanthocephalan infection in Babolsar district, southern coastal of Caspian Sea, Northern Iran. Materials and Methods: Between September 2012 and August 2014, a total of 360 G. aculeatus fishes were randomly collected by drift nets from coastal regions in Babolsar and then examined the intestine and body cavity for worm infections. Results: A total of 360 G. aculeatus fishes, 109 (30.3%) were found infected with at least one Corynosoma capsicum, and there was no significant association between genders and the prevalence infection of acanthocephalan. Moreover, there was a significant difference in infected rate between summer (79%, 86/109) and spring (21%, 23/109) (p<0.05). Conclusion: The high occurrence of Corynosoma infection in G. aculeatus indicates the enzootic constancy status of the infection in the southern coastal of Caspian Sea, Northern Iran. PMID:29062206

  20. Developmental toxicity of p,p'-dichlorodiphenyltrichloroethane, 2,4,6-trinitrotoluene, their metabolites, and benzo[a]pyrene in Xenopus laevis embryos.

    PubMed

    Saka, Masahiro

    2004-04-01

    Since 1995, high incidences of deformed frogs have been documented in Kitakyushu, Japan. In this area, relatively high concentrations of DDT, trinitrotoluene (TNT), their metabolites (p,p'-dichlorodiphenyldichloroethylene [DDE], p,p'-dichlorodiphenyldichloroethane [DDD], 2-amino-4,6-dinitrotoluene [2ADNT], and 4-amino-2,6-dinitrotoluene [4ADNT]), and benzo[a]pyrene [BaP]) have been identified from field samples. I used a standardized assay with Xenopus laevis embryos (frog embryo teratogenesis assay--Xenopus, FETAX) to examine the developmental toxicity of these compounds. Both DDE and BaP were considered nearly nontoxic in embryonic development because they induced low (< 10%) mortality and malformation incidence even at the highest concentrations tested (DDE, 393 microM; BaP, 13.2 microM). The DDD (96-h median lethal concentration [LC50] = 44.1 microM, 96-h median effective concentration [EC50] for malformation = 14.9 microM) was more lethal and teratogenic than its parent compound, DDT (96-h LC50 = 101 microM, 96-h EC50 = 41.5 microM). Predominant symptoms observed were axial malformations (DDT and DDD) and irregular gut coiling (DDT). However, DDT and DDD should not act as major lethal or teratogenic toxicants in the aquatic environment within a short-term exposure via water because their 96-h LC50 and 96-h EC50 values were extremely high, considering their low solubility in water. The TNT (96-h LC50 = 16.7 microM) was more lethal than 2ADNT (96-h LC50 = 166 microM) or 4ADNT (96-h LC50 = 115 microM). Although 4ADNT (96-h EC50 = 85.8 microM) induced various tadpole malformations, it was a weak teratogen compared with TNT (96-h EC50 = 9.78 microM) and 2ADNT (96-h EC50 = 16.9 microM). The most typical malformations observed were axial malformations, eye abnormalities (TNT), edema, and irregular gut coiling (2ADNT and 4ADNT). The 96-h LC50 and 96-h EC50 values of TNT, 2ADNT, and 4ADNT were lower than their saturated concentrations in water. Therefore, these

  1. New genus, new species of Acanthocephala (Echinorhynchidae) from the Brazilian frog Hylodes phyllodes (Anura: Leptodactylidae).

    PubMed

    Bursey, Charles R; Vrcibradic, Davor; Hatano, Fabio H; Rocha, Carlos Frederico D

    2006-04-01

    Anuracanthorhynchus tritaxisentis n. gen., n. sp. from the intestines of the Brazilian frog Hylodes phyllodes (Leptodactylidae) is described and illustrated. Anuracanthorhynchus tritaxisentis n. gen., n. sp., is unique among the echinorhynchid Acanthocephala by possessing a spherical proboscis supporting a small number of hooks of equal size. It is the sixth acanthocephalan species reported from South American anurans.

  2. Lueheia inscripta (Westrumb, 1821) (Acanthocephala: Plagiorhynchidae) in anurans (Leptodactylidae: Bufonidae) from Mexico.

    PubMed

    Salgado-Maldonado, G; Caspeta-Mandujano, J M

    2010-06-01

    Juveniles of Lueheia inscripta (Westrumb, 1821 Travassos, 1919 (Acanthocephala: Plagiorhynchidae), an acanthocephalan with six lemnisci, are reported and described from mesenteries of frogs Leptodactylus fragilis Brochi, 1877 and a toad Bufo marinus (Linnaeus, 1758) from Morelos state, Mexico. These are new host records extending the known geographical distribution of this species from Brazil and Puerto Rico to Mexico.

  3. Physiological effects and bioconcentration of triclosan on amphibian larvae.

    PubMed

    Palenske, Nicole M; Nallani, Gopinath C; Dzialowski, Edward M

    2010-08-01

    We examined the acute effects of triclosan (TCS) exposure, a common antimicrobial found as a contaminant in the field, on survival and physiology of amphibian larvae. LC50 values were determined after 96h for North American larval species: Acris crepitans blanchardii, Bufo woodhousii woodhousii, Rana sphenocephala, and for a developmental model: Xenopus laevis. Amphibian larvae were most sensitive to TCS exposure during early development based upon 96-h LC50 values. Heart rates for X. laevis and North American larvae exposed to TCS were variable throughout development. Metabolic rates of X. laevis and R. sphenocephala larvae exposed to TCS were significantly affected in larvae exposed to [50% LC50] and [LC50]. Tissue uptake and tissue bioconcentration factor (BCF) of TCS were investigated in X. laevis, B. woodhousii woodhousii, and R. sphenocephala. In general, a significant increase was observed as exposure concentration increased. Tissue BCF values were dependent upon stage and species. While TCS concentrations used here are higher than environmental concentrations, exposure to TCS was dependent upon species and developmental stage, with early developmental stages being most sensitive to TCS exposure. Copyright 2010 Elsevier Inc. All rights reserved.

  4. Effects of photoinduced toxicity of fluoranthene on amphibian embryos and larvae

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hatch, A.C.; Burton, G.A. Jr.

    1998-09-01

    Embryos and newly hatched larvae of three amphibian species, the spotted salamander (Ambystoma maculatum), the northern leopard frog (Rana pipiens), and the African clawed frog (Xenopus laevis), were exposed to fluoranthene and ultraviolet (UV) light in two scenarios. Embryos were exposed in a laboratory setting from an early developmental stage through hatching under artificial UV light, and newly hatched larvae were exposed outdoors in varying sunlight intensity levels. Outdoor exposures indicated greater sensitivity in the toxic response than did laboratory exposures. In the laboratory, mortality and malformation of X. laevis were the most sensitive indicators of exposure. Xenopus laevis wasmore » also the most sensitive species tested to the effects of UV light alone. Hatching success of R. pipiens was monitored outdoors and was not a useful predictive endpoint in the determination of photoinduced toxicity; however, newly hatched larvae were sensitive to the effects of photoinduced toxicity. Amybstoma maculatum and X. laevis larvae were affected by low ({micro}g/L) concentrations of fluoranthene in sunlight. These findings suggest that low levels of polycyclic aromatic hydrocarbons could be acting synergistically with environmental factors such as UV light to place young amphibians at risk.« less

  5. Comparative toxicity of ammonium and nitrate compounds to Pacific treefrog and African clawed frog tadpoles

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Schuytema, G.S.; Nebeker, A.V.

    1999-10-01

    The effects of ammonium nitrate, ammonium chloride, ammonium sulfate, and sodium nitrate on survival and growth of Pacific treefrog (Pseudacris regilla [Baird and Girard]) and African clawed frog (Xenopus laevis [Daudin]) tadpoles were determined in static-renewal tests. The 10-d ammonium nitrate and ammonium sulfate LC50s for P. regilla were 55.2 and 89.7 mg/L NH{sub 4}-N, respectively. The 10-d LC50s for X. laevis for the three ammonium compounds ranged from 45 to 64 mg/L NH{sub 4}-N. The 10-d sodium nitrate LC50s were 266.2 mg/L NO{sub 3}-N for P. regilla and 1,236.2 mg/L NO{sub 3}-N for X. laevis. The lowest observed adversemore » effect level (LOAEL) of ammonium compound based on reduced length or weight was 24.6 mg/L NH{sub 4}-N for P. regilla and 99.5 mg/L NH{sub 4}-N for X. laevis. The lowest sodium nitrate LOAELs based on reduced length or weight were {lt}30.1 mg/L NO{sub 3}-N for P. regilla and 126.3 mg/L NO{sub 3}-N for X. laevis. Calculated un-ionized NH{sub 3} comprised 0.3 to 1.0% of measured NH{sub 4}-N concentrations. Potential harm to amphibians could occur if sensitive life stages were impacted by NH{sub 4}-N and NO{sub 3}-N in agricultural runoff or drainage for a sufficiently long period.« less

  6. Effects of a commonly used glyphosate-based herbicide formulation on early developmental stages of two anuran species.

    PubMed

    Wagner, Norman; Müller, Hendrik; Viertel, Bruno

    2017-01-01

    Environmental contamination, especially due to the increasing use of pesticides, is suggested to be one out of six main reasons for the global amphibian decline. Adverse effects of glyphosate-based herbicides on amphibians have been already discussed in several studies with different conclusions, especially regarding sublethal effects at environmentally relevant concentrations. Therefore, we studied the acute toxic effects (mortality, growth, and morphological changes) of the commonly used glyphosate-based herbicide formulation Roundup® UltraMax on early aquatic developmental stages of two anuran species with different larval types (obligate vs. facultative filtrating suspension feeders), the African clawed frog (Xenopus laevis) and the Mediterranean painted frog (Discoglossus pictus). While X. laevis is an established anuran model organism in amphibian toxicological studies, we aim to establish D. pictus as another model for species with facultative filtrating larvae. A special focus of the present study lies on malformations in X. laevis embryos, which were investigated using histological preparations. In general, embryos and larvae of X. laevis reacted more sensitive concerning lethal effects compared to early developmental stages of D. pictus. It was suggested, that especially the different morphology of their filter apparatus and the higher volume of water pumped through the buccopharynx of X. laevis larvae lead to higher exposure to the formulation. The test substance induced similar lethal effects in D. pictus larvae as it does in the teleost standard test organism used in pesticide approval, the rainbow trout (Oncorhynchus mykiss), whereas embryos of both species are apparently more tolerant and, conversely, X. laevis larvae about two times more sensitive. In both species, early larvae always reacted significantly more sensitive than embryos. Exposure to the test substance increased malformation rates in embryos of both species in a concentration

  7. DEVELOPMENTAL DIVERSITY OF AMPHIBIANS

    PubMed Central

    Elinson, Richard P.; del Pino, Eugenia M.

    2011-01-01

    The current model amphibian, Xenopus laevis, develops rapidly in water to a tadpole which metamorphoses into a frog. Many amphibians deviate from the X. laevis developmental pattern. Among other adaptations, their embryos develop in foam nests on land or in pouches on their mother’s back or on a leaf guarded by a parent. The diversity of developmental patterns includes multinucleated oogenesis, lack of RNA localization, huge non-pigmented eggs, and asynchronous, irregular early cleavages. Variations in patterns of gastrulation highlight the modularity of this critical developmental period. Many species have eliminated the larva or tadpole and directly develop to the adult. The wealth of developmental diversity among amphibians coupled with the wealth of mechanistic information from X. laevis permit comparisons that provide deeper insights into developmental processes. PMID:22662314

  8. [Corynosoma hannae Zdzitowiecki, 1984--a parasite of sea leopard from the Pacific sector of Antarctic].

    PubMed

    Striukov, A A; Iurakhno, M V

    2007-01-01

    Specimens of the acanthocephalan Corynosoma hannae Zdzitowiecki, 1984 from the Pacific (Balleni islands) and Atlantic (South Shetland) sectors of Antarctic are compared with those from the Pacific sector of Subantarctic (Auckland and Campbell islands). Probably New Zealand sea lion Phocarctos hookeri (Gray, 1844) is the secondary definitive host for Corynosoma hannae. Description and figures of the specimens examined are provided.

  9. Microgravity-induced modifications of the vestibuloocular reflex in Xenopus laevis tadpoles are related to development and the occurrence of tail lordosis.

    PubMed

    Horn, Eberhard R

    2006-08-01

    During space flights, tadpoles of the clawed toad Xenopus laevis occasionally develop upward bended tails (tail lordosis). The tail lordosis disappears after re-entry to 1g within a couple of days. The mechanisms responsible for the induction of the tail lordosis are unknown; physical conditions such as weight de-loading or physiological factors such as decreased vestibular activity in microgravity might contribute. Microgravity (microg) also exerts significant effects on the roll-induced vestibuloocular reflex (rVOR). The rVOR was used to clarify whether tail lordosis is caused by physiological factors, by correlating the occurrence of microg-induced tail lordosis with the extent of microg-induced rVOR modifications. Post-flight recordings from three space flights (D-2 Spacelab mission, STS-55 in 1993; Shuttle-to-Mir mission SMM-06, STS-84 in 1997; French Soyuz taxi flight Andromède to ISS in 2001) were analyzed in these experiments. At onset of microgravity, tadpoles were at stages 25-28, 33-36 or 45. Parameters tested were rVOR gain (ratio between the angular eye movement and the lateral 30 degrees roll) and rVOR amplitude (maximal angular postural change of the eyes during a 360 degrees lateral roll). A ratio of 22-84% of tadpoles developed lordotic tails, depending on the space flight. The overall observation was that the rVOR of tadpoles with normal tails was either not affected by microgravity, or it was enhanced. In contrast, the rVOR of lordotic animals always revealed a depression. In particular, during post-flight days 1-11, tadpoles with lordotic tails from all three groups (25-28, 33-36 and 45) showed a lower rVOR gain and amplitude than the 1g-controls. The rVOR gain and amplitude of tadpoles from the groups 25-28 and 33-36 that developed normal tails was not affected by microgravity while the rVOR of microg-tadpoles from the stage-45 group with normal tails revealed a significant rVOR augmentation. (1) the vestibular system of tadpoles with lordotic

  10. Evaluation, grafting success and field establishment of cashew rootstock as influenced by VAM fungi.

    PubMed

    Lakshmipathy, R; Balakrishna, A N; Bagyaraj, D J; Sumana, D A; Kumar, D P

    2004-11-01

    Seven isolates of vesicular arbuscular mycorrhizal (VAM) fungi were isolated from cashew rhizosphere soil of different cashew growing regions of South India. These seven isolates along with two more VAM fungi namely Acaulospora laevis and Glomus mosseae, which were found to be better symbionts for cashew during our earlier study were used to study their effectiveness on the growth and nutrition of cashew rootstock Ullal-1. Four promising VAM fungi were selected based on this study. Rootstocks inoculated with these four fungi were evaluated for their vigour through grafting success, using Ullal-3 cashew variety as scion. Grafting success was more in rootstocks inoculated with A. laevis and one of local isolates Glomus etunicatum. Grafts with rootstock treated with G. etunicatum and A. laevis survived and performed better when planted in the field compared to the uninoculated and other VAM fungal treatments.

  11. Parasitation of sea trout (Salmo trutta trutta L.) from the spawning ground and German coastal waters off Mecklenburg-Western Pomerania, Baltic Sea.

    PubMed

    Unger, Patrick; Palm, Harry W

    2016-01-01

    A total of 52 sea trouts, Salmo trutta trutta, were studied for parasites, originating from German freshwater streams and coastal waters of the Baltic Sea. While 35 specimens were caught mainly close to the shoreline in the Baltic Sea, 17 were sampled during their spawning migration in Warnow River and other neighboring rivers in Mecklenburg-Western Pomerania. A total of 12 different metazoan parasite species were found in sea trout originating from the Baltic Sea, including five digeneans, two cestodes, three nematodes, and two acanthocephalans. Marine and freshwater species were found. In the migratory trout, seven different parasite species were recorded (one digenean, two cestodes, one nematode, and one acanthocephalan), demonstrating lower parasite diversity and load during the spawning migration compared with the fish from the Baltic Sea. The anisakid nematodes Anisakis simplex (sensu stricto), Contraceacum rudolphii, and Hysterothylacium aduncum were identified by molecular analyses of the ITS-1, 5.8S, ITS-2, and flanking sequences of the rDNA. Together with the digenean Derogenes varicus, Hemiurus communis and H. luehei, and the cestode Diphyllobothrium dendriticum seven new host records for sea trout from the German part of the Baltic Sea are made.

  12. Parasites in harbour seals ( Phoca vitulina) from the German Wadden Sea between two Phocine Distemper Virus epidemics

    NASA Astrophysics Data System (ADS)

    Lehnert, K.; Raga, J. A.; Siebert, U.

    2007-12-01

    Parasites were collected from 107 harbour seals ( Phoca vitulina) found on the coasts of Schleswig-Holstein, Germany, between 1997 and 2000. The prevalence of the parasites and their associated pathology were investigated. Eight species of parasites, primarily nematodes, were identified from the examined organs: two anisakid nematodes ( Pseudoterranova decipiens (sensu lato) , Contracaecum osculatum (sensu lato)) from the stomach, Otostrongylus circumlitus (Crenosomatidae) and Parafilaroides gymnurus (Filaroididae) from the respiratory tract, one filarioid nematode ( Acanthocheilonema spirocauda) from the heart, two acanthocephalans, Corynosoma strumosum and C. semerme (Polymorphidae), from the intestine and an ectoparasite, Echinophthirius horridus (Anoplura, Insecta). Lungworm infection was the most prominent parasitological finding and secondary bacterial bronchopneumonia the most pathogenic lesion correlated with the parasites. Heavy nematode burdens in the respiratory tract were highly age-related and more frequent in young seals. A positive correlation was observed between high levels of pulmonary infection and severity of bronchopneumonia. The prevalence of lungworms in this study was higher than in seals that died during the 1988/1989 Phocine Distemper Virus epidemic, and the prevalence of acanthocephalans and heartworms had decreased compared to findings from the first die-off.

  13. Bioaccumulation of six PCB indicator congeners in a heavily polluted water reservoir in Eastern Slovakia: tissue-specific distribution in fish and their parasites.

    PubMed

    Brázová, Tímea; Hanzelová, Vladimíra; Miklisová, Dana

    2012-08-01

    Concentrations of six indicator PCB congeners (IUPAC nos. 28, 52, 101, 138, 153, and 180) were measured in several organs and adipose tissue of a freshwater predatory fishes (European perch, northern pike, pike perch, wels catfish) as well as in nonpredators (common carp, freshwater bream, goldfish, white bream) and in acanthocephalan Acanthocephalus lucii from the water reservoir Zemplínska šírava (Eastern Slovakia), which is considered to be one of the most PCB-contaminated places in Europe. Concentration of PCBs was determined by capillary gas chromatography in samples from May to September 2009. The two-way main-effect ANOVA confirmed that feeding habits of fish (P < 0.00001) and peculiarity of individual fish organs (P < 0.01) affect PCB bioaccumulation. The total amount of PCBs was significantly higher (P < 0.05) in predators compared to nonpredators. Tissue-specific differences were found in PCB accumulation in both fish groups. PCBs were predominantly accumulated in the liver and hard roe. Individual congeners were not distributed homogeneously within the investigated organs and adipose tissue. PCB 153 was present in higher concentrations than the other congeners in all fish organs as well as in adipose tissue comprising an average 31 and 34 % of ΣPCB in predators and nonpredators, respectively. Acanthocephalans, attached to the intestine of perch, absorbed significantly higher concentrations of PCBs (P < 0.001) than the muscles, liver, kidney, brain, and adipose tissue of their host. About 20 times lower amount of PCBs was detected in the liver and almost 3 times in muscles of infected perch. Data on PCB accumulation in perch infected with acanthocephalans demonstrated a decline of PCB values in all organs as well as in adipose tissue compared to noninfected fish. About 20 times lower amount of PCBs was detected in the liver and almost 3 times in muscles of infected perch. Present results could indicate that some parasitic organisms may

  14. Comparative Histopathology of Acanthocephalan Infections in Some Freshwater Fishes

    DTIC Science & Technology

    1980-08-01

    Zool. 48:19-42. Choquette, L. P. E. 1951. Parasites of fresh-vater fish. V. Parasitic helminths of the muskellunge, Estox in. niasqu ltoni MitchllI. ill...G. W. and W. J. lluffines. 1973. Histopatiology assoa-iated with endoparasimic helminths in bass. .1. Parasit . 59:306-313. Jansson, B. and R. Olsson...R. 1966. Some helminths of Itasca Park fishes. J. M’linnesota Acad. Sci. 33:99-101. Meyer, M. C. 1954. The larger animial parasites of the freshwater

  15. A method for hormonal induction of sperm release in anurans (eight species) and in vitro fertilization in lepidobatrachus species.

    PubMed

    Waggener, W L; Carroll, E J

    1998-02-01

    Injections of synthetic human gonadotropin releasing hormone (GnRH) into the dorsal pelvic area were used in an attempt to stimulate sperm release in isolated males of eight anuran species including Xenopus laevis, Rana pipiens and Lepidobatrachus laevis. Sperm were obtained within 1-5 h post injection either by mechanical stimulation or by cloacal lavage. Sperm suspensions varied from 8 microL to 7 mL and the cell densities ranged from 4 x 10(5) to 4 x 10(7) sperm/mL. The sperm obtained from seven species using GnRH-induced release were viable based on light microscopic observations of motility. In addition, sperm preparations fertilized eggs in vitro and produced normal tadpoles in the case of L. laevis and L. Ilanensis. This hormonal method of anuran sperm collection will provide a convenient non-injurious way to obtain anuran sperm for basic studies of reproduction and development.

  16. Quantitative comparative analysis of the nasal chemosensory organs of anurans during larval development and metamorphosis highlights the relative importance of chemosensory subsystems in the group.

    PubMed

    Jungblut, Lucas David; Reiss, John O; Paz, Dante A; Pozzi, Andrea G

    2017-09-01

    The anuran peripheral olfactory system is composed of a number of subsystems, represented by distinct neuroepithelia. These include the main olfactory epithelium and vomeronasal organ (found in most tetrapods) and three specialized epithelia of anurans: the buccal-exposed olfactory epithelium of larvae, and the olfactory recess and middle chamber epithelium of postmetamorphic animals. To better characterize the developmental changes in these subsystems across the life cycle, morphometric changes of the nasal chemosensory organs during larval development and metamorphosis were analyzed in three different anuran species (Rhinella arenarum, Hypsiboas pulchellus, and Xenopus laevis). We calculated the volume of the nasal chemosensory organs by measuring the neuroepithelial area from serial histological sections at four different stages. In larvae, the vomeronasal organ was relatively reduced in R. arenarum compared with the other two species; the buccal-exposed olfactory epithelium was absent in X. laevis, and best developed in H. pulchellus. In postmetamorphic animals, the olfactory epithelium (air-sensitive organ) was relatively bigger in terrestrial species (R. arenarum and H. pulchellus), whereas the vomeronasal and the middle chamber epithelia (water-sensitive organs) was best developed in X. laevis. A small olfactory recess (likely homologous with the middle chamber epithelium) was found in R. arenarum juveniles, but not in H. pulchellus. These results support the association of the vomeronasal and middle chamber epithelia with aquatic olfaction, as seen by their enhanced development in the secondarily aquatic juveniles of X. laevis. They also support a role for the larval buccal-exposed olfactory epithelium in assessment of oral contents: it was absent in X. laevis, an obligate suspension feeder, while present in the two grazing species. These initial quantitative results give, for the first time, insight into the functional importance of the peripheral

  17. Replication and Transcription of Eukaryotic DNA in Esherichia coli

    PubMed Central

    Morrow, John F.; Cohen, Stanley N.; Chang, Annie C. Y.; Boyer, Herbert W.; Goodman, Howard M.; Helling, Robert B.

    1974-01-01

    Fragments of amplified Xenopus laevis DNA, coding for 18S and 28S ribosomal RNA and generated by EcoRI restriction endonuclease, have been linked in vitro to the bacterial plasmid pSC101; and the recombinant molecular species have been introduced into E. coli by transformation. These recombinant plasmids, containing both eukaryotic and prokaryotic DNA, replicate stably in E. coli. RNA isolated from E. coli minicells harboring the plasmids hybridizes to amplified X. laevis rDNA. Images PMID:4600264

  18. Manipulative parasites may not alter intermediate host distribution but still enhance their transmission: field evidence for increased vulnerability to definitive hosts and non-host predator avoidance.

    PubMed

    Lagrue, C; Güvenatam, A; Bollache, L

    2013-02-01

    Behavioural alterations induced by parasites in their intermediate hosts can spatially structure host populations, possibly resulting in enhanced trophic transmission to definitive hosts. However, such alterations may also increase intermediate host vulnerability to non-host predators. Parasite-induced behavioural alterations may thus vary between parasite species and depend on each parasite definitive host species. We studied the influence of infection with 2 acanthocephalan parasites (Echinorhynchus truttae and Polymorphus minutus) on the distribution of the amphipod Gammarus pulex in the field. Predator presence or absence and predator species, whether suitable definitive host or dead-end predator, had no effect on the micro-distribution of infected or uninfected G. pulex amphipods. Although neither parasite species seem to influence intermediate host distribution, E. truttae infected G. pulex were still significantly more vulnerable to predation by fish (Cottus gobio), the parasite's definitive hosts. In contrast, G. pulex infected with P. minutus, a bird acanthocephalan, did not suffer from increased predation by C. gobio, a predator unsuitable as host for P. minutus. These results suggest that effects of behavioural changes associated with parasite infections might not be detectable until intermediate hosts actually come in contact with predators. However, parasite-induced changes in host spatial distribution may still be adaptive if they drive hosts into areas of high transmission probabilities.

  19. Correlations Between Emerita analoga and Profilicollis spp. as Influenced by Environmental Factors at Ocean Beach, San Francisco CA

    NASA Astrophysics Data System (ADS)

    Fang, E.; Le, A.

    2014-12-01

    Since 2002 the Careers in Science (CiS) intern program has monitored Ocean Beach in San Francisco, California for the population of Emerita analoga (Pacific Mole Crab) as part of a partnership program with the Long-term Monitoring Program and Experiential Training for Students (LiMPETS). LiMPETS is an organization that conducts citizen science with Bay Area youth such as the CiS interns. We specifically assist in the collection of Pacific Mole Crab population statistics at Ocean Beach during the summer from June through August. The purpose of collecting Pacific Mole Crabs is to monitor Profilicollis spp. (Acanthocephalan parasites) - to which Pacific Mole Crabs serve as intermediate hosts - and to learn more about our environment as Pacific Mole Crabs are indicator species. During our collections at Ocean Beach we record size, sex, and number of individuals at specific transects. We then take a random sample from the day, return to the lab, and record their sizes, sexes, and Acanthocephalan parasite load. The results of the collection and dissections are then entered into the LiMPETS online database for scientist and researchers to use. Our project will focus on correlations relating to the data collected (Pacific Mole Crab population, parasite load, abiotic and biotic factors, et cetera).

  20. Correlations Between Emerita analoga and Profilicollis spp. as Influenced by Environmetal Factors at Ocean Beach, San Francisco CA

    NASA Astrophysics Data System (ADS)

    Wright, S.; Garza, F.; Zhang, P.

    2015-12-01

    Since 2002 the Careers in Science (CiS) intern program has monitored Ocean Beach in San Francisco, California for the population of Emerita analoga (Pacific Mole Crab) as part of a partnership program with the Long-term Monitoring Program and Experiential Training for Students (LiMPETS). LiMPETS is an organization that conducts citizen science with Bay Area youth such as the CiS interns. We specifically assist in the collection of Pacific Mole Crab population statistics at Ocean Beach during the summer from June through August. The purpose of collecting Pacific Mole Crabs is to monitor Acanthocephalan parasites (Profilicollis spp.) - to which Pacific Mole Crabs serve as intermediate hosts - and to learn more about our environment as Pacific Mole Crabs are indicator species. During our collections at Ocean Beach we record size, sex, and number of individuals at specific transects. We then take a random sample from the day, return to the lab, and record their sizes, sexes, and Acanthocephalan parasite load. The results of the collection and dissections are then entered into the LiMPETS online database for scientist and researchers to use. Our project will focus on correlations relating to the data collected (Pacific Mole Crab population, parasite load, abiotic and biotic factors, et cetera).

  1. Correlations Between Emerita analoga and Profilicollis spp. as Influenced by Environmental Factors at Ocean Beach, San Francisco CA

    NASA Astrophysics Data System (ADS)

    Ma, J. H.; Hackett, C.; Lucana, F.; Esquivel, A.

    2016-12-01

    Since 2002 the Careers in Science (CiS) intern program has monitored Ocean Beach in San Francisco, California for the population of Emerita analoga (Pacific Mole Crab) as part of a partnership program with the Long-term Monitoring Program and Experiential Training for Students (LiMPETS). LiMPETS is an organization that conducts citizen science with Bay Area youth such as the CiS interns. We specifically assist in the collection of Pacific Mole Crab population statistics at Ocean Beach during the summer from June through August. The purpose of collecting Pacific Mole Crabs is to monitor Profilicollis spp. (Acanthocephalan parasites) - to which Pacific Mole Crabs serve as intermediate hosts - and to learn more about our environment, as Pacific Mole Crabs are indicator species. During our collections at Ocean Beach we record size, sex, and number of individuals at specific transects. We then take a random sample from the day, return to the lab, and record their sizes, sexes, and Acanthocephalan parasite load. The results of the collection and dissections are then entered into the LiMPETS online database for scientist and researchers to use. Our project will focus on correlations relating to the data collected (Pacific Mole Crab population, parasite load, abiotic and biotic factors, et cetera).

  2. Intestinal parasites of the Arctic fox in relation to the abundance and distribution of intermediate hosts.

    PubMed

    Stien, A; Voutilainen, L; Haukisalmi, V; Fuglei, E; Mørk, T; Yoccoz, N G; Ims, R A; Henttonen, H

    2010-01-01

    The intestinal parasite community of Arctic foxes (Vulpes lagopus) on the Svalbard archipelago in the High Arctic was investigated in relation to the abundance and distribution of intermediate hosts. Five species of cestodes (Echinococcus multilocularis, Taenia crassiceps, Taenia polyacantha, Taenia krabbei and Diphyllobothrium sp.), ascaridoid nematodes and one unidentified acanthocephalan species were found. The cestodes E. multilocularis, T. crassiceps and T. polyacantha all showed a decreasing prevalence in the fox population with increasing distance from their spatially restricted intermediate host population of sibling voles (Microtus levis). In addition, the prevalence of E. multilocularis in a sample from the vole population was directly related to the local vole abundance. The cestode T. krabbei uses reindeer as intermediate host, and its prevalence in female foxes was positively related to the density of reindeer (Rangifer tarandus platyrhyncus). Finally, the prevalence of the ascaridoid nematodes also decreased with increasing distance from the vole population, a finding that is consistent with the idea that voles are involved in transmission, most likely as paratenic hosts. The prevalence of the remaining species (Diphyllobothrium sp. and an unidentified acanthocephalan) was very low. We conclude that the distribution and abundance of intermediate host structure the gastrointestinal parasite community of the Arctic fox on the Svalbard archipelago.

  3. Molluscs of an intertidal soft-sediment area in China: Does overfishing explain a high density but low diversity community that benefits staging shorebirds?

    NASA Astrophysics Data System (ADS)

    Yang, Hong-Yan; Chen, Bing; Piersma, Theunis; Zhang, Zhengwang; Ding, Changqing

    2016-03-01

    The Yellow Sea is a key staging ground for shorebirds that migrate from Australasia to the Arctic each spring. A lot of attention has been paid to the impact of habitat loss due to land reclamation on shorebird survival, but any effects of overfishing of coastal resources are unclear. In this study, the abundance of molluscs in the intertidal mudflats of northern Bohai Bay on the Chinese Yellow Sea was investigated in 2008-2014 from the perspective of their importance as food for northward migrating shorebirds, especially Red Knots Calidris canutus. Numerically contributing 96% to the numbers of 17 species found in spring 2008, the bivalve Potamocorbula laevis (the staple food of Red Knots and other shorebirds) dominated the intertidal mollusc community. In the spring of 2008-2014, the densities of P. laevis were surprisingly high, varying between 3900 and 41,000 individuals/m2 at distinctly small sizes (average shell lengths of 1.1 to 4.8 mm), and thus reaching some of the highest densities of marine bivalves recorded worldwide and providing good food for shorebirds. The distribution of P. laevis was associated with relatively soft sediments in close proximity to the recently built seawalls. A monthly sampling programme showed steep seasonal changes in abundance and size. P. laevis were nearly absent in winter, each year settling on the intertidal mudflats anew. Peak densities were reached in spring, when 0-age P. laevis were 1-3 mm long. The findings point to a highly unusual demographic structure of the species, suggesting that some interfering factors are at play. We hypothesise that the current dominance of young P. laevis in Bohai Bay reflects the combined pressures of a nearly complete active removal of adult populations from mid-summer to autumn for shrimp farming (this clearing of adults may offer space for recruitment during the next spring) and low numbers of epibenthic predators of bivalves, such as shrimps and crabs, due to persistent overfishing in

  4. Morphology and life history of Petasiger variospinosus (trematoda: echinostomatidae) in the Free State, South Africa.

    PubMed

    King, P H; Van As, J G

    2000-04-01

    Specimens of the freshwater snail Bulinus tropicus (Krauss, 1848) collected in the Free State, South Africa shed cercariae with an oral collar bearing 27 spines. Tadpoles of the African clawed toad Xenopus laevis laevis Daudin, 1802 collected from the same waters harbored metacercariae with a similar collar of spines. Adults were obtained after feeding infected tadpoles to laboratory-reared reed cormorants, Phalacrocorax africanus (Gmelin, 1789). The parasite was identified as Petasiger variospinosus (Odhner, 1910), the life cycle was experimentally completed, and stages described by the use of light and scanning electron microscopy.

  5. Liquid chromatography tandem mass spectrometry method using solid-phase extraction and bead-beating-assisted matrix solid-phase dispersion to quantify the fungicide tebuconazole in controlled frog exposure study: analysis of water and animal tissue.

    PubMed

    Hansen, Martin; Poulsen, Rikke; Luong, Xuan; Sedlak, David L; Hayes, Tyrone

    2014-11-01

    This paper presents the development, optimization, and validation of a LC-MS/MS methodology to determine the concentration of the antifungal drug and fungicide tebuconazole in a controlled exposure study of African clawed frogs (Xenopus laevis). The method is validated on animal tank water and on tissue from exposed and non-exposed adult X. laevis. Using solid-phase extraction (SPE), the analytical method allows for quantification of tebuconazole at concentrations as low as 3.89 pg mL(-1) in 10 mL water samples. Using bead-beating-assisted matrix solid-phase dispersion (MSPD), it was possible to quantify tebuconazole down to 0.63 pg mg(-1) wet weight liver using 150 mg tissue. The deuterated analogue of tebuconazole was used as internal standard, and ensured method accuracy in the range 80.6-99.7% for water and 68.1-109% for tissue samples. The developed method was successfully applied in a 4-week X. laevis repeated-exposure study, revealing high levels of tebuconazole residues in adipose and liver tissue, and with experimental bioconcentration factors up to 18,244 L kg(-1).

  6. Development of the Larval Amphibian Growth and Development Assay: effects of chronic 4-tert-octylphenol or 17β-trenbolone exposure in Xenopus laevis from embryo to juvenile.

    PubMed

    Haselman, Jonathan T; Kosian, Patricia A; Korte, Joseph J; Olmstead, Allen W; Iguchi, Taisen; Johnson, Rodney D; Degitz, Sigmund J

    2016-12-01

    The Larval Amphibian Growth and Development Assay (LAGDA) is a globally harmonized test guideline developed by the U.S. Environmental Protection Agency in collaboration with Japan's Ministry of the Environment. The LAGDA was designed to evaluate apical effects of chronic chemical exposure on growth, thyroid-mediated amphibian metamorphosis and reproductive development. During the validation phase, two well-characterized endocrine-disrupting chemicals were tested to evaluate the performance of the initial assay design: xenoestrogen 4-tert-octylphenol (tOP) and xenoandrogen 17β-trenbolone (TB). Xenopus laevis embryos were exposed, in flow-through conditions, to tOP (nominal concentrations: 0.0, 6.25, 12.5, 25 and 50 µg l -1 ) or TB (nominal concentrations: 0.0, 12.5, 25, 50 and 100 ng l -1 ) until 8 weeks post-metamorphosis, at which time growth measurements were taken, and histopathology assessments were made of the gonads, reproductive ducts, liver and kidneys. There were no effects on growth in either study and no signs of overt toxicity, sex reversal or gonad dysgenesis. Exposure to tOP caused a treatment-related decrease in circulating thyroxine and an increase in thyroid follicular cell hypertrophy and hyperplasia (25 and 50 µg l -1 ) during metamorphosis. Müllerian duct development was affected after exposure to both chemicals; tOP exposure caused dose-dependent maturation of oviducts in both male and female frogs, whereas TB exposure caused accelerated Müllerian duct regression in males and complete regression in >50% of the females in the 100 ng l -1 treatment. Based on these results, the LAGDA performed adequately to evaluate apical effects of chronic exposure to two endocrine-active compounds and is the first standardized amphibian multiple life stage toxicity test to date. Published 2016. This article is a U.S. Government work and is in the public domain in the USA. Published 2016. This article is a U.S. Government work and is in

  7. Parasitic helminths of black bear cubs (Ursus americanus) from Florida.

    PubMed

    Foster, Garry W; Cunningham, Mark W; Kinsella, John M; Forrester, Donald J

    2004-02-01

    Twenty-two Florida black bear (Ursus americanus floridanus) cubs (< or = 12 mo old) from Florida were examined for endoparasites between 1998 and 2003. Eleven species of helminths were found (8 nematodes, 2 trematodes, and 1 acanthocephalan). The most prevalent helminths were Ancylostoma caninum (64%), Macracanthorhynchus ingens (36%), Strongyloides sp. (27%), Baylisascaris transfuga (23%), and A. tubaeforme (14%). New host records include A. tubaeforme and Brachylaima virginianum. This is the first report of B. transfuga in black bears from Florida.

  8. Introduced brown trout alter native acanthocephalan infections in native fish.

    PubMed

    Paterson, Rachel A; Townsend, Colin R; Poulin, Robert; Tompkins, Daniel M

    2011-09-01

    1. Native parasite acquisition provides introduced species with the potential to modify native host-parasite dynamics by acting as parasite reservoirs (with the 'spillback' of infection increasing the parasite burdens of native hosts) or sinks (with the 'dilution' of infection decreasing the parasite burdens of native hosts) of infection. 2. In New Zealand, negative correlations between the presence of introduced brown trout (Salmo trutta) and native parasite burdens of the native roundhead galaxias (Galaxias anomalus) have been observed, suggesting that parasite dilution is occurring. 3. We used a multiple-scale approach combining field observations, experimental infections and dynamic population modelling to investigate whether native Acanthocephalus galaxii acquisition by brown trout alters host-parasite dynamics in native roundhead galaxias. 4. Field observations demonstrated higher infection intensity in introduced trout than in native galaxias, but only small, immature A. galaxii were present in trout. Experimental infections also demonstrated that A. galaxii does not mature in trout, although parasite establishment and initial growth were similar in the two hosts. Taken together, these results support the hypothesis that trout may serve as an infection sink for the native parasite. 5. However, dynamic population modelling predicts that A. galaxii infections in native galaxias should at most only be slightly reduced by dilution in the presence of trout. Rather, model exploration indicates parasite densities in galaxias are highly sensitive to galaxias predation on infected amphipods, and to relative abundances of galaxias and trout. Hence, trout presence may instead reduce parasite burdens in galaxias by either reducing galaxias density or by altering galaxias foraging behaviour. © 2011 The Authors. Journal of Animal Ecology © 2011 British Ecological Society.

  9. Comparative skull anatomy of terrestrial and crevice-dwelling Trachylepis skinks (Squamata: Scincidae) with a survey of resources in scincid cranial osteology.

    PubMed

    Paluh, Daniel J; Bauer, Aaron M

    2017-01-01

    Skinks account for more than 25% of all lizard species; however, representatives of fewer than a quarter of all species have been characterized osteologically. All but a few of the available cranial descriptions concentrate solely on characters that can be seen externally on the intact skull. Mabuyid skinks of the genus Trachylepis are the dominant, fully limbed skinks in Sub-Saharan Africa, and nearly all species have the same generalized body plan. Although a few rock crevice-dwelling species possess slight body depression, extreme dorsoventral depression is observed only in Trachylepis laevis. We investigated the detailed skull anatomy of three Trachylepis skinks (T. laevis, T. sulcata, and T. gonwouoi, a recently described species allied to T. affinis) using high-resolution X-ray micro-computed tomography. Our goals were to review the scincid cranial osteology literature in a phylogenetic context, provide a detailed anatomical atlas for the mabuyid lineage, and investigate the morphological adaptations of the highly modified T. laevis. Our results demonstrate that there is significant morphological variation between these three taxa, including the loss and fusion of structures, as well as changes in the shape, scale, and relationship between individual elements. Trachylepis laevis possesses several osteological modifications that have produced a reducton in head depth that are likely functional consequences of extreme rupicolous habits, including a flat skull roof, many strongly recumbent elements, and a depressed neurocranium.We hypothesize these modifications may correspond to descreased bite force and increased capabilities of cranial kinesis. Our study is the first element-by-element description of a skink using computed tomography technology.

  10. Transcription factors Mix1 and VegT, relocalization of vegt mRNA, and conserved endoderm and dorsal specification in frogs

    PubMed Central

    Sudou, Norihiro; Garcés-Vásconez, Andrés; López-Latorre, María A.; Taira, Masanori

    2016-01-01

    Protein expression of the transcription factor genes mix1 and vegt characterized the presumptive endoderm in embryos of the frogs Engystomops randi, Epipedobates machalilla, Gastrotheca riobambae, and Eleutherodactylus coqui, as in Xenopus laevis embryos. Protein VegT was detected in the animal hemisphere of the early blastula in all frogs, and only the animal pole was VegT-negative. This finding stimulated a vegt mRNA analysis in X. laevis eggs and embryos. vegt mRNA was detected in the animal region of X. laevis eggs and early embryos, in agreement with the VegT localization observed in the analyzed frogs. Moreover, a dorso-animal relocalization of vegt mRNA occurred in the egg at fertilization. Thus, the comparative analysis indicated that vegt may participate in dorsal development besides its known roles in endoderm development, and germ-layer specification. Zygotic vegt (zvegt) mRNA was detected as a minor isoform besides the major maternal (mvegt) isoform of the X. laevis egg. In addition, α-amanitin–insensitive vegt transcripts were detected around vegetal nuclei of the blastula. Thus, accumulation of vegt mRNA around vegetal nuclei was caused by relocalization rather than new mRNA synthesis. The localization of vegt mRNA around vegetal nuclei may contribute to the identity of vegetal blastomeres. These and previously reportedly localization features of vegt mRNA and protein derive from the master role of vegt in the development of frogs. The comparative analysis indicated that the strategies for endoderm, and dorsal specification, involving vegt and mix1, have been evolutionary conserved in frogs. PMID:27140624

  11. Comparative skull anatomy of terrestrial and crevice-dwelling Trachylepis skinks (Squamata: Scincidae) with a survey of resources in scincid cranial osteology

    PubMed Central

    Bauer, Aaron M.

    2017-01-01

    Skinks account for more than 25% of all lizard species; however, representatives of fewer than a quarter of all species have been characterized osteologically. All but a few of the available cranial descriptions concentrate solely on characters that can be seen externally on the intact skull. Mabuyid skinks of the genus Trachylepis are the dominant, fully limbed skinks in Sub-Saharan Africa, and nearly all species have the same generalized body plan. Although a few rock crevice-dwelling species possess slight body depression, extreme dorsoventral depression is observed only in Trachylepis laevis. We investigated the detailed skull anatomy of three Trachylepis skinks (T. laevis, T. sulcata, and T. gonwouoi, a recently described species allied to T. affinis) using high-resolution X-ray micro-computed tomography. Our goals were to review the scincid cranial osteology literature in a phylogenetic context, provide a detailed anatomical atlas for the mabuyid lineage, and investigate the morphological adaptations of the highly modified T. laevis. Our results demonstrate that there is significant morphological variation between these three taxa, including the loss and fusion of structures, as well as changes in the shape, scale, and relationship between individual elements. Trachylepis laevis possesses several osteological modifications that have produced a reducton in head depth that are likely functional consequences of extreme rupicolous habits, including a flat skull roof, many strongly recumbent elements, and a depressed neurocranium.We hypothesize these modifications may correspond to descreased bite force and increased capabilities of cranial kinesis. Our study is the first element-by-element description of a skink using computed tomography technology. PMID:28902864

  12. Helminths of three species of opossums (Mammalia, Didelphidae) from Mexico

    PubMed Central

    Acosta-Virgen, Karla; López-Caballero, Jorge; García-Prieto, Luis; Mata-López, Rosario

    2015-01-01

    Abstract From August 2011 to November 2013, 68 opossums (8 Didelphis sp., 40 Didelphis virginiana, 15 Didelphis marsupialis, and 5 Philander opossum) were collected in 18 localities from 12 Mexican states. A total of 12,188 helminths representing 21 taxa were identified (6 trematodes, 2 cestodes, 3 acanthocephalans and 10 nematodes). Sixty-six new locality records, 9 new host records, and one species, the trematode Brachylaima didelphus, is added to the composition of the helminth fauna of the opossums in Mexico. These data, in conjunction with previous records, bring the number of taxa parasitizing the Mexican terrestrial marsupials to 41. Among these species, we recognized a group of helminths typical of didelphids in other parts of the Americas. This group is constituted by the trematode Rhopalias coronatus, the acanthocephalan Oligacanthorhynchus microcephalus and the nematodes Cruzia tentaculata, Gnathostoma turgidum, and Turgida turgida. In general, the helminth fauna of each didelphid species showed a stable taxonomic composition with respect to previously sampled sites. This situation suggests that the rate of accumulation of helminth species in the inventory of these 3 species of terrestrial marsupials in the Neotropical portion of Mexico is decreasing; however, new samplings in the Nearctic portion of this country will probably increase the richness of the helminthological inventory of this group of mammals. PMID:26257556

  13. Phylogeny of Syndermata (syn. Rotifera): Mitochondrial gene order verifies epizoic Seisonidea as sister to endoparasitic Acanthocephala within monophyletic Hemirotifera.

    PubMed

    Sielaff, Malte; Schmidt, Hanno; Struck, Torsten H; Rosenkranz, David; Mark Welch, David B; Hankeln, Thomas; Herlyn, Holger

    2016-03-01

    A monophyletic origin of endoparasitic thorny-headed worms (Acanthocephala) and wheel-animals (Rotifera) is widely accepted. However, the phylogeny inside the clade, be it called Syndermata or Rotifera, has lacked validation by mitochondrial (mt) data. Herein, we present the first mt genome of the key taxon Seison and report conflicting results of phylogenetic analyses: while mt sequence-based topologies showed monophyletic Lemniscea (Bdelloidea+Acanthocephala), gene order analyses supported monophyly of Pararotatoria (Seisonidea+Acanthocephala) and Hemirotifera (Bdelloidea+Pararotatoria). Sequence-based analyses obviously suffered from substitution saturation, compositional bias, and branch length heterogeneity; however, we observed no compromising effects in gene order analyses. Moreover, gene order-based topologies were robust to changes in coding (genes vs. gene pairs, two-state vs. multistate, aligned vs. non-aligned), tree reconstruction methods, and the treatment of the two monogonont mt genomes. Thus, mt gene order verifies seisonids as sister to acanthocephalans within monophyletic Hemirotifera, while deviating results of sequence-based analyses reflect artificial signal. This conclusion implies that the complex life cycle of extant acanthocephalans evolved from a free-living state, as retained by most monogononts and bdelloids, via an epizoic state with a simple life cycle, as shown by seisonids. Hence, Acanthocephala represent a rare example where ancestral transitional stages have counterparts amongst the closest relatives. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Helminth parasites of the osprey, Pandion haliaetus, in North America

    USGS Publications Warehouse

    Kinsella, J.M.; Cole, Rebecca A.; Forrester, Donald J.; Roderick, Constance L.

    1996-01-01

    A total of 28 species of helminths (17 trematodes, 3 cestodes, 7 nematodes, and 1 acanthocephalan) was recovered from 17 ospreys (Pandion haliaetus) from the United States. Intensities of infection were low and no lesions were attributed to the parasites. Seven species appear to be specialists in ospreys, 2 species generalists in raptors, and the remainder generalists in other orders of fish-eating birds. Pandiontrema rjikovi, Diasiella diasi, and Contracaecum pandioni are reported for the first time from North America.

  15. Regulation of nuclear factor of activated T cells (NFAT) and downstream myogenic proteins during dehydration in the African clawed frog.

    PubMed

    Zhang, Yichi; English, Simon G; Storey, Kenneth B

    2018-06-19

    Xenopus laevis, otherwise known as the African clawed frog, undergoes natural dehydration of up to 30% of its total body water during the dry season in sub-Saharan Africa. To survive under these conditions, a variety of physiological and biochemical changes take place in X. laevis. We were interested in understanding the role that the calcineurin-NFAT pathway plays during dehydration stress response in the skeletal muscles of X. laevis. Immunoblotting was performed to characterize the protein levels of NFATc1-4, calcium signalling proteins, in addition to myogenic proteins (MyoD, MyoG, myomaker). In addition, DNA-protein interaction ELISAs were used to assess the binding of NFATs to their consensus binding sequence, and to identify the effect of urea on NFAT-binding. Our results showed that NFATc1 and c4 protein levels decreased during dehydration, and there were no changes in NFATc2, c3, and calcium signalling proteins. However, MyoG and myomaker both showed increases in protein levels during dehydration, thus indicating that the late myogenic program involving myoblast differentiation, but not satellite cell activation and myoblast proliferation, could be involved in preserving the skeletal muscle of X. laevis during dehydration. In addition, we observed that urea seems to reduce NFATc3-binding to DNA during control, but not during dehydration, possibly indicating that NFATc3 is protected from the denaturing effects of urea as it accumulates during dehydration. These findings expand upon our knowledge of adaptive responses to dehydration, and they identify specific protein targets that could be used to protect the skeletal muscle from damage during stress.

  16. Evolution of vocal patterns: tuning hindbrain circuits during species divergence.

    PubMed

    Barkan, Charlotte L; Zornik, Erik; Kelley, Darcy B

    2017-03-01

    The neural circuits underlying divergent courtship behaviors of closely related species provide a framework for insight into the evolution of motor patterns. In frogs, male advertisement calls serve as unique species identifiers and females prefer conspecific to heterospecific calls. Advertisement calls of three relatively recently (∼8.5 Mya) diverged species - Xenopus laevis , X. petersii and X. victorianus - include rapid trains of sound pulses (fast trills). We show that while fast trills are similar in pulse rate (∼60 pulses s -1 ) across the three species, they differ in call duration and period (time from the onset of one call to the onset of the following call). Previous studies of call production in X. laevis used an isolated brain preparation in which the laryngeal nerve produces compound action potentials that correspond to the advertisement call pattern (fictive calling). Here, we show that serotonin evokes fictive calling in X. petersii and X. victorianus as it does in X. laevis As in X. laevis , fictive fast trill in X. petersii and X. victorianus is accompanied by an N -methyl-d-aspartate receptor-dependent local field potential wave in a rostral hindbrain nucleus, DTAM. Across the three species, wave duration and period are strongly correlated with species-specific fast trill duration and period, respectively. When DTAM is isolated from the more rostral forebrain and midbrain and/or more caudal laryngeal motor nucleus, the wave persists at species-typical durations and periods. Thus, intrinsic differences within DTAM could be responsible for the evolutionary divergence of call patterns across these related species. © 2017. Published by The Company of Biologists Ltd.

  17. Properties of a U1 RNA enhancer-like sequence.

    PubMed Central

    Ciliberto, G; Palla, F; Tebb, G; Mattaj, I W; Philipson, L

    1987-01-01

    The properties of a X.laevis U1B snRNA gene enhancer have been studied by microinjection in Xenopus oocytes. The enhancer-like sequence, defined as a short DNA stretch that is able to activate transcription in an orientation independent manner, is interchangeable between different U snRNA genes. The enhancer sequence alone does not, however, efficiently activate transcription from an SV40 pol II promoter but regains its activity when combined with the U-gene specific proximal sequence element. DNase I protection experiments show that the X.laevis U1B enhancer can interact specifically with a nuclear factor present in mammalian cells. Images PMID:3031597

  18. Label-free Quantification of Proteins in Single Embryonic Cells with Neural Fate in the Cleavage-Stage Frog (Xenopus laevis) Embryo using Capillary Electrophoresis Electrospray Ionization High-Resolution Mass Spectrometry (CE-ESI-HRMS).

    PubMed

    Lombard-Banek, Camille; Reddy, Sushma; Moody, Sally A; Nemes, Peter

    2016-08-01

    Quantification of protein expression in single cells promises to advance a systems-level understanding of normal development. Using a bottom-up proteomic workflow and multiplexing quantification by tandem mass tags, we recently demonstrated relative quantification between single embryonic cells (blastomeres) in the frog (Xenopus laevis) embryo. In this study, we minimize derivatization steps to enhance analytical sensitivity and use label-free quantification (LFQ) for single Xenopus cells. The technology builds on a custom-designed capillary electrophoresis microflow-electrospray ionization high-resolution mass spectrometry platform and LFQ by MaxLFQ (MaxQuant). By judiciously tailoring performance to peptide separation, ionization, and data-dependent acquisition, we demonstrate an ∼75-amol (∼11 nm) lower limit of detection and quantification for proteins in complex cell digests. The platform enabled the identification of 438 nonredundant protein groups by measuring 16 ng of protein digest, or <0.2% of the total protein contained in a blastomere in the 16-cell embryo. LFQ intensity was validated as a quantitative proxy for protein abundance. Correlation analysis was performed to compare protein quantities between the embryo and n = 3 different single D11 blastomeres, which are fated to develop into the nervous system. A total of 335 nonredundant protein groups were quantified in union between the single D11 cells spanning a 4 log-order concentration range. LFQ and correlation analysis detected expected proteomic differences between the whole embryo and blastomeres, and also found translational differences between individual D11 cells. LFQ on single cells raises exciting possibilities to study gene expression in other cells and models to help better understand cell processes on a systems biology level. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  19. Thermal ecological physiology of native and invasive frog species: do invaders perform better?

    PubMed

    Cortes, Pablo A; Puschel, Hans; Acuña, Paz; Bartheld, José L; Bozinovic, Francisco

    2016-01-01

    Biological invasions are recognized as an important biotic component of global change that threatens the composition, structure and functioning of ecosystems, resulting in loss of biodiversity and displacement of native species. Although ecological characteristics facilitating the establishment and spread of non-native species are widely recognized, little is known about organismal attributes underlying invasion success. In this study, we tested the effect of thermal acclimation on thermal tolerance and locomotor performance in the invasive Xenopus laevis and the Chilean native Calyptocephalella gayi . In particular, the maximal righting performance (μ MAX ), optimal temperature ( T O ), lower (CT min ) and upper critical thermal limits (CT max ), thermal breadth ( T br ) and the area under the performance curve (AUC) were studied after 6 weeks acclimation to 10 and 20°C. We observed higher values of μ max and AUC in X. laevis in comparison to C. gayi . On the contrary, the invasive species showed lower values of CT min in comparison to the native one. In contrast, CT max , T O and T br showed no inter-specific differences. Moreover, we found that both species have the ability to acclimate their locomotor performance and lower thermal tolerance limit at low temperatures. Our results demonstrate that X. laevis is a better performer than C. gayi . Although there were differences in CT min , the invasive and native frogs did not differ in their thermal tolerance. Interestingly, in both species the lower and upper critical thermal limits are beyond the minimal and maximal temperatures encountered in nature during the coldest and hottest month, respectively. Overall, our findings suggest that both X. laevis and C. gayi would be resilient to climate warming expectations in Chile.

  20. Ecotoxicoparasitology of the gastrointestinal tracts of pinnipeds: the effect of parasites on the potential bioavailability of total mercury (THg).

    PubMed

    McGrew, Ashley K; O'Hara, Todd M; Stricker, Craig A; Salman, Mo D; Van Bonn, William; Gulland, Frances M D; Whiting, Alex; Ballweber, Lora R

    2018-08-01

    Acanthocephalans, cestodes, and some species of nematodes acquire nutrients from the lumen contents in the gastrointestinal (GI) tract of their definitive host. These parasites are exposed to toxicants, such as mercury (Hg), through passive or active feeding mechanisms; therefore, the focus of this study was to determine if there is an effect of parasites on the dietary availability of total mercury (THg) within piscivorous pinniped hosts. THg concentrations ([THg]) in selected host tissues, parasites, and GI lumen contents from 22 California sea lions (Zalophus californianus), 15 ringed seals (Phoca hispida), and 4 spotted seals (Phoca largha) were determined. Among all pinnipeds, [THg] in acanthocephalans of the large intestine were significantly higher than concentrations in other samples (host lumen contents, other parasites and host intestinal wall), irrespective of location within the host GI tract. δ 15 N values of parasites depended both on parasite group and location within the GI tract. δ 15 N values were consistently higher in parasites inhabiting the large intestine, compared to elsewhere in the GI tract, for both sea lions and seals. δ 13 C values in parasites did not differ significantly from host GI tissues. Based on both [THg] and stable isotope values, parasites are likely affecting the Hg bioavailability within the GI lumen contents and host tissues, and toxicant-parasite interactions appear to depend on both parasitic taxon as well as their location within the host intestine. Copyright © 2018. Published by Elsevier B.V.

  1. Parasite Manipulation of Host Behaviour: Acanthocephalans and Shrimps in the Laboratory.

    ERIC Educational Resources Information Center

    Brown, A. F.; Thompson, D. B. A.

    1986-01-01

    Describes three experiments for undergraduates which illustrate associations of parasites with their host. Includes a table of parasite-induced alterations of selected host species. Instructional suggestions are also provided. (ML)

  2. Functional Characterization of the 1,5-Benzodiazepine Clobazam and Its Major Active Metabolite N-Desmethylclobazam at Human GABAA Receptors Expressed in Xenopus laevis Oocytes

    PubMed Central

    Hammer, Harriet; Ebert, Bjarke; Jensen, Henrik Sindal; Jensen, Anders A.

    2015-01-01

    The 1,5-benzodiazepine clobazam is indicated for the adjunctive treatment of seizures associated with Lennox-Gastaut syndrome in patients 2 years of age or older in the United States, and for treatment of anxiety and various forms of epilepsy elsewhere. Clobazam has been reported to exhibit different in vivo adverse effects and addiction liability profile than the classic 1,4-benzodiazepines. In this study, it was investigated whether the in vitro pharmacological properties of clobazam and its major active metabolite N-desmethylclobazam could explain some of these clinical differences. The functional properties of the two 1,5-benzodiazepines were characterized at the human γ-aminobutyric acid type A receptor (GABAAR) subtypes α1β2γ2S, α2β2γ2S, α3β2γ2S, α5β2γ2S and α6β2δ expressed in Xenopus laevis oocytes by use of two-electrode voltage-clamp electrophysiology and compared to those exhibited by the 1,4-benzodiazepine clonazepam. All three compounds potentiated GABA EC20-evoked responses through the α1,2,3,5β2γ2S GABAARs in a reversible and concentration-dependent manner, with each displaying similar EC50 values at the four subtypes. Furthermore, the degrees of potentiation of the GABA EC20 currents through the four receptors mediated by saturating modulator concentrations did not differ substantially for any of the three benzodiazepines. The three compounds were substantially less potent (200-3900 fold) as positive allosteric modulators at the α6β2δ GABAAR than at the α1,2,3,5β2γ2S receptors. Interestingly, however, clobazam and especially N-desmethylclobazam were highly efficacious potentiators of α6β2δ receptor signaling. Although this activity component is unlikely to contribute to the in vivo effects of clobazam/N-desmethylclobazam, the 1,5-benzodiazepine could constitute an interesting lead for novel modulators targeting this low-affinity binding site in GABAARs. In conclusion, the non-selective modulation exerted by clobazam, N

  3. Metazoan Parasites of Antarctic Fishes.

    PubMed

    Oğuz, Mehmet Cemal; Tepe, Yahya; Belk, Mark C; Heckmann, Richard A; Aslan, Burçak; Gürgen, Meryem; Bray, Rodney A; Akgül, Ülker

    2015-06-01

    To date, there have been nearly 100 papers published on metazoan parasites of Antarctic fishes, but there has not yet been any compilation of a species list of fish parasites for this large geographic area. Herein, we provide a list of all documented occurrences of monogenean, cestode, digenean, acanthocephalan, nematode, and hirudinean parasites of Antarctic fishes. The list includes nearly 250 parasite species found in 142 species of host fishes. It is likely that there are more species of fish parasites, which are yet to be documented from Antarctic waters.

  4. Economic design in a long-distance migrating molluscivore: how fast-fuelling red knots in Bohai Bay, China, get away with small gizzards.

    PubMed

    Yang, Hong-Yan; Chen, Bing; Ma, Zhi-Jun; Hua, Ning; van Gils, Jan A; Zhang, Zheng-Wang; Piersma, Theunis

    2013-10-01

    We carried out an observational and experimental study to decipher how resource characteristics, in interaction with the predator's phenotype, constrain a fitness-determining performance measure, i.e. refuelling in a migrant bird. Two subspecies of red knot (Calidris canutus rogersi and C. c. piersmai) use northern Bohai Bay, Yellow Sea, China, for the final prebreeding stopover, during their 10,000-15,000 km long migrations between wintering and breeding areas. Here, they feed on small bivalves, especially 2-7 mm long Potamocorbula laevis. With an average stay of 29 days, and the need to store 80 g of fat for the onward flights to high-Arctic breeding grounds, red knots need to refuel fast. Using existing knowledge, we expected them to achieve this on the basis of (1) prey with high flesh to shell mass ratios, (2) large gizzards to crush the ingested molluscs, or (3) a combination of the two. Rejecting all three predictions, we found that red knots staging in Bohai Bay had the smallest gizzards on record (4.9 ± 0.8 g, mean ± s.e.m., N = 27), and also found that prey quality of P. laevis is much lower than predicted for the measured gizzard size (i.e. 1.3 rather than the predicted 4.5 kJ g(-1) dry shell mass, DM(shell)). The estimated handling time of P. laevis (0.2 s) is much shorter than the observed time between two prey ingestions (0.7 s), indicating that prey handling time is no constraint. Based on field observations of dropping rates and on indoor digestion trails, the shell processing rate was estimated at 3.9 mg DM(shell) s(-1), i.e. three times higher the rate previously predicted for red knots eating as fast as they can with the measured gizzard size. This is explained by the small and easily crushed P. laevis enabling high processing rates. As P. laevis also occurred in high densities, the metabolizable energy intake rate of red knots with small gizzards at 5 J s(-1) was as high as at northward staging sites elsewhere in the world. Currently

  5. The influence of clinostat rotation on the fertilized amphibian egg.

    NASA Technical Reports Server (NTRS)

    Tremor, J. W.; Souza, K. A.

    1972-01-01

    Study in which unrestrained, fertilized eggs of Rana pipiens and Xenopus laevis were rotated in a plane parallel to the normal gravity vector. In R. pipiens rotation at 1/4 rpm for five days at 18 C produced a significantly increased number of commonly occurring abnormalities. Rotation at 1/15, 1/8, 1, 2, 5 and 10 rpm did not significantly affect normal development. X. laevis eggs reacted similarly. R. pipiens eggs were most sensitive to rotation at 1/4 rpm when exposure was initiated before first cleavage. Mixing of intracellular constituents apparently occurred only at 1/4 rpm in R. pipiens (of the clinostat speeds studied), and may have been the cause of the increased abnormality observed at this rate.

  6. Acanthocephala Larvae parasitizing Ameiva ameiva ameiva (Linnaeus, 1758) (Squamata: Teiidae).

    PubMed

    Macedo, Lilian Cristina; Melo, Francisco Tiago de Vasconcelos; Ávila-Pires, Teresa Cristina Sauer; Giese, Elane Guerreiro; dos Santos, Jeannie Nascimento

    2016-01-01

    Knowledge concerning the taxonomy and biology of species of Acanthocephala, helminth parasites of the helminth species of the phylum Acanthocephala, parasites of lizards in Brazilian Amazonia, is still insufficient, but reports of Acanthocephala in reptiles are becoming increasingly common in the literature. Cystacanth-stage Acanthocephalan larvae have been found in the visceral peritoneum during necropsy of Ameiva ameiva ameivalizards from the "Osvaldo Rodrigues da Cunha" Herpetology Collection of the Emílio Goeldi Museum, Belém, Pará, Brazil. The aim of this study was to present the morphological study of the Acanthocephala larvae found in A. ameiva ameiva lizard.

  7. [Parasite fauna of the European river lamprey Lampetra fluviatilis (L.) from Lake Onega].

    PubMed

    Evseeva, N V

    2007-01-01

    Data on the parasite fauna of the adult European river lamprey Lampetra fluviatilis (L.) from Lake Onega are reported. Ten parasite species are found, including trematodes Diplostomum petromyzifluviatilis and D. spathaceum (metacercariae), cestode Proteocephalus longicollis, nematodes Cucullanus truttae and Raphidascaris acus, acanthocephalan Echinorhynchus salmonis, ectoparasitic infusoria Chilodonella hexastica, Trichodina tenuidens, and Trichodinella epizootica, and fungus Saprolegnia parasitica. Three species are found to be dominate, namely D. petromyzifluviatilis, Cucullanus truttae, and P. longicollis. Comparative analysis of the parasite faunas of the lampreys from other basins is carried out. Some similarity in the parasite faunas of lampreys and salmonids is discovered.

  8. Parasitic helminths in the digestive tract of six species of owls (Strigiformes) in Spain.

    PubMed

    Ferrer, David; Molina, Rafael; Castellà, Joaquim; Kinsella, John M

    2004-03-01

    A survey was carried out in Catalonia, Spain to determine the prevalence of parasitic helminths infesting the digestive tract of nocturnal raptors (Strigiformes). One hundred birds belonging to six owl species were examined. The number of birds examined ranged between three and 30. The overall level of infestation was 65% and the most frequent helminths present were nematodes, followed by trematodes, cestodes and acanthocephalans. Strigiformes showing the highest levels of infestation were little owls (Athene noctua), 86.7% of which were infested. Helminths with the highest prevalence were Synhimantus spp., intestinal capillarids, Brachylaima spp., cestodes and Centrorhynchus aluconis.

  9. Suitability of thirteen different host species for elm leaf beetle, Xanthogaleruca luteola (Coleoptera: Chrysomelidae)

    Treesearch

    Richard W. Hall; Alden M. Townsend; Jack H. Barger

    1987-01-01

    Thirteen different host species for elm leaf beetle, Xanthogaleruca luteola (Muller), were assayed to determine their relative suitability. Species examined were Ulmus parvifolia Jacq., U. thomasii Sarg., U. laevis Pall., U. wilsoniana Schneid., U....

  10. Impact of tire debris on in vitro and in vivo systems

    PubMed Central

    Gualtieri, Maurizio; Andrioletti, Manuela; Mantecca, Paride; Vismara, Claudio; Camatini, Marina

    2005-01-01

    Background It is estimated that over 80% of respirable particulate matter (PM10) in cities comes from road transport and that tire and brake wear are responsible for the 3–7% emission of it. Data on the indicators of environmental impact of tire debris (TD), originated from the tire abrasion on roads, are extremely scarce, even though TD contains chemicals (zinc and organic compounds) which can be released in the environment. Methods TD particle morphology was analysed with SEM, TEM and FIB instruments. TD eluates and TD organic extracts were tested at dilution series on human cell lines and Xenopus laevis embryos. 50 and 100 g/L TD were used for the eluates obtained after 24 h at pH 3 and the quantity of zinc present was measured with a ICP-AES. Eluates diluted to 1%, 10%, 50% in culture media and undiluted were used on X. laevis embryos in the FETAX test. HepG2 cells were exposed for 24 h to 0.05 – 50 μg/ml of zinc salt while A549 cells were exposed for 24, 48 and 72 h to 10, 50, 60, or 75 μg/ml of TD extract. X. laevis embryos were exposed to 50, 80, 100, or 120 μg/ml TD extract. Results The solution of undiluted 50 g/L TD produced 80.2% mortality (p < 0.01) in X. laevis embryos and this toxic effect was three times greater than that produced by 100 g/L TD. Zn accumulation in HepG2 cells was evident after 4 h exposure. A549 cells exposed to TD organic extract for 72 h presented a modified morphology, a decrease in cell proliferation and an increase in DNA damage as shown by comet assay. The dose 80 μg/ml of TD extract produced 14.6% mortality in X. laevis embryos and 15.9% mortality at 120 μg/ml. Treatment with 80, 100, or 120 μg/ml TD organic extract increased from 14.8% to 37.8% malformed larvae percentages compared to 5.6% in the control. Conclusion Since the amount of Zn leached from TD is related to pH, aggregation of particles and elution process, the quantity of TD present in the environment has to be taken into account. Moreover the atmospheric

  11. Variability of Rheotaxis Behaviors in Larval Bullfrogs Highlights Species Diversity in Lateral Line Function

    PubMed Central

    Brown, Erika E. A.

    2016-01-01

    The morphology and distribution of lateral line neuromasts vary between ecomorphological types of anuran tadpoles, but little is known about how this structural variability contributes to differences in lateral-line mediated behaviors. Previous research identified distinct differences in one such behavior, positive rheotaxis towards the source of a flow, in two tadpole species, the African clawed frog (Xenopus laevis; type 1) and the American bullfrog (Rana catesbeiana; type 4). Because these two species had been tested under different flow conditions, we re-evaluated these findings by quantifying flow-sensing behaviors of bullfrog tadpoles in the same flow field in which X. laevis tadpoles had been tested previously. Early larval bullfrog tadpoles were exposed to flow in the dark, in the presence of a discrete light cue, and after treatment with the ototoxin gentamicin. In response to flow, tadpoles moved downstream, closer to a side wall, and higher in the water column, but they did not station-hold. Tadpoles exhibited positive rheotaxis, but with long latencies, low to moderate accuracy, and considerable individual variability. This is in contrast to the robust, stereotyped station-holding and accurate rheotaxis of X. laevis tadpoles. The presence of a discrete visual cue and gentamicin treatment altered spatial positioning and disrupted rheotaxis in both tadpole species. Species differences in lateral-line mediated behaviors may reflect differences in neuromast number and distribution, life history, or perceptual salience of other environmental cues. PMID:27870909

  12. The extraordinary biology and development of marsupial frogs (Hemiphractidae) in comparison with fish, mammals, birds, amphibians and other animals.

    PubMed

    Del Pino, Eugenia M

    2018-01-03

    The study of oogenesis and early development of frogs belonging to the family Hemiphractidae provide important comparison to the aquatic development of other frogs, such as Xenopus laevis, because reproduction on land characterizes the Hemiphractidae. In this review, the multinucleated oogenesis of the marsupial frog Flectonotus pygmaeus (Hemiphractidae) is analyzed and interpreted. In addition, the adaptations associated with the incubation of embryos in the pouch of the female marsupial frog Gastrotheca riobambae (Hemiphractidae) and the embryonic development of this frog are summarized. Moreover, G. riobambae gastrulation is compared with the gastrulation modes of Engystomops randi and Engystomops coloradorum (Leptodactylidae); Ceratophrys stolzmanni (Ceratophryidae); Hyalinobatrachium fleischmanni and Espadarana callistomma (Centrolenidae); Ameerega bilinguis, Dendrobates auratus, Epipedobates anthonyi, Epipedobates machalilla, Epipedobates tricolor, and Hyloxalus vertebralis (Dendrobatidae); Eleutherodactylus coqui (Terrarana: Eleutherodactylidae), and X. laevis (Pipidae). The comparison indicated two modes of frog gastrulation. In X. laevis and in frogs with aquatic reproduction, convergent extension begins during gastrulation. In contrast, convergent extension occurs in the post-gastrula of frogs with terrestrial reproduction. These two modes of gastrulation resemble the transitions toward meroblastic cleavage found in ray-finned fishes (Actinopterygii). In spite of this difference, the genes that guide early development seem to be highly conserved in frogs. I conclude that the shift of convergent extension to the post-gastrula accompanied the diversification of frog egg size and terrestrial reproductive modes. Copyright © 2018 Elsevier B.V. All rights reserved.

  13. Comparison of amphibian and mammalian thyroperoxidase ...

    EPA Pesticide Factsheets

    Thyroperoxidase (TPO) catalyzes the production of thyroid hormones in the vertebrate thyroid gland by oxidizing iodide (I- ) to produce iodinated tyrosines on thyroglobulin, and further coupling of specific mono- or di-iodinated tyrosines to generate the triiodo- and tetra-iodothyronine, precursors to thyroid hormone. This enzyme is a target for thyroid disrupting chemicals. TPO-inhibition by xenobiotics is a molecular initiating event that is known to perturb the thyroid axis by preventing synthesis of thyroid hormone. Previous work on TPO-inhibition has been focused on mammalian TPO; specifically, the rat and pig. A primary objective of this experiment was to directly measure TPO activity in a non-mammalian system, in this case a thyroid gland homogenate from Xenopus laevis; as well as compare chemical inhibition from past mammalian studies to the amphibian data generated. Thyroid glands obtained from X. laevis tadpoles at NF stages 58-60, were pooled and homogenized by sonication in phosphate buffer. This homogenate was then used to test 24 chemicals for inhibition of TPO as measured by conversion of Amplex UltraRed (AUR) substrate to its fluorescent product. The test chemicals were selected based upon previous results from rat in vitro TPO assays, and X. laevis in vitro and in vivo studies for thyroid disrupting endpoints, and included both positive and negative chemicals in these assays. An initial screening of the chemicals was done at a single high con

  14. Environmental parasitology: Parasites as accumulation bioindicators in the marine environment

    NASA Astrophysics Data System (ADS)

    Nachev, Milen; Sures, Bernd

    2016-07-01

    Parasites can be used as effective monitoring tools in environmental impact studies as they are able to accumulate certain pollutants (e.g. metals) at levels much higher than those of their ambient environment and of free-living sentinels. Thus, they provide valuable information not only about the chemical conditions of their and their hosts' environment but also deliver insights into the biological availability of allochthonous substances. While a large number of different freshwater parasites (mainly acanthocephalans and cestodes) were investigated in terms of pollutant bioaccumulation, studies based on marine host-parasites systems remain scarce. However, available data show that different marine parasite taxa such as nematodes, cestodes and acanthocephalans exhibit also an excellent metal accumulation capacity. The biological availability of metals and their uptake routes in marine biota and parasites differ from those of freshwater organisms. We assume that a large part of metals and other pollutants are also taken up via the digestive system of the host. Therefore, in addition to environmental conditions the physiology of the host also plays an important role for the accumulation process. Additionally, we highlight some advantages in using parasites as accumulation indicators in marine ecosystems. As parasites occur ubiquitously in marine food webs, the monitoring of metals in their tissues can deliver information about the spatial and trophic distribution of pollutants. Accordingly, parasites as indicators offer an ecological assessment on a broader scale, in contrast to established free-living marine indicators, which are mostly benthic invertebrates and therefore limited in habitat distribution. Globally distributed parasite taxa, which are highly abundant in a large number of host species, are suggested as worldwide applicable sentinels.

  15. Neoandracantha peruensis n. gen. n. sp. (Acanthocephala, Polymorphidae) described from cystacanths infecting the ghost crab Ocypode gaudichaudii on the Peruvian coast

    PubMed Central

    2017-01-01

    The cystacanths of Neoandracantha peruensis n. gen. n. sp. are described from the ghost crab Ocypode gaudichaudii collected from the Pacific coast of Peru. While it is uncommon to describe acanthocephalan taxa from immature stages, the presence of clear-cut distinguishing features separating the present material from its nearest congeneric taxa, and the absence of adults, justifies the erection N. peruensis. The new genus is distinguished by having three separate fields of trunk spines. Specimens of N. peruensis have a slender trunk with two anterior swellings, 3 separate fields of spines on the foretrunk swelling, and no genital spines on the hindtrunk. The proboscis of the new species is heavily armored with 21–22 longitudinal rows of 22 hooks each. Hook no. 14 is more robust ventrally than dorsally. Cystacanths of N. peruensis also have a long tubular hindtrunk and the males have diagonal testes in the midtrunk swelling. Specimens of the closely related Andracantha Schmidt, 1975 have anteriorly enlarged pear-shaped Corynosoma-like trunks, only two fields of anterior trunk spines with occasional genital spines, and bilateral or tandem testes. Proboscides of species of Andracantha have considerably fewer hooks that gradually decrease in size posteriorly. The taxonomic component of this work is amplified by metal analysis of hooks and spines that shows a marked amount of magnesium (Mg) in hooks but not in spines. The highest level of sulfur (S) was found in the outer layer of hooks and anterior spines. The metal footprint of hooks and spines varies in different species of acanthocephalans and has an interspecific diagnostic value. PMID:29072573

  16. DEVELOPMENT OF AN AMPHIBIAN METAMORPHOSIS MODEL FOR DETECTING THYROID AXIS DISRUPTION

    EPA Science Inventory

    Metamorphosis in Xenopus laevis represents an elaborate process of post-embryonic development which is thyroid hormone (TH) dependent. The development of a functional thyroid axis and the responses of tissues to different TH concentrations are well defined in this species, provid...

  17. ADVANCED PROTEOMICS AND BIOINFORMATICS TOOLS IN TOXICOLOGY RESEARCH: OVERCOMING CHALLENGES TO PROVIDE SIGNIFICANT RESULTS

    EPA Science Inventory

    This presentation specifically addresses the advantages and limitations of state of the art gel, protein arrays and peptide-based labeling proteomic approaches to assess the effects of a suite of model T4 inhibitors on the thyroid axis of Xenopus laevis.

  18. Saduria entomon infected with Hysterothylacium aduncum found in situ in the stomach of cod (Gadus morhua) from the Baltic Sea.

    PubMed

    Pawlak, J; Nadolna-Ałtyn, K; Szostakowska, B; Pachur, M; Podolska, M

    2017-10-12

    The parasite fauna of cod (Gadus morhus) is well described, but the life cycles of Baltic cod parasites are known only in general terms. Invertebrates commonly found in the stomach of cod are recognized as intermediate hosts in the life cycles of nematodes or acanthocephalans. The aim of this study was to determine the source of infection of Baltic cod with parasites found in situ in invertebrates present in the cod stomach. Our results indicate that Saduria entomon is both a source of infection of Baltic cod with parasites and an intermediate host in the life cycle of Hysterothylacium aduncum in the Baltic Sea.

  19. Kinematics and hydrodynamics analysis of swimming anurans reveals striking inter-specific differences in the mechanism for producing thrust.

    PubMed

    Richards, Christopher T

    2010-02-15

    This study aimed to compare the swimming kinematics and hydrodynamics within and among aquatic and semi-aquatic/terrestrial frogs. High-speed video was used to obtain kinematics of the leg joints and feet as animals swam freely across their natural range of speeds. Blade element analysis was then used to model the hydrodynamic thrust as a function of foot kinematics. Two purely aquatic frogs, Xenopus laevis and Hymenochirus boettgeri, were compared with two semi-aquatic/terrestrial frogs, Rana pipiens and Bufo americanus. The four species performed similarly. Among swimming strokes, peak stroke velocity ranged from 3.3+/-1.1 to 20.9+/-2.5, from 6.8+/-2.1 to 28.6+/-3.7 and from 4.9+/-0.5 to 20.9+/-4.1 body lengths per second (BL s(-1)) in X. laevis, H. boettgeri and R. pipiens, respectively (means +/- s.d.; N=4 frogs for each). B. americanus swam much more slowly at 3.1+/-0.3 to 7.0+/-2.0 BL s(-1) (N=3 frogs). Time-varying joint kinematics patterns were superficially similar among species. Because foot kinematics result from the cumulative motion of joints proximal to the feet, small differences in time-varying joint kinematics among species resulted in species-specific foot kinematics (therefore hydrodynamics) patterns. To obtain a simple measure of the hydrodynamically useful motion of the foot, this study uses 'effective foot velocity' (EFV): a measure of the component of foot velocity along the axis of swimming. Resolving EFV into translational and rotational components allows predictions of species-specific propulsion strategies. Additionally, a novel kinematic analysis is presented here that enables the partitioning of translational and rotational foot velocity into velocity components contributed by extension at each individual limb joint. Data from the kinematics analysis show that R. pipiens and B. americanus translated their feet faster than their body moved forward, resulting in positive net translational EFV. Conversely, translational EFV was slower than

  20. The kinematic determinants of anuran swimming performance: an inverse and forward dynamics approach.

    PubMed

    Richards, Christopher T

    2008-10-01

    The aims of this study were to explore the hydrodynamic mechanism of Xenopus laevis swimming and to describe how hind limb kinematics shift to control swimming performance. Kinematics of the joints, feet and body were obtained from high speed video of X. laevis frogs (N=4) during swimming over a range of speeds. A blade element approach was used to estimate thrust produced by both translational and rotational components of foot velocity. Peak thrust from the feet ranged from 0.09 to 0.69 N across speeds ranging from 0.28 to 1.2 m s(-1). Among 23 swimming strokes, net thrust impulse from rotational foot motion was significantly higher than net translational thrust impulse, ranging from 6.1 to 29.3 N ms, compared with a range of -7.0 to 4.1 N ms from foot translation. Additionally, X. laevis kinematics were used as a basis for a forward dynamic anuran swimming model. Input joint kinematics were modulated to independently vary the magnitudes of foot translational and rotational velocity. Simulations predicted that maximum swimming velocity (among all of the kinematics patterns tested) requires that maximal translational and maximal rotational foot velocity act in phase. However, consistent with experimental kinematics, translational and rotational motion contributed unequally to total thrust. The simulation powered purely by foot translation reached a lower peak stroke velocity than the pure rotational case (0.38 vs 0.54 m s(-1)). In all simulations, thrust from the foot was positive for the first half of the power stroke, but negative for the second half. Pure translational foot motion caused greater negative thrust (70% of peak positive thrust) compared with pure rotational simulation (35% peak positive thrust) suggesting that translational motion is propulsive only in the early stages of joint extension. Later in the power stroke, thrust produced by foot rotation overcomes negative thrust (due to translation). Hydrodynamic analysis from X. laevis as well as forward

  1. Influences of disturbance and vegetation on abundance of native and exotic detritivores in a southwestern riparian forest

    Treesearch

    D. Max Smith; Jeffrey F. Kelly; Deborah M. Finch

    2006-01-01

    Detritivores play important roles in energy and nutrient flow in riparian ecosystems. Endemic crickets (Gryllus alogus Rehn) and exotic isopods (Armadillidium vulagare Latreille and Porcellio laevi Latreille.) are abundant detritivores in riparian forest floors of central New Mexico. To determine how disturbance...

  2. Two New Disease-Resistant Hibiscus: 'Lufkin Red' and 'Lufkin White'

    USDA-ARS?s Scientific Manuscript database

    The Agricultural Research Service, United States Department of Agriculture, has released Hibiscus laevis ‘Lufkin Red’ and ‘Lufkin White’, two new hardy native ornamental hibiscus. These cultivars are recommended for trial by gardeners and horticulturists as summer flowering perennial landscape pl...

  3. Tumor immunology viewed from alternative animal models—the Xenopus story

    PubMed Central

    Banach, Maureen; Robert, Jacques

    2017-01-01

    a) Purpose of review Nonmammalian comparative animal models are important not only to gain fundamental evolutionary understanding of the complex interactions of tumors with the immune system, but also to better predict the applicability of novel immunotherapeutic approaches to humans. After reviewing recent advances in developing alternative models, we focus on the amphibian Xenopus laevis and its usefulness in deciphering the perplexing roles of MHC class I-like molecules and innate (i)T cells in tumor immunity. b) Recent findings Experiments using MHC-defined inbred and cloned animals, tumor cell lines, effective reagents, sequenced genomes, and adapted gene editing techniques in Xenopus, have revealed that the critical involvement of class I-like molecules and iT cells in tumor immunity has been conserved during evolution. c) Summary Comparative studies with the X. laevis tumor immunity model can contribute to the development of better and more efficient cancer immunotherapies. PMID:28944105

  4. Evolution of Heat Sensors Drove Shifts in Thermosensation between Xenopus Species Adapted to Different Thermal Niches*

    PubMed Central

    Saito, Shigeru; Ohkita, Masashi; Saito, Claire T.; Takahashi, Kenji; Tominaga, Makoto; Ohta, Toshio

    2016-01-01

    Temperature is one of the most critical environmental factors affecting survival, and thus species that inhabit different thermal niches have evolved thermal sensitivities suitable for their respective habitats. During the process of shifting thermal niches, various types of genes expressed in diverse tissues, including those of the peripheral to central nervous systems, are potentially involved in the evolutionary changes in thermosensation. To elucidate the molecular mechanisms behind the evolution of thermosensation, thermal responses were compared between two species of clawed frogs (Xenopus laevis and Xenopus tropicalis) adapted to different thermal environments. X. laevis was much more sensitive to heat stimulation than X. tropicalis at the behavioral and neural levels. The activity and sensitivity of the heat-sensing TRPA1 channel were higher in X. laevis compared with those of X. tropicalis. The thermal responses of another heat-sensing channel, TRPV1, also differed between the two Xenopus species. The species differences in Xenopus TRPV1 heat responses were largely determined by three amino acid substitutions located in the first three ankyrin repeat domains, known to be involved in the regulation of rat TRPV1 activity. In addition, Xenopus TRPV1 exhibited drastic species differences in sensitivity to capsaicin, contained in chili peppers, between the two Xenopus species. Another single amino acid substitution within Xenopus TRPV1 is responsible for this species difference, which likely alters the neural and behavioral responses to capsaicin. These combined subtle amino acid substitutions in peripheral thermal sensors potentially serve as a driving force for the evolution of thermal and chemical sensation. PMID:27022021

  5. Chromosome Banding in Amphibia. XXXII. The Genus Xenopus (Anura, Pipidae).

    PubMed

    Schmid, Michael; Steinlein, Claus

    2015-01-01

    Mitotic chromosomes of 16 species of the frog genus Xenopus were prepared from kidney and lung cell cultures. In the chromosomes of 7 species, high-resolution replication banding patterns could be induced by treating the cultures with 5-bromodeoxyuridine (BrdU) and deoxythymidine (dT) in succession, and in 6 of these species the BrdU/dT-banded chromosomes could be arranged into karyotypes. In the 3 species of the clade with 2n = 20 and 4n = 40 chromosomes (X. tropicalis, X. epitropicalis, X. new tetraploid 1), as well as in the 3 species with 4n = 36 chromosomes (X. laevis, X. borealis, X. muelleri), the BrdU/dT-banded karyotypes show a high degree of homoeology, though differences were detected between these groups. Translocations, inversions, insertions or sex-specific replication bands were not observed. Minor replication asynchronies found between chromosomes probably involve heterochromatic regions. BrdU/dT replication banding of Xenopus chromosomes provides the landmarks necessary for the exact physical mapping of genes and repetitive sequences. FISH with an X. laevis 5S rDNA probe detected multiple hybridization sites at or near the long-arm telomeric regions in most chromosomes of X. laevis and X. borealis, whereas in X. muelleri, the 5S rDNA sequences are located exclusively at the long-arm telomeres of a single chromosome pair. Staining with the AT base pair-specific fluorochrome quinacrine mustard revealed brightly fluorescing heterochromatic regions in the majority of X. borealis chromosomes which are absent in other Xenopus species.

  6. Notice to nurserymen of the nameing and release for propagation of lufin white an american hardy hibiscus cultivar

    USDA-ARS?s Scientific Manuscript database

    Hibiscus laevis ‘LUFKIN WHITE’, a new hardy native ornamental hibiscus cultivar is recommended for trial by nurserymen and horticulturists as a summer flowering perennial landscape plant nationwide but is particularly adapted to conditions in the South. ‘LUFKIN WHITE’ was selected for its exception...

  7. Mycobacterial infections in striped bass from Delaware Bay

    USGS Publications Warehouse

    Ottinger, C.A.; Brown, J.J.; Densmore, Christine L.; Starliper, C.E.; Blazer, V.S.; Weyers, H.S.; Beauchamp, K.A.; Rhodes, M.W.; Kator, H.; Gauthier, David T.; Vogelbein, W.K.

    2007-01-01

    Eighty striped bass Morone saxatilis were obtained from Delaware Bay using commercial gill nets set adjacent to Woodland Beach (n = 70) and Bowers Beach (n = 10) in December 2003. Fish were examined for gross lesions. Total lengths (TLs) and eviscerated weights were determined to calculate condition factors (K). Portions of spleens were aseptically harvested for bacterial culture, and portions of spleens, kidneys (anterior and posterior), livers, and gonads were obtained for histological examination. The size distribution of the striped bass was relatively homogeneous; the mean TL was about 600 mm for all samples. Mean K exceeded 0.95 in all samples and was not significantly different (P > 0.05) among samples. Significant differences in mycobacterial infection prevalence (P ??? 0.05) were observed among samples; samples obtained at Woodland Beach (WB) on December 10 (53.8%, n = 13) and December 17 (7.1%, n = 42) exhibited the most striking differences in prevalence. Mycobacterial infection intensity ranged from 1 ?? 102 to 1 ?? 107 colony-forming units per gram of spleen. Acanthocephalan infection prevalence and intensity, non-acid-fast bacterial infection prevalence, and fish sex ratio were also significantly different among the samples (P ??? 0.05). Similar to the mycobacterial infections, differences in sex ratio, acanthocephalan infection, and non-acid-fast bacterial infection were observed between the WB samples taken on December 10 and 17. However, no significant associations (P > 0.05) were observed between sex ratio or these infections and mycobacterial infection. The differences in bacterial and parasite infection prevalence and intensity and fish sex ratio in some samples indicate that these fish had a different history and that the epizootiology of mycobacterial infection in striped bass from Delaware Bay may be relatively complex. ?? Copyright by the American Fisheries Society 2007.

  8. Prevalence and molecular characterisation of Acanthocephala in pinnipedia of the North and Baltic Seas.

    PubMed

    Waindok, Patrick; Lehnert, Kristina; Siebert, Ursula; Pawliczka, Iwona; Strube, Christina

    2018-04-01

    Harbour seals ( Phoca vitulina ) and grey seals ( Halichoerus grypus ) are final hosts of acanthocephalans in the German North and Baltic Seas. Parasitic infections in seals can cause pathological changes, which may result in deteriorated health of the host. Common gastrointestinal parasites of harbour and grey seals are acanthocephalans and a number of 275 of 2460 (11.2%) investigated seals from 1996 to 2013 were infected with Corynosoma spp. (Acanthocephala, Polymorphidae). The prevalence showed a wave-like pattern: it increased from 1.2% and 0.4% in 1996 and 1997, respectively, to 23.9% during the second phocine distemper epizootic in 2002 and decreased to 6.2% in 2004. In 2005, prevalence peaked again with 25.0% followed by a decrease to 9.3% in 2009 and an increase to 38.5% in 2012. Statistical analysis revealed that harbour seals originating from the North Sea showed a higher prevalence than grey seals, whereas no significant difference between grey and harbour seals from the Baltic Sea was observed. Furthermore, juvenile pinnipedia from the North Sea were significantly less infected with Corynosoma spp. than seals older than seven month. Molecular species identification as well as phylogenetic relationship analysis among the detected Corynosoma species were achieved by sequencing and comparisons of the ribosomal ITS1-5.8S-ITS2-complex and cytochrome-c-oxidase I gene. Molecular analysis resulted in a newly arranged distribution of Acanthocephala in the North Sea as in contrast to previous studies, C. strumosum could not be confirmed as predominant species. Instead, C. magdaleni and a C. magdaleni isolate (isolate Pv1NS) with an atypical number of longitudinal rows of hooks at the proboscis were detected. Furthermore, morphological and molecular analyses indicate the possible finding of a cryptic species (Candidatus Corynosoma nortmeri sp. nov.).

  9. Mycobacterial infections in striped bass from Delaware Bay.

    PubMed

    Ottinger, C A; Brown, J J; Densmore, C L; Starliper, C E; Blazer, V S; Weyers, H S; Beauchamp, K A; Rhodes, M W; Kator, H; Gauthier, D T; Vogelbein, W K

    2007-06-01

    Eighty striped bass Morone saxatilis were obtained from Delaware Bay using commercial gill nets set adjacent to Woodland Beach (n = 70) and Bowers Beach (n = 10) in December 2003. Fish were examined for gross lesions. Total lengths (TLs) and eviscerated weights were determined to calculate condition factors (K). Portions of spleens were aseptically harvested for bacterial culture, and portions of spleens, kidneys (anterior and posterior), livers, and gonads were obtained for histological examination. The size distribution of the striped bass was relatively homogeneous; the mean TL was about 600 mm for all samples. Mean K exceeded 0.95 in all samples and was not significantly different (P > 0.05) among samples. Significant differences in mycobacterial infection prevalence (P < or = 0.05) were observed among samples; samples obtained at Woodland Beach (WB) on December 10 (53.8%, n = 13) and December 17 (7.1%, n = 42) exhibited the most striking differences in prevalence. Mycobacterial infection intensity ranged from 1 X 10(2) to 1 X 10(7) colony-forming units per gram of spleen. Acanthocephalan infection prevalence and intensity, non-acid-fast bacterial infection prevalence, and fish sex ratio were also significantly different among the samples (P < or = 0.05). Similar to the mycobacterial infections, differences in sex ratio, acanthocephalan infection, and non-acid-fast bacterial infection were observed between the WB samples taken on December 10 and 17. However, no significant associations (P > 0.05) were observed between sex ratio or these infections and mycobacterial infection. The differences in bacterial and parasite infection prevalence and intensity and fish sex ratio in some samples indicate that these fish had a different history and that the epizootiology of mycobacterial infection in striped bass from Delaware Bay may be relatively complex.

  10. Intestinal helminths of golden jackals and red foxes from Tunisia.

    PubMed

    Lahmar, Samia; Boufana, Belgees; Ben Boubaker, Sarra; Landolsi, Faouzi

    2014-08-29

    Forty wild canids including 31 golden jackals (Canis aureus Linné, 1758) and 9 red foxes (Vulpes vulpes Linné, 1758) collected between 2008 and 2011 in the northeast, northwest and center of Tunisia were necropsied and examined for intestinal helminth parasites. All jackals and foxes were found infected with a prevalence rate of 95% for cestodes, 82.5% for nematodes and 7.5% for acanthocephalans. A total of twelve helminth species were recorded in red foxes: cestodes, Dipylidium caninum (55.6%), Diplopylidium noelleri (55.6%), Mesocestoïdes lineatus (55.6%), Mesocestoïdes litteratus (33%), Mesocestoïdes corti (22%); nematodes, Ancylostoma caninum (11%), Uncinaria stenocephala (44%), Spirura rytipleurites (11%), Trichuris vulpis (33%), Pterygodermatites affinis (67%), Oxynema linstowi (33%) and the acanthocephalan Macracanthorhynchus hirudinaceus (22%). The fifteen recovered helminth species in jackals were Echinococcus granulosus (9.7%), D. caninum (16%), D. noelleri (16%), M. lineatus (74%), M. litteratus (23%), M. corti (12.9%), Taenia pisiformis (3.2%), Taenia spp. (19%), Toxocara canis (16%), Toxascaris leonina (6.5%), A. caninum (9.7%), U. stenocephala (68%), P. affinis (6.5%), O. linstowi (3.2%) and Macracanthorhynchus hirudinaceus (3.2%). This is the first report on the presence of P. affinis, D. noelleri and O. linstowi in Tunisia. E. granulosus was found in young jackals, aged less than 4 years old, with a higher abundance in females (8.9 worms). M. lineatus presented the highest mean intensity of 231.86 and 108.8 tapeworms respectively in jackals and foxes. Canids from the northwest region had the highest prevalence (77.5%) and highest intensity (243.7) of helminth species compared to those from the northeast and central areas. U. stenocephala and O. linstowi had the highest mean intensity for nematodes in both jackals and foxes at 14.3 and 88 worms respectively. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Evaluation of architectural and histopathological biomarkers in the intestine of brown trout (Salmo trutta Linnaeus, 1758) challenged with environmental pollution.

    PubMed

    Barišić, Josip; Filipović Marijić, Vlatka; Mijošek, Tatjana; Čož-Rakovac, Rozelindra; Dragun, Zrinka; Krasnići, Nesrete; Ivanković, Dušica; Kružlicová, Dáša; Erk, Marijana

    2018-06-14

    In the present study novel histopathological approach, using fish intestine as a sensitive bioindicator organ of pollution impact in the freshwater ecosystem, was proposed. Histopathological alterations were compared between native brown trout (Salmo trutta Linnaeus, 1758) from the reference (Krka River spring) and pollution impacted location (influence of technological/municipal wastewaters and agricultural runoff near the Town of Knin) of the karst Krka River in Croatia. In brown trout from both locations, severe parasitic infestation with acanthocephalan species Dentitruncus trutae was found, enabling evaluation of acanthocephalan infestation histopathology, which indicated parasite tissue reaction in a form of inflammatory, necrotic and hyperplastic response that extended throughout lamina epithelialis mucosae, lamina propria, and lamina muscularis mucosae. New semi-quantitative histological approach was proposed in order to foresee alterations classified in three reaction patterns: control tissue appearance, moderate (progressive) tissue impairment and severe (regressive and inflammatory) tissue damage. The most frequent progressive alteration was hyperplasia of epithelium on the reference site, whereas the most frequent regressive alterations were atrophy and necrosis seen on the polluted site. Furthermore, histopathological approach was combined with micromorphological and macromorphological assessment as an additional indicator of pollution impact. Among 15 observed intestinal measures, two biomarkers of intestinal tissue damage were indicated as significant, height of supranuclear space (hSN) and number of mucous cells over 100 μm fold distance of intestinal mucosa (nM), which measures were significantly lower in fish from polluted area compared to the reference site. Obtained results indicated that combined histological and morphological approach on fish intestinal tissue might be used as a valuable biological tool for assessing pollution impact on

  12. Phosphorylation at S384 regulates the activity of the TaALMT1 malate transporter that underlies aluminum resistance in wheat

    USDA-ARS?s Scientific Manuscript database

    In this study we examined the role of protein phosphorylation & dephosphorylation in the transport properties of the wheat root malate efflux transporter underlying Al resistance, TaALMT1. Preincubation of Xenopus laevis oocytes expressing TaALMT1 with protein kinase inhibitors (K252a and staurospo...

  13. Notice to nurserymen of the naming and release for propagation of lufkin red an american hardy hibiscus

    USDA-ARS?s Scientific Manuscript database

    Hibiscus laevis ‘Lufkin Red’, a new hardy native ornamental hibiscus is recommended for trial by nurserymen and horticulturists as a summer flowering perennial landscape plant nationwide but is particularly adapted to conditions in the South. ‘Lufkin Red’ was selected for its outstanding red flowe...

  14. Cloning and functional characterization of the Xenopus orthologue of the Treacher Collins syndrome (TCOF1) gene product.

    PubMed

    Gonzales, Bianca; Yang, Hushan; Henning, Dale; Valdez, Benigno C

    2005-10-10

    Treacher Collins syndrome (TCS) is an autosomal dominant disorder of craniofacial development caused by mutations in the TCOF1 gene, which encodes the nucleolar phosphoprotein treacle. We previously reported a function for mammalian treacle in ribosomal DNA gene transcription by its interaction with upstream binding factor. As an initial step in the development of a TCS model for frog the cDNA that encodes the Xenopus laevis treacle was cloned. Although the derived amino acid sequence shows a poor homology with its mammalian orthologues, Xenopus treacle has 11 highly homologous direct repeats near the center of the protein molecule similar to those present in its human, dog and mouse orthologues. Comparison of their amino acid compositions indicates conservation of predominant specific amino acid residues. Antisense-mediated down-regulation of treacle expression in X. laevis oocytes resulted in inhibition of rDNA gene transcription. The results suggest evolutionary conservation of the function of treacle in ribosomal RNA biogenesis in higher eukaryotes.

  15. Seed germination of three Ulmus species from Turkey as influenced by temperature and light.

    PubMed

    Cicek, Emrah; Tilki, Fahrettin

    2007-04-01

    The effect of temperature and light on the germination performance of Ulmus minor, Ulmus glabra and Ulmus laevis were studied in this research. Seeds were germinated under constant temperatures of 20 and 25 degrees C and alternating temperatures of 25/15 and 30/20 degrees C. Within each temperature regime, seeds were subjected daily to the following photoperiods: total darkness and 8 hr photoperiod. Temperature and light affected seed germination percentage (GP) and germination rate expressed as peak value (PV) in Ulmus minor and 25 and 30/20 degrees C under light gave the highest GP (>95%) and PV (>23). The temperatures of 25/15 and 30/20 degrees C gave the highest GP (>89%) in Ulmus glabra and light did not significantly affect GP. But the highest PV in Ulmus glabra was found at these temperatures under light. Germination percentage of Ulmus laevis was not affected by temperature and light, but the alternating temperature of 30/20 degrees C produced the highest germination rate under darkness.

  16. [A comparative analysis of notochord formation in amphibian embryos].

    PubMed

    Novoselov, V V

    1992-01-01

    We studied the origin, structure, and development of the notochord in Pleurodeles waltlii (Urodela) and Xenopus laevis (Anura) embryos. The notochord rudiment is formed in both species at the early gastrula stage as a cluster of polarized chorda-mesoderm cells located along the sagittal plane of the embryo. In Pl. waltlii the notochord rudiment is separated from the gastrocoele roof as a result of contraction of apical cell surfaces. The contraction wave spreads forward and backward along craniocaudal axis, i.e., segregation of the notochord rudiment progresses in two directions simultaneously. Similar process takes place in X. laevis embryos; however, propagation of the contraction wave in the anterior part of the body somewhat differs from that in the posterior part. While the "anterior" contraction wave resembles that in Pl. waltlii embryos, progression of the wave in the posterior part of the body is distinguished by a closer association of the notochord rudiment with ectoderm and the presence of its delamination boundaries with the somite mesoderm.

  17. Membrane currents in the oocyte of the toad Bufo arenarum.

    PubMed

    Kotsias, Basilio A; Damiano, Alicia E; Godoy, Sebastian; Assef, Yanina; Ibarra, Cristina; Cantiello, Horacio F

    2002-03-01

    The amphibian oocyte cell model is widely used for heterologous expression of ionic channels and receptors. Little is known, however, about the physiology of oocyte cell models other than Xenopus laevis. In this study, the two-electrode voltage clamp technique was used to assess the most common electrical patterns of oocytes of the South American toad Bufo arenarum. Basal membrane resistance, resting potential, and ionic currents were determined in this cell model. The oocyte transmembrane resistance was 0.35 M(Omega), and the resting potential in normal saline was about -33 mV with a range between -20 mV and -50 mV. This is, to our knowledge, the first attempt to begin an understanding of the ion transport mechanisms of Bufo arenarum oocytes. This cell model may provide a viable alternative to the expression of ion channels, in particular those endogenously observed in Xenopus laevis oocytes. Copyright 2002 Wiley-Liss, Inc.

  18. Age, composition, and stand structure of old-growth oak sites in the Florida high pine landscape: implications for ecosystem management and restoration

    Treesearch

    Cathryn H. Greenberg; Robert W. Simons

    1999-01-01

    The authors sampled tree age, species composition, and stand structure of four high pine sites composed of old-growth sand post oak (Q. margaretta Ashe), old-growth turkey oak (Quercus laevis Walt.), and young longleaf pine (Pinus palustris Mill.) in north and central peninsular Florida. The oldest turkey oak...

  19. Taxonomy and Distribution of Freshwater Pearl Mussels (Unionoida: Margaritiferidae) of the Russian Far East

    PubMed Central

    Bolotov, Ivan N.; Bespalaya, Yulia V.; Vikhrev, Ilya V.; Aksenova, Olga V.; Aspholm, Paul E.; Gofarov, Mikhail Y.; Klishko, Olga K.; Kolosova, Yulia S.; Kondakov, Alexander V.; Lyubas, Artyom A.; Paltser, Inga S.; Konopleva, Ekaterina S.; Tumpeesuwan, Sakboworn; Bolotov, Nikita I.; Voroshilova, Irina S.

    2015-01-01

    The freshwater pearl mussel family Margaritiferidae includes 13 extant species, which are all listed by IUCN as endangered or vulnerable taxa. In this study, an extensive spatial sampling of Margaritifera spp. across the Russian Far East (Amur Basin, Kamchatka Peninsula, Kurile Archipelago and Sakhalin Island) was conducted for a revision of their taxonomy and distribution ranges. Based on their DNA sequences, shell and soft tissue morphology, three valid species were identified: Margaritifera dahurica (Middendorff, 1850), M. laevis (Haas, 1910) and M. middendorffi (Rosén, 1926). M. dahurica ranges across the Amur basin and some of the nearest river systems. M. laevis is distributed in Japan, Sakhalin Island and the Kurile Archipelago. M. middendorffi was previously considered an endemic species of the Kamchatka. However, it is widespread in the rivers of Kamchatka, Sakhalin Island, the Kurile Islands (across the Bussol Strait, which is the most significant biogeographical boundary within the archipelago), and, likely, in Japan. The Japanese species M. togakushiensis Kondo & Kobayashi, 2005 seems to be conspecific with M. middendorffi because of similar morphological patterns, small shell size (<100 mm long) and overlapped ranges, but it is in need of a separate revision. Phylogenetic analysis reveals that two NW Pacific margaritiferid species, M. laevis and M. middendorffi, formed a monophyletic 18S rDNA clade together with the North American species M. marrianae and M. falcata. The patterns that were found in these Margaritifera spp. are similar to those of freshwater fishes, indicating multiple colonizations of Eastern Asia by different mitochondrial lineages, including an ancient Beringian exchange between freshwater faunas across the Pacific. PMID:26011762

  20. Vitelline envelope of Bufo arenarum: biochemical and biological characterization.

    PubMed

    Barisone, Gustavo A; Hedrick, Jerry L; Cabada, Marcelo O

    2002-04-01

    Vitelline envelopes (VEs) of Bufo arenarum were isolated in order to study their composition and their role in fertilization. VEs are composed of four glycoproteins, with molecular masses of 120, 75, 41, and 38 kDa. To characterize its biological properties, we quantitatively determined sperm-VE binding and the induction of the acrosome reaction. Heterologous binding of B. arenarum sperm to Xenopus laevis VE components was observed with about one-third the efficiency of homologous binding. Equivalent binding of X. laevis sperm to the B. arenarum VE was observed. When B. arenarum sperm were incubated with fluorescein isothiocyanate-labeled VE, the labeled glycoproteins bound to the anterior end of the sperm head, showing a lateral distribution. Induction of the acrosome reaction was evaluated by incubating sperm in hypotonic saline media with VE glycoproteins. VEs induced the acrosome reaction in a time- and concentration-dependent manner. The acrosome reaction was maximal after 10 min. The half-maximal effect was obtained at a glycoprotein concentration of 1 microg/ml. Specificity was determined using fertilization envelope glycoproteins, which failed to induce the acrosome reaction. The B. arenarum VE is biochemically similar to other egg envelopes. It also seems that its biological properties are similar to other species in regard to sperm binding and induction of the acrosome reaction. However, as far as we are aware, this is the first observation of the VE inducing the sperm acrosome reaction in amphibians. The relatively small differences observed in heterologous sperm-VE binding in X. laevis and B. arenarum are inconsistent with the current paradigm that species specificity in fertilization is regulated at the sperm-VE binding step.

  1. Determination of notochord cells of Xenopus laevis.

    PubMed

    Zeng, M B

    1993-12-01

    In amphibians, numerous works of influences of the notochord on neighbouring tissues have been accumulated. However, on the contrary, scarcely any work is known about how the notochord is influenced by its neighbouring tissues and how it is determined. By using the experimental method of explantation and culturing in vitro, how the notochord is determined in the early development and whether the neighbouring tissues exert influences on it have been investigated. The results showed that the determination of notochord is a progressive process and the presumptive notochord of Xenopus appears to be a very good material to study influences of neighbouring tissues on the determination of the notochord.

  2. Regulative development of Xenopus laevis in microgravity

    NASA Technical Reports Server (NTRS)

    Black, S.; Larkin, K.; Jacqmotte, N.; Wassersug, R.; Pronych, S.; Souza, K.

    1996-01-01

    To test whether gravity is required for normal amphibian development, Xenopus leavis females were induced to ovulate aboard the orbiting Space Shuttle. Eggs were fertilized in vitro, and although early embryonic stages showed some abnormalities, the embryos were able to regulate and produce nearly normal larvae. These results demonstrate for the first time that a vertebrate can ovulate in the virtual absence of gravity, and that the eggs can develop to a free-living stage.

  3. Response of amphibian egg non-yolk cytoplasm to gravity orientation

    NASA Technical Reports Server (NTRS)

    Smith, R. C.; Neff, A. W.; Malacinski, G. M.

    1985-01-01

    In order to study amphibian egg cytoplasmic organization and egg symmetrization at the molecular level, a library of seventeen monoclonal antibodies (MoAbs) against Xenopus laevis non-yolk egg proteins was produced. Several of these MoAbs react with non-yolk cytoplasmic antigens which are unevenly distributed in the fertile Xenopus egg.

  4. Introducing Environmental Toxicology in Instructional Labs: The Use of a Modified Amphibian Developmental Toxicity Assay to Support Inquiry-Based Student Projects

    ERIC Educational Resources Information Center

    Sauterer, Roger; Rayburn, James R.

    2012-01-01

    Introducing students to the process of scientific inquiry is a major goal of high school and college labs. Environmental toxins are of great concern and public interest. Modifications of a vertebrate developmental toxicity assay using the frog Xenopus laevis can support student-initiated toxicology experiments that are relevant to humans. Teams of…

  5. Phosphorylation by Cdk1 Increases the Binding of Eg5 to Microtubules In Vitro and in Xenopus Egg Extract Spindles

    PubMed Central

    Cahu, Julie; Olichon, Aurelien; Hentrich, Christian; Schek, Henry; Drinjakovic, Jovana; Zhang, Cunjie; Doherty-Kirby, Amanda; Lajoie, Gilles; Surrey, Thomas

    2008-01-01

    Background Motor proteins from the kinesin-5 subfamily play an essential role in spindle assembly during cell division of most organisms. These motors crosslink and slide microtubules in the spindle. Kinesin-5 motors are phosphorylated at a conserved site by Cyclin-dependent kinase 1 (Cdk1) during mitosis. Xenopus laevis kinesin-5 has also been reported to be phosphorylated by Aurora A in vitro. Methodology/Principal Findings We investigate here the effect of these phosphorylations on kinesin-5 from Xenopus laevis, called Eg5. We find that phosphorylation at threonine 937 in the C-terminal tail of Eg5 by Cdk1 does not affect the velocity of Eg5, but strongly increases its binding to microtubules assembled in buffer. Likewise, this phosphorylation promotes binding of Eg5 to microtubules in Xenopus egg extract spindles. This enhancement of binding elevates the amount of Eg5 in spindles above a critical level required for bipolar spindle formation. We find furthermore that phosphorylation of Xenopus laevis Eg5 by Aurora A at serine 543 in the stalk is not required for spindle formation. Conclusions/Significance These results show that phosphorylation of Eg5 by Cdk1 has a direct effect on the interaction of this motor with microtubules. In egg extract, phosphorylation of Eg5 by Cdk1 ensures that the amount of Eg5 in the spindle is above a level that is required for spindle formation. This enhanced targeting to the spindle appears therefore to be, at least in part, a direct consequence of the enhanced binding of Eg5 to microtubules upon phosphorylation by Cdk1. These findings advance our understanding of the regulation of this essential mitotic motor protein. PMID:19079595

  6. The ontogeny of the olfactory system in ceratophryid frogs (Anura, Ceratophryidae).

    PubMed

    Quinzio, Silvia I; Reiss, John O

    2018-01-01

    The aquatic-to-terrestrial shift in the life cycle of most anurans suggests that the differences between the larval and adult morphology of the nose are required for sensory function in two media with different physical characteristics. However, a better controlled test of specialization to medium is to compare adult stages of terrestrial frogs with those that remain fully aquatic as adults. The Ceratophryidae is a monophyletic group of neotropical frogs whose diversification from a common terrestrial ancestor gave rise to both terrestrial (Ceratophrys, Chacophrys) and aquatic (Lepidobatrachus) adults. So, ceratophryids represent an excellent model to analyze the morphology and possible changes related to a secondary aquatic life. We describe the histomorphology of the nose during the ontogeny of the Ceratophryidae, paying particular attention to the condition in adult stages of the recessus olfactorius (a small area of olfactory epithelium that appears to be used for aquatic olfaction) and the eminentia olfactoria (a raised ridge on the floor of the principal cavity correlated with terrestrial olfaction). The species examined (Ceratophrys cranwelli, Chacophrys pierottii, Lepidobatrachus laevis, and L. llanensis) share a common larval olfactory organ composed by the principal cavity, the vomeronasal organ and the lateral appendix. At postmetamorphic stages, ceratophryids present a common morphology of the nose with the principal, middle, and inferior cavities with characteristics similar to other neobatrachians at the end of metamorphosis. However, in advanced adult stages, Lepidobatrachus laevis presents a recessus olfactorius with a heightened (peramorphic) development and a rudimentary (paedomorphic) eminentia olfactoria. Thus, the adult nose in Lepidobatrachus laevis arises from a common developmental 'terrestrial' pathway up to postmetamorphic stages, when its ontogeny leads to a distinctive morphology related to the evolutionarily derived, secondarily aquatic

  7. Analysis of cutin and suberin biomarker patterns in alluvial sedi-ments

    NASA Astrophysics Data System (ADS)

    Herschbach, Jennifer; Sesterheim, Anna; König, Frauke; Fuchs, Elmar

    2015-04-01

    Cutin and suberin are the primary source of hydrolysable aliphatic lipid polyesters in soil organic matter (SOM). They are known as geochemical biomarkers to estimate the contribution of different plant species and tissues to SOM. Despite their potential as biomarkers, cutin and suberin have received less attention as flood plain sediment biomarkers. The objectives of this study were to investigate the efficiency of cutin and suberin as biomarkers in floodplains. Therefore similarities between the lipid pattern in alluvial sediments and in the actual vegetation were considered. Lipids of plant tissues (roots, twigs, leaves) from different species (reed (e.g. Phalaris arun-diacea), Salix alba, Ulmus laevis and grassland (e.g. Carex spec.)) and of the un-derlying soils and sediments were obtained and investigated at four sites in the nature reserve Knoblauchsaue (Hessen, Germany). The four sampling sites differ not only with respect to their vegetation, but also within their distance to the river Rhine. Cutin and suberin monomers of plants and soils were analysed by alkaline hydrolysis, methylation and acetylation and subsequent gas chromatography-mass spectrometry. Resulting lipid patterns were specific for the appropriate plant species and tissues. However, the traceability of single selected lipids was decreasing alongside the soil profile, with the exception of monomers in softwood floodplain soils. Selected tissue specific lipid ratios showed a higher traceability due to strong attributions of lipid ratios in soils and roots of U. laevis and Carex spec. and in leaves of U. laevis and S. alba. In contrast, there was no accordance between the suberin specific lipid ratios in soils and roots of S. alba and P. arundiacea. The most robust interpretations were afforded when a set of multiple biomarkers (i.e. a combination of free lipid ratios and ratios of hydrolysable lipids) was used to collectively reconstruct the source vegetation of different sediment layers.

  8. Sucrose Transporter Localization and Function in Phloem Unloading in Developing Stems.

    PubMed

    Milne, Ricky J; Perroux, Jai M; Rae, Anne L; Reinders, Anke; Ward, John M; Offler, Christina E; Patrick, John W; Grof, Christopher P L

    2017-02-01

    How sucrose transporters (SUTs) regulate phloem unloading in monocot stems is poorly understood and particularly so for species storing high Suc concentrations. To this end, Sorghum bicolor SUTs SbSUT1 and SbSUT5 were characterized by determining their transport properties heterologously expressed in yeast or Xenopus laevis oocytes, and their in planta cellular and subcellular localization. The plasma membrane-localized SbSUT1 and SbSUT5 exhibited a strong selectivity for Suc and high Suc affinities in X. laevis oocytes at pH 5-SbSUT1, 6.3 ± 0.7 mm, and SbSUT5, 2.4 ± 0.5 mm Suc. The Suc affinity of SbSUT1 was dependent on membrane potential and pH. In contrast, SbSUT5 Suc affinity was independent of membrane potential and pH but supported high transport rates at neutral pH. Suc transport by the tonoplast localized SbSUT4 could not be detected using yeast or X. laevis oocytes. Across internode development, SUTs, other than SbSUT4, were immunolocalized to sieve elements, while for elongating and recently elongated internodes, SUTs also were detected in storage parenchyma cells. We conclude that apoplasmic Suc unloading from de-energized protophloem sieve elements in meristematic zones may be mediated by reversal of SbSUT1 and/or by uniporting SWEETs. Storage parenchyma localized SbSUT1 and SbSUT5 may accumulate Suc from the stem apoplasms of elongating and recently elongated internodes, whereas SbSUT4 may function to release Suc from vacuoles. Transiting from an apoplasmic to symplasmic unloading pathway as the stem matures, SbSUT1 and SbSUT5 increasingly function in Suc retrieval into metaphloem sieve elements to maintain a high turgor to drive symplasmic unloading by bulk flow. © 2017 American Society of Plant Biologists. All Rights Reserved.

  9. The history and development of FETAX (ASTM standard guide, E-1439 on conducting the frog embryo teratogenesis Assay-Xenopus)

    USGS Publications Warehouse

    Dumont, J.N.; Bantle, J.A.; Linder, G.; ,

    2003-01-01

    The energy crisis of the 1970's and 1980's prompted the search for alternative sources of fuel. With development of alternate sources of energy, concerns for biological resources potentially adversely impacted by these alternative technologies also heightened. For example, few biological tests were available at the time to study toxic effects of effluents on surface waters likely to serve as receiving streams for energy-production facilities; hence, we began to use Xenopus laevis embryos as test organisms to examine potential toxic effects associated with these effluents upon entering aquatic systems. As studies focused on potential adverse effects on aquatic systems continued, a test procedure was developed that led to the initial standardization of FETAX. Other .than a limited number of aquatic toxicity tests that used fathead minnows and cold-water fishes such as rainbow trout, X. laevis represented the only other aquatic vertebrate test system readily available to evaluate complex effluents. With numerous laboratories collaborating, the test with X. laevis was refined, improved, and developed as ASTM E-1439, Standard Guide for the Conducting Frog Embryo Teratogenesis Assay-Xenopus (FETAX). Collabrative work in the 1990s yielded procedural enhancements, for example, development of standard test solutions and exposure methods to handle volatile organics and hydrophobic compounds. As part of the ASTM process, a collaborative interlaboratory study was performed to determine the repeatability and reliability of FETAX. Parallel to these efforts, methods were also developed to test sediments and soils, and in situ test methods were developed to address "lab-to-field extrapolation errors" that could influence the method's use in ecological risk assessments. Additionally, a metabolic activation system composed of rat liver microsomes was developed which made FETAX more relevant to mammalian studies.

  10. Microinjection of recombinant O-GlcNAc transferase potentiates Xenopus oocytes M-phase entry

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dehennaut, Vanessa; EA 4020, Laboratoire de Regulation des Signaux de Division, USTL, IFR147, Villeneuve d'Ascq; Hanoulle, Xavier

    2008-05-02

    In order to understand the importance of the cytosolic and nuclear-specific O-linked N-acetylglucosaminylation (O-GlcNAc) on cell cycle regulation, we recently reported that inhibition of O-GlcNAc transferase (OGT) delayed or blocked Xenopus laevis oocyte germinal vesicle breakdown (GVBD). Here, we show that increased levels of the long OGT isoform (ncOGT) accelerate X. laevis oocyte GVBD. A N-terminally truncated isoform (sOGT) with a similar in vitro catalytic activity towards a synthetic CKII-derived peptide had no effect, illustrating the important role played by the N-terminal tetratrico-peptide repeats. ncOGT microinjection in the oocytes increases both the speed and extent of O-GlcNAc addition, leads tomore » a quicker activation of the MPF and MAPK pathways and finally results in a faster GVBD. Microinjection of anti-OGT antibodies leads to a delay of the GVBD kinetics. Our results hence demonstrate that OGT is a key molecule for the timely progression of the cell cycle.« less

  11. High diadenosine tetraphosphate (Ap4A) level in germ cells and embryos of sea urchin and Xenopus and its effect on DNA synthesis.

    PubMed

    Weinmann-Dorsch, C; Grummt, F

    1985-09-01

    Ap4A levels in sperms, eggs and different developmental stages of sea urchin (Psammechinus miliaris) and (Xenopus laevis) were determined by a method based on ATP measurement with luciferin/luciferase after splitting diadenosine 5',5'''-P1,P4-tetraphosphate (Ap4A) into ATP and AMP. Appreciable storage pools of Ap4A were found in unfertilized eggs of Psammechinus and Xenopus as well as in sea urchin sperms. The actual Ap4A concentration of 28 microM in sperm represents the highest Ap4A level so far observed in eukaryotic cells. Upon fertilization an instant onset of de novo synthesis of Ap4A was demonstrated. Ap4A levels during early embryogenesis of P. miliaris and X. laevis (2.5-4 microM) are higher than those in exponentially growing mammalian culture cells and mammalian fetuses. Microinjection of Ap4A into unfertilized eggs of Psammechinus miliaris caused a 3-7 fold increase of DNA synthesis in comparison with mock-injected eggs.

  12. The mammalian retina as a clock

    NASA Technical Reports Server (NTRS)

    Tosini, Gianluca; Fukuhara, Chiaki

    2002-01-01

    Many physiological, cellular, and biochemical parameters in the retina of vertebrates show daily rhythms that, in many cases, also persist under constant conditions. This demonstrates that they are driven by a circadian pacemaker. The presence of an autonomous circadian clock in the retina of vertebrates was first demonstrated in Xenopus laevis and then, several years later, in mammals. In X. laevis and in chicken, the retinal circadian pacemaker has been localized in the photoreceptor layer, whereas in mammals, such information is not yet available. Recent advances in molecular techniques have led to the identification of a group of genes that are believed to constitute the molecular core of the circadian clock. These genes are expressed in the retina, although with a slightly different 24-h profile from that observed in the central circadian pacemaker. This result suggests that some difference (at the molecular level) may exist between the retinal clock and the clock located in the suprachiasmatic nuclei of hypothalamus. The present review will focus on the current knowledge of the retinal rhythmicity and the mechanisms responsible for its control.

  13. Subcellular components of the amphibian egg - Insights provided by gravitational studies

    NASA Technical Reports Server (NTRS)

    Neff, A. W.; Ritzenthaler, J. D.; Rosenbaum, J. F.

    1989-01-01

    The variability in the response of Xenopus laevis eggs to a given force environment is studied. The roles of cytoplasmic organelle, the yolk platelets, and cytoskeletal components in varying in cytoplasmic mobility are examined. The data reveal that the packing of yolk platelets is not a major factor in causing cytoplasmic mobility differences and microtubules may affect cytoplasmic mobility.

  14. Intestinal helminth fauna of the South American sea lion Otaria flavescens and fur seal Arctocephalus australis from northern Patagonia, Argentina.

    PubMed

    Hernández-Orts, J S; Montero, F E; Juan-García, A; García, N A; Crespo, E A; Raga, J A; Aznar, F J

    2013-09-01

    We report on the intestinal helminth fauna of 56 South American sea lions, Otaria flavescens, and 5 South American fur seals, Arctocephalus australis, from northern Patagonia, Argentina. A total of 97,325 helminth specimens were collected from sea lions. Gravid individuals were represented by 6 species of parasites: 1 digenean (Ascocotyle (Ascocotyle) patagoniensis), 1 cestode (Diphyllobothrium spp.), 3 nematodes (Uncinaria hamiltoni, Contracaecum ogmorhini s.s., Pseudoterranova cattani) and 1 acanthocephalan (Corynosoma australe). In addition, third-stage larvae of 2 nematodes (Contracaecum sp. and Anisakis sp. type I) and 3 juvenile acanthocephalans (Andracantha sp., Profilicollis chasmagnathi and Corynosoma cetaceum) were also collected. Andracantha sp., C. ogmorhini s.s. and P. chasmagnathi represent new host records. A total of 1516 helminth specimens were collected from fur seals. Gravid individuals were represented by three species of parasites, namely, Diphyllobothrium spp., C. ogmorhini s.s. and C. australe. In addition, larvae of Contracaecum sp. and P. cattani, juveniles of C. cetaceum and immature cestodes (Tetrabothriidae gen. sp.) were also collected. Corynosoma australe was the most prevalent and abundant parasite in both hosts, accounting for >90% of all specimens. Sea lions and furs seals from northern Patagonia harbour the intestinal helminth communities that could be predicted for otariids, i.e. the combination of species of the genera Corynosoma, Diphyllobothrium, Pseudoterranova, Contracaecum and, in pups, Uncinaria. Additionally, both species of otariid are apparently unsuitable hosts (i.e. non-hosts) for as many as five parasite taxa. The inclusion or exclusion of these species affects estimation of species richness at both component community (11 versus 6 species in sea lions; 7 versus 3 species in fur seals) and infracommunity (mean: 3.1 versus 2.6 in sea lions; 2.2 versus 1.7 species) levels. Information about the reproductive status of

  15. Metazoan fish parasites of Macrourus berglax Lacepède, 1801 and other macrourids of the North Atlantic: Invasion of the deep sea from the continental shelf

    NASA Astrophysics Data System (ADS)

    Palm, H. W.; Klimpel, S.

    2008-01-01

    Between 2001 and 2003, a total of 105 Macrourus berglax Lacepède, 1801 were collected from Irminger Sea, at depths between 250 and 450 m, and were studied for parasites. Twenty-four different parasite species were identified. Nematodes (eight spp.) and digeneans (six spp.) were the most species rich, followed by crustaceans (four spp.), cestodes (three spp.), acanthocephalans (two spp.) and monogeneans (one sp.). Core species included the digenean Gonocerca phycidis (maximum 62.9% prevalence), the nematodes Anisakis sp. (62.9%), Capillaria gracilis (65.8%), Hysterothylacium aduncum (60.0%), Neoascarophis macrouri (88.6%), Spinitectus oviflagellis (82.9%), and the acanthocephalan Echinorhynchus gadi (97.1%). M. berglax was the final host for most of the parasites, and was infested with only a few larval forms. The species composition, diversity as well as the prevalence and intensity of infestation for most parasite species collected, were similar in the different years, indicating no distinct interannual variation at this deep-sea locality. Comparisons of the parasite fauna of M. berglax with macrourids belonging to the genera Macrourus, Coryphaenoides, Coelorhynchus and Nezumia revealed a similar infestation pattern amongst these deep-sea fish, consisting of the same or closely related species. This observation suggests that the parasite life-cycles in these benthopelagic deep-sea fishes follow similar pathways independent of geographical location. A similar habitat and food preference of macrourids results in a similar parasite fauna. The host specificity of some of the parasites is low, with most species (nine) infesting Teleostei, Gadiformes (six), Macrouridae (two), Macrourus spp. (one) and specifically M. berglax (five). Overlapping infestation patterns of M. berglax parasites with phylogenetically related gadiform fish from the continental shelf region suggest that the deep-sea parasite fauna in macrourids has evolved along with their hosts from parasite

  16. Helminths in an intensively stocked population of lake trout, Salvelinus namaycush, from Lake Huron

    USGS Publications Warehouse

    Muzzall, Patrick M.; Bowen, Charles A.

    2000-01-01

    Eighty stocked lake trout Salvelinus namaycush (Salmonidae), collected from 2 locations in Lake Huron in May 1995, were examined for parasites. The parasite fauna of this top predator in Lake Huron was characterized by only 6 helminth species. Echinorhynchus salmonis infected all lake trout with a mean intensity of 163.9. The intensity of this acanthocephalan species significantly increased with host length and weight. Eubothrium salvelini infected 78 lake trout with a maximum number of 81 scoleces counted. Diplostomum sp., Cyathocephalus truncatus, Capillaria salvelini, and Neoechinorhynchus sp. infrequently infected lake trout. The low parasite species richness in these lake trout is believed to be due to their large size at stocking and to the loss of historical enzootic host-parasite relationships that followed the absence of this fish species in Lake Huron for 26 yr.

  17. Community structure of helminth parasites of the "Cururu" toad, Rhinella icterica (Anura: Bufonidae) from southern Brazil.

    PubMed

    Dos Santos, Viviane Gularte Tavares; Amato, Suzana B; Borges-Martins, Márcio

    2013-03-01

    Sixty specimens of the "cururu" toad, Rhinella icterica (Spix 1824) (Bufonidae), were collected in Campo Belo do Sul, State of Santa Catarina, Brazil, between May 2009 and January 2011, and were examined for the presence of helminth parasites. Nine species of adult helminths were found: Catadiscus cohni, Rudolphitrema rudolphii, Cylindrotaenia sp., Rhabdias fuelleborni, Strongyloides sp., Cosmocerca rara, Cosmocerca brasiliensis, Aplectana elenae, and Oxyascaris sp., in addition to an unidentified adult nematode species. Females of cosmocercid nematodes, proteocephalan plerocercoid, and acanthocephalan cystacanth were found but not identified for lack absolute of taxonomic characters. The sex of the anurans had no influence on prevalence, abundance, and richness of helminth species. Length and body mass of hosts did not influence the prevalence and richness of helminths, while the abundance of R. fuelleborni was significantly correlated with both parameters.

  18. [Intercellular relationship of notochord determination of Xenopus laevis].

    PubMed

    Zeng, M B; Zhou, M Y; Wang, Y

    1995-09-01

    During the process of determination, the presumptive notochord is situated beneath neuroepithelium, flanked at two sides by presumptive somites and underlain with archenteron roof ventrally. Among these neighbouring embryonic tissues, presumptive somites were found to exert the main influence on notochord determination. By electron microscopic observations, the presumptive notochord and somite cells were seen to situate either close to each other (plate I, Fig. 1) or connected by cytoplasmic processes forming intercellular lumen (plate I, Fig. 5). Coated pits and coated vesicles appeared at the outer surface of both types of cells (plate I, Figs. 1-4). For the presumptive somite cells, spherical bodies of different sizes and variable contents were observed either near or protruding from the outer surface (plate II, Figs. 6-10). The spherical bodies were also found in the intercellular lumen (plate III, Fig. 11). These spherical bodies were mainly composed of granules, loosely scattered or densely packed. The granules were of similar size and similar shade of electron staining as those of ribosomes of the presumptive somite cells. For the presumptive notochord cells, no spherical bodies of the above mentioned type were found, but phenomenon of engulfing luminal material was observed (plate III, Fig. 12). The significance of the appearance of these spherical bodies in the determination of notochord cells has been discussed.

  19. Intravascular Neural Interface with Nanowire Electrode

    PubMed Central

    Watanabe, Hirobumi; Takahashi, Hirokazu; Nakao, Masayuki; Walton, Kerry; Llinás, Rodolfo R.

    2010-01-01

    Summary A minimally invasive electrical recording and stimulating technique capable of simultaneously monitoring the activity of a significant number (e.g., 103 to 104) of neurons is an absolute prerequisite in developing an effective brain–machine interface. Although there are many excellent methodologies for recording single or multiple neurons, there has been no methodology for accessing large numbers of cells in a behaving experimental animal or human individual. Brain vascular parenchyma is a promising candidate for addressing this problem. It has been proposed [1, 2] that a multitude of nanowire electrodes introduced into the central nervous system through the vascular system to address any brain area may be a possible solution. In this study we implement a design for such microcatheter for ex vivo experiments. Using Wollaston platinum wire, we design a submicron-scale electrode and develop a fabrication method. We then evaluate the mechanical properties of the electrode in a flow when passing through the intricacies of the capillary bed in ex vivo Xenopus laevis experiments. Furthermore, we demonstrate the feasibility of intravascular recording in the spinal cord of Xenopus laevis. PMID:21572940

  20. Notch activates Wnt-4 signalling to control medio-lateral patterning of the pronephros.

    PubMed

    Naylor, Richard W; Jones, Elizabeth A

    2009-11-01

    Previous studies have highlighted a role for the Notch signalling pathway during pronephrogenesis in the amphibian Xenopus laevis, and in nephron development in the mammalian metanephros, yet a mechanism for this function remains elusive. Here, we further the understanding of how Notch signalling patterns the early X. laevis pronephros anlagen, a function that might be conserved in mammalian nephron segmentation. Our results indicate that early phase pronephric Notch signalling patterns the medio-lateral axis of the dorso-anterior pronephros anlagen, permitting the glomus and tubules to develop in isolation. We show that this novel function acts through the Notch effector gene hrt1 by upregulating expression of wnt4. Wnt-4 then patterns the proximal pronephric anlagen to establish the specific compartments that span the medio-lateral axis. We also identified pronephric expression of lunatic fringe and radical fringe that is temporally and spatially appropriate for a role in regulating Notch signalling in the dorso-anterior region of the pronephros anlagen. On the basis of these results, along with data from previous publications, we propose a mechanism by which the Notch signalling pathway regulates a Wnt-4 function that patterns the proximal pronephric anlagen.

  1. Temperature-dependent regulation of vocal pattern generator.

    PubMed

    Yamaguchi, Ayako; Gooler, David; Herrold, Amy; Patel, Shailja; Pong, Winnie W

    2008-12-01

    Vocalizations of Xenopus laevis are generated by central pattern generators (CPGs). The advertisement call of male X. laevis is a complex biphasic motor rhythm consisting of fast and slow trills (a train of clicks). We found that the trill rate of these advertisement calls is sensitive to temperature and that this rate modification of the vocal rhythms originates in the central pattern generators. In vivo the rates of fast and slow trills increased linearly with an increase in temperature. In vitro a similar linear relation between temperature and compound action potential frequency in the laryngeal nerve was found when fictive advertisement calls were evoked in the isolated brain. Temperature did not limit the contractile properties of laryngeal muscles within the frequency range of vocalizations. We next took advantage of the temperature sensitivity of the vocal CPG in vitro to localize the source of the vocal rhythms. We focused on the dorsal tegmental area of the medulla (DTAM), a brain stem nucleus that is essential for vocal production. We found that bilateral cooling of DTAM reduced both fast and slow trill rates. Thus we conclude that DTAM is a source of biphasic vocal rhythms.

  2. Echinoderms from Middle and Upper Ordovician rocks of Kentucky

    USGS Publications Warehouse

    Parsley, R.L.

    1981-01-01

    The Middle and Upper Ordovician limestones of Kentucky, especially the Lexington Limestone, have yielded a diverse silicified echinoderm fauna, including: Stylophora-Enoploura cf. E. punctata; Paracrinoidea-A mygdalocystites; Crinoidea, Inadunata-Hybocrir/us tumidus, Hybocystites problem,aticus, Carabocrinus sp., Cupulocrinus sp., Heterocrinus sp.; Cyclocystoidea-Cyclocystoides sp. A rhombiferan cystoid, A mecystis laevis, from the Edinburg Formation, Virginia, is also discussed. No new taxa are introduced.

  3. Annexin IV (Xanx-4) has a functional role in the formation of pronephric tubules.

    PubMed

    Seville, Rachel A; Nijjar, Sarbjit; Barnett, Mark W; Massé, Karine; Jones, Elizabeth A

    2002-04-01

    Vertebrate kidney organogenesis is characterised by the successive formation of the pronephros, the mesonephros and the metanephros. The pronephros is the first to form and is the functional embryonic kidney of lower vertebrates; although it is vestigial in higher vertebrates, it is a necessary precursor for the other kidney types. The Xenopus pronephros is a simple paired organ; each nephron consists of a single large glomus, one set of tubules and a single duct. The simple organisation of the pronephros and the amenability of Xenopus laevis embryos to manipulation make the Xenopus pronephros an attractive system in which to study organogenesis. It has been shown that pronephric tubules can be induced to form in presumptive ectodermal tissue by treatment with RA and activin. We have used this system in a subtractive hybridisation screen that resulted in the cloning of Xenopus laevis annexin IV (Xanx-4). Xanx-4 transcripts are specifically located to the developing pronephric tubules, and the protein to the luminal surface of these tubules. Temporal expression shows zygotic transcription is upregulated at the time of pronephric tubule specification and persists throughout pronephric development. The temporal and spatial expression pattern of Xanx-4 suggests it may have a role in pronephric tubule development. Overexpression of Xanx-4 yields no apparent phenotype, but Xanx-4 depletion, using morpholinos, produces a shortened, enlarged tubule phenotype. The phenotype observed can be rescued by co-injection of Xanx-4 mRNA. Although the function of annexins is not yet clear, studies have suggested a role for annexins in a number of cellular processes. Annexin IV has been shown to have an inhibitory role in the regulation of epithelial calcium-activated chloride ion conductance. The enlarged pronephric tubule phenotype observed may be attributed to incorrect modulation of exocytosis, membrane plasticity or ion channels and/or water homeostasis. In this study, we

  4. Intestinal parasites of raccoons (Procyon lotor) from southwest British Columbia.

    PubMed Central

    Ching, H L; Leighton, B J; Stephen, C

    2000-01-01

    This is the first extensive survey of metazoan parasites (particularly of the roundworm Baylisascaris procyonis) from the intestines of raccoons in British Columbia. The sample collected in 1997-1998 consisted of 82 raccoons that had been sick or had been killed accidentally by automobiles. Fifteen parasite taxa were found: 3 nematodes, 9 digenetic trematodes, 2 acanthocephalans and 1 cestode. Ten of these parasites constitute new host records for raccoons, including 4 digenetic trematodes that have been reported in marine birds and mammals on the Pacific Coast of North America. Baylisascaris procyonis infected 61% of the raccoons with a mean intensity of 27. The high rate of infection indicates a large potential for environmental contamination and, thus, human and animal exposure to infectious eggs. Prevention of larva migrans is discussed, particularly for people in contact with raccoons in wildlife rehabilitation centers. PMID:10805249

  5. Eggs: The role of gravity in the establishment of the dorso-ventral axis in the amphibian embryo (7-IML-1)

    NASA Technical Reports Server (NTRS)

    Ubbels, Geertje A.

    1992-01-01

    The purpose of this experiment is to fertilize frog (Xenopus laevis) eggs under microgravity, to perform histological fixations at two different programmed times, and after return to Earth, to determine whether timing and pattern of egg cleavages and axis formation are normal. Because of the limited viability of the gametes, this experiment will be the very first to be activated in the Biorack. Various aspects of this investigation are discussed.

  6. Tumor Suppression by BRCA-1: A Critical Role at DNA Replication Forks

    DTIC Science & Technology

    2006-10-01

    replication defect. We wished to test the hypothesis that BRCA1/BARD1 function during DNA replication supporting DNA transactions at replication forks. We...are using cell-free extracts derived from Xenopus laevis eggs that support: 1. Semi-conservative, cell-cycle regulated DNA replication ; 2. Many facets...complex assembles to chromatin in a DNA replication -dependent manner. Finally, we show that BRCA1/BARD1 loading to chromatin does not dramatically

  7. Amphibian Development in the Virtual Absence of Gravity

    NASA Technical Reports Server (NTRS)

    Souza, Kenneth A.; Black, Steven D.; Wassersug, Richard J.

    1995-01-01

    To test whether gravity is required for normal amphibian development, Xenopus laevis females were induced to ovulate aboard the orbiting Space Shuttle. Eggs were fertilized in vitro, and although early embryonic stages showed some abnormalities, the embryos were able to regulate and produce nearly normal larvae. These results demonstrate that a vertebrate can ovulate in the virtual absence of gravity and that the eggs can develop to a free-living stage.

  8. Evolutionary conservativeness of electric field in the Cu,Zn superoxide dismutase active site. Evidence for co-ordinated mutation of charged amino acid residues.

    PubMed

    Desideri, A; Falconi, M; Polticelli, F; Bolognesi, M; Djinovic, K; Rotilio, G

    1992-01-05

    Equipotential lines were calculated, using the Poisson-Boltzmann equation, for six Cu,Zn superoxide dismutases with different protein electric charge and various degrees of sequence homology, namely those from ox, pig, sheep, yeast, and the isoenzymes A and B from the amphibian Xenopus laevis. The three-dimensional structures of the porcine and ovine superoxide dismutases were obtained by molecular modelling reconstruction using the structure of the highly homologous bovine enzyme as a template. The three-dimensional structure of the evolutionary distant yeast Cu,Zn superoxide dismutase was recently resolved by us, while computer-modelled structures are available for X. laevis isoenzymes. The six proteins display large differences in the net protein charge and distribution of electrically charged surface residues but the trend of the equipotential lines in the proximity of the active sites was found to be constant in all cases. These results are in line with the very similar catlytic rate constants experimentally measured for the corresponding enzyme activities. This analysis shows that electrostatic guidance for the enzyme-substrate interaction in Cu,Zn superoxide dismutases is related to a spatial distribution of charges, arranged so as to maintain, in the area surrounding the active sites, an identical electrostatic potential distribution, which is conserved in the evolution of this protein family.

  9. Transcriptome profiles of metamorphosis in the ornamented pygmy frog Microhyla fissipes clarify the functions of thyroid hormone receptors in metamorphosis.

    PubMed

    Zhao, Lanying; Liu, Lusha; Wang, Shouhong; Wang, Hongyuan; Jiang, Jianping

    2016-06-02

    Anuran metamorphosis is an excellent system in which to study postembryonic development. Studies on Xenopus (Mesobatrachia) show that thyroid hormone receptors (TRs) regulate metamorphosis in a ligand-dependent manner by coordinating the action of hundreds of genes. However, whether this mechanism is conserved among amphibians is still unknown. To understand the molecular mechanism of this universal phenomenon, we report the transcriptional profiles of the three key developmental stages in Microhyla fissipes (Neobatrachia): premetamorphosis (PM), metamorphic climax (MC) and completion of metamorphosis (CM). In total, 2,293 differentially expressed genes were identified from comparisons of transcriptomes, and these genes showed stage-specific expression patterns. Unexpectedly, we found that TRα was highly expressed in Xenopus laevis and Bufo gargarizans at premetamorphosis but showed low expression in M. fissipes. In contrast, TRβ was highly expressed during metamorphosis in M. fissipes and X. laevis. This result may imply that TRβ is essential for initiating metamorphosis, at least in M. fissipes. Thus, our work not only identifies genes that are likely to be involved in Neobatrachia metamorphosis but also clarifies the roles of unliganded TRα in regulating tadpole growth and timing of metamorphosis, which may be conserved in anurans, and the role of liganded TRβ in launching metamorphosis.

  10. Transcriptome profiles of metamorphosis in the ornamented pygmy frog Microhyla fissipes clarify the functions of thyroid hormone receptors in metamorphosis

    PubMed Central

    Zhao, Lanying; Liu, Lusha; Wang, Shouhong; Wang, Hongyuan; Jiang, Jianping

    2016-01-01

    Anuran metamorphosis is an excellent system in which to study postembryonic development. Studies on Xenopus (Mesobatrachia) show that thyroid hormone receptors (TRs) regulate metamorphosis in a ligand-dependent manner by coordinating the action of hundreds of genes. However, whether this mechanism is conserved among amphibians is still unknown. To understand the molecular mechanism of this universal phenomenon, we report the transcriptional profiles of the three key developmental stages in Microhyla fissipes (Neobatrachia): premetamorphosis (PM), metamorphic climax (MC) and completion of metamorphosis (CM). In total, 2,293 differentially expressed genes were identified from comparisons of transcriptomes, and these genes showed stage-specific expression patterns. Unexpectedly, we found that TRα was highly expressed in Xenopus laevis and Bufo gargarizans at premetamorphosis but showed low expression in M. fissipes. In contrast, TRβ was highly expressed during metamorphosis in M. fissipes and X. laevis. This result may imply that TRβ is essential for initiating metamorphosis, at least in M. fissipes. Thus, our work not only identifies genes that are likely to be involved in Neobatrachia metamorphosis but also clarifies the roles of unliganded TRα in regulating tadpole growth and timing of metamorphosis, which may be conserved in anurans, and the role of liganded TRβ in launching metamorphosis. PMID:27254593

  11. Tissue-Specific Upregulation of MDS/EVI Gene Transcripts in the Intestine by Thyroid Hormone during Xenopus Metamorphosis

    PubMed Central

    Hasebe, Takashi; Fu, Liezhen; Heimeier, Rachel A.; Das, Biswajit; Ishizuya-Oka, Atsuko; Shi, Yun-Bo

    2013-01-01

    Background Intestinal remodeling during amphibian metamorphosis resembles the maturation of the adult intestine during mammalian postembryonic development when the adult epithelial self-renewing system is established under the influence of high concentrations of plasma thyroid hormone (T3). This process involves de novo formation and subsequent proliferation and differentiation of the adult stem cells. Methodology/Principal Findings The T3-dependence of the formation of adult intestinal stem cell during Xenopus laevis metamorphosis offers a unique opportunity to identify genes likely important for adult organ-specific stem cell development. We have cloned and characterized the ectopic viral integration site 1 (EVI) and its variant myelodysplastic syndrome 1 (MDS)/EVI generated via transcription from the upstream MDS promoter and alternative splicing. EVI and MDS/EVI have been implicated in a number of cancers including breast, leukemia, ovarian, and intestinal cancers. We show that EVI and MDS/EVI transcripts are upregulated by T3 in the epithelium but not the rest of the intestine in Xenopus laevis when adult stem cells are forming in the epithelium. Conclusions/Significance Our results suggest that EVI and MDS/EVI are likely involved in the development and/or proliferation of newly forming adult intestinal epithelial cells. PMID:23383234

  12. Frogs as integrative models for understanding digestive organ development and evolution.

    PubMed

    Womble, Mandy; Pickett, Melissa; Nascone-Yoder, Nanette

    2016-03-01

    The digestive system comprises numerous cells, tissues and organs that are essential for the proper assimilation of nutrients and energy. Many aspects of digestive organ function are highly conserved among vertebrates, yet the final anatomical configuration of the gut varies widely between species, especially those with different diets. Improved understanding of the complex molecular and cellular events that orchestrate digestive organ development is pertinent to many areas of biology and medicine, including the regeneration or replacement of diseased organs, the etiology of digestive organ birth defects, and the evolution of specialized features of digestive anatomy. In this review, we highlight specific examples of how investigations using Xenopus laevis frog embryos have revealed insight into the molecular and cellular dynamics of digestive organ patterning and morphogenesis that would have been difficult to obtain in other animal models. Additionally, we discuss recent studies of gut development in non-model frog species with unique feeding strategies, such as Lepidobatrachus laevis and Eleutherodactylous coqui, which are beginning to provide glimpses of the evolutionary mechanisms that may generate morphological variation in the digestive tract. The unparalleled experimental versatility of frog embryos make them excellent, integrative models for studying digestive organ development across multiple disciplines. Copyright © 2016 Elsevier Ltd. All rights reserved.

  13. AmphiBase: A new genomic resource for non-model amphibian species.

    PubMed

    Kwon, Taejoon

    2017-01-01

    More than five thousand genes annotated in the recently published Xenopus laevis and Xenopus tropicalis genomes do not have a candidate orthologous counterpart in other vertebrate species. To determine whether these sequences represent genuine amphibian-specific genes or annotation errors, it is necessary to analyze them alongside sequences from other amphibian species. However, due to large genome sizes and an abundance of repeat sequences, there are limited numbers of gene sequences available from amphibian species other than Xenopus. AmphiBase is a new genomic resource covering non-model amphibian species, based on public domain transcriptome data and computational methods developed during the X. laevis genome project. Here, I review the current status of AmphiBase, including amphibian species with available transcriptome data or biological samples, and describe the challenges of building a comprehensive amphibian genomic resource in the absence of genomes. This mini-review will be informative for researchers interested in functional genomic experiments using amphibian model organisms, such as Xenopus and axolotl, and will assist in interpretation of results implicating "orphan genes." Additionally, this study highlights an opportunity for researchers working on non-model amphibian species to collaborate in their future efforts and develop amphibian genomic resources as a community. © 2017 Wiley Periodicals, Inc.

  14. Histological development of the gonad in juvenile Xenopus laevis

    EPA Science Inventory

    As directed by the Food Quality Protection Act, the US Environmental Protection Agency is developing a screening program for endocrine disrupting compounds. The Larval Amphibian Growth and Development Assay (LAGDA) is a tier II test intended to identify and characterize the adver...

  15. Modeling cell-cycle synchronization during embryogenesis in Xenopus laevis

    NASA Astrophysics Data System (ADS)

    McIsaac, R. Scott; Huang, K. C.; Sengupta, Anirvan; Wingreen, Ned

    2010-03-01

    A widely conserved aspect of embryogenesis is the ability to synchronize nuclear divisions post-fertilization. How is synchronization achieved? Given a typical protein diffusion constant of 10 μm^2sec, and an embryo length of 1mm, it would take diffusion many hours to propagate a signal across the embryo. Therefore, synchrony cannot be attained by diffusion alone. We hypothesize that known autocatalytic reactions of cell-cycle components make the embryo an ``active medium'' in which waves propagate much faster than diffusion, enforcing synchrony. We report on robust spatial synchronization of components of the core cell cycle circuit based on a mathematical model previously determined by in vitro experiments. In vivo, synchronized divisions are preceded by a rapid calcium wave that sweeps across the embryo. Experimental evidence supports the hypothesis that increases in transient calcium levels lead to derepression of a negative feedback loop, allowing cell divisions to start. Preliminary results indicate a novel relationship between the speed of the initial calcium wave and the ability to achieve synchronous cell divisions.

  16. The nervus terminalis in larval and adult Xenopus laevis.

    PubMed

    Hofmann, M H; Meyer, D L

    1989-09-25

    Nervus terminalis (nt) projections were studied by HRP injections into one nostril in adult Xenopus and in Xenopus tadpoles. Central nt targets are: medial septum, preoptic nucleus, nucleus of the anterior commissure, and hypothalamus (mainly ipsilaterally). In Xenopus tadpoles, additional fibers reach the ipsilateral dorsal thalamus and the mesencephalic tegmentum, bilaterally; furthermore, hypothalamic projections are bilateral. Xenopus tadpole nt connections resemble those of adult urodeles more closely than the projections of frogs. However, Xenopus tadpoles lack nt innervation of the medial septum.

  17. REGULATION OF THE THYROID AXIS IN DEVELOPING XENOPUS LAEVIS

    EPA Science Inventory

    The focus of the research presented here is the development of an in vitro pituitary gland culture system to test the effect of chemicals directly on the gland without influence of other parts of the HPT axis.

  18. Expression of membrane targeted aequorin in Xenopus laevis oocytes.

    PubMed

    Daguzan, C; Nicolas, M T; Mazars, C; Leclerc, C; Moreau, M

    1995-08-01

    We described here a system for high level of expression of the calcium activated photoprotein aequorin. This protein has been targeted to the plasma membrane of Xenopus oocyte by nuclear microinjection of a plasmid containing a construction of a chimeric cDNA encoding a fusion protein composed of the photoprotein aequorin and the 5-HT1A receptor. The expression of this fusion protein is placed under the control of RSV promoter. Functional photoprotein was reconstituted in the oocyte by incubation with coelenterazine. The amount of photoprotein 24 h after nuclear microinjection of the plasmid was sufficient to trigger a detectable light emission following calcium entry. The efficiency of the expression is correlated with the dose of plasmid injected. Intracytoplasmic injection of the plasmid always failed in photoprotein expression. Targeting of the apoprotein was demonstrated by immunolocalization under confocal microscopy. In our experimental conditions, the apoprotein was always localized at the animal pole above the nucleus. We never observed expression and targeting to the plasma membrane of the vegetal pole. WE suggest that such expression might be of great interest for the study of numerous problems of developmental biology, in which calcium-dependent pathways are involved.

  19. Host sharing and host manipulation by larval helminths in shore crabs: cooperation or conflict?

    PubMed

    Poulin, Robert; Nichol, Katherine; Latham, A David M

    2003-04-01

    Larval helminths of different species that share the same intermediate host and are transmitted by predation to the same definitive host may cooperate in their attempts to manipulate the behaviour of the intermediate host, while at the same time having conflicts of interests over the use of host resources. A few studies have indicated that intermediate hosts harbouring larval helminths have altered concentrations of neurotransmitters in their nervous system, and thus measuring levels of neurotransmitters in host brains could serve to assess the respective and combined effect of different helminth species on host behaviour. Here, we investigate potential cooperation and conflict among three helminths in two species of crab intermediate hosts. The acanthocephalan Profilicollis spp., the trematode Maritrema sp. and an acuariid nematode, all use Macrophthalmus hirtipes (Ocypodidae) as intermediate host, whereas Profilicollis and Maritrema also use Hemigrapsus crenulatus (Grapsidae). All three helminths mature inside gulls or other shore birds. There was a significant decrease in the mean volume of Profilicollis cystacanths as the intensity of infection by this parasite increased in H. crenulatus, the only host in which this was investigated; however, there was no measurable effect of other helminth species on the size of acanthocephalans, suggesting no interspecific conflict over resource use within crabs. There was, in contrast, evidence of a positive interspecific association between the two most common helminth species: numbers of Profilicollis and Maritrema were positively correlated among crabs, independently of crab size, in M. hirtipes but not H. crenulatus. More importantly, we found that the total number of larval helminths per crab correlated significantly, and negatively, with concentrations of serotonin in crab brains, again only in M. hirtipes; numbers of each parasite species separately did not covary in either crab species with serotonin or dopamine, the

  20. Contrasting parasite communities among allopatric colour morphs of the Lake Tanganyika cichlid Tropheus

    PubMed Central

    2013-01-01

    Background Adaptation to different ecological environments is thought to drive ecological speciation. This phenomenon culminates in the radiations of cichlid fishes in the African Great Lakes. Multiple characteristic traits of cichlids, targeted by natural or sexual selection, are considered among the driving factors of these radiations. Parasites and pathogens have been suggested to initiate or accelerate speciation by triggering both natural and sexual selection. Three prerequisites for parasite-driven speciation can be inferred from ecological speciation theory. The first prerequisite is that different populations experience divergent infection levels. The second prerequisite is that these infection levels cause divergent selection and facilitate adaptive divergence. The third prerequisite is that parasite-driven adaptive divergence facilitates the evolution of reproductive isolation. Here we investigate the first and the second prerequisite in allopatric chromatically differentiated lineages of the rock-dwelling cichlid Tropheus spp. from southern Lake Tanganyika (Central Africa). Macroparasite communities were screened in eight populations belonging to five different colour morphs. Results Parasite communities were mainly composed of acanthocephalans, nematodes, monogeneans, copepods, branchiurans, and digeneans. In two consecutive years (2011 and 2012), we observed significant variation across populations for infection with acanthocephalans, nematodes, monogeneans of the genera Gyrodactylus and Cichlidogyrus, and the copepod Ergasilus spp. Overall, parasite community composition differed significantly between populations of different colour morphs. Differences in parasite community composition were stable in time. The genetic structure of Tropheus populations was strong and showed a significant isolation-by-distance pattern, confirming that spatial isolation is limiting host dispersal. Correlations between parasite community composition and Tropheus genetic

  1. Pathosphere.org: Pathogen Detection and Characterization Through a Web-based, Open-source Informatics Platform

    DTIC Science & Technology

    2015-12-29

    human), Homo sapiens chromosome (human), Mus_musculus ( rodent ), Sus scrofa (pig), mitochondrion genome, and Xenopus laevis (frog) . The taxonomy... Amazon Web Services. PLoS Comput Biol 2011, 7:e1002147. 10. Briese T, Paweska JT, McMullan LK, Hutchison SK, Street C, Palacios G, Khristova ML...human enterovirus C genotypes found in respiratory samples from Peru . J Gen Virol 2013, 94(Pt 1):120–7. 54. Jacob ST, Crozier I, Schieffelin JS

  2. Development of the Zebra Danio Model: Carcinogenesis and Gene Transfer Studies

    DTIC Science & Technology

    1996-09-01

    J., and Enomoto, M. (1988). Liver cell carcinomas in the medaka (Oryzias latipes) induced by methylazoxymethanol-acetate. J. Comp. Path. 98, 441-452...accelerate steroid- induced cell division in Xenopus oocytes (Sadler et al., 1986). More recently, ras p21 has been implicated in the transduction of a... induced cell division in Xenopus laevis oocytes. Mol Cell Biol 6:719-722. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989). Molecular Cloning: A

  3. New species of Oswaldocruzia (Nematoda: Molineidae) and other helminths in Bolitoglossa subpalmata (Caudata:Plethodontidae) from Costa Rica.

    PubMed

    Bursey, Charles R; Goldberg, Stephen R

    2011-04-01

    Oswaldocruzia cartagoensis n. sp. (Strongylida: Molineidae) from the intestines of Bolitoglossa subpalmata (Caudata: Plethodontidae) is described and illustrated. Oswaldocruzia cartagoensis n. sp. represents the 86 th species assigned to the genus and the 39th species from the Neotropical region. It is most similar to the Neotropical species of the genus that possess type I bursa, i.e., Oswaldocruzia bonsi , Oswaldocruzia brasiliensis , Oswaldocruzia lopesi , Oswaldocruzia neghmei , and Oswaldocruzia vitti . Of these, O. bonsi, O. brasiliensis, and O. neghmei lack cervical alae, rib 4 in individuals of O. vitti reaches the edge of the bursal membrane, species of O. lopesi and O. cartagoensis can be separated on the basis of spicule structure, the blade in O. lopesi is bifurcate, and that of O. cartagoensis terminates in 6-8 fine points. In addition to the new species of Oswaldocruzia, Cosmocera parva, Cosmocera podicipinus, and acanthocephalan cystacanths were also found.

  4. Endo-parasite fauna of rodents caught in five wet markets in Kuala Lumpur and its potential zoonotic implications.

    PubMed

    Paramasvaran, S; Sani, R A; Hassan, L; Hanjeet, K; Krishnasamy, M; John, J; Santhana, R; Sumarni, M G; Lim, K H

    2009-04-01

    Rodents were collected from five wet markets (Chow Kit, Dato Keramat, Setapak, Jinjang and Kepong) in Kuala Lumpur, Federal Territory between March to April 2006. Ninety seven rats were trapped using wire traps measuring 29 x 22 x 50 cm baited with fruits, coconuts, dried fish or sweet potatoes. A total of 17 different species of parasites were identified from three species of rats out of which 11 (65%) were identified to be zoonotic. The helminths identified from the urban rats were nematodes- Capillaria hepatica, Gongylonema neoplasticum, Heterakis spumosa, Heterakis sp., Masterphorus muris, Nippostrongylus brasiliensis, Physolaptera sp., Pterogodermatis sp., Rictularia tani and Syphacia muris; cestodes- Hymenolepis nana, Hymenolepis diminuta, Hymenolepis sabnema, Hymenolepis sp., Raillietina sp. and Taenia taeniaeformis, and acanthocephalan- Moniliformis moniliformis. The following parasites are of potential medical importance: C. hepatica, G. neoplasticum, R. tani, S. muris, H. diminuta, H. nana, Raillietina sp. and T. taeniaeformis.

  5. Checklists of Parasites of Farm Fishes of Babylon Province, Iraq

    PubMed Central

    Al-Rubaie, Abdul-Razzak L.

    2016-01-01

    Literature reviews of all references concerning the parasitic fauna of fishes in fish farms of Babylon province, middle of Iraq, showed that a total of 92 valid parasite species are so far known from the common carp (Cyprinus carpio), the grass carp (Ctenopharyngodon idella), and the silver carp (Hypophthalmichthys molitrix) as well as from three freshwater fish species (Carassius auratus, Liza abu, and Heteropneustes fossilis) which were found in some fish farms of the same province. The parasitic fauna included one mastigophoran, three apicomplexans, 13 ciliophorans, five myxozoans, five trematodes, 45 monogeneans, five cestodes, three nematodes, two acanthocephalans, nine arthropods, and one mollusc. The common carp was found to harbour 81 species of parasites, the grass carp 30 species, the silver carp 28 species, L. abu 13 species, C. auratus one species, and H. fossilis one species. A host-parasite list for each fish species was also provided. PMID:27559480

  6. Host-defense peptides from skin secretions of Fraser's clawed frog Xenopus fraseri (Pipidae): Further insight into the evolutionary history of the Xenopodinae.

    PubMed

    Conlon, J Michael; Mechkarska, Milena; Kolodziejek, Jolanta; Nowotny, Norbert; Coquet, Laurent; Leprince, Jérôme; Jouenne, Thierry; Vaudry, Hubert

    2014-12-01

    Peptidomic analysis of norepinephrine-stimulated skin secretions of the tetraploid frog Xenopus fraseri Boulenger, 1905 (Pipidae) led to identification of 13 host-defense peptides. The primary structures of the peptides demonstrate that they belong to the magainin (3 peptides), peptide glycine-leucine-amide, PGLa (4 peptides), and xenopsin-precursor fragment, XPF (2 peptides) families, first identified in Xenopus laevis, together with caerulein precursor fragment-related peptides, CPF-RP (4 peptides), first identified in Silurana tropicalis. In addition, the secretions contain a molecular variant of xenopsin displaying the substitution Arg(4)→Lys compared with X. laevis xenopsin and peptide glycine-tyrosine-amide (PGYa) (GRIIPIYPEFERVFA KKVYPLY.NH2) whose function is unknown. The most potent antimicrobial peptide identified is CPF-RP-F1 (GFGSVLGKALKFGANLL.NH2) with MIC=12.5μM against Staphylococcus aureus and 50μM against Escherichia coli. On the basis of similarities in morphology and advertisement calls, X. fraseri has been placed in a species group that includes the octoploids Xenopus amieti and Xenopus andrei, and the tetraploid Xenopus pygmaeus. Cladistic analyses based upon the primary structures of magainin, PGLa, and CPF-RP peptides support a close evolutionary relationship between X. fraseri, X. amieti and X. andrei but suggest a more distant relationship with X. pygmaeus. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. An Aryl Hydrocarbon Receptor from the Salamander Ambystoma mexicanum Exhibits Low Sensitivity to 2,3,7,8-Tetrachlorodibenzo-p-dioxin.

    PubMed

    Shoots, Jenny; Fraccalvieri, Domenico; Franks, Diana G; Denison, Michael S; Hahn, Mark E; Bonati, Laura; Powell, Wade H

    2015-06-02

    Structural features of the aryl hydrocarbon receptor (AHR) can underlie species- and population-specific differences in its affinity for 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). These differences often explain variations in TCDD toxicity. Frogs are relatively insensitive to dioxin, and Xenopus AHRs bind TCDD with low affinity. Weak TCDD binding results from the combination of three residues in the ligand-binding domain: A354 and A370, and N325. Here we sought to determine whether this mechanism of weak TCDD binding is shared by other amphibian AHRs. We isolated an AHR cDNA from the Mexican axolotl (Ambystoma mexicanum). The encoded polypeptide contains identical residues at positions that confer low TCDD affinity to X. laevis AHRs (A364, A380, and N335), and homology modeling predicts they protrude into the binding cavity. Axolotl AHR bound one-tenth the TCDD of mouse AHR in velocity sedimentation analysis, and in transactivation assays, the EC50 for TCDD was 23 nM, similar to X. laevis AHR1β (27 nM) and greater than AHR containing the mouse ligand-binding domain (0.08 nM). Sequence, modeled structure, and function indicate that axolotl AHR binds TCDD weakly, predicting that A. mexicanum lacks sensitivity toTCDD toxicity. We hypothesize that this characteristic of axolotl and Xenopus AHRs arose in a common ancestor of the Caudata and Anura.

  8. Quantifying the relative contributions of different solute carriers to aggregate substrate transport

    PubMed Central

    Taslimifar, Mehdi; Oparija, Lalita; Verrey, Francois; Kurtcuoglu, Vartan; Olgac, Ufuk; Makrides, Victoria

    2017-01-01

    Determining the contributions of different transporter species to overall cellular transport is fundamental for understanding the physiological regulation of solutes. We calculated the relative activities of Solute Carrier (SLC) transporters using the Michaelis-Menten equation and global fitting to estimate the normalized maximum transport rate for each transporter (Vmax). Data input were the normalized measured uptake of the essential neutral amino acid (AA) L-leucine (Leu) from concentration-dependence assays performed using Xenopus laevis oocytes. Our methodology was verified by calculating Leu and L-phenylalanine (Phe) data in the presence of competitive substrates and/or inhibitors. Among 9 potentially expressed endogenous X. laevis oocyte Leu transporter species, activities of only the uniporters SLC43A2/LAT4 (and/or SLC43A1/LAT3) and the sodium symporter SLC6A19/B0AT1 were required to account for total uptake. Furthermore, Leu and Phe uptake by heterologously expressed human SLC6A14/ATB0,+ and SLC43A2/LAT4 was accurately calculated. This versatile systems biology approach is useful for analyses where the kinetics of each active protein species can be represented by the Hill equation. Furthermore, its applicable even in the absence of protein expression data. It could potentially be applied, for example, to quantify drug transporter activities in target cells to improve specificity. PMID:28091567

  9. acr-23 Encodes a Monepantel-Sensitive Channel in Caenorhabditis elegans

    PubMed Central

    Rufener, Lucien; Bedoni, Nicola; Baur, Roland; Rey, Samantha; Glauser, Dominique A.; Bouvier, Jacques; Beech, Robin; Sigel, Erwin; Puoti, Alessandro

    2013-01-01

    Monepantel is a member of the recently identified class of anthelmintics known as the amino-acetonitrile derivatives (AADs). Monepantel controls all major gastro-intestinal nematodes in sheep including those that are resistant to the classical anthelmintics. Previous studies have shown that the Caenorhabditis elegans acr-23 and the Haemonchus contortus Hco-mptl-1 genes may be prominent targets of monepantel. With this discovery it became possible to investigate the mode of action of monepantel in nematodes at the molecular level. In the present study, we show that a C. elegans mutant acr-23 strain is fully rescued by expressing the wild-type acr-23 gene. Moreover, we present a new mutant allele, and characterize acr-23 alleles genetically. We also show that acr-23 is expressed in body wall muscle cells, and provide therefore a possible explanation for the paralysis caused by monepantel. Furthermore, genetic evidence suggests that the chaperone RIC-3 is required for expression of full monepantel resistance. Finally, we present reconstitution of the C. elegans ACR-23 receptor in Xenopus laevis oocytes and provide direct evidence of its modulation by monepantel. Conversely, co-injection of the chaperone RIC-3 had no impact for channel reconstitution in X. laevis oocytes. These results reinforce the involvement of the ACR-23 family in the mode of action of monepantel and advance our understanding of this new class of anthelmintics. PMID:23950710

  10. Developmental origins of a novel gut morphology in frogs.

    PubMed

    Bloom, Stephanie; Ledon-Rettig, Cris; Infante, Carlos; Everly, Anne; Hanken, James; Nascone-Yoder, Nanette

    2013-05-01

    Phenotypic variation is a prerequisite for evolution by natural selection, yet the processes that give rise to the novel morphologies upon which selection acts are poorly understood. We employed a chemical genetic screen to identify developmental changes capable of generating ecologically relevant morphological variation as observed among extant species. Specifically, we assayed for exogenously applied small molecules capable of transforming the ancestral larval foregut of the herbivorous Xenopus laevis to resemble the derived larval foregut of the carnivorous Lepidobatrachus laevis. Appropriately, the small molecules that demonstrate this capacity modulate conserved morphogenetic pathways involved in gut development, including downregulation of retinoic acid (RA) signaling. Identical manipulation of RA signaling in a species that is more closely related to Lepidobatrachus, Ceratophrys cranwelli, yielded even more similar transformations, corroborating the relevance of RA signaling variation in interspecific morphological change. Finally, we were able to recover the ancestral gut phenotype in Lepidobatrachus by performing a reverse chemical manipulation to upregulate RA signaling, providing strong evidence that modifications to this specific pathway promoted the emergence of a lineage-specific phenotypic novelty. Interestingly, our screen also revealed pathways that have not yet been implicated in early gut morphogenesis, such as thyroid hormone signaling. In general, the chemical genetic screen may be a valuable tool for identifying developmental mechanisms that underlie ecologically and evolutionarily relevant phenotypic variation. © 2013 Wiley Periodicals, Inc.

  11. Developmental origins of novel gut morphology in frogs

    PubMed Central

    Bloom, Stephanie; Ledon-Rettig, Cris; Infante, Carlos; Everly, Anne; Hanken, James; Nascone-Yoder, Nanette

    2013-01-01

    SUMMARY Phenotypic variation is a prerequisite for evolution by natural selection, yet the processes that give rise to the novel morphologies upon which selection acts are poorly understood. We employed a chemical genetic screen to identify developmental changes capable of generating ecologically relevant morphological variation as observed among extant species. Specifically, we assayed for exogenously applied small molecules capable of transforming the ancestral larval foregut of the herbivorous Xenopus laevis to resemble the derived larval foregut of the carnivorous Lepidobatrachus laevis. Appropriately, the small molecules that demonstrate this capacity modulate conserved morphogenetic pathways involved in gut development, including downregulation of retinoic acid (RA) signaling. Identical manipulation of RA signaling in a species that is more closely related to Lepidobatrachus, Ceratophrys cranwelli, yielded even more similar transformations, corroborating the relevance of RA signaling variation in interspecific morphological change. Finally, we were able to recover the ancestral gut phenotype in Lepidobatrachus by performing a reverse chemical manipulation to upregulate RA signaling, providing strong evidence that modifications to this specific pathway promoted the emergence of a lineage-specific phenotypic novelty. Interestingly, our screen also revealed pathways that have not yet been implicated in early gut morphogenesis, such as thyroid hormone signaling. In general, the chemical genetic screen may be a valuable tool for identifying developmental mechanisms that underlie ecologically and evolutionarily relevant phenotypic variation. PMID:23607305

  12. Jeltraxin, a frog egg jelly glycoprotein, has calcium-dependent lectin properties and is related to human serum pentraxins CRP and SAP.

    PubMed

    Peavy, Thomas R; Hernandez, Cesar; Carroll, Edward J

    2003-11-11

    The egg jelly that encapsulates amphibian eggs is essential for fertilization, but its molecular composition and roles remain largely unknown. We identified a calcium-dependent lectin from the pentraxin superfamily in the egg jelly coat from the South American burrowing frog, Lepidobatrachus laevis. This lectin, jeltraxin, was related to the host-response acute phase serum proteins C-reactive P component (CRP) and serum amyloid P component (SAP). The amino acid sequence of jeltraxin is 44% identical to that of Xenopus laevis CRP, 31-35% identical to those of mammalian CRP and SAP, and 21-27% identical to those of the large fusion pentraxins. Expression of jeltraxin mRNA was restricted to the oviduct, which distinguishes it as the first serum-related pentraxin not expressed in the liver. Purified jeltraxin was previously shown to exist in an oligomeric complex of approximately 250 kDa comprised of self-associating subunits. We have demonstrated by MALDI-TOF that this configuration is due to a decameric complex of 27.7 kDa subunits. Biotinylated jeltraxin bound to the high-molecular mass components of the egg jelly in a calcium-dependent manner with specificity for beta-galactose residues. On the basis of homology modeling, we predict that jeltraxin will coordinate two calcium ions. The function of jeltraxin will likely be related to its calcium-dependent lectin properties.

  13. The effect of long-term corticosterone treatment on blood cell differentials and function in laboratory and wild-caught amphibian models.

    PubMed

    Falso, Paul G; Noble, Christopher A; Diaz, Jesus M; Hayes, Tyrone B

    2015-02-01

    The effect of long-term stress on amphibian immunity is not well understood. We modeled a long-term endocrine stress scenario by elevating plasma corticosterone in two species of amphibians and examined effects on white blood cell differentials and innate immune activity. Plasma corticosterone was elevated in American bullfrogs (Lithobates catesbeianus) by surgically implanting corticosterone capsules and in African clawed frogs (Xenopus laevis) by immersion in corticosterone-treated water. To provide a context for our results within endogenous corticosterone fluctuations, diurnal plasma corticosterone cycles were determined. A daily low of corticosterone was observed in X. laevis at 12:00, while a significant pattern was not observed in L. catesbeianus. Elevated plasma corticosterone levels increased the ratio of peripheral neutrophils to lymphocytes, in both species, and decreased eosinophil concentrations in L. catesbeianus over a long-term period. Whole blood oxidative burst generally correlated with neutrophil concentrations, and thus was increased with corticosterone treatment, significantly in L. catesbeianus. In L. catesbeianus, an endogenous response of eosinophils and lymphocytes to implanted empty (sham) capsules was observed, but this effect was attenuated by corticosterone. Peripheral monocyte and basophil concentrations were not significantly altered by corticosterone treatment in either species. Our results show that long-term stress can alter amphibian immune parameters for extended periods and may play a role in susceptibility to disease. Copyright © 2015 Elsevier Inc. All rights reserved.

  14. Conodont biostratigraphy from the upper San Juan Formation (Middle Ordovician) at Niquivil, Argentine Precordillera

    NASA Astrophysics Data System (ADS)

    Mango, M. J.; Albanesi, G. L.

    2018-07-01

    The present work deals with the conodont biostratigraphy from the upper San Juan Formation in the section of Niquivil, Central Precodillera of San Juan, Argentina. We study the upper 129.45 m of the San Juan Formation, starting from the upper strata of the second reef horizon up to the top of the formation. Digested limestone samples yielded 20 conodont species. The presence of Tripodus laevis Bradshaw not associated to Baltoniodus navis (Lindström) allows the recognition of the Baltoniodus triangularis-Tripodus laevis Zone, which is interpreted as correlative with the "Parapanderodus" nogamii/Parapanderodus gracilis/Ansella jemtlandica Association of Lehnert (1993, 1995; Lehnert and Keller, 1993), conversely to previous interpretations that suggested the latter as correlative to the Baltoniodus navis Zone. The zonal identification is supported by the associated conodonts Protopanderodus rectus (Lindström), Juanognathus jaanussoni Serpagli, Juanognathus n. sp., Protopanderodus gradatus (Serpagli), Rossodus barnesi Albanesi, Paltodus subaequalis Pander, Drepanodus arcuatus (Pander), Cornuodus longibasis (Lindström), Protopanderodus elongatus Serpagli, Oistodus lanceolatus Pander, Periodon flabellum (Lindström), Semiacontiodus potrerillensis Albanesi, Triangulodus brevibasis (Sergeeva), Paroistodus originalis (Sergeeva), Drepanoistodus forceps (Lindström), Oistodus multicorrugatus Harris, Parapanderodus paracornuformis (Ethington and Clark), Anodontus longus Stouge and Bagnoli, and Pteracontiodus cryptodens Mound. The petrographic microscope analysis of carbonate rocks thin sections refer to proximal middle ramp deposits.

  15. A nuclear F-actin scaffold stabilizes RNP droplets against gravity in large cells

    PubMed Central

    Feric, Marina; Brangwynne, Clifford P.

    2013-01-01

    The size of a typical eukaryotic cell is on the order of ≈10 μm. However, some cell types grow to very large sizes, including oocytes (immature eggs) of organisms from humans to starfish. For example, oocytes of the frog X. laevis grow to a diameter ≥1 mm. They contain a correspondingly large nucleus (germinal vesicle, GV) of ≈450 μm in diameter, which is similar to smaller somatic nuclei, but contains a significantly higher concentration of actin. The form and structure of this nuclear actin remain controversial, and its potential mechanical role within these large nuclei is unknown. Here, we use a microrheology and quantitative imaging approach to show that GVs contain an elastic F-actin scaffold that mechanically stabilizes these large nuclei against gravitational forces, which are usually considered negligible within cells. We find that upon actin disruption, RNA/protein droplets, including nucleoli and histone locus bodies (HLBs), undergo gravitational sedimentation and fusion. We develop a model that reveals how gravity becomes an increasingly potent force as cells and their nuclei grow larger than ≈10 μm, explaining the requirement for a stabilizing nuclear F-actin scaffold in large X. laevis ooctyes. All life forms are subject to gravity, and our results may have broad implications for cell growth and size control. PMID:23995731

  16. Methodological issues affecting the study of fish parasites. III. Effect of fish preservation method.

    PubMed

    Kvach, Yuriy; Ondračková, Markéta; Janáč, Michal; Jurajda, Pavel

    2018-03-05

    The aim of this study was to evaluate the influence of preservation method on the results of parasite community studies. Two host species, European perch Perca fluviatilis and European bitterling Rhodeus amarus, were examined for parasites after having been subjected to 4 different storage treatments: freezing, preservation in 4% formaldehyde or 70% ethanol and transportation of live (fresh) fish as a control. Preservation prior to dissection resulted in a loss of information, leading to incomplete quantitative data (all preservation treatments), qualitative data (ethanol and formaldehyde preservation) and a lowered ability to determine parasites to species level based on morphology compared to dissecting fresh fish. Of the more abundant taxa, only crustaceans and acanthocephalans provided relatively even results between treatments. We conclude that preservation media, such as ethanol or formaldehyde, significantly affects the ability to obtain precise parasite community data; hence, we recommend the use of freshly sacrificed fish for parasite community studies whenever possible. Alternatively, freezing may prove acceptable for evaluating parasite community taxonomic composition.

  17. Helminth parasites of the raccoon (Procyon lotor) from north-central Arkansas.

    PubMed

    Richardson, D J; Owen, W B; Snyder, D E

    1992-02-01

    Twenty-three species of helminths (4 trematodes, 2 cestodes, 14 nematodes, and 3 acanthocephalans) were found in the gastrointestinal tract, lungs, pancreas, tongue, urinary bladder, and subcutaneous tissues of 30 live-trapped or hunter-shot raccoons from north-central Arkansas between November 1989 and April 1990. Helminths were not detected in the brain, diaphragm, gallbladder, heart, liver, reproductive system, or trachea. Each raccoon examined was infected with 3 or more of the following helminths: Brachylaima virginiana, Eurytrema procyonis, Fibricola cratera, Pharyngostomoides procyonis, Atriotaenia procyonis, Mesocestoides spp., Arthrocephalus lotoris, Capillaria aerophila, Capillaria plica, Capillaria procyonis, Capillaria putorii, Crenosoma goblei, Cruzia americana, Dirofilaria tenuis, Dracunculus insignis, Enterobius sp., Gnathostoma procyonis, Molineus barbatus, Physaloptera rara, Trichinella spiralis, Centrorhynchus wardae, Macracanthorhynchus ingens, and Oligacanthorhynchus tortuosa. All helminths collected with the exception of D. insignis constitute new geographic distribution records. Occurrences of C. aerophila, C. wardae, and O. tortuosa are new host records. One nymphal pentastome, Porocephalus crotali, was found in the liver of 1 raccoon, constituting a new host record.

  18. Reef fishes have higher parasite richness at unfished Palmyra Atoll compared to fished Kiritimati Island

    USGS Publications Warehouse

    Lafferty, Kevin D.; Shaw, Jenny C.; Kuris, Armand M.

    2008-01-01

    We compared parasite communities at two coral atolls in the Line Islands chain of the central Pacific (Kiritimati Island and Palmyra Atoll). Palmyra Atoll is relatively pristine while Kiritimati Island is heavily fished. At each island, we sampled five fish species for helminth and arthropod endoparasites: Chromis margaritifer, Plectroglyphidodon dickii,Paracirrhites arcatus, Acanthurus nigricans, and Lutjanus bohar. The surveys found monogeneans, digeneans, cestodes, nematodes, acanthocephalans, and copepods. Parasite richness was higher at Palmyra compared to Kiritimati for all five fish species. Fishes from Palmyra also tended to have more parasites species per host, higher parasite prevalence, and higher parasite abundance than did fishes from Kiritimati. The lower parasitism at Kiritimati may result from a simplified food web due to over fishing. Low biodiversity could impair parasite transmission by reducing the availability of hosts required by parasites with complex life cycles. Most notably, the lower abundances of larval shark tapeworms at Kiritimati presumably reflect the fact that fishing has greatly depleted sharks there in comparison to Palmyra.

  19. The morphology and histopathology of Nephridiacanthus major (Acanthocephala: Oligacanthorhynchidae) from hedgehogs in Iran.

    PubMed

    Heckmann, Richard A; Amin, Omar M; Halajian, Ali; El-Naggar, Atif M

    2013-02-01

    The morphology of Nephridiacanthus major (Bremser 1811 in Westrumb 1821) Golvan, 1962 collected from the long-eared hedgehog Hemiechinus auritus (Gmelin 1770) and the Eastern European hedgehog Erinaceus concolor Martin, 1838 (Erinaceidae) is described using SEM for the first time. This acanthocephalan was previously described from hedgehogs in Europe, Asia, and Africa. Measurements of specimens from Iran, Bulgaria, Germany, Central Asia, Morocco, and Egypt show considerable variations in the size of the trunk, proboscis, proboscis hooks and receptacle, and eggs. The SEM studies add new perspectives to its morphology. Features observed for the first time include the near terminal position and shape of the female gonopore and orifice, among others. Histopathological studies for this species are reported for the first time. Tissue sections show extensive damage near the proboscis with hemorrhaging and formation of collagenous connective tissue, compression of the intestinal mucosa, obstruction of intestinal lumen, and extensive necrosis of host epithelial tissue.

  20. Helminths of foxes and other wild carnivores from rural areas in Greece.

    PubMed

    Papdopoulos, H; Himonas, C; Papazahariadou, M; Antoniadou-Sotiriadou, K

    1997-09-01

    Twenty species of helminth parasites were identified from fox, wolf, jackal and wild cat material collected in Greece. Of the 314 foxes (Vulpes vulpes) examined, 18 helminth species were recovered comprising one trematode, eight cestodes, seven nematodes and two acanthocephalans, with the cestode species Mesocestoides sp. (73.2%), Joyeuxiella echinorhynchoides (24.5%) and the nematode species Uncinaria stenocephala (43.9%), and Toxara canis (28.6%) being the most prevalent. Five cestode and three nematode species were reported from six wolves (CaniS lupus), together with one trematode, three cestode and four nematode species from five jackals (Canis aureus) and two cestode and three nematode species from four wild cats (Felis silvestris) examined. The species J. echinorhynchoides, Taenia crassiceps and Onicola canis and the genera Spirometra, Rictularia and Pachysentis are reported here for the first time in Greece. The results are discussed in the light of the feeding characteristics of wild carnivores in rural areas of Greece.