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1

Tracking and predation on earthworms by the invasive terrestrial planarian Bipalium adventitium (Tricladida, Platyhelminthes).  

PubMed

The potential ecological impact of exotic terrestrial planarians will be determined in part by their sensory abilities and predatory behavior. It has been suggested that these flatworms may only encounter their earthworm prey by chance, hence restricting the breadth of species they will feed upon and the number of microhabitats in which predator-prey interactions occur. We hypothesized that those flatworms that have already successfully invaded North America (genus Bipalium) actually detect and follow chemical trails of earthworms and possess the behavioral repertoire needed to feed on the prey in a range of microhabitats. We examined: (1) the tendency of Bipalium adventitium to follow chemical trails left by injured and un-injured earthworms; (2) the behavioral repertoire and predatory success of B. adventitium feeding on three earthworm species in subterranean tunnels; and (3) the response of flatworms to the reportedly defensive secretions of the earthworm Eisenia fetida in tunnels. B. adventitium detected and followed trails of earthworm mucus and secretions left by injured and un-injured earthworms. Flatworms followed trails on a range of substrates and pursued and captured three species of earthworms in subterranean tunnels, including individuals many times their mass. Although most behavior exhibited during underground attacks was similar to that reported for surface encounters, the flatworms also behaved in ways that blocked earthworm escape from tunnels. The flatworms were less successful at preying on E. fetida than on Lumbricus rubellus and Lumbricus terrestris in underground tunnels and showed some aversion to the secretions from E. fetida. PMID:15518983

Fiore, Cara; Tull, Jamie L; Zehner, Sean; Ducey, Peter K

2004-11-30

2

Occurrence and abundance of a mariner-like element in freshwater and terrestrial planarians (Platyhelminthes, Tricladida) from southern Brazil  

PubMed Central

Transposable elements are DNA sequences present in all the large phylogenetic groups, both capable of changing position within the genome and constituting a significant part of eukaryotic genomes. The mariner family of transposons is one of the few which occurs in a wide variety of taxonomic groups, including freshwater planarians. Nevertheless, so far only five planarian species have been reported to carry mariner-like elements (MLEs), although several different species have been investigated. Regarding the number of copies of MLEs, Girardia tigrina is the only planarian species in which this has been evaluated, with an estimation of 8,000 copies of the element per haploid genome. Preliminary results obtained in our laboratory demonstrated that MLE is found in a large number of different species of planarians, including terrestrial. With this in mind, the aim was to evaluate the occurrence and estimate the number of MLE copies in different planarian species collected in south Brazil. Twenty-eight individuals from 15 planarian species were analyzed. By using PCR and the hybridization of nucleic acids, it was found that MLE was present in all the analyzed species, the number of copies being high, probably over 103 per haploid genome.

2009-01-01

3

Biodiversity of Australian freshwater planarians (Platyhelminthes: Tricladida: Paludicola): New species and localities, and a review of paludicolan distribution in Australia  

Microsoft Academic Search

On the basis of one extensive collection, the study provides new information on the diversity, taxonomy, anatomy and geographic distribution of 21 species of Australian freshwater planarians, including 7 species that are described as new. The material includes old type specimens of three species that have remained enigmatic since their collection and description almost 100 years ago, viz. Planaria rava

Lauryne J. Grant; Ronald Sluys; David Blair

2006-01-01

4

Global diversity of land planarians (Platyhelminthes, Tricladida, Terricola): a new indicator-taxon in biodiversity and conservation studies  

Microsoft Academic Search

Biodiversity conservation requires prioritization of areas for in situ conservation. In that perspective, the present study documents the global diversity of a component of the soil macrofauna, the land planarians, and concerns an exploratory analysis of their possible role as indicators of biodiversity. Diversity is described by three quantitative methods: (1) hotspots of species richness, selecting areas richest in species,

Ronald Sluys

1999-01-01

5

Integrative taxonomy of a new species of planarian from the Lake Ohrid basin, including an analysis of biogeographical patterns in freshwater triclads from the Ohrid region (Platyhelminthes, Tricladida, Dugesiidae)  

PubMed Central

Abstract A new species of the genus Dugesia is described from the Lake Ohrid region in the western part of the Balkan Peninsula, forming the first fully documented species description for this genus in the Ohrid area. The morphological species delimitation is supported by complementary molecular, karyological, and cytogenetic data available from the literature. Therefore, species delineation is based on a truly integrative approach. Further, a short account on the degree of freshwater planarian endemicity in the Ohrid region is provided.

Stocchino, Giacinta Angela; Sluys, Ronald; Deri, Paolo; Manconi, Renata

2013-01-01

6

Ultrastructural and cytochemical aspects of the female gonad of Geoplana burmeisteri (Platyhelminthes, Tricladida, Terricola).  

PubMed

The ultrastructure of the female gonad of the land planarian Geoplana burmeisteri was investigated by means of electron microscopy and cytochemical techniques. It consists of two small germaria located ventral to the intestine and of two irregular, lateral rows of vitelline follicles, both enveloped by a tunica composed of an extracellular lamina and an inner sheath of accessory cells. Accessory cell projections completely surround developing oocytes and vitellocytes. The main feature of oocyte maturation is the appearance of chromatoid bodies and the development of the rough endoplasmic reticulum (RER) and Golgi complexes. These organelles appear to be correlated with the production of egg inclusions of medium electron density, about 1.5-1.8 microm in diameter, which remain scattered in the ooplasm of mature oocytes. On the basis of cytochemical tests demonstrating their glycoprotein composition, these inclusions were interpreted as residual yolk globules. Vitellocytes are typical secretory cells with well-developed RER and Golgi complexes that are mainly involved in the production of yolk globules and eggshell globules, respectively. Eggshell globules appear to arise from repeated coalescence of small Golgi-derived vesicles and, at an intermediate stage of maturation, show a multigranular pattern. Later, after vesicle fusion, they reach a diameter of 1.3-1.6 microm when completely mature and show a meandering/concentric pattern, as is typical of the situation seen in most Proseriata and Tricladida. The content of yolk globules is completely digested by pronase, while the content of eggshell globules is unaffected. Mature vitellocytes contain, in addition, a large quantity of glycogen and lipid droplets as further reserve material. On the basis of the ultrastructural characteristics of the female gonad described above and in relation to the current literature, we conclude that G. burmeisteri appears to be more closely related to the freshwater triclads, in particular to members of the Dugesiidae, than to the marine triclads. PMID:16323219

Falleni, Alessandra; Lucchesi, Paolo; Ghezzani, Claudio; Silveira, Marina; Gremigni, Vittorio

2006-03-01

7

The power of regeneration and the stem-cell kingdom: freshwater planarians (Platyhelminthes).  

PubMed

The great powers of regeneration shown by freshwater planarians, capable of regenerating a complete organism from any tiny body fragment, have attracted the interest of scientists throughout history. In 1814, Dalyell concluded that planarians could "almost be called immortal under the edge of the knife". Equally impressive is the developmental plasticity of these platyhelminthes, including continuous growth and fission (asexual reproduction) in well-fed organisms, and shrinkage (degrowth) during prolonged starvation. The source of their morphological plasticity and regenerative capability is a stable population of totipotent stem cells--"neoblasts"; this is the only cell type in the adult that has mitotic activity and differentiates into all cell types. This cellular feature is unique to planarians in the Bilateria clade. Over the last fifteen years, molecular studies have begun to reveal the role of developmental genes in regeneration, although it would be premature to propose a molecular model for planarian regeneration. Genomic and proteomic data are essential in answering some of the fundamental questions concerning this remarkable morphological plasticity. Such information should also pave the way to understanding the genetic pathways associated with metazoan somatic stem-cell regulation and pattern formation. PMID:16615086

Saló, Emili

2006-05-01

8

[Telomere length and phylogenetic relationship of Baikal and Siberian planarians (Turbellaria, Tricladida)].  

PubMed

Dynamics of the telomeric DNA (tDNA) and the phylogeny of the Baikal and Siberian planarians have been studied based on the analysis of the 18S rDNA and beta-actin gene fragments. A relationship between tDNA and the planarians size has been demonstrated. Giant planarians with a minor exception have longer tDNA than little planarians. Phylogenetic affinity between the species that have the stretched tracks of tDNA, big size and similar habitats may indicate possible role of tDNA in the development of the indefinite regenerative capacity of planarians. PMID:23875458

Koroleva, A G; Evtushenko, E V; Timoshkin, O A; Vershinin, A V; Kiril'chik, S V

2013-01-01

9

Chromosomal polymorphism in planarians (Turbellaria, Tricladida) and the plate tectonics of the western Mediterranean  

Microsoft Academic Search

In two populations of Dugesia (Schmidtea) mediterranea the third chromosome pair consists of two heteromorphous elements. This chromosomal polymorphism, which is common in other animal groups, was hitherto unknown in planarians.

E. J. De Vries; J. Baguńŕ; I. R. Ball

1984-01-01

10

Fine-scale differences in diel activity among nocturnal freshwater planarias (Platyhelminthes: Tricladida)  

PubMed Central

Background Although most freshwater planarias are well known photonegative organisms, their diel rhythms have never been quantified. Differences in daily activity rhythms may be particularly important for temperate-climate, freshwater planarias, which tend to overlap considerably in spatial distribution and trophic requirements. Methods Activity of stress-free, individually tested young adults of three common planarian species was recorded at 3-h intervals in a 10-d experiment under natural sunlight and photoperiod during autumnal equinox (D:L ~12:12). Individual activity status was averaged over the 10-d experiment, each tested individual thus serving as a true replicate. Twelve individuals per species were tested. Food was provided every 36 h, resulting in alternating day- and nighttime feeding events. Activity during the first post-feeding h was recorded and analyzed separately. Statistical procedures included ANOVAs, correlations, and second-order analyses of angles. Results Dugesia (= Girardia) tigrina Girard 1850 exhibited clear nocturnal behavior, Dugesia (= Schmidtea) polychroa Schmidt 1861 was predominantly but not exclusively nocturnal, and Polycelis tenuis Ijima 1884 was relatively more active from midnight through noon. Species-specific activity peaks were statistically similar, with peaks at dawn for P. tenuis and just before midnight for the two dugesiids; however, D. tigrina was comparatively more active in the early night hours, while D. polychroa was more active than D. tigrina during daytime. D. tigrina also responded less readily to daytime food addition. P. tenuis remained poorly active and unresponsive throughout the experiment. Individual variability in diel behavior was highest for D. polychroa and lowest for D. tigrina. P. tenuis's general low degree of activity and late activity peak in the experiment may be related to a strong reliance on external stimuli. Conclusions The tested species are mainly nocturnal, consistent with their photonegative characteristics. The fine-scale differences in diel behavior among these three triclad species may not be sufficient to allow coexistence in the wild, with the nonnative D. tigrina eventually displacing D. polychroa and P. tenuis in many European waters. The link between planarian diel rhythms and ecological characteristics are worth of further, detailed investigation.

2011-01-01

11

Prey-tracking behavior in the invasive terrestrial planarian Platydemus manokwari (Platyhelminthes, Tricladida)  

NASA Astrophysics Data System (ADS)

Platydemus manokwari is a broadly distributed invasive terrestrial flatworm that preys heavily on land snails and has been credited with the demise of numerous threatened island faunas. We examined whether P. manokwari tracks the mucus trails of land snail prey, investigated its ability to determine trail direction, and evaluated prey preference among various land snail species. A plastic treatment plate with the mucus trail of a single species and a control plate without the trail were placed side by side at the exit of cages housing P. manokwari. All trials were then videotaped overnight. The flatworms moved along plates with mucus trails, but did not respond to plates without trails, blank control (distilled water), or with conspecific flatworm trails. When presented at the midpoint of a snail mucus trail, the flatworms followed the trail in a random direction. The flatworms showed a preference when choosing between two plates, each with a mucus trail of different land snail species. Our results suggest that P. manokwari follows snail mucus trails based on chemical cues to increase the chance of encountering prey; however, trail-tracking behavior showed no directionality.

Iwai, Noriko; Sugiura, Shinji; Chiba, Satoshi

2010-11-01

12

Prey-tracking behavior in the invasive terrestrial planarian Platydemus manokwari (Platyhelminthes, Tricladida).  

PubMed

Platydemus manokwari is a broadly distributed invasive terrestrial flatworm that preys heavily on land snails and has been credited with the demise of numerous threatened island faunas. We examined whether P. manokwari tracks the mucus trails of land snail prey, investigated its ability to determine trail direction, and evaluated prey preference among various land snail species. A plastic treatment plate with the mucus trail of a single species and a control plate without the trail were placed side by side at the exit of cages housing P. manokwari. All trials were then videotaped overnight. The flatworms moved along plates with mucus trails, but did not respond to plates without trails, blank control (distilled water), or with conspecific flatworm trails. When presented at the midpoint of a snail mucus trail, the flatworms followed the trail in a random direction. The flatworms showed a preference when choosing between two plates, each with a mucus trail of different land snail species. Our results suggest that P. manokwari follows snail mucus trails based on chemical cues to increase the chance of encountering prey; however, trail-tracking behavior showed no directionality. PMID:20853096

Iwai, Noriko; Sugiura, Shinji; Chiba, Satoshi

2010-09-19

13

Freshwater planarians from artesian springs in Queensland, Australia (Platyhelminthes, Tricladida, Paludicola)  

Microsoft Academic Search

Two new species of triclad flatworm are described from artesian springs in Queensland, Australia, viz. Dugesia artesiana Sluys and Grant, sp. nov. and Weissius capaciductus Sluys, gen. et sp. nov. Some historical biogeographic scenarios are discussed that may explain the occurrence of the new species and their close relatives in Australia.

R. Sluys; L. J. Grant; D. Blair

2007-01-01

14

Ultrastructure of spermatogenesis and mature spermatozoon of the freshwater planarian Schmidtea mediterranea (Platyhelminthes, Paludicola).  

PubMed

The aim of this study was to characterize for the first time spermatogenesis and spermiogenesis in the freshwater planarian Schmidtea mediterranea using both light and electron microscopy. Starting from the border towards the testis lumen we found types I and II spermatogonia, clusters of primary and secondary spermatocytes, spermatids and free spermatozoa. Light microscope observations show that type I spermatogonia have a large and pale nucleus whereas type II spermatogonia are significantly smaller than the one of type I, and show a darker and central bulky nucleus. At the ultrastructure level, both type I and type II spermatogonia are characterized by a wide nucleus with scanty cytoplasm containing free ribosomes, mitochondria and a dense chromatoid body whereas endoplasmic reticulum and Golgi complex were not observed. The cytoplasm of primary and secondary spermatocytes displays numerous free ribosomes and many endoplasmic reticulum cisternae and Golgi complexes, suggesting that the development of these organelles during spermatogenesis might contribute to the synthesis of hormones and proteins such as testosterone, transcription factors and tubulin. Mature spermatozoa structure closely resembles those of other freshwater triclads with a nucleus, a single fused mitochondrion, a row of cortical microtubules and a pair of flagella conforming to the 9+'1' microtubule pattern described for other Platyhelminthes. PMID:22325561

Harrath, Abdul Halim; Alwasel, Saleh; Zghal, Fathia; Tekaya, Saďda

2012-01-09

15

Comparative transcriptome analysis between planarian Dugesia japonica and other platyhelminth species  

PubMed Central

Background Planarians are considered to be among the extant animals close to one of the earliest groups of organisms that acquired a central nervous system (CNS) during evolution. Planarians have a bilobed brain with nine lateral branches from which a variety of external signals are projected into different portions of the main lobes. Various interneurons process different signals to regulate behavior and learning/memory. Furthermore, planarians have robust regenerative ability and are attracting attention as a new model organism for the study of regeneration. Here we conducted large-scale EST analysis of the head region of the planarian Dugesia japonica to construct a database of the head-region transcriptome, and then performed comparative analyses among related species. Results A total of 54,752 high-quality EST reads were obtained from a head library of the planarian Dugesia japonica, and 13,167 unigene sequences were produced by de novo assembly. A new method devised here revealed that proteins related to metabolism and defense mechanisms have high flexibility of amino-acid substitutions within the planarian family. Eight-two CNS-development genes were found in the planarian (cf. C. elegans 3; chicken 129). Comparative analysis revealed that 91% of the planarian CNS-development genes could be mapped onto the schistosome genome, but one-third of these shared genes were not expressed in the schistosome. Conclusions We constructed a database that is a useful resource for comparative planarian transcriptome studies. Analysis comparing homologous genes between two planarian species showed that the potential of genes is important for accumulation of amino-acid substitutions. The presence of many CNS-development genes in our database supports the notion that the planarian has a fundamental brain with regard to evolution and development at not only the morphological/functional, but also the genomic, level. In addition, our results indicate that the planarian CNS-development genes already existed before the divergence of planarians and schistosomes from their common ancestor.

2012-01-01

16

Sex-inducing effect of a hydrophilic fraction on reproductive switching in the planarian Dugesia ryukyuensis (Seriata, Tricladida)  

PubMed Central

Background The mechanisms underlying the switching from an asexual to a sexual mode of reproduction, and vice versa, remain unknown in metazoans. In planarians, asexual worms acquire cryptic sexuality when fed with sexual worms, indicating that sexual worms contain a sex-inducing substance. Although such a chemical compound will provide clues about the mechanisms underlying the switching, information on the sex-inducing substance is poor. In order to identify this substance, we have established an assay system for sexual induction in asexual worms of Dugesia ryukyuensis by feeding them with sexual worms. Here, we carried out an isolation study on the sex-inducing substance using this assay system. Results After centrifugation of sexual worms homogenised in saline solution, we found that not only did the precipitate have a sex-inducing effect on the asexual worms, which has been shown previously, but the cytosolic fraction did as well. We confirmed that the sex-inducing activity in the cytosolic fraction was recovered in a hydrophilic fraction separated on an octadecylsilane (ODS) column. We showed that the sex-inducing substance in the hydrophilic fraction is papain-resistant and a putative low-molecular-weight compound of less than 500. We also suggest the presence of an enhancer of sexual induction with a molecular weight (MW) of more than 5 K in the hydrophilic fraction. Conclusion Our experiments showed the existence of a sex-inducing substance and an enhancer of sex-induction in a hydrophilic fraction, and a putative hydrophobic sex-inducing substance in the precipitate. Sexual induction in the asexual worms might be triggered by additive or synergistic effects of these chemical compounds.

2011-01-01

17

Planarian homeobox genes: cloning, sequence analysis, and expression.  

PubMed Central

Freshwater planarians (Platyhelminthes, Turbellaria, and Tricladida) are acoelomate, triploblastic, unsegmented, and bilaterally symmetrical organisms that are mainly known for their ample power to regenerate a complete organism from a small piece of their body. To identify potential pattern-control genes in planarian regeneration, we have isolated two homeobox-containing genes, Dth-1 and Dth-2 [Dugesia (Girardia) tigrina homeobox], by using degenerate oligonucleotides corresponding to the most conserved amino acid sequence from helix-3 of the homeodomain. Dth-1 and Dth-2 homeodomains are closely related (68% at the nucleotide level and 78% at the protein level) and show the conserved residues characteristic of the homeodomains identified to data. Similarity with most homeobox sequences is low (30-50%), except with Drosophila NK homeodomains (80-82% with NK-2) and the rodent TTF-1 homeodomain (77-87%). Some unusual amino acid residues specific to NK-2, TTF-1, Dth-1, and Dth-2 can be observed in the recognition helix (helix-3) and may define a family of homeodomains. The deduced amino acid sequences from the cDNAs contain, in addition to the homeodomain, other domains also present in various homeobox-containing genes. The expression of both genes, detected by Northern blot analysis, appear slightly higher in cephalic regions than in the rest of the intact organism, while a slight increase is detected in the central period (5 days) or regeneration. Images

Garcia-Fernandez, J; Baguna, J; Salo, E

1991-01-01

18

Planarian Hox genes: novel patterns of expression during regeneration.  

PubMed

Platyhelminthes are widely considered to be the sister group of coelomates (Philippe, H., Chenuil, A. and Adoutte, A. (1994)Development 1994 Supplement, 15-24) and the first organisms to show bilateral symmetry and cephalization. Within this phylum, the freshwater planarians (Turbellaria, Tricladida) have been used as model systems for studying bidirectional regeneration (Slack, J. M. W. (1980) J. Theor. Biol 82, 105-140). We have been attempting to identify potential pattern-control genes involved in the regeneration of planarian heads and tails after amputation. Since Hox cluster genes determine positional identity along the anteroposterior axis in a wide range of animals (Slack, J. M. W., Holland, P. W. H. and Graham, C. F. (1993) Nature 361,490-492), we performed an extensive search for Hox-related genes in the planarian Dugesia(G)tigrina. Sequence analyses of seven planarian Dthox genes (Dthox-A to Dthox-G) reveal high similarities with the homeodomain region of the Hox cluster genes, allowing us to assign planarian Dthox genes to anterior and medial Hox cluster paralogous groups. Whole-mount in situ hybridization studies in regenerating adults showed very early, synchronous and colocalized activation of Dthox-D, Dthox-A, Dthox-C, Dthox-E, Dthox-G and Dthox-F. After one hour of regeneration a clear expression was observed in all Dthox genes studied. In addition, all seemed to be expressed in the same regenerative tissue, although in the last stages of regeneration (9 to 15 days) a differential timing of deactivation was observed. The same Dthox genes were also expressed synchronously and were colocalized during intercalary regeneration, although their expression was delayed. Terminal regeneration showed identical Dthox gene expression in anterior and posterior blastemas, which may prevent these genes from directing the distinction between head and tail. Finally, continuous expression along the whole lateral blastema in sagittal regenerates reflected a ubiquitous Dthox response in all types of regeneration that was not related specifically with the anteroposterior polarity. PMID:9006075

Bayascas, J R; Castillo, E; Muńoz-Mármol, A M; Saló, E

1997-01-01

19

Phylogenetic analysis of the Argonaute protein family in platyhelminths.  

PubMed

Argonaute proteins (AGOs) are mediators of gene silencing via recruitment of small regulatory RNAs to induce translational regression or degradation of targeted molecules. Platyhelminths have been reported to express microRNAs but the diversity of AGOs in the phylum has not been explored. Phylogenetic relationships of members of this protein family were studied using data from six platyhelminth genomes. Phylogenetic analysis showed that all cestode and trematode AGOs, along with some triclad planarian AGOs, were grouped into the Ago subfamily and its novel sister clade, here referred to as Cluster 1. These were very distant from Piwi and Class 3 subfamilies. By contrast, a number of planarian Piwi-like AGOs formed a novel sister clade to the Piwi subfamily. Extensive sequence searching revealed the presence of an additional locus for AGO2 in the cestode Echinococcus granulosus and exon expansion in this species and E. multilocularis. The current study suggests the absence of the Piwi subfamily and Class 3 AGOs in cestodes and trematodes and the Piwi-like AGO expansion in a free-living triclad planarian and the occurrence of exon expansion prior to or during the evolution of the most-recent common ancestor of the Echinococcus species studied. PMID:23211720

Zheng, Yadong

2012-12-02

20

Comparative analysis of known miRNAs across platyhelminths.  

PubMed

MicroRNAs (miRNAs) are a subtype of small regulatory RNAs that are involved in numerous biological processes through small RNA-induced silencing networks. In an attempt to explore the phylogeny of miRNAs across five platyhelminths, we integrated annotated miRNAs and their full genomes. We identified conserved miRNA clusters and, in particular, miR-71/2 was conserved from planarian to parasitic flatworms and was expanded in free-living Schmidtea mediterranea. Analysis of 22 miRNA loci provided compelling evidence that most known miRNAs are conserved across platyhelminths. Meanwhile, we also observed alterations of known protein-coding genes flanking miRNA(s), such as transcriptional direction conversion and locus relocation, in around ~ 41% of 22 known miRNA loci. Compared with Echinococcus multilocularis, the majority of these events occurred in evolution-distant Hymenolepis microstoma, Schistosoma japonicum or/and S. mediterranea. These results imply rearrangement events occurred near the known miRNA loci. PMID:23777576

Jin, Xiaoliang; Lu, Lixia; Su, Hailong; Lou, Zhongzi; Wang, Fang; Zheng, Yadong; Xu, Guo-Tong

2013-07-10

21

Alterations in polyamine levels of nematode, earthworm, leech and planarian during regeneration, temperature and osmotic stresses.  

PubMed

Free-living nematodes, Caenorhabditis elegans and Dorylaimus fodori, contain putrescine and spermidine. Putrescine, spermidine and spermine occur in the parasitic Nematoda, Ascaris suum, Anisakis simplex and Dirofilaria immitis. Earthworms, Eisenia foetida, Tubifex hattai and Pheretima communissima and the leech, Hirudo nipponia (belonging to Annelida) and the planarian, Dugesia japonica (belonging to Platyhelminthes) contain homospermidine and spermine in addition to putrescine and spermidine. Regenerated heads of E. foetida and D. japonica are rich in putrescine indicating the stimulation of its synthesis during regeneration. Putrescine and spermidine levels temporarily increase after heat shock in C. elegans, E. foetida and D. japonica and cold shock and hypertonic osmotic shock treatments in D. japonica. PMID:7749639

Hamana, K; Hamana, H; Shinozawa, T

1995-05-01

22

Nitric oxide synthase in the pharynx of the planarian Dugesia tigrina.  

PubMed

The distribution of the nicotinamide adenine dinucleotide phosphate-(NADPH) diaphorase reaction, an indicator of nitric oxide synthase activity, was studied in the freshwater planarian Dugesia tigrina (Platyhelminthes). The reaction was restricted to the pharynx, where the inner epithelium was intensely stained and the outer epithelium moderately stained. Neurons that innervated the pharynx were also stained. The enzyme activity was studied by high pressure liquid chromatographic quantitation of the formed citrulline. The presumed nitric oxide synthase was dependent on NADPH, whereas no dependency on Ca2+ and calmodulin could be detected. Tetrahydrobiopterin increased the activity about fivefold to 218.2+/-24.9 fmol/mg protein per min. Nomega-nitro-l-arginine depressed the enzyme activity by about 80%. The results indicate that nitric oxide has a role in the feeding behavior of planarians. PMID:8929343

Eriksson, K S

1996-12-01

23

Transgenic planarian lines obtained by electroporation using transposon-derived vectors and an eye-specific GFP marker  

PubMed Central

To generate transgenic planarians we used a set of versatile vectors for animal transgenesis based on the promiscuous transposons, mariner, Hermes and piggyBac, and a universal enhanced GFP (EGFP) marker system with three Pax6 dimeric binding sites, the 3xP3-EGFP developed by Berghammer et al. [Berghammer, A. J., Klinger, M. & Wimmer, E. A. (1999) Nature 402, 370–371]. This marker is expressed specifically in the eyes of various arthropod taxa. Upon microinjection into the parenchyma of adult planarians and subsequent electroporation, these vectors transpose efficiently into the planarian genome. One of the cell types transformed are the totipotent “neoblast” stem cells present in the adults, representing 30% of total cells. The neoblast represents a unique cell type with the capacity to proliferate and to differentiate into all somatic cell types as well as into germ cells. All three transposon vectors have high transformation efficiency, but only Hermes and piggyBac show stable integration. The mariner vector is frequently lost presumably because of the presence of active mariner-type transposons in the genome of the Girardia tigrina. Transformed animals are mosaics containing both transformed and untransformed neoblasts. These differentiate to form EGFP-positive and -negative photoreceptor cells. Such mosaicism is maintained through several cycles of regeneration induced by decapitation or asexual reproduction. Transformed neoblasts also contribute to the germ line, and can give rise to pure transgenic planarian lines in which EGFP is expressed in all photoreceptor cells after sexual reproduction. The presence of the transgenes was confirmed by PCR, plasmid rescue assay, inverse PCR, and Southern blotting. Our results with the 3xP3-EGFP marker confirm the presence of Pax6 activity in the differentiated photoreceptor cells of planarian eyes. Transgenesis will be an important tool to dissect developmental molecular mechanisms in planarian regeneration, development and stem cell biology, and may also be an entry point to analyze the biology of parasitic Platyhelminthes.

Gonzalez-Estevez, C.; Momose, T.; Gehring, W. J.; Salo, E.

2003-01-01

24

Stem cells and neural signalling: the case of neoblast recruitment and plasticity in low dose X-ray treated planarians.  

PubMed

Planarians (Platyhelminthes) possess an abundant population of adult stem cells, the neoblasts, capable to give rise to both somatic and germ cells. Although neoblasts share similar morphological features, several pieces of evidence suggest that they constitute a heterogeneous population of cells with distinct ultrastructural and molecular features. We found that in planarians treated with low X-ray doses (5 Gy), only a few neoblasts survive. Among these cells, those located close to the nervous system activate an intense proliferation program and migrate to reconstitute the whole complex neoblast population. This phenomenon is inhibited by the substance P receptor antagonist spantide, and accompanied by the up-regulation of a number of genes implicated in neuronal signalling and plasticity, suggesting that signals of neural origin modulate neoblast proliferation and/or migration. Here, we review these findings and the literature available on the influence of the nervous system on stem cell activity, both in planarians and vertebrates, and we propose 5 Gy-treated planarians as a unique model system to study the influence of neural signalling on stem cell biology. PMID:22451001

Rossi, Leonardo; Iacopetti, Paola; Salvetti, Alessandra

2012-01-01

25

Autophagy and apoptosis in planarians  

Microsoft Academic Search

Adult planarians are capable of undergoing regeneration and body remodelling in order to adapt to physical damage or extreme\\u000a environmental conditions. Moreover, most planarians can tolerate long periods of starvation and during this time, they shrink\\u000a from an adult size to, and sometimes beyond, the initial size at hatching. Indeed, these properties have made them a classic\\u000a model to study

Cristina Gonzalez-EstevezEmili; Emili Saló

2010-01-01

26

Identification of members of several homeobox genes in a planarian using a ligation-mediated polymerase chain reaction technique.  

PubMed Central

I have used a novel single-sided specific polymerase chain reaction (PCR) strategy inspired by ligation-mediated PCR to clone fragments of divergent homeobox genes from a flatworm, the planarian Polycelis nigra. Eight homeobox-containing fragments were amplified, belonging to the Hox, msh, NK-1 and NK-2 classes. Together with the results obtained from several genomes of platyhelminths, my screening shows the presence of the same array of homeodomain developmental regulators in planarians, traditionally regarded as primitive metazoans in terms of body plan, as in coelomate organisms. However, the presence of a Ubx/abd-A homolog may indicate that platyhelminths are more closely related to protostomes than to deuterostomes and supports the idea that flatworms have inherited an elaborate HOX cluster (seven or eight genes) from their ancestor. Likely homologs of the fly genes tinman, bagpipe and S59 suggest that the mesoderm might be patterned by the same genes in all bilaterally symmetrical animals. Finally, a msh-like gene, a family known to be involved in inductive mechanisms in vertebrates, has been found. These results support the hypothesis that the tremendous diversity of metazoan body plans is specified by a largely conserved array of homeobox-containing developmental genes.

Balavoine, G

1996-01-01

27

[Morphogenesis in planarians Dugesia tigrina].  

PubMed

We carried out computer morphometry in regenerates of planarians Dugesia tigrina. The blastema growth was analyzed in fragments of planarians after their fission and after transverse transection at different body levels. The blastema was growing at a higher rate on tail fragments than on the head fragments and the growth rate was the higher, the closer the transection was to the head end. After fission, the blastema was growing at a slower rate than after transection in the fission zone. The growth of adjacent blastemas formed on both sides after fission or transection proceeded at different rates as a function of new body polarity. PMID:15487347

She?man, I M; Kreshchenko, N D; Sedel'nikov, Z V; Grozny?, A V

28

Determinants of planarian aggregation behavior  

Microsoft Academic Search

Using a filter technique, five experiments examined the role of chemical and visual cues in recruiting individual planarians\\u000a to established aggregations. The results indicated thatCura foremani depend upon a combination of chemical and visual cues whilePlanaria dactyligera depend almost entirely on chemical cues for aggregation formation.

James H. Reynierse; Kathryn Gleason

1975-01-01

29

The body margin of the planarian Dugesia japonica: characterization by the expression of an intermediate filament gene.  

PubMed

We have cloned and sequenced a cDNA encoding an intermediate filament protein (IF) from the planarian Dugesia japonica named DjIFb. The deduced amino acid sequence of DjIFb has similarity to those of protostomic IFs and lamins, supporting a previous hypothesis that the protostomic IFs, including DjIFb, are evolutionarily closer to lamins than to vertebrate cytoplasmic IFs. In addition, analysis of the exon/intron organization revealed that 8 out of 10 introns of DjIFb were coincident in their position, even in the codon phase, with those of the non-neuronal IF of the snail Helix aspersa. This suggests that the Platyhelminthes are not the most primitive Bilateria but instead are evolutionarily close to the Mollusca. The DjIFb gene was expressed in particular cells, probably a kind of adhesive gland cell, which were present in the marginal region encircling the planarian body. The localization of DjIFb protein suggests that it plays an important role in the secretion of an adhesive substance. The specific expression pattern of the DjIFb gene enabled us to monitor how the body margin forms during planarian regeneration. PMID:12203092

Tazaki, Akira; Kato, Kentaro; Orii, Hidefumi; Agata, Kiyokazu; Watanabe, Kenji

2002-07-13

30

Complete Functional Segregation of Planarian ?-Catenin-1 and -2 in Mediating Wnt Signaling and Cell Adhesion*  

PubMed Central

?-Catenin is a bifunctional protein participating in both cell adhesion and canonical Wnt signaling. In cell adhesion, it bridges the transmembrane cadherin and the actin-binding protein ?-catenin and is essential for adherens junction formation, whereas in canonical Wnt signaling, it shuttles between the cytosol and nucleus and functions as an essential transcriptional activator. Schmidtea mediterranea ?-catenin-1 was identified as a determinant of antero-posterior polarity during body regeneration by mediating Wnt signaling. Here we show that S. mediterranea ?-catenin-2 is specifically expressed in epithelial cells in the gut and pharynx, where it has a putative role in mediating cell adhesion. We show evidence that planarian ?-catenin-1 and -2 have distinct biochemical properties. ?-Catenin-1 can interact with the components of the canonical Wnt signaling pathway but not with ?-catenin, whereas ?-catenin-2 interacts with cell adhesion molecules, including E-cadherin and ?-catenin, but not with Wnt signaling components. Consistent with their specific function, ?-catenin-1 is a potent transcriptional activator, whereas ?-catenin-2 has no transcriptional activity. Protein sequence alignment also indicates that the planarian ?-catenin-1 and -2 retain distinct critical residues and motifs, which are in agreement with the differences in their biochemical properties. At last, phylogenetic analysis reveals a probable Platyhelminthes- specific structural and functional segregation from which the monofunctional ?-catenins evolved. Our results thus identify the first two monofunctional ?-catenins in metazoans.

Chai, Guoliang; Ma, Changxin; Bao, Kai; Zheng, Liang; Wang, Xinquan; Sun, Zhirong; Salo, Emili; Adell, Teresa; Wu, Wei

2010-01-01

31

Planarian Regeneration and Stem Cells  

NSDL National Science Digital Library

A mini-documentary discussing the remarkable regenerative capabilities of the planarian, and how HHMI researcher Alejandro Snchez Alvarado uses them to study the biology of stem cells. This presentation is also featured on the DVD Potent Biology: Stem Cells, Cloning, and Regeneration, available for free from HHMI. This video is 11 minutes and 46 seconds in length, and available for download in Quicktime (114 MB) and Windows Media (156 MB) formats. All Stem Cell videos are located at: http://www.hhmi.org/biointeractive/stemcells/video.html.

Alejandro SĂ¡nchez Alvarado (Howard Hughes Medical Institute;)

2007-03-31

32

Dynamics of asexual reproduction in planarians  

NASA Astrophysics Data System (ADS)

Planaria research is experiencing a resurgence due to the development of molecular tools, the Planarian genome project and database resources. Despite the resulting progress in planarian biology research, an extensive study of their physical properties remains to be undertaken. We developed a method to collect a large amount of data on the dynamics of clonal reproduction in the freshwater planarian S.mediterranea. The capability of planarians to regenerate an entire organism from a minuscule body part is based on a homogeneously distributed stem cell population that comprises 25-30% of all cells. Due to this stem cell contingent, planarians can reproduce spontaneously by dividing into a larger head and a smaller tail piece, which then will rebuild the missing body parts, including a central nervous system, within about a week. Time-lapse imaging allows us to characterize the fission process in detail, revealing the stages of the process as well as capturing the nature of the rupture itself. A traction force measurement setup is being developed to allow us to quantify the forces planarians exert on the substrate during reproduction, a macroscopic analog to the Traction Force Microscopy setups used to determine local cellular forces. We are particularly interested in the molecular processes during division and the interplay between tissue mechanics and cell signaling.

Schoetz, Eva-Maria; Lincoln, Bryan; Quinodoz, Sofia

2011-03-01

33

Nicotine behavioral pharmacology: clues from planarians  

PubMed Central

Background Nicotine is one of the world’s most addictive substances and the primary reason that humans inhale tobacco smoke. The pharmacological effects of nicotine can be investigated in planarians, aquatic flatworms that possess an integrated neural network including cephalic ganglia that some consider the earliest “brain” and spinal cord. Here, we tested the hypothesis that nicotine exposure elicits mammalian-like behaviors in planarians. Methods Planarian motility and stereotypy (C-shape hyperkinesias) were quantified following acute nicotine exposure. During repeated nicotine exposure, we investigated the presence of withdrawal, tolerance, behavioral sensitization, and environmental place conditioning. Results Acute nicotine exposure increased stereotypical activity and elicited biphasic effects on motility. A low concentration (0.01 mM) increased motility whereas higher concentrations (0.3 – 10 mM) elicited the opposite effect. Planarians exposed to nicotine (0.03 mM) for 60 min and then tested in water displayed reduced motility that was not observed during exposure to water, acute nicotine, or continuous nicotine. Nicotine-treated planarians withdrawn from the drug for 3 days before being challenged with nicotine displayed behavioral sensitization at low concentrations (0.1, 0.3 mM) but tolerance at higher concentrations (1, 3 mM). Planarians conditioned with nicotine in the ambient light (non-preferred environment) displayed a reduction in their natural preference for a dark environment. Conclusions The present results suggest nicotine elicits mammalian-like effects in planarians, including decreased motility and increased stereotypy following acute administration and abstinence-induced withdrawal, behavioral sensitization, tolerance, and place conditioning during repeated exposure.

Rawls, Scott M.; Patil, Tanvi; Tallarida, Christopher S.; Baron, Steven; Kim, Myongji; Song, Kevin; Ward, Sara; Raffa, Robert B.

2011-01-01

34

Centrosome loss in the evolution of planarians.  

PubMed

The centrosome, a cytoplasmic organelle formed by cylinder-shaped centrioles surrounded by a microtubule-organizing matrix, is a hallmark of animal cells. The centrosome is conserved and essential for the development of all animal species described so far. Here, we show that planarians, and possibly other flatworms, lack centrosomes. In planarians, centrioles are only assembled in terminally differentiating ciliated cells through the acentriolar pathway to trigger the assembly of cilia. We identified a large set of conserved proteins required for centriole assembly in animals and note centrosome protein families that are missing from the planarian genome. Our study uncovers the molecular architecture and evolution of the animal centrosome and emphasizes the plasticity of animal cell biology and development. PMID:22223737

Azimzadeh, Juliette; Wong, Mei Lie; Downhour, Diane Miller; Sánchez Alvarado, Alejandro; Marshall, Wallace F

2012-01-05

35

Centrosome Loss in the Evolution of Planarians  

PubMed Central

The centrosome, a cytoplasmic organelle formed by cylinder-shaped centrioles surrounded by a microtubule-organizing matrix, is a hallmark of animal cells. The centrosome is conserved and essential for the development of all animal species described so far. Here, we show that, unlike the rest of animals, planarians and possibly other flatworms as well completely lack centrosomes. We found that in planarians, centrioles are only assembled in terminally differentiating ciliated cells through a so-called acentriolar pathway to trigger the assembly of cilia. This unique characteristic allowed us to identify a large set of conserved proteins required for centriole assembly in animals, as well as the centrosome signature proteins missing from the planarian genome. Our study uncovers the molecular architecture and evolution of the animal centrosome and emphasizes the plasticity of animal cell biology and development.

Wong, Mei Lie; Downhour, Diane Miller; Alvarado, Alejandro Sanchez

2012-01-01

36

Muscle cells provide instructions for planarian regeneration.  

PubMed

Regeneration requires both potential and instructions for tissue replacement. In planarians, pluripotent stem cells have the potential to produce all new tissue. The identities of the cells that provide regeneration instructions are unknown. Here, we report that position control genes (PCGs) that control regeneration and tissue turnover are expressed in a subepidermal layer of nonneoblast cells. These subepidermal cells coexpress many PCGs. We propose that these subepidermal cells provide a system of body coordinates and positional information for regeneration, and identify them to be muscle cells of the planarian body wall. Almost all planarian muscle cells express PCGs, suggesting a dual function: contraction and control of patterning. PCG expression is dynamic in muscle cells after injury, even in the absence of neoblasts, suggesting that muscle is instructive for regeneration. We conclude that planarian regeneration involves two highly flexible systems: pluripotent neoblasts that can generate any new cell type and muscle cells that provide positional instructions for the regeneration of any body region. PMID:23954785

Witchley, Jessica N; Mayer, Mirjam; Wagner, Daniel E; Owen, Jared H; Reddien, Peter W

2013-08-15

37

Planarian immobilization, partial irradiation, and tissue transplantation.  

PubMed

The planarian, a freshwater flatworm, has proven to be a powerful system for dissecting metazoan regeneration and stem cell biology. Planarian regeneration of any missing or damaged tissues is made possible by adult stem cells termed neoblasts. Although these stem cells have been definitively shown to be pluripotent and singularly capable of reconstituting an entire animal, the heterogeneity within the stem cell population and the dynamics of their cellular behaviors remain largely unresolved. Due to the large number and wide distribution of stem cells throughout the planarian body plan, advanced methods for manipulating subpopulations of stem cells for molecular and functional study in vivo are needed. Tissue transplantation and partial irradiation are two methods by which a subpopulation of planarian stem cells can be isolated for further study. Each technique has distinct advantages. Tissue transplantation allows for the introduction of stem cells, into a naďve host, that are either inherently genetically distinct or have been previously treated pharmacologically. Alternatively, partial irradiation allows for the isolation of stem cells within a host, juxtaposed to tissue devoid of stem cells, without the introduction of a wound or any breech in tissue integrity. Using these two methods, one can investigate the cell autonomous and non-autonomous factors that control stem cell functions, such as proliferation, differentiation, and migration. Both tissue transplantation and partial irradiation have been used historically in defining many of the questions about planarian regeneration that remain under study today. However, these techniques have remained underused due to the laborious and inconsistent nature of previous methods. The protocols presented here represent a large step forward in decreasing the time and effort necessary to reproducibly generate large numbers of grafted or partially irradiated animals with efficacies approaching 100 percent. We cover the culture of large animals, immobilization, preparation for partial irradiation, tissue transplantation, and the optimization of animal recovery. Furthermore, the work described here demonstrates the first application of the partial irradiation method for use with the most widely studied planarian, Schmidtea mediterranea. Additionally, efficient tissue grafting in planaria opens the door for the functional testing of subpopulations of naďve or treated stem cells in repopulation assays, which has long been the gold-standard method of assaying adult stem cell potential in mammals. Broad adoption of these techniques will no doubt lead to a better understanding of the cellular behaviors of adult stem cells during tissue homeostasis and regeneration. PMID:23007410

Guedelhoefer, Otto C; Sánchez Alvarado, Alejandro

2012-08-06

38

Parthenogenesis and asexual multiplication among parasitic platyhelminths.  

PubMed

Among flatworms with parasitic and commensal modes of existence, parthenogenesis and asexual multiplication appear to be largely confined to the Digenea and Cestoda, the only parasitic platyhelminths that routinely utilize indirect life-cycles. Parthenogenesis is apparently restricted to a minority of adult digeneans and cestodes inhabiting their final hosts, and a survey is made of the particular modes of parthenogenesis (i.e. apomictic, automictic and generative) which are employed by such adults. Asexual (amictic) multiplication, in the form of fissioning, is demonstrated by young adults of the cyclophyllidean cestode, Mesocestoides corti, but is otherwise not exhibited by adult cestodes or digeneans, other than in the perplexing phenomenon of proglottid formation in polyzoic tapeworms. Secondary multiplication is of ubiquitous occurrence in digenean life-cycles in the form of the proliferation which takes place within sporocysts and rediae (germinal sacs) located in the first intermediate host. The controversy concerning the nature of this multiplication is reconsidered in the context of recent findings which have centred on cellular aspects. On the basis of present evidence germinal sac multiplication should be regarded as an asexual rather than a parthenogenetic process. The cestode asexual multiplication which occurs in intermediate hosts is a function of the metacestode stage of development. Metacestode proliferation is only known from about 20 species and 6 families of polyzoic cestodes with approximately half the described instances occurring in the family Taeniidae. The organization of these proliferative metacestodes, findings concerning their totipotent stem cells and the ontogeny of buds and new scolices are all reviewed. Finally, the capacity for population expansion of multiplicative larval digeneans and metacestodes are compared, while the ecological roles and the genetical consequences of both parthenogenesis and amictic multiplication in the two taxa are also examined. PMID:6346231

Whitfield, P J; Evans, N A

1983-04-01

39

Identification of planarian homeobox sequences indicates the antiquity of most Hox/homeotic gene subclasses.  

PubMed Central

The homeotic gene complex (HOM-C) is a cluster of genes involved in the anteroposterior axial patterning of animal embryos. It is composed of homeobox genes belonging to the Hox/HOM superclass. Originally discovered in Drosophila, Hox/HOM genes have been identified in organisms as distantly related as arthropods, vertebrates, nematodes, and cnidarians. Data obtained in parallel from the organization of the complex, the domains of gene expression during embryogenesis, and phylogenetic relationships allow the subdivision of the Hox/HOM superclass into five classes (lab, pb/Hox3, Dfd, Antp, and Abd-B) that appeared early during metazoan evolution. We describe a search for homologues of these genes in platyhelminths, triploblast metazoans emerging as an outgroup to the great coelomate ensemble. A degenerate PCR screening for Hox/HOM homeoboxes in three species of triclad planarians has revealed 10 types of Antennapedia-like genes. The homeobox-containing sequences of these PCR fragments allowed the amplification of the homeobox-coding exons for five of these genes in the species Polycelis nigra. A phylogenetic analysis shows that two genes are clear orthologues of Drosophila labial, four others are members of a Dfd/Antp superclass, and a seventh gene, although more difficult to classify with certainty, may be related to the pb/Hox3 class. Together with previously identified Hox/HOM genes in other flatworms, our analyses demonstrate the existence of an elaborate family of Hox/HOM genes in the ancestor of all triploblast animals. Images Fig. 4

Balavoine, G; Telford, M J

1995-01-01

40

Sex in Parthenogenetic Planarians: Phylogenetic Relic or Evolutionary Resurrection?  

Microsoft Academic Search

\\u000a Besides their remarkable capability of regeneration, planarian flatworms are well known for their wide range of reproductive\\u000a modes. In this chapter, we elucidate the evolutionary significance of sexual and parthenogenetic reproduction in the freshwater\\u000a planarian Schmidtea polychroa. In accordance with the major theories of sex, parthenogenetic S. polychroa seem to suffer from both a higher mutation load and parasite load.

Thomas G. D’Souza; Nico K. Michiels

41

The Retinoblastoma pathway regulates stem cell proliferation in freshwater planarians.  

PubMed

Freshwater planarians are flatworms of the Lophotrochozoan superphylum and are well known for their regenerative abilities, which rely on a large population of pluripotent adult stem cells. However, the mechanisms by which planarians maintain a precise population of adult stem cells while balancing proliferation and cell death, remain to be elucidated. Here we have identified, characterized, and functionally tested the core Retinoblastoma (Rb) pathway components in planarian adult stem cell biology. The Rb pathway is an ancient and conserved mechanism of proliferation control from plants to animals and is composed of three core components: an Rb protein, and a transcription factor heterodimer of E2F and DP proteins. Although the planarian genome contains all components of the Rb pathway, we found that they have undergone gene loss from the ancestral state, similar to other species in their phylum. The single Rb homolog (Smed-Rb) was highly expressed in planarian stem cells and was required for stem cell maintenance, similar to the Rb-homologs p107 and p130 in vertebrates. We show that planarians and their phylum have undergone the most severe reduction in E2F genes observed thus far, and the single remaining E2F was predicted to be a repressive-type E2F (Smed-E2F4-1). Knockdown of either Smed-E2F4-1 or its dimerization partner Dp (Smed-Dp) by RNAi resulted in temporary hyper-proliferation. Finally, we showed that known Rb-interacting genes in other systems, histone deacetylase 1 and cyclinD (Smed-HDAC1; Smed-cycD), were similar to Rb in expression and phenotypes when knocked down by RNAi, suggesting that these established interactions with Rb may also be conserved in planarians. Together, these results showed that planarians use the conserved components of the Rb tumor suppressor pathway to control proliferation and cell survival. PMID:23123964

Zhu, Shu Jun; Pearson, Bret J

2012-11-01

42

Planarians in pharmacology: parthenolide is a specific behavioral antagonist of cocaine in the planarian Girardia tigrina.  

PubMed

Planarians are traditional animal models in developmental and regeneration biology. Recently, these organisms are arising as vertebrate-relevant animal models in neuropharmacology. Using an adaptation of published behavioral protocols, we have described the alleviation of cocaine-induced planarian seizure-like movements (pSLM) by a naturally-occurring sesquiterpene lactone, parthenolide. Interestingly, parthenolide does not prevent the expression of pSLM induced by amphetamines; in vertebrates, amphetamines interact with the same protein target as cocaine. Parthenolide is also unable to prevent pSLM elicited by the cholinergic com-pounds nicotine and cytisine or by the glutamatergic agents L- or D- glutamic acid or NMDA. Thus, we conclude that parthenolide is a specific anti-cocaine agent in this experimental organism. PMID:22451007

Pagán, Oné R; Baker, Debra; Deats, Sean; Montgomery, Erica; Tenaglia, Matthew; Randolph, Clinita; Kotturu, Dharini; Tallarida, Christopher; Bach, Daniel; Wilk, Galia; Rawls, Scott; Raffa, Robert B

2012-01-01

43

Stem cells and the Planarian Schmidtea mediterranea  

PubMed Central

In recent years, somatic stem cells have been heralded as potential therapeutic agents to address a large number of degenerative diseases. Yet, in order to rationally utilize these cells as effective therapeutic agents, and/or improve treatment of stem-cell-associated malignancies such as leukemias and carcinomas, a better understanding of the basic biological properties of stem cells needs to be acquired. A major limitation in the study of somatic stem cells lies in the difficulty of accessing and studying these cells in vivo. This barrier is further compounded by the limitations of in vitro culture systems, which are unable to emulate the microenvironments in which stem cells reside and which are known to provide critical regulatory signals for their proliferation and differentiation. Given the complexity of vertebrate adult somatic stem cell populations and their relative inaccessibility to in vivo molecular analyses, the study of somatic stem cells should benefit from analyzing their counterparts in simpler model organisms. In the past, the use of Drosophila or C. elegans has provided invaluable contributions to our understanding of genes and pathways involved in a variety of human diseases. However, stem cells in these organisms are mostly restricted to the gonads, and more importantly neither Drosophila, nor C. elegans are capable of regenerating body parts lost to injury. Therefore, a simple animal with experimentally accessible stem cells playing a role in tissue maintenance and/or regeneration should be very useful in identifying and functionally testing the mechanisms regulating stem cell activities. The planarian Schmidtea mediterranea is poised to fill this experimental gap. S. mediterranea displays robust regenerative properties driven by an adult, somatic stem cell population capable of producing the ?40 different cell types found in this organism, including the germ cells. Given that all known metazoans depend on stem cells for their survival, it is extremely likely that the molecular events regulating stem cell biology would have been conserved throughout evolution, and that the knowledge derived from studying planarian stem cells could be vertically integrated to the study of vertebrate somatic stem cells. Current efforts, therefore, are aimed at further characterizing the somatic population of planarian stem cells in order to define its suitability as a model system in which to mechanistically dissect the basic biological attributes of metazoans stem cells.

Sanchez Alvarado, Alejandro

2007-01-01

44

Establishing and maintaining a colony of planarians.  

PubMed

INTRODUCTIONTo provide sufficient material for experimentation, a laboratory needs to expand and maintain a colony of planarians. It is crucial to keep a stable, healthy population of animals in a consistent environment to avoid inter-animal variability and modifier effects that can mask true phenotypes from experimental perturbation. In this protocol, we describe basic procedures for establishing and maintaining healthy colonies of Dugesia japonica, Schmidtea mediterranea, and Girardia tigrina (commonly found in the wild and commercially available in the United States). Although the recommendations are based on our optimization of conditions for G. tigrina, many of the procedures (such as food preparation and feeding strategy) can be applied to other species. For best results, the culture water must be carefully monitored and adjusted for each species. PMID:21356691

Oviedo, Néstor J; Nicolas, Cindy L; Adams, Dany S; Levin, Michael

2008-10-01

45

Synchronous and early activation of planarian Hox genes and the re-specification of body axes during regeneration in Dugesia (G.) tigrina (Turbellaria; Tricladida)  

Microsoft Academic Search

Seven Hox cluster-related genes (Dthox-A to -G) have been isolated from the freshwater triclad Dugesia (G.) tigrina, their\\u000a sequence compared to other Hox genes and their expression in intact and regenerating organisms analyzed by whole mount in\\u000a situ hybridization. Sequence comparison analyses show high similarities of D. tigrina Hox genes to anterior and medial groups\\u000a of coelomate Hox genes. Expression

J. R. Bayascas; E. Castillo; A. M. Muńoz-Mármol; J. Baguńŕ; E. Saló

1998-01-01

46

Novel RNAi-Mediated Approach to G Protein-Coupled Receptor Deorphanization: Proof of Principle and Characterization of a Planarian 5-HT Receptor  

PubMed Central

G protein-coupled receptors (GPCRs) represent the largest known superfamily of membrane proteins extending throughout the Metazoa. There exists ample motivation to elucidate the functional properties of GPCRs given their role in signal transduction and their prominence as drug targets. In many target organisms, these efforts are hampered by the unreliable nature of heterologous receptor expression platforms. We validate and describe an alternative loss-of-function approach for ascertaining the ligand and G protein coupling properties of GPCRs in their native cell membrane environment. Our efforts are focused on the phylum Platyhelminthes, given the heavy health burden exacted by pathogenic flatworms, as well as the role of free-living flatworms as model organisms for the study of developmental biology. RNA interference (RNAi) was used in conjunction with a biochemical endpoint assay to monitor cAMP modulation in response to the translational suppression of individual receptors. As proof of principle, this approach was used to confirm the neuropeptide GYIRFamide as the cognate ligand for the planarian neuropeptide receptor GtNPR-1, while revealing its endogenous coupling to G?i/o. The method was then extended to deorphanize a novel G?s-coupled planarian serotonin receptor, DtSER-1. A bioinformatics protocol guided the selection of receptor candidates mediating 5-HT-evoked responses. These results provide functional data on a neurotransmitter central to flatworm biology, while establishing the great potential of an RNAi-based deorphanization protocol. Future work can help optimize and adapt this protocol for higher-throughput platforms as well as other phyla.

Zamanian, Mostafa; Agbedanu, Prince N.; Wheeler, Nicolas J.; McVeigh, Paul; Kimber, Michael J.; Day, Tim A.

2012-01-01

47

High-resolution profiling and discovery of planarian small RNAs  

PubMed Central

Freshwater planarian flatworms possess uncanny regenerative capacities mediated by abundant and collectively totipotent adult stem cells. Key functions of these cells during regeneration and tissue homeostasis have been shown to depend on PIWI, a molecule required for Piwi-interacting RNA (piRNA) expression in planarians. Nevertheless, the full complement of piRNAs and microRNAs (miRNAs) in this organism has yet to be defined. Here we report on the large-scale cloning and sequencing of small RNAs from the planarian Schmidtea mediterranea, yielding altogether millions of sequenced, unique small RNAs. We show that piRNAs are in part organized in genomic clusters and that they share characteristic features with mammalian and fly piRNAs. We further identify 61 novel miRNA genes and thus double the number of known planarian miRNAs. Sequencing, as well as quantitative PCR of small RNAs, uncovered 10 miRNAs enriched in planarian stem cells. These miRNAs are down-regulated in animals in which stem cells have been abrogated by irradiation, and thus constitute miRNAs likely associated with specific stem-cell functions. Altogether, we present the first comprehensive small RNA analysis in animals belonging to the third animal superphylum, the Lophotrochozoa, and single out a number of miRNAs that may function in regeneration. Several of these miRNAs are deeply conserved in animals.

Friedlander, Marc R.; Adamidi, Catherine; Han, Ting; Lebedeva, Svetlana; Isenbarger, Thomas A.; Hirst, Martin; Marra, Marco; Nusbaum, Chad; Lee, William L.; Jenkin, James C.; Alvarado, Alejandro Sanchez; Kim, John K.; Rajewsky, Nikolaus

2009-01-01

48

Reactivating head regrowth in a regeneration-deficient planarian species.  

PubMed

Species capable of regenerating lost body parts occur throughout the animal kingdom, yet close relatives are often regeneration incompetent. Why in the face of 'survival of the fittest' some animals regenerate but others do not remains a fascinating question. Planarian flatworms are well known and studied for their ability to regenerate from minute tissue pieces, yet species with limited regeneration abilities have been described even amongst planarians. Here we report the characterization of the regeneration defect in the planarian Dendrocoelum lacteum and its successful rescue. Tissue fragments cut from the posterior half of the body of this species are unable to regenerate a head and ultimately die. We find that this defect originates during the early stages of head specification, which require inhibition of canonical Wnt signalling in other planarian species. Notably, RNA interference (RNAi)-mediated knockdown of Dlac-?-catenin-1, the Wnt signal transducer, restored the regeneration of fully functional heads on tail pieces, rescuing D. lacteum's regeneration defect. Our results demonstrate the utility of comparative studies towards the reactivation of regenerative abilities in regeneration-deficient animals. Furthermore, the availability of D. lacteum as a regeneration-impaired planarian model species provides a first step towards elucidating the evolutionary mechanisms that ultimately determine why some animals regenerate and others do not. PMID:23883932

Liu, S-Y; Selck, C; Friedrich, B; Lutz, R; Vila-Farré, M; Dahl, A; Brandl, H; Lakshmanaperumal, N; Henry, I; Rink, J C

2013-07-24

49

Bromodeoxyuridine specifically labels the regenerative stem cells of planarians.  

PubMed

The singular regenerative abilities of planarians require a population of stem cells known as neoblasts. In response to wounding, or during the course of cell turnover, neoblasts are signaled to divide and/or differentiate, thereby replacing lost cell types. The study of these pluripotent stem cells and their role in planarian regeneration has been severely hampered by the reported inability of planarians to incorporate exogenous DNA precursors; thus, very little is known about the mechanisms that control proliferation and differentiation of this stem cell population within the planarian. Here we show that planarians are, in fact, capable of incorporating the thymidine analogue bromodeoxyuridine (BrdU), allowing neoblasts to be labeled specifically during the S phase of the cell cycle. We have used BrdU labeling to study the distribution of neoblasts in the intact animal, as well as to directly demonstrate the migration and differentiation of neoblasts. We have examined the proposal that a subset of neoblasts is arrested in the G2 phase of the cell cycle by double-labeling with BrdU and a mitosis-specific marker; we find that the median length of G2 (approximately 6 h) is sufficient to account for the initial mitotic burst observed after feeding or amputation. Continuous BrdU-labeling experiments also suggest that there is not a large, slow-cycling population of neoblasts in the intact animal. The ability to label specifically the regenerative stem cells, combined with the recently described use of double-stranded RNA to inhibit gene expression in the planarian, should serve to reignite interest in the flatworm as an experimental model for studying the problems of metazoan regeneration and the control of stem cell proliferation. PMID:10753506

Newmark, P A; Sánchez Alvarado, A

2000-04-15

50

A regulatory program for excretory system regeneration in planarians  

PubMed Central

Planarians can regenerate any missing body part, requiring mechanisms for the production of organ systems in the adult, including their prominent tubule-based filtration excretory system called protonephridia. Here, we identify a set of genes, Six1/2-2, POU2/3, hunchback, Eya and Sall, that encode transcription regulatory proteins that are required for planarian protonephridia regeneration. During regeneration, planarian stem cells are induced to form a cell population in regeneration blastemas expressing Six1/2-2, POU2/3, Eya, Sall and Osr that is required for excretory system formation. POU2/3 and Six1/2-2 are essential for these precursor cells to form. Eya, Six1/2-2, Sall, Osr and POU2/3-related genes are required for vertebrate kidney development. We determined that planarian and vertebrate excretory cells express homologous proteins involved in reabsorption and waste modification. Furthermore, we identified novel nephridia genes. Our results identify a transcriptional program and cellular mechanisms for the regeneration of an excretory organ and suggest that metazoan excretory systems are regulated by genetic programs that share a common evolutionary origin.

Scimone, M. Lucila; Srivastava, Mansi; Bell, George W.; Reddien, Peter W.

2011-01-01

51

Planarian embryology in the era of comparative developmental biology.  

PubMed

During the last decade, the field of evolutionary developmental biology (evo-devo) has emerged as a major research discipline in modern biology and an essential approach to understanding evolutionary relationships in the animal kingdom. At the same time, planarians have become a useful and important model with which to address basic questions regarding the molecular and cellular basis of regeneration, tissue repair and stem cells in adult organisms. Nevertheless, little attention has been paid to their embryonic development, even though this provides a unique opportunity for studying how molecular developmental mechanisms are re-deployed during adult regeneration or the independent losses of spiral cleavage that took place in different lophotrochozoan lineages. In this paper, we review the most relevant works on planarian embryos from a historical point of view. In doing so, we highlight the questions that have recurrently intrigued researchers, most of which remain unanswered. Finally, we present a comprehensive scenario for planarian embryogenesis in an attempt to provide a testable hypothesis that will help to bridge the gap between this divergent mode of development, the ancestral canonical spiral cleavage, and adult planarian regeneration. PMID:22450993

Martín-Durán, José M; Monjo, Francisco; Romero, Rafael

2012-01-01

52

A regulatory program for excretory system regeneration in planarians.  

PubMed

Planarians can regenerate any missing body part, requiring mechanisms for the production of organ systems in the adult, including their prominent tubule-based filtration excretory system called protonephridia. Here, we identify a set of genes, Six1/2-2, POU2/3, hunchback, Eya and Sall, that encode transcription regulatory proteins that are required for planarian protonephridia regeneration. During regeneration, planarian stem cells are induced to form a cell population in regeneration blastemas expressing Six1/2-2, POU2/3, Eya, Sall and Osr that is required for excretory system formation. POU2/3 and Six1/2-2 are essential for these precursor cells to form. Eya, Six1/2-2, Sall, Osr and POU2/3-related genes are required for vertebrate kidney development. We determined that planarian and vertebrate excretory cells express homologous proteins involved in reabsorption and waste modification. Furthermore, we identified novel nephridia genes. Our results identify a transcriptional program and cellular mechanisms for the regeneration of an excretory organ and suggest that metazoan excretory systems are regulated by genetic programs that share a common evolutionary origin. PMID:21937596

Scimone, M Lucila; Srivastava, Mansi; Bell, George W; Reddien, Peter W

2011-10-01

53

The early emergence of platyhelminths is contradicted by the agreement between 18S rRNA and Hox genes data  

Microsoft Academic Search

The phylogenetic position of thé platyhelminths within the metazoan tree is examined using two independent sets of molecular characters, the evolution of 18S ribosomal RNA sequences and the diversity of the genes belonging to the HOX cluster. Among the various hypotheses that have been considered by zoologists, a position of the platyhelminths within the protostomes, related to the phyla with

Guillaume Balavoine

1997-01-01

54

First evidence that drugs of abuse produce behavioral sensitization and cross sensitization in planarians.  

PubMed

Behavioral sensitization in mammals, including humans, is sensitive to factors such as administration route, testing environment, and pharmacokinetic confounds, unrelated to the drugs themselves that are difficult to eliminate. Simpler animals less susceptible to these confounding influences may be advantageous substitutes for studying sensitization. We tested this hypothesis by determining whether planarians display sensitization and cross sensitization to cocaine and glutamate. Planarian hyperactivity was quantified as the number of C-like hyperkinesias during a 1-min drug exposure. Planarians exposed initially to cocaine (or glutamate) on day 1 were challenged with cocaine (or glutamate) after 2 or 6 days of abstinence. Acute cocaine or glutamate produced concentration-related hyperactivity. Cocaine or glutamate challenge after 2 and 6 days of abstinence enhanced the hyperactivity, indicating the substances produced planarian behavioral sensitization. Cross-sensitization experiments showed that cocaine produced greater hyperactivity in planarians earlier exposed to glutamate than in glutamate-naive planarians, and vice versa. Behavioral responses were pharmacologically selective because neither scopolamine nor caffeine produced planarian behavioral sensitization despite causing hyperactivity after initial administration, and acute gamma-aminobutyric acid did not cause hyperactivity. Demonstration of pharmacologically selective behavioral sensitization in planarians suggests that these flatworms represent a sensitive in-vivo model to study cocaine behavioral sensitization and to screen potential abuse-deterrent therapeutics. PMID:20512030

Rawls, Scott M; Patil, Tavni; Yuvasheva, Ekaternia; Raffa, Robert B

2010-07-01

55

Distribution of the stem cells (neoblasts) in the planarian Dugesia japonica  

Microsoft Academic Search

It has been postulated that the high regeneration ability of planarians is supported by totipotent stem cells, called neoblasts. There have been a few reports showing the distribution of neoblasts in planarians. However, the findings were not completely consistent. To determine the distribution of neoblasts, we focused on proliferating cell nuclear antigen (PCNA), which is present in proliferative cells. We

Hidefumi Orii; Takashige Sakurai; Kenji Watanabe

2005-01-01

56

Wnt signaling in planarians: new answers to old questions.  

PubMed

Wnts are secreted glycoproteins involved in a broad range of essential cell functions, including proliferation, migration and cell-fate determination. Recent years have seen substantial research effort invested in elucidating the role of the Wnt signaling pathway in planarians, flatworms with incredible regenerative capacities. In this review, we summarize current knowledge on the role of canonical (?-catenin-dependent) and non-canonical (?-catenin-independent) Wnt signaling in planarians, not only during regeneration, but also during normal homeostasis. We also describe some of the preliminary data that has been obtained regarding the role of these pathways during embryogenesis. Models are proposed to integrate the different results which have been obtained to date and highlight those questions that still remain to be answered. PMID:22450995

Almuedo-Castillo, Maria; Sureda-Gómez, Miquel; Adell, Teresa

2012-01-01

57

Chemo-klino-kinesis in planarian food location.  

PubMed

The effects of a homologous series of buffered fatty acid solutions on planarian behaviour were measured as the change in klino-kinetic activity. On the basis of the results obtained a model demonstrating the function of this behaviour pattern in the food-locating response was proposed. The model suggested that qualitative rather than quantitative gradients were effective in stimulating klino-kinetic orientation. PMID:1163855

Mason, P R

1975-05-01

58

Resistance of two planarian species to UV-irradiation.  

PubMed

The aim of this work was to determine the effects of 20, 25 and 30 minute UV-irradiation periods lambda = 253.5 nm to two planarian species Dugesia tigrina (Gir.) and Polycelis felina (Daly.). In vivo, UV light effects have been reported to affect intracellular receptors and disrupt simple behaviour. The effects of UV-rays on mortality and behavior as well as morphological, cytological and histological changes in the two planarian species were assessed, and the course and the dynamics of regenerative processes were compared between them. Experimental populations of Dugesia tigrina and Polycelis felina species were maintained in laboratory conditions at room temperature. Mortality, behavioral and morphological changes were monitored daily by means of a light stereomicroscope. For cytological and histopathological analysis, planarians were fixed in Bouine fixative on the first, second, third, fifth and seventh day after exposure to UV-irradiation, respectively. They were embedded in paraffin, cut on a microtome, stained with toluidin blue and embedded in Canada-balsam. UV-rays caused mortality, behavioral, morphological, cytological and histological changes in each planarian species. In regeneration of damaged body parts reticular cells and neoblasts played the main role. Neoblasts as totipotent cells extremely increased in number in the area of damaged tissue, immediately after UV-exposure. Dugesia tigrina was more sensitive to UV-rays than Polycelis felina due to possession of less pigmented cells. The course of regeneration in both species was similar. Most individuals of both species regenerated in 5 to 12 days after UV-irradiation. PMID:17220004

Kalafati?, Mirjana; Kovacevi?, Goran; Franjevi?, Damjan

2006-01-01

59

Effects of dimethylsulfoxide on behavior and antioxidant enzymes response of planarian Dugesia japonica.  

PubMed

In this study, the toxicity, behavioral and antioxidant activity effects of dimethylsulfoxide (DMSO) on planarian Dugesia japonica were investigated. The results showed that the mortality was directly proportional to the DMSO concentration, and planarian locomotor velocity decreased as the concentration of DMSO increased. The recovery of the motility for planarians pre-exposed to DMSO was found to be time- and dose-dependent, and only those pre-exposed to 0.1-3% DMSO resulted in full recovery. The antioxidant enzymes of planarians in response to long-term DMSO stress was also altered in a time- and dose-dependent manner. Planarians revealed more tolerance to DMSO toxicity at low DMSO (0.1%) level in short- and long-term DMSO stress, in which an efficient antioxidant system was involved and the motility was not affected. PMID:21976142

Yuan, Zuoqing; Zhao, Bosheng; Zhang, Yu

2011-10-05

60

Study of planarian stem cell proliferation by means of flow cytometry.  

PubMed

The stem cells in freshwater flatworms (planarian) are called neoblasts. Neoblasts are capable of proliferation and differentiation into every cell type, including the gametes. For the investigation of the mechanisms of stem cells proliferation and differentiation the proper evaluation of changes in the cell cycle of neoblasts in different physiological conditions of planarian is necessary. In the present study the possibility of qualitative and quantitative characteristics of the neoblasts population were investigated using flow cytometry. In the cell suspension prepared from planarian tissue proliferating neoblasts have been observed in heterogenic cell population. Quantitative estimation of the cell cycle related changes of planarian stem cells system have been performed in various physiological conditions (intact and regenerating animals) and under the influence of physical (ionizing radiation) and chemical (melatonin and colchicine) factors. The modified protocol for planarian stem cells isolation proved to be effective and reproducible and can be recommended for flow cytometry analyses of human and animal proliferating cells. PMID:21688150

Ermakov, Artem M; Ermakova, Olga N; Kudravtsev, Andrei A; Kreshchenko, Natalia D

2011-06-19

61

Cell Death and Tissue Remodeling in Planarian Regeneration  

PubMed Central

Many long-lived organisms, including humans, can regenerate some adult tissues lost to physical injury or disease. Much of the previous research on mechanisms of regeneration has focused on adult stem cells, which give rise to new tissue necessary for the replacement of missing body parts. Here we report that apoptosis of differentiated cells complements stem cell division during regeneration in the planarian Schmidtea mediterranea. Specifically, we developed a whole-mount TUNEL assay that allowed us to document two dramatic increases in the rate of apoptosis following amputation – an intial localized response near the wound site and a subsequent systemic response that varies in magnitude depending on the type of fragment examined. The latter cell death response can be induced in uninjured organs, occurs in the absence of planarian stem cells, and can also be triggered by prolonged starvation. Taken together, our results implicate apoptosis in the restoration of proper anatomical scale and proportion through remodeling of existing tissues. We also report results from initial mechanistic studies of apoptosis in planarians, which revealed that a S. mediterranea homolog of the antiapoptotic gene BCL2 is required for cell survival in adult animals. We propose that apoptosis is a central mechanism working in concert with stem cell division to restore anatomical form and function during metazoan regeneration.

Pellettieri, Jason; Fitzgerald, Patrick; Watanabe, Shigeki; Mancuso, Joel; Green, Douglas R.; Alvarado, Alejandro Sanchez

2010-01-01

62

pbx is required for pole and eye regeneration in planarians.  

PubMed

Planarian regeneration involves regionalized gene expression that specifies the body plan. After amputation, planarians are capable of regenerating new anterior and posterior poles, as well as tissues polarized along the anterior-posterior, dorsal-ventral and medial-lateral axes. Wnt and several Hox genes are expressed at the posterior pole, whereas Wnt inhibitory genes, Fgf inhibitory genes, and prep, which encodes a TALE-family homeodomain protein, are expressed at the anterior pole. We found that Smed-pbx (pbx for short), which encodes a second planarian TALE-family homeodomain transcription factor, is required for restored expression of these genes at anterior and posterior poles during regeneration. Moreover, pbx(RNAi) animals gradually lose pole gene expression during homeostasis. By contrast, pbx was not required for initial anterior-posterior polarized responses to wounds, indicating that pbx is required after wound responses for development and maintenance of poles during regeneration and homeostatic tissue turnover. Independently of the requirement for pbx in pole regeneration, pbx is required for eye precursor formation and, consequently, eye regeneration and eye replacement in homeostasis. Together, these data indicate that pbx promotes pole formation of body axes and formation of regenerative progenitors for eyes. PMID:23318641

Chen, Chun-Chieh G; Wang, Irving E; Reddien, Peter W

2013-01-14

63

pbx is required for pole and eye regeneration in planarians  

PubMed Central

Planarian regeneration involves regionalized gene expression that specifies the body plan. After amputation, planarians are capable of regenerating new anterior and posterior poles, as well as tissues polarized along the anterior-posterior, dorsal-ventral and medial-lateral axes. Wnt and several Hox genes are expressed at the posterior pole, whereas Wnt inhibitory genes, Fgf inhibitory genes, and prep, which encodes a TALE-family homeodomain protein, are expressed at the anterior pole. We found that Smed-pbx (pbx for short), which encodes a second planarian TALE-family homeodomain transcription factor, is required for restored expression of these genes at anterior and posterior poles during regeneration. Moreover, pbx(RNAi) animals gradually lose pole gene expression during homeostasis. By contrast, pbx was not required for initial anterior-posterior polarized responses to wounds, indicating that pbx is required after wound responses for development and maintenance of poles during regeneration and homeostatic tissue turnover. Independently of the requirement for pbx in pole regeneration, pbx is required for eye precursor formation and, consequently, eye regeneration and eye replacement in homeostasis. Together, these data indicate that pbx promotes pole formation of body axes and formation of regenerative progenitors for eyes.

Chen, Chun-Chieh G.; Wang, Irving E.; Reddien, Peter W.

2013-01-01

64

Comparative study of adaptive radiations with an example using parasitic flatworms (Platyhelminthes): Cercomeria  

Microsoft Academic Search

Studies of adaptive radiations require robust phylogenies, estimates of species numbers for monophyletic groups within clades, assessments of the adaptive value of putative key innovations, and estimates of the frequency of speciation modes. Four criteria are necessary to identify an adaptive radiation within the parasitic platyhelminths: (1) a group contains significantly more species than its sister group, (2) species richness

Daniel R. Brooks; Deborah A. McLennan

1993-01-01

65

Atypical properties of a conventional calcium channel ? subunit from the platyhelminth Schistosoma mansoni  

Microsoft Academic Search

BACKGROUND: The function of voltage-gated calcium (Cav) channels greatly depends on coupling to cytoplasmic accessory ? subunits, which not only promote surface expression, but also modulate gating and kinetic properties of the ?1 subunit. Schistosomes, parasitic platyhelminths that cause schistosomiasis, express two ? subunit subtypes: a structurally conventional ? subunit and a variant ? subunit with unusual functional properties. We

Vicenta Salvador-Recatalŕ; Toni Schneider; Robert M Greenberg

2008-01-01

66

Correlating early evolution of parasitic platyhelminths to Gondwana breakup.  

PubMed

Investigating patterns and processes of parasite diversification over ancient geological periods should involve comparisons of host and parasite phylogenies in a biogeographic context. It has been shown previously that the geographical distribution of host-specific parasites of sarcopterygians was guided, from Palaeozoic to Cainozoic times, mostly by evolution and diversification of their freshwater hosts. Here, we propose phylogenies of neobatrachian frogs and their specific parasites (Platyhelminthes, Monogenea) to investigate coevolutionary processes and historical biogeography of polystomes and further discuss all the possible assumptions that may account for the early evolution of these parasites. Phylogenetic analyses of concatenated rRNA nuclear genes (18S and partial 28S) supplemented by cophylogenetic and biogeographic vicariance analyses reveal four main parasite lineages that can be ascribed to centers of diversity, namely Australia, India, Africa, and South America. In addition, the relationships among these biogeographical monophyletic groups, substantiated by molecular dating, reflect sequential origins during the breakup of Gondwana. The Australian polystome lineage may have been isolated during the first stages of the breakup, whereas the Indian lineage would have arisen after the complete separation of western and eastern Gondwanan components. Next, polystomes would have codiverged with hyloid sensu stricto and ranoid frog lineages before the completion of South American and African plate separation. Ultimately, they would have undergone an extensive diversification in South America when their ancestral host families diversified. Therefore, the presence of polystome parasites in specific anuran host clades and in discrete geographic areas reveals the importance of biogeographic vicariance in diversification processes and supports the occurrence and radiation of amphibians over ancient and recent geological periods. PMID:21856629

Badets, Mathieu; Whittington, Ian; Lalubin, Fabrice; Allienne, Jean-Francois; Maspimby, Jean-Luc; Bentz, Sophie; Du Preez, Louis H; Barton, Diane; Hasegawa, Hideo; Tandon, Veena; Imkongwapang, Rangpenyuba; Imkongwapang, Rangpenyubai; Ohler, Annemarie; Combes, Claude; Verneau, Olivier

2011-08-18

67

Sucrose produces withdrawal and dopamine-sensitive reinforcing effects in planarians.  

PubMed

Sucrose produces physical dependence and reinforcing effects in rats. We hypothesized that similar effects could be demonstrated in planarians, the earliest animal with a centralized nervous system. We used two assays, one that quantifies withdrawal responses during drug absence as a reduction in motility and another that quantifies reinforcing effects using a conditioned place preference (CPP) design. In withdrawal experiments, planarians exposed to sucrose (1%) for 60 min and then tested in water for 5 min displayed reduced motility compared to water controls. Acute or continuous sucrose (1%) exposure did not affect motility. CPP experiments used a biased design to capitalize upon planarians' natural preference for the dark (pretest, sucrose conditioning in the light, posttest). Planarians conditioned with sucrose (1%) displayed a greater preference shift than sucrose-naďve planarians. Glucose (0.1, 1%), but not the non-digestible disaccharide lactulose (0.1, 1%), also produced a greater preference shift than water-exposed planarians. Development of sucrose-induced CPP was inhibited when sucrose (1%) conditioning was conducted in combination with dopamine receptor antagonists SCH 23390 (1 ?M) or sulpiride (1 ?M). These results suggest that the rewarding and reinforcing effects of sugar are highly conserved across species and that planarians offer an invertebrate model to provide insight into the pharmacological effects of sucrose and related sweeteners. PMID:23415661

Zhang, Charlie; Tallarida, Christopher S; Raffa, Robert B; Rawls, Scott M

2013-02-13

68

Interrelationships and evolution of the tapeworms (Platyhelminthes: Cestoda).  

PubMed

Interrelationships of the tapeworms (Platyhelminthes: Cestoda) were examined by use of small (SSU) and large (LSU) subunit ribosomal DNA sequences and morphological characters. Fifty new complete SSU sequences were added to 21 sequences previously determined, and 71 new LSU (D1-D3) sequences were determined for the complementary set of taxa representing each of the major lineages of cestodes as currently understood. New sequences were determined for three amphilinidean taxa, but were removed from both alignments due to their excessively high degree of divergence from other cestode sequences. A morphological character matrix coded for supraspecific taxa was constructed by the modification of matrices from recently published studies. Maximum-parsimony (MP) analyses were performed on the LSU, SSU, LSU+SSU, and morphological data partitions, and minimum-evolution (ME) analyses utilizing a general time reversible model of nucleotide substitution including estimates of among-site rate heterogeneity were performed on the molecular data partitions. Resulting topologies were rooted at the node separating the Gyrocotylidea from the Eucestoda. The LSU data were found to be more informative than the SSU data and were more consistent with inferences from morphology, although nodal support was generally weak for most basal nodes. One class of transitions was found to be saturated for comparisons between the most distantly related taxa (gyrocotylideans vs cyclophyllideans and tetrabothriideans). Differences in the topologies resulting from MP and ME analyses were not statistically significant. Nonstrobilate orders formed the basal lineages of trees resulting from analysis of LSU data and morphology. Difossate orders were basal to tetrafossate orders, the latter of which formed a strongly supported clade. A clade including the orders Cyclophyllidea, Nippotaeniidea, and Tetrabothriidea was supported by all data partitions and methods of analysis. Paraphyly of the orders Pseudophyllidea, Tetraphyllidea, and Trypanorhyncha was consistent among the molecular data partitions. Inferences are made regarding a monozoic (nonsegmented) origin of the Eucestoda as represented by the Caryophyllidea and for the evolution of the strobilate and acetabulate/tetrafossate conditions having evolved in a stepwise pattern. PMID:11399152

Olson, P D; Littlewood, D T; Bray, R A; Mariaux, J

2001-06-01

69

Constitutive gene expression and specification of tissue identity in adult planarian biology  

PubMed Central

Planarians are flatworms that constitutively maintain adult tissues through cell turnover and can regenerate entire organisms from tiny body fragments. In addition to requiring new cells (from neoblasts), these feats require mechanisms that specify tissue identity in the adult. Critical roles for Wnt and BMP signaling in regeneration and maintenance of the body axes have been uncovered, among other regulatory factors. Available data indicate that genes involved in positional identity regulation at key embryonic stages in other animals display persisting regionalized expression in adult planarians. These expression patterns suggest that a constitutively active gene expression map exists for maintenance of the planarian body. Planarians therefore present a fertile ground for identification of factors regulating regionalization of the metazoan body plan and for study of the attributes of these factors that can lead to maintenance and regeneration of adult tissues.

Reddien, Peter W.

2011-01-01

70

Transcriptome analysis of the planarian eye identifies ovo as a specific regulator of eye regeneration.  

PubMed

Among the millions of invertebrate species with visual systems, the genetic basis of eye development and function is well understood only in Drosophila melanogaster. We describe an eye transcriptome for the planarian Schmidtea mediterranea. Planarian photoreceptors expressed orthologs of genes required for phototransduction and microvillus structure in Drosophila and vertebrates, and optic pigment cells expressed solute transporters and melanin synthesis enzymes similar to those active in the vertebrate retinal pigment epithelium. Orthologs of several planarian eye genes, such as bestrophin-1 and Usher syndrome genes, cause eye defects in mammals when perturbed and were not previously described to have roles in invertebrate eyes. Five previously undescribed planarian eye transcription factors were required for normal eye formation during head regeneration. In particular, a conserved, transcription-factor-encoding ovo gene was expressed from the earliest stages of eye regeneration and was required for regeneration of all cell types of the eye. PMID:22884275

Lapan, Sylvain W; Reddien, Peter W

2012-08-02

71

Transcriptome analysis of the planarian eye identifies ovo as a specific regulator of eye regeneration  

PubMed Central

Summary Among the millions of invertebrate species with visual systems, the genetic basis of eye development and function is well understood only in Drosophila melanogaster. We describe an eye transcriptome for the planarian Schmidtea mediterranea. Planarian photoreceptors expressed orthologs of genes required for phototransduction and microvillus structure in Drosophila and vertebrates, and optic pigment cells expressed solute transporters and melanin synthesis enzymes similar to those active in the vertebrate retinal pigment epithelium. Orthologs of several planarian eye genes, such as bestrophin-1 and Usher syndrome genes, cause eye defects in mammals when perturbed and were not previously described to have roles in invertebrate eyes. Five previously undescribed planarian eye transcription factors were required for normal eye formation during head regeneration. In particular, a conserved, transcription factor-encoding ovo gene was expressed from the earliest stages of eye regeneration and was required for regeneration of all cell types of the eye.

Lapan, Sylvain W.; Reddien, Peter W.

2013-01-01

72

The history and enduring contributions of planarians to the study of animal regeneration.  

PubMed

Having an almost unlimited capacity to regenerate tissues lost to age and injury, planarians have long fascinated naturalists. In the Western hemisphere alone, their documented history spans more than 200 years. Planarians were described in the early 19th century as being 'immortal under the edge of the knife', and initial investigation of these remarkable animals was significantly influenced by studies of regeneration in other organisms and from the flourishing field of experimental embryology in the late 19th and early 20th centuries. This review strives to place the study of planarian regeneration into a broader historical context by focusing on the significance and evolution of knowledge in this field. It also synthesizes our current molecular understanding of the mechanisms of planarian regeneration uncovered since this animal's relatively recent entrance into the molecular-genetic age. PMID:23799578

Elliott, Sarah A; Sánchez Alvarado, Alejandro

2012-07-23

73

Cell, tissue-, and position-specific monoclonal antibodies against the planarian Dugesia (Girardia) tigrina  

Microsoft Academic Search

To obtain specific immunological probes for studying molecular mechanisms involved in cell renewal, cell differentiation,\\u000a and pattern formation in intact and regenerating planarians, we have produced a hybridoma library specific for the asexual\\u000a race of the freshwater planarian Dugesia (Girardia) tigrina. Among the 276 monoclonal antibodies showing tissue-, cell-, cell subtype-, subcellular- and position-specific staining,\\u000a we have found monoclonal antibodies

D. Bueno; Jaume Baguńŕ; Rafael Romero

1997-01-01

74

Epimorphic regeneration of the distal part of the planarian pharynx.  

PubMed

The totipotent stem cells called neoblasts seem to be concerned with the remarkable regeneration ability of planarians. However, the pharynx is able to regenerate after the amputation of its distal part, in spite of a lack of neoblasts in the pharynx. The process of regeneration has been referred to as morphallaxis, based on conventional histochemical observations. We examined it again immuno-histochemically using anti-Dugesia japonica proliferating cell nuclear antigen (DjPCNA) antibody for neoblasts and anti-D. japonica myosin heavy chain-A (DjMHC-A) antibody for pharynx muscle fibers. This immuno-histochemical study, together with observations of the regeneration process of planarians irradiated with X-rays in particular regions, revealed that after the amputation, neoblasts from outside the pharynx entered that organ, moved through the mesenchyme of the pharynx to the wounded area, and differentiated into the cells that had been lost there. We show here that the regeneration after amputation of the distal part of the pharynx is an 'epimorphic' process. PMID:11277401

Ito, H; Saito, Y; Watanabe, K; Orii, H

2001-01-01

75

Signification of the sexualizing substance produced by the sexualized planarians.  

PubMed

Asexual worms of an exclusively fissiparous strain (the OH strain) of the planarian Dugesia ryukyuensis keep developing hermaphroditic reproductive organs and eventually undergo sexual reproduction instead of asexual reproduction, namely fission, if they are fed with sexually mature worms of an exclusively oviparous planarian, Bdellocephala brunnea, suggesting that the sexually mature worms has a sexualizing substance(s). The fully sexualized worms no longer need the feeding on sexual worms to maintain the sexuality. Here, we demonstrate that the sexualized worms produce enough of their own sexualizing substance similar to that contained in B. brunnea. In case of surgical ablation of the sexualized worms, the fragments with sexual organs regenerate to become sexual, while those without sexual organs, namely head fragments, regenerate to return to the asexual state. The asexual regenerants from the sexualized worms are also fully sexualized by being fed with B. brunnea. Additionally, it was reported that head region in sexually mature worms lacks the putative sexualizing substance necessary for complete sexualization (Sakurai, 1981). These results suggest that the fragments without sexual organ lack enough of an amount of the putative sexualizing substance and the sexuality is maintained by the sexualizing substance contained in the sexualized worms. PMID:12130794

Kobayashi, Kazuya; Arioka, Sachiko; Hase, Sumitaka; Hoshi, Motonori

2002-06-01

76

Role of c-Jun N-terminal kinase activation in blastema formation during planarian regeneration.  

PubMed

The robust regenerative abilities of planarians absolutely depend on a unique population of pluripotent stem cells called neoblasts, which are the only mitotic somatic cells in adult planarians and are responsible for blastema formation after amputation. Little is known about the molecular mechanisms that drive blastema formation during planarian regeneration. Here we found that treatment with the c-Jun N-terminal kinase (JNK) inhibitor SP600125 blocked the entry of neoblasts into the M-phase of the cell cycle, while allowing neoblasts to successfully enter S-phase in the planarian Dugesia japonica. The rapid and efficient blockage of neoblast mitosis by treatment with the JNK inhibitor provided a method to assess whether temporally regulated cell cycle activation drives blastema formation during planarian regeneration. In the early phase of blastema formation, activated JNK was detected prominently in a mitotic region (the "postblastema") proximal to the blastema region. Furthermore, we demonstrated that undifferentiated mitotic neoblasts in the postblastema showed highly activated JNK at the single cell level. JNK inhibition by treatment with SP600125 during this period caused a severe defect of blastema formation, which accorded with a drastic decrease of mitotic neoblasts in regenerating animals. By contrast, these animals still retained many undifferentiated neoblasts near the amputation stump. These findings suggest that JNK signaling plays a crucial role in feeding into the blastema neoblasts for differentiation by regulating the G2/M transition in the cell cycle during planarian regeneration. PMID:21447099

Tasaki, Junichi; Shibata, Norito; Sakurai, Toshihide; Agata, Kiyokazu; Umesono, Yoshihiko

2011-03-30

77

The use of lectins as markers for differentiated secretory cells in planarians  

PubMed Central

Freshwater planarians have reemerged as excellent models to investigate mechanisms underlying regeneration. The introduction of molecular tools has facilitated the study of planarians, but cell- and tissue-specific markers are still needed to examine differentiation of most cell types. Here we report the utility of fluorescent lectin-conjugates to label tissues in the planarian Schmidtea mediterranea. We show that 16 lectin-conjugates stain planarian cells or tissues; 13 primarily label the secretory cells, their cytoplasmic projections, and terminal pores. Thus, we examined regeneration of the secretory system using lectin markers and functionally characterized two genes expressed in the secretory cells: marginal adhesive gland-1 (mag-1) and Smed-reticulocalbin1 (Smed-rcn1). RNAi knockdown of these genes caused a dramatic reduction of secretory cell lectin staining, suggesting a role for mag-1 and Smed-rcn1 in secretory cell differentiation. Our results provide new insights into planarian secretory system regeneration and add new markers for labeling several planarian tissues.

Zayas, Ricardo M.; Cebria, Francesc; Guo, Tingxia; Feng, Junjie; Newmark, Phillip A.

2010-01-01

78

A cembranoid from tobacco prevents the expression of nicotine-induced withdrawal behavior in planarian worms  

PubMed Central

Using an adaptation of published behavioral protocols, we determined that acute exposure to the cholinergic compounds nicotine and carbamylcholine decreased planarian motility in a concentration-dependent manner. A tobacco cembranoid (1S,2E,4R,6R,7E,11E)-cembra-2,7,11-triene-4,6-diol (4R-cembranoid), also decreased planarian motility. Experiments in the presence of 1 ?M 4R-cembranoid did increase the IC50 for nicotine- but not carbamylcholine-induced decrease in planarian motility. When planarians were exposed for 24 h to either nicotine or carbamylcholine at concentrations near their respective IC50 values and then transferred to plain media, nicotine-exposed, but not carbamylcholine- or cembranoid-exposed worms displayed withdrawal-like distress behaviors. In experiments where planarians were pre-exposed to 100 ?M nicotine for 24 h in the presence of 1 ?M 4R-cembranoid, the withdrawal-like effects were significantly reduced. These results indicate that the 4R-cembranoid might have valuable applications for tobacco abuse research. This experimental approach using planarians is useful for the initial screening of compounds relevant to drug abuse and dependence.

Pagan, One R.; Rowlands, Amanda L.; Fattore, Angela L.; Coudron, Tamara; Urban, Kimberly R.; Bidja, Apurva H.; Eterovic, Vesna A.

2010-01-01

79

A cembranoid from tobacco prevents the expression of nicotine-induced withdrawal behavior in planarian worms.  

PubMed

Using an adaptation of published behavioral protocols, we determined that acute exposure to the cholinergic compounds nicotine and carbamylcholine decreased planarian motility in a concentration-dependent manner. A tobacco cembranoid (1S,2E,4R,6R,7E,11E)-cembra-2,7,11-triene-4,6-diol (4R-cembranoid), also decreased planarian motility. Experiments in the presence of 1 microM 4R-cembranoid did increase the IC50 for nicotine- but not carbamylcholine-induced decrease in planarian motility. When planarians were exposed for 24 h to either nicotine or carbamylcholine at concentrations near their respective IC50 values and then transferred to plain media, nicotine-exposed, but not carbamylcholine- or cembranoid-exposed worms displayed withdrawal-like distress behaviors. In experiments where planarians were pre-exposed to 100 microM nicotine for 24 h in the presence of 1 microM 4R-cembranoid, the withdrawal-like effects were significantly reduced. These results indicate that the 4R-cembranoid might have valuable applications for tobacco abuse research. This experimental approach using planarians is useful for the initial screening of compounds relevant to drug abuse and dependence. PMID:19490913

Pagán, Oné R; Rowlands, Amanda L; Fattore, Angela L; Coudron, Tamara; Urban, Kimberly R; Bidja, Apurva H; Eterovi?, Vesna A

2009-05-30

80

Voltage-gated calcium channel subunits from platyhelminths: Potential role in praziquantel action?  

PubMed Central

Voltage-gated calcium (Ca2+) channels provide the pathway for Ca2+ influxes that underlie Ca2+-dependent responses in muscles, nerves and other excitable cells. They are also targets of a wide variety of drugs and toxins. Ca2+ channels are multisubunit protein complexes consisting of a pore-forming ?1 subunit and other modulatory subunits, including the ? subunit. Here, we review the structure and function of schistosome Ca2+ channel subunits, with particular emphasis on variant Ca2+ channel ? subunits (Cav?var) found in these parasites. In particular, we examine the role these ? subunits may play in the action of praziquantel, the current drug of choice against schistosomiasis. We also present evidence that Cav?var homologs are found in other praziquantel-sensitive platyhelminths such as the pork tapeworm, Taenia solium, and that these variant ? subunits may thus represent a platyhelminth-specific gene family.

Jeziorski, Michael C.; Greenberg, Robert M.

2013-01-01

81

Mystery tubes coiled around deep-water tropical gorgonians: fecampiid cocoons (Platyhelminthes: Fecampiida) resembling Solenogastres (Mollusca).  

PubMed

During the examination of a large suite of tropical deep-water molluscs, a number of Solenogastres were found, some of them typically curled around gorgonian stems. A subsequent closer examination of the Solenogastres revealed another type of object also curled around the gorgonians, which strongly resembled Solenogastres but lacked their external features. These objects proved to be cocoons with egg capsules, each containing two eggs or young larvae, typical of the parasitic platyhelminth group Fecampiida. PMID:17143569

Handl, Claudia; Bouchet, Philippe

2006-12-02

82

5-HT1A-like receptor activation inhibits abstinence-induced methamphetamine withdrawal in planarians  

PubMed Central

No pharmacological therapy is approved to treat methamphetamine physical dependence, but it has been hypothesized that serotonin (5-HT)-enhancing drugs might limit the severity of withdrawal symptoms. To test this hypothesis, we used a planarian model of physical dependence that quantifies withdrawal as a reduction in planarian movement. Planarians exposed to methamphetamine (10 µM) for 60 min, and then placed (tested) into drug-free water for 5 min, displayed less movement (i.e., withdrawal) than either methamphetamine-naďve planarians tested in water or methamphetamine-exposed planarians tested in methamphetamine. A concentration-related inhibition of withdrawal was observed when methamphetamine-exposed planarians were placed into a solution containing either methamphetamine and 5-HT (0.1 – 100 µM) or methamphetamine and the 5-HT1A receptor agonist 8-Hydroxy-N,N-dipropyl-2-aminotetralin (8-OH-DPAT) (10, 20 µM). Planarians with prior methamphetamine exposure displayed enhanced withdrawal when tested in a solution of the 5-HT1A receptor antagonist N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]- N-(2-pyridyl)cyclohexanecarboxamide (WAY 100635) (1 µM). Methamphetamine-induced withdrawal was not affected by the 5-HT2B/2C receptor agonist meta-chlorophenylpiperazine (m-CPZ) (0.1 – 20 µM). These results provide pharmacological evidence that serotonin-enhancing drugs inhibit expression of methamphetamine physical dependence in an invertebrate model of withdrawal, possibly through a 5-HT1A-like receptor-dependent mechanism.

Rawls, Scott M.; Shah, Hardik; Ayoub, George; Raffa, Robert B.

2010-01-01

83

Bioelectric signaling regulates head and organ size during planarian regeneration.  

PubMed

A main goal of regenerative medicine is to replace lost or damaged tissues and organs with functional parts of the correct size and shape. But the proliferation of new cells is not sufficient; we will also need to understand how the scale and ultimate form of newly produced tissues are determined. Using the planarian model system, we report that membrane voltage-dependent bioelectric signaling determines both head size and organ scaling during regeneration. RNA interference of the H(+),K(+)-ATPase ion pump results in membrane hyperpolarization, which has no effect on the amount of new tissue (blastema) that is regenerated yet produces regenerates with tiny 'shrunken' heads and proportionally oversized pharynges. Our data show that this disproportionality results from a lack of the apoptosis required to adjust head and organ size and placement, highlighting apoptotic remodeling as the link between bioelectric signaling and the establishment of organ size during regeneration. PMID:23250205

Beane, Wendy Scott; Morokuma, Junji; Lemire, Joan M; Levin, Michael

2013-01-15

84

Follistatin antagonizes Activin signaling and acts with Notum to direct planarian head regeneration  

PubMed Central

Animals establish their body plans in embryogenesis, but only a few animals can recapitulate this signaling milieu for regeneration after injury. In planarians, a pluripotent stem cell population and perpetual signaling of polarity axes collaborate to direct a steady replacement of cells during homeostasis and to power robust regeneration after even severe injuries. Several studies have documented the roles of conserved signaling pathways in maintaining and resetting axial polarity in planarians, but it is unclear how planarians reestablish polarity signaling centers after injury and whether these centers serve to influence identity decisions of stem cell progeny during their differentiation. Here we find that a planarian Follistatin homolog directs regeneration of anterior identity by opposing an Activin/ActR-1/Smad2/3 signaling pathway. Follistatin and Notum, a Wnt inhibitor, are mutually required to reestablish an anterior signaling center that expresses both cues. Furthermore, we show that the direction of cells down particular differentiation paths requires regeneration of this anterior signaling center. Just as its amphibian counterpart in the organizer signals body plan and cell fate during embryogenesis, planarian Follistatin promotes reestablishment of anterior polarity during regeneration and influences specification of cell types in the head and beyond.

Roberts-Galbraith, Rachel H.; Newmark, Phillip A.

2013-01-01

85

Ingestion of bacterially expressed double-stranded RNA inhibits gene expression in planarians  

PubMed Central

Freshwater planarian flatworms are capable of regenerating complete organisms from tiny fragments of their bodies; the basis for this regenerative prowess is an experimentally accessible stem cell population that is present in the adult planarian. The study of these organisms, classic experimental models for investigating metazoan regeneration, has been revitalized by the application of modern molecular biological approaches. The identification of thousands of unique planarian ESTs, coupled with large-scale whole-mount in situ hybridization screens, and the ability to inhibit planarian gene expression through double-stranded RNA-mediated genetic interference, provide a wealth of tools for studying the molecular mechanisms that regulate tissue regeneration and stem cell biology in these organisms. Here we show that, as in Caenorhabditis elegans, ingestion of bacterially expressed double-stranded RNA can inhibit gene expression in planarians. This inhibition persists throughout the process of regeneration, allowing phenotypes with disrupted regenerative patterning to be identified. These results pave the way for large-scale screens for genes involved in regenerative processes.

Newmark, Phillip A.; Reddien, Peter W.; Cebria, Francesc; Alvarado, Alejandro Sanchez

2003-01-01

86

Regeneration of dopaminergic neurons after 6-hydroxydopamine-induced lesion in planarian brain.  

PubMed

Planarians have robust regenerative ability dependent on X-ray-sensitive pluripotent stem cells, called neoblasts. Here, we report that planarians can regenerate dopaminergic neurons after selective degeneration of these neurons caused by treatment with a dopaminergic neurotoxin (6-hydroxydopamine; 6-OHDA). This suggests that planarians have a system to sense the degeneration of dopaminergic neurons and to recruit stem cells to produce dopaminergic neurons to recover brain morphology and function. We confirmed that X-ray-irradiated planarians do not regenerate brain dopaminergic neurons after 6-OHDA-induced lesioning, suggesting that newly generated dopaminergic neurons are indeed derived from pluripotent stem cells. However, we found that the majority of regenerated dopaminergic neurons were 5-bromo-2'-deoxyuridine-negative cells. Therefore, we carefully analyzed when proliferating stem cells became committed to become dopaminergic neurons during regeneration by a combination of 5-bromo-2'-deoxyuridine pulse-chase experiments, immunostaining/in situ hybridization, and 5-fluorouracil treatment. The results strongly suggested that G(2) -phase stem cells become committed to dopaminergic neurons in the mesenchymal space around the brain, after migration from the trunk region following S-phase. These new findings obtained from planarian regeneration provide hints about how to conduct cell-transplantation therapy for future regenerative medicine. PMID:21985107

Nishimura, Kaneyasu; Inoue, Takeshi; Yoshimoto, Kanji; Taniguchi, Takashi; Kitamura, Yoshihisa; Agata, Kiyokazu

2011-11-03

87

A molecular wound response program associated with regeneration initiation in planarians  

PubMed Central

Planarians are capable of regenerating any missing body part and present an attractive system for molecular investigation of regeneration initiation. The gene activation program that occurs at planarian wounds to coordinate regenerative responses remains unknown. We identified a large set of wound-induced genes during regeneration initiation in planarians. Two waves of wound-induced gene expression occurred in differentiated tissues. The first wave includes conserved immediate early genes. Many second-wave genes encode conserved patterning factors required for proper regeneration. Genes of both classes were generally induced by wounding, indicating that a common initial gene expression program is triggered regardless of missing tissue identity. Planarian regeneration uses a population of regenerative cells (neoblasts), including pluripotent stem cells. A class of wound-induced genes was activated directly within neoblasts, including the Runx transcription factor-encoding runt-1 gene. runt-1 was required for specifying different cell types during regeneration, promoting heterogeneity in neoblasts near wounds. Wound-induced gene expression in neoblasts, including that of runt-1, required SRF (serum response factor) and sos-1. Taken together, these data connect wound sensation to the activation of specific cell type regeneration programs in neoblasts. Most planarian wound-induced genes are conserved across metazoans, and identified genes and mechanisms should be important broadly for understanding wound signaling and regeneration initiation.

Wenemoser, Danielle; Lapan, Sylvain W.; Wilkinson, Alex W.; Bell, George W.; Reddien, Peter W.

2012-01-01

88

Effects of N,N-dimethylformamide on behaviour and regeneration of planarian Dugesia japonica.  

PubMed

In this study, the toxicity, behavioural and regeneration effects of dimethylformamide (DMF) on planarian Dugesia japonica were investigated. One control and six different concentrations of DMF (10 ppm, 100 ppm, 500 ppm, 1000 ppm, 5000 ppm and 10,000 ppm) were used in triplicate. The results showed that the mortality was directly proportional to the DMF concentration and planarian locomotor velocity (pLMV) was significantly reduced by increasing the exposure time and DMF concentration. pLMV of D. japonica was significantly reduced at a lower concentration of 10 ppm after 7 days of continuous exposure to DMF. The recovery of the motility of planarians pretreated with DMF was found to be time- and dose dependent, all planarians had complete recovery in their motility after 48 h. The appearance of auricles in regenerating animals was easily affected by DMF exposure in comparison with the appearance of eyespot. The present results suggest that the intact adult mobility in the aquatic planarian D. japonica is a more sensitive biomarker than mortality, and the appearance of auricles in regenerating animals is a more sensitive biomarker than eyespot. PMID:22495519

Zhang, Jianyong; Yuan, Zuoqing; Zheng, Mingyue; Sun, Yuqian; Wang, Youjun; Yang, Shudong

2012-04-10

89

A molecular wound response program associated with regeneration initiation in planarians.  

PubMed

Planarians are capable of regenerating any missing body part and present an attractive system for molecular investigation of regeneration initiation. The gene activation program that occurs at planarian wounds to coordinate regenerative responses remains unknown. We identified a large set of wound-induced genes during regeneration initiation in planarians. Two waves of wound-induced gene expression occurred in differentiated tissues. The first wave includes conserved immediate early genes. Many second-wave genes encode conserved patterning factors required for proper regeneration. Genes of both classes were generally induced by wounding, indicating that a common initial gene expression program is triggered regardless of missing tissue identity. Planarian regeneration uses a population of regenerative cells (neoblasts), including pluripotent stem cells. A class of wound-induced genes was activated directly within neoblasts, including the Runx transcription factor-encoding runt-1 gene. runt-1 was required for specifying different cell types during regeneration, promoting heterogeneity in neoblasts near wounds. Wound-induced gene expression in neoblasts, including that of runt-1, required SRF (serum response factor) and sos-1. Taken together, these data connect wound sensation to the activation of specific cell type regeneration programs in neoblasts. Most planarian wound-induced genes are conserved across metazoans, and identified genes and mechanisms should be important broadly for understanding wound signaling and regeneration initiation. PMID:22549959

Wenemoser, Danielle; Lapan, Sylvain W; Wilkinson, Alex W; Bell, George W; Reddien, Peter W

2012-05-01

90

Follistatin antagonizes activin signaling and acts with notum to direct planarian head regeneration.  

PubMed

Animals establish their body plans in embryogenesis, but only a few animals can recapitulate this signaling milieu for regeneration after injury. In planarians, a pluripotent stem cell population and perpetual signaling of polarity axes collaborate to direct a steady replacement of cells during homeostasis and to power robust regeneration after even severe injuries. Several studies have documented the roles of conserved signaling pathways in maintaining and resetting axial polarity in planarians, but it is unclear how planarians reestablish polarity signaling centers after injury and whether these centers serve to influence identity decisions of stem cell progeny during their differentiation. Here we find that a planarian Follistatin homolog directs regeneration of anterior identity by opposing an Activin/ActR-1/Smad2/3 signaling pathway. Follistatin and Notum, a Wnt inhibitor, are mutually required to reestablish an anterior signaling center that expresses both cues. Furthermore, we show that the direction of cells down particular differentiation paths requires regeneration of this anterior signaling center. Just as its amphibian counterpart in the organizer signals body plan and cell fate during embryogenesis, planarian Follistatin promotes reestablishment of anterior polarity during regeneration and influences specification of cell types in the head and beyond. PMID:23297191

Roberts-Galbraith, Rachel H; Newmark, Phillip A

2013-01-07

91

Morphological and molecular development of the eyes during embryogenesis of the freshwater planarian Schmidtea polychroa.  

PubMed

Photoreception is one of the most primitive sensory functions in metazoans. Despite the diversity of forms and components of metazoan eyes, many studies have demonstrated the existence of a common cellular and molecular basis for their development. Genes like pax6, sine oculis, eyes absent, dachshund, otx, Rx and atonal are known to be associated with the specification and development of the eyes. In planarians, sine oculis, eyes absent and otxA play an essential role during the formation of the eye after decapitation, whereas pax6, considered by many authors as a master control gene for eye formation, does not seem to be involved in adult eye regeneration. Whether this is a peculiarity of adult planarians or, on the contrary, is also found in embryogenesis remains unknown. Herein, we characterize embryonic eye development in the planarian species Schmidtea polychroa using histological sections and molecular markers. Additionally, we analyse the expression pattern of the pax6-sine oculis-eyes absent-dachshund network, and the genes Rx, otxA, otxB and atonal. We demonstrate that eye formation in planarian embryos shows great similarities to adult eye regeneration, both at the cellular and molecular level. We thus conclude that planarian eyes exhibit divergent molecular patterning mechanisms compared to the prototypic ancestral metazoan eye. PMID:22327190

Martín-Durán, José María; Monjo, Francisco; Romero, Rafael

2012-02-12

92

Early planarian brain regeneration is independent of blastema polarity mediated by the Wnt/?-catenin pathway.  

PubMed

Analysis of anteroposterior (AP) axis specification in regenerating planarian flatworms has shown that Wnt/?-catenin signaling is required for posterior specification and that the FGF-like receptor molecule nou-darake (ndk) may be involved in restricting brain regeneration to anterior regions. The relationship between re-establishment of AP identity and correct morphogenesis of the brain is, however, still poorly understood. Here we report the characterization of two axin paralogs in the planarian Schmidtea mediterranea. Although Axins are well known negative regulators of Wnt/?-catenin signaling, no role in AP specification has previously been reported for axin genes in planarians. We show that silencing of Smed-axin genes by RNA interference (RNAi) results in two-tailed planarians, a phenotype previously reported after silencing of Smed-APC-1, another ?-catenin inhibitor. More strikingly, we show for the first time that while early brain formation at anterior wounds remains unaffected, subsequent development of the brain is blocked in the two-tailed planarians generated after silencing of Smed-axin genes and Smed-APC-1. These findings suggest that the mechanisms underlying early brain formation can be uncoupled from the specification of AP identity by the Wnt/?-catenin pathway. Finally, the posterior expansion of the brain observed following Smed-ndk RNAi is enhanced by silencing Smed-APC-1, revealing an indirect relationship between the FGFR/Ndk and Wnt/?-catenin signaling systems in establishing the posterior limits of brain differentiation. PMID:21806978

Iglesias, Marta; Almuedo-Castillo, Maria; Aboobaker, A Aziz; Saló, Emili

2011-07-20

93

Identification of a receptor tyrosine kinase involved in germ cell differentiation in planarians.  

PubMed

To investigate external signals involved in germ cell differentiation from somatic stem cells, we have tried to identify protein kinases whose expression is regulated during the process of sexualization of asexual-state planarians. It is known that in planarians germ cells differentiate from totipotent somatic stem cells called "neoblasts" during sexualization. As a first step, we have isolated twelve protein kinase genes from cDNAs of sexual-state planarians, including three non-receptor tyrosine kinases, three receptor-tyrosine kinases and three non-receptor serine/threonine kinases, and then analyzed their expression patterns during sexualization. One of them, the DjPTK1 gene, is specifically expressed in germ cells of sexual-state planarians. DjPTK1-positive cells were also detected in the mesenchymal space during the process of sexualization, and it appears that these cells migrate to the dorsal side and then differentiate into spermatogonia/spermatocytes in testis. Sequence analysis indicated that the DjPTK1 gene encodes a receptor protein tyrosine kinase belonging to the FGFR/PDGF family. These results suggest that a receptor tyrosine kinase system may be involved both at an early stage of germ cell differentiation and in a step of germ cell maturation in planarians. PMID:9675112

Ogawa, K; Wakayama, A; Kunisada, T; Orii, H; Watanabe, K; Agata, K

1998-07-01

94

Regeneration and maintenance of the planarian midline is regulated by a slit orthologue  

PubMed Central

Several families of evolutionarily conserved axon guidance cues orchestrate the precise wiring of the nervous system during embryonic development. The remarkable plasticity of freshwater planarians provides the opportunity to study these molecules in the context of neural regeneration and maintenance. Here we characterize a homologue of the Slit family of guidance cues from the planarian Schmidtea mediterranea. Smed-slit is expressed along the planarian midline, in both dorsal and ventral domains. RNA interference targeting Smed-slit results in the collapse of many newly regenerated tissues at the midline; these include the cephalic ganglia, ventral nerve cords, photoreceptors, and the posterior digestive system. Surprisingly, Smed-slit RNAi knockdown animals also develop morphologically distinguishable, ectopic neural structures near the midline in uninjured regions of intact and regenerating planarians. These results suggest that Smed-slit acts not only as a repulsive cue required for proper midline formation during regeneration but that it may also act to regulate the behavior of neural precursors at the midline in intact planarians.

Cebria, Francesc; Guo, Tingxia; Jopek, Jessica; Newmark, Phillip A.

2007-01-01

95

Expression of hsp90 mediates cytoprotective effects in the gastrodermis of planarians  

PubMed Central

Heat shock proteins (HSPs) play a crucial role in the protection of cells. In the present study, we have identified an hsp90-related gene (Djhsp90) encoding a cytosolic form of HSP90 that is primarily expressed in gastrodermis of the planarian Dugesia japonica. Djhsp90 becomes significantly induced after traumatic amputation or other stress stimuli, such as exposure to X-ray or ultraviolet radiations, heat shock, or prolonged starvation. When Djhsp90 is silenced by ribonucleic acid interference (RNAi), planarians dramatically decrease in size, becoming unable to eat, and die in a few weeks. Our results indicate that this gene plays an essential cytoprotective role in the gastrodermis of planarians and suggest that this chaperone can be involved in autophagic processes that are activated by this tissue.

Conte, Maria; Isolani, Maria Emilia; Deri, Paolo; Mannini, Linda

2010-01-01

96

A Formaldehyde-based Whole-Mount In Situ Hybridization Method for Planarians  

PubMed Central

Whole-mount in situ hybridization (WISH) is a powerful tool for visualizing gene expression patterns in specific cell and tissue types. Each model organism presents its own unique set of challenges for achieving robust and reproducible staining with cellular resolution. Here we describe a formaldehyde-based WISH method for the freshwater planarian Schmidtea mediterranea developed by systematically comparing and optimizing techniques for fixation, permeabilization, hybridization and post-processing. The new method gives robust, high-resolution labeling in fine anatomical detail, allows co-labeling with fluorescent probes, and is sufficiently sensitive to resolve the expression pattern of a microRNA in planarians. Our WISH methodology not only provides significant advancements over current protocols that make it a valuable asset for the planarian community, but should also find wide applicability in WISH methods used in other systems.

Pearson, Bret J.; Eisenhoffer, George T.; Gurley, Kyle A.; Rink, Jochen C.; Miller, Diane E.; Alvarado, Alejandro Sanchez

2009-01-01

97

The maintenance and regeneration of the planarian excretory system are regulated by EGFR signaling  

PubMed Central

The maintenance of organs and their regeneration in case of injury are crucial to the survival of all animals. High rates of tissue turnover and nearly unlimited regenerative capabilities make planarian flatworms an ideal system with which to investigate these important processes, yet little is known about the cell biology and anatomy of their organs. Here we focus on the planarian excretory system, which consists of internal protonephridial tubules. We find that these assemble into complex branching patterns with a stereotyped succession of cell types along their length. Organ regeneration is likely to originate from a precursor structure arising in the blastema, which undergoes extensive branching morphogenesis. In an RNAi screen of signaling molecules, we identified an EGF receptor (Smed-EGFR-5) as a crucial regulator of branching morphogenesis and maintenance. Overall, our characterization of the planarian protonephridial system establishes a new paradigm for regenerative organogenesis and provides a platform for exploring its functional and evolutionary homologies with vertebrate excretory systems.

Rink, Jochen C.; Vu, Hanh Thi-Kim; Sanchez Alvarado, Alejandro

2011-01-01

98

An RNAi screen reveals intestinal regulators of branching morphogenesis, differentiation, and stem cell proliferation in planarians.  

PubMed

Planarians grow and regenerate organs by coordinating proliferation and differentiation of pluripotent stem cells with remodeling of postmitotic tissues. Understanding how these processes are orchestrated requires characterizing cell-type-specific gene expression programs and their regulation during regeneration and homeostasis. To this end, we analyzed the expression profile of planarian intestinal phagocytes, cells responsible for digestion and nutrient storage/distribution. Utilizing RNA interference, we identified cytoskeletal regulators required for intestinal branching morphogenesis and a modulator of bioactive sphingolipid metabolism, ceramide synthase, required for the production of functional phagocytes. Additionally, we found that a gut-enriched homeobox transcription factor, nkx-2.2, is required for somatic stem cell proliferation, suggesting a niche-like role for phagocytes. Identification of evolutionarily conserved regulators of intestinal branching, differentiation, and stem cell dynamics demonstrates the utility of the planarian digestive system as a model for elucidating the mechanisms controlling postembryonic organogenesis. PMID:23079596

Forsthoefel, David J; James, Noëlle P; Escobar, David J; Stary, Joel M; Vieira, Ana P; Waters, Forrest A; Newmark, Phillip A

2012-10-16

99

An RNAi screen reveals intestinal regulators of branching morphogenesis, differentiation, and stem cell proliferation in planarians  

PubMed Central

SUMMARY Planarians grow and regenerate organs by coordinating proliferation and differentiation of pluripotent stem cells with remodeling of post-mitotic tissues. Understanding how these processes are orchestrated requires characterizing cell type-specific gene expression programs and their regulation during regeneration and homeostasis. To this end, we analyzed the expression profile of planarian intestinal phagocytes, cells responsible for digestion and nutrient storage/distribution. Utilizing RNA interference, we identified cytoskeletal regulators required for intestinal branching morphogenesis, and a modulator of bioactive sphingolipid metabolism, ceramide synthase, required for the production of functional phagocytes. Additionally, we found that a gut-enriched homeobox transcription factor, nkx-2.2, is required for somatic stem cell proliferation, suggesting a niche-like role for phagocytes. Identification of evolutionarily conserved regulators of intestinal branching, differentiation, and stem cell dynamics demonstrates the utility of the planarian digestive system as a model for elucidating the mechanisms controlling post-embryonic organogenesis.

Forsthoefel, David J.; James, Noelle P.; Escobar, David J.; Stary, Joel M.; Vieira, Ana P.; Waters, Forrest A.; Newmark, Phillip A.

2012-01-01

100

Heterogeneity of planarian stem cells in the S/G2/M phase.  

PubMed

The planarian adult stem cell (pASC) population has a specific molecular signature and can be easily visualized and isolated by flow cytometry. However, the lack of antibodies against specific surface markers for planarian cells prevents a deeper analysis of specific cell populations. Here, if we describe the results of the immunoscreening of pASC plasma membrane proteins (PMPs). A novel papain-based method for planarian cell dissociation enabling both high yield and improved cell viability was used to generate single cell preparations for PMP purification. PMPs were used for intraperitoneal immunization of mice and thus about 1000 hybridoma clones were generated and screened. Supernatants collected from the hybridoma clones were first screened by ELISA and then by live immuno-staining. About half of these supernatants stained all the planarian cells, whereas the other half specifically labeled a subfraction thereof. A detailed analysis of two hybridoma supernatants revealed that large subfractions of the X1, X2 and Xin populations differentially express specific membrane markers. Quantitative PCR data disclosed a correlation between the immunostaining results and the expression of markers of the early and late progeny, also for those pASCs in the S/G2/M phase of the cell cycle (X1 population). Thus, about two thirds of the cycling pASCs showed a specific membrane signature coupled with the expression of markers hitherto considered to be restricted to differentiating, post-mitotic progeny. In summary, a library of 66 monoclonal antibodies against planarian PMPs was generated. The analysis of two of the clones generated revealed that a subset of cells of the X1 population expresses early and late progeny markers, which might indicate that these cells are committed while still proliferating. The findings demonstrate the usefulness of our PMP antibody library for planarian research. PMID:22450999

Moritz, Sören; Stöckle, Franziska; Ortmeier, Claudia; Schmitz, Henning; Rodríguez-Esteban, Gustavo; Key, Göran; Gentile, Luca

2012-01-01

101

The molecular logic for planarian regeneration along the anterior-posterior axis.  

PubMed

The planarian Dugesia japonica can regenerate a complete individual from a head, trunk or tail fragment via activation of somatic pluripotent stem cells. About a century ago, Thomas Hunt Morgan attempted to explain the extraordinary regenerative ability of planarians by positing two opposing morphogenetic gradients of formative "head stuff" and "tail stuff" along the anterior-posterior axis. However, Morgan's hypothesis remains open to debate. Here we show that extracellular signal-related kinase (ERK) and Wnt/?-catenin signalling pathways establish a solid framework for planarian regeneration. Our data suggest that ERK signalling forms a spatial gradient in the anterior region during regeneration. The fibroblast growth factor receptor-like gene nou-darake (which serves as an output of ERK signalling in the differentiating head) and posteriorly biased ?-catenin activity negatively regulate ERK signalling along the anterior-posterior axis in distinct manners, and thereby posteriorize regenerating tissues outside the head region to reconstruct a complete head-to-tail axis. On the basis of this knowledge about D. japonica, we proposed that ?-catenin signalling is responsible for the lack of head-regenerative ability of tail fragments in the planarian Phagocata kawakatsui, and our confirmation thereof supports the notion that posterior ?-catenin signalling negatively modulates the ERK signalling involved in anteriorization across planarian species. These findings suggest that ERK signalling has a pivotal role in triggering globally dynamic differentiation of stem cells in a head-to-tail sequence through a default program that promotes head tissue specification in the absence of posteriorizing signals. Thus, we have confirmed the broad outline of Morgan's hypothesis, and refined it on the basis of our proposed default property of planarian stem cells. PMID:23883928

Umesono, Yoshihiko; Tasaki, Junichi; Nishimura, Yui; Hrouda, Martina; Kawaguchi, Eri; Yazawa, Shigenobu; Nishimura, Osamu; Hosoda, Kazutaka; Inoue, Takeshi; Agata, Kiyokazu

2013-07-24

102

Content of mitochondrial protein in the planarian Polycelis nigra during starvation and feeding.  

PubMed

To discover the possible mechanisms determining the level of energy metabolism during changes in the body size of animals, the content of mitochondrial protein was studied in planarians during starvation and feeding. In the course of starvation, the relative content of mitochondrial protein decreases, whereas during feeding it increases. Comparison of the experimental results with previous observations on respiration of planarians under similar conditions shows that changes in the level of respiration during starvation and feeding correlate with the content of mitochondrial protein. However, the degree of the functional load on the mitochondria must be taken into account under these circumstances. PMID:549686

Grudnitskii, V A; Dontsova, G V

103

FACS analysis of the planarian stem cell compartment as a tool to understand regenerative mechanisms.  

PubMed

Planarians provide a relatively simple model system in which to study stem cell dynamics and regenerative phenomena. As with other systems understanding the dynamics of stem cell and stem cell progeny is crucial in order to get at the molecular mechanisms orchestrating stem cell biology. Planarians have an abundant adult stem cell population that can be observed using Fluorescence-Activated Cell Sorting (FACS). This approach allows different subpopulations of stem cells and their progeny to be monitored and sorted for downstream studies in response to different regenerative scenarios, drug treatments, or RNAi knockdown of genes required for regenerative events. PMID:22914940

Romero, Belen Tejada; Evans, Deborah J; Aboobaker, A Aziz

2012-01-01

104

Diplostomum ( Austrodiplostomum ) compactum (Lutz, 1928) (Platyhelminthes, Digenea) metacercariae in fish from the floodplain of the Upper Paraná River, Brazil  

Microsoft Academic Search

Diplostomum (Austrodiplostomum) compactum (Lutz, 1928) metacercariae (Platyhelminthes, Digenea) were found in six fish species, belonging to two orders (Characiformes and Perciformes) and three families (Erythrinidae, Sciaenidae and Cichlidae). A total of 477 individuals were collected, from August 1999 to May 2001, in the upper Paraná River floodplain, Brazil. The metacercariae were infecting the eyes of six host species and the

P. M. Machado; R. M. Takemoto; G. C. Pavanelli

2005-01-01

105

Smithsoniarhynches, a new genus of interstitial Gnathorhynchidae (Platyhelminthes: Kalyptorhynchia) from Mosquito Lagoon and Indian River Lagoon, Florida  

Microsoft Academic Search

A new genus of marine, interstitial schizorhynch (Platyhelminthes: Kalyptorhynchia) is described from sediment collected in Mosquito Lagoon and Indian River Lagoon, Florida, USA. Smithsoniarhynches is char- acterized by the presence of proboscis hooks constructed of ten individual spine-like teeth that emerge separately from a basal plate. Histology and confocal laser scanning microscopy are used to reveal addi- tional details of

Rick Hochberg

2004-01-01

106

The past and present of planarians--an interview with Vittorio Gremigni. Interview by Alessandra Salvetti and Leonardo Rossi.  

PubMed

Vittorio Gremigni is a scientific leader in the field of planarian biology with a very long historical perspective. By using electron microscopy, he contributed to the reconstruction of the phylogenesis of free living "Turbellaria", and he pioneered the study of the origin of blastema cells by using chromosomal markers. In this interview, Professor Gremigni describes the steps that moved his career towards the planarian field, the main scientific achievements he obtained and the changes that are taking place in the field. He also discusses recent progress and unanswered questions on planarian neoblasts and regeneration. PMID:22450994

Gremigni, Vittorio

2012-01-01

107

Seasonal changes in the sexualization of the planarian Dugesia ryukyuensis.  

PubMed

Asexual individuals in a fissiparous clone of the planarian Dugesia ryukyuensis develop hermaphroditic sexual organs and eventually undergo sexual reproduction instead of asexual reproduction if they are fed with the adults of Bdellocephala brunnea, an oviparous planaria. The experimental sexualization means that the adults of B. brunnea contain a putative sexualizing substance(s), which is the first candidate for the chemical(s) responsible for switching from asexual to sexual reproduction in metazoans. In the present study, the feeding experiment over two consecutive years revealed that the experimental sexualization has seasonal changes. In summer, the asexual individuals were not fully sexualized, though they developed a pair of ovaries. The developing ovaries degenerate if the feeding is stopped. On the contrary, in winter, they developed all the sexual organs. The sexual organs keep developing even if the feeding is stopped after a certain critical point named the point-of-no-return. It was demonstrated that the extreme difference of the sexualization was attributed to the seasonal change of the quality and/or quantity of the sexualizing substance contained in B. brunnea, as well as the minor change of the susceptibility to the sexualizing substance in the asexual individuals. On the other hand, the histological research of B. brunnea revealed that the degree of the maturation of the sexual organs varied extremely through a year. Taking these results into account, we suggest that the production of the sexualizing substance has no direct relation to any particular mature sexual organs. PMID:12499671

Kobayashi, Kazuya; Arioka, Sachiko; Hoshi, Motonori

2002-11-01

108

Discovery of Platyhelminth-Specific ?/?-Integrin Families and Evidence for Their Role in Reproduction in Schistosoma mansoni  

PubMed Central

In all metazoa, the response of cells to molecular stimuli from their environment represents a fundamental principle of regulatory processes controlling cell growth and differentiation. Among the membrane-linked receptors mediating extracellular communication processes are integrin receptors. Besides managing adhesion to the extracellular matrix or to other cells, they arrange information flow into the cells by activating intracellular signaling pathways often acting synergistically through cooperation with growth factor receptors. Although a wealth of information exists on integrins in different model organisms, there is a big gap of knowledge for platyhelminths. Here we report on the in silico detection and reconstruction of ? and ? integrins from free-living and parasitic platyhelminths, which according to structural and phylogenetic analyses form specific clades separate from each other and from further metazoan integrins. As representative orthologs of parasitic platyhelminths we have cloned one beta-integrin (Sm?-Int1) and four alpha-integrins (Sm?-Int1 - Sm?-Int4) from Schistosoma mansoni; they were characterized by molecular and biochemical analyses. Evidence is provided that Sm?-Int1 interacts and co-localizes in the reproductive organs with known schistosome cellular tyrosine kinases (CTKs), of which the Syk kinase SmTK4 appeared to be the strongest interaction partner as shown by yeast two-hybrid analyses and coimmunoprecipitation experiments. By a novel RNAi approach with adult schistosomes in vitro we demonstrate for the first time multinucleated oocytes in treated females, indicating a decisive role Sm?-Int1 during oogenesis as phenotypically analyzed by confocal laser scanning microscopy (CLSM). Our findings provide a first comprehensive overview about platyhelminth integrins, of which the parasite group exhibits unique features allowing a clear distinction from the free-living groups. Furthermore, we shed first lights on the functions of integrins in a trematode model parasite, revealing the complexity of molecular processes involved in its reproductive biology, which may be representative for other platyhelminths.

Beckmann, Svenja; Quack, Thomas; Dissous, Colette; Cailliau, Katia; Lang, Gabriele; Grevelding, Christoph G.

2012-01-01

109

Discovery of platyhelminth-specific ?/?-integrin families and evidence for their role in reproduction in Schistosoma mansoni.  

PubMed

In all metazoa, the response of cells to molecular stimuli from their environment represents a fundamental principle of regulatory processes controlling cell growth and differentiation. Among the membrane-linked receptors mediating extracellular communication processes are integrin receptors. Besides managing adhesion to the extracellular matrix or to other cells, they arrange information flow into the cells by activating intracellular signaling pathways often acting synergistically through cooperation with growth factor receptors. Although a wealth of information exists on integrins in different model organisms, there is a big gap of knowledge for platyhelminths. Here we report on the in silico detection and reconstruction of ? and ? integrins from free-living and parasitic platyhelminths, which according to structural and phylogenetic analyses form specific clades separate from each other and from further metazoan integrins. As representative orthologs of parasitic platyhelminths we have cloned one beta-integrin (Sm?-Int1) and four alpha-integrins (Sm?-Int1 - Sm?-Int4) from Schistosoma mansoni; they were characterized by molecular and biochemical analyses. Evidence is provided that Sm?-Int1 interacts and co-localizes in the reproductive organs with known schistosome cellular tyrosine kinases (CTKs), of which the Syk kinase SmTK4 appeared to be the strongest interaction partner as shown by yeast two-hybrid analyses and coimmunoprecipitation experiments. By a novel RNAi approach with adult schistosomes in vitro we demonstrate for the first time multinucleated oocytes in treated females, indicating a decisive role Sm?-Int1 during oogenesis as phenotypically analyzed by confocal laser scanning microscopy (CLSM). Our findings provide a first comprehensive overview about platyhelminth integrins, of which the parasite group exhibits unique features allowing a clear distinction from the free-living groups. Furthermore, we shed first lights on the functions of integrins in a trematode model parasite, revealing the complexity of molecular processes involved in its reproductive biology, which may be representative for other platyhelminths. PMID:23300694

Beckmann, Svenja; Quack, Thomas; Dissous, Colette; Cailliau, Katia; Lang, Gabriele; Grevelding, Christoph G

2012-12-27

110

Germ layer specification and axial patterning in the embryonic development of the freshwater planarian Schmidtea polychroa.  

PubMed

Although patterning during regeneration in adult planarians has been studied extensively, very little is known about how the initial planarian body plan arises during embryogenesis. Herein, we analyze the process of embryo patterning in the species Schmidtea polychroa by comparing the expression of genes involved in the establishment of the metazoan body plan. Planarians present a derived ectolecithic spiralian development characterized by dispersed cleavage within a yolk syncytium and an early transient embryo capable of feeding on the maternally supplied yolk cells. During this stage of development, we only found evidence of canonical Wnt pathway, mostly associated with the development of its transient pharynx. At these stages, genes involved in gastrulation (snail) and germ layer determination (foxA and twist) are specifically expressed in migrating blastomeres and those giving rise to the temporary gut and pharyngeal muscle. After yolk ingestion, the embryo expresses core components of the canonical Wnt pathway and the BMP pathway, suggesting that the definitive axial identities are established late. These data support the division of planarian development into two separate morphogenetic stages: a highly divergent gastrulation stage, which segregates the three germ layers and establishes the primary organization of the feeding embryo; and subsequent metamorphosis, based on totipotent blastomeres, which establishes the definitive adult body plan using mechanisms that are similar to those used during regeneration and homeostasis in the adult. PMID:20100474

Martín-Durán, José María; Amaya, Enrique; Romero, Rafael

2010-01-25

111

MicroRNAs from the Planarian Schmidtea mediterranea: A model system for stem cell biology  

PubMed Central

MicroRNAs (miRNAs) are ?22-nt RNA molecules that typically bind to the 3? untranslated regions of target mRNAs and function to either induce mRNA degradation or repress translation. miRNAs have been shown to play important roles in the function of stem cells and cell lineage decisions in a variety of organisms, including humans. Planarians are bilaterally symmetric metazoans that have the unique ability to completely regenerate lost tissues or organs. This regenerative capacity is facilitated by a population of stem cells known as neoblasts. Planarians are therefore an excellent model system for studying many aspects of stem cell biology. Here we report the cloning and initial characterization of 71 miRNAs from the planarian Schmidtea mediterranea. While several of the S. mediterranea miRNAs are members of miRNA families identified in other species, we also identified a number of planarian-specific miRNAs. This work lays the foundation for functional studies aimed at addressing the role of these miRNAs in regeneration, cell lineage decisions, and basic stem cell biology.

Palakodeti, Dasaradhi; Smielewska, Magda; Graveley, Brenton R.

2006-01-01

112

Expression of secreted Wnt pathway components reveals unexpected complexity of the planarian amputation response  

PubMed Central

Regeneration is widespread throughout the animal kingdom, but our molecular understanding of this process in adult animals remains poorly understood. Wnt/?-catenin signaling plays crucial roles throughout animal life from early development to adulthood. In intact and regenerating planarians, the regulation of Wnt/?-catenin signaling functions to maintain and specify anterior/posterior (A/P) identity. Here, we explore the expression kinetics and RNAi phenotypes for secreted members of the Wnt signaling pathway in the planarian Schmidtea mediterranea. Smed-wnt and sFRP expression during regeneration is surprisingly dynamic and reveals fundamental aspects of planarian biology that have been previously unappreciated. We show that after amputation, a wounding response precedes rapid reorganization of the A/P axis. Furthermore, cells throughout the body plan can mount this response and reassess their new A/P location in the complete absence of stem cells. While initial stages of the amputation response are stem cell independent, tissue remodeling and the integration of new A/P address with anatomy are stem cell dependent. We also show that WNT5 functions in a reciprocal manner with SLIT to pattern the planarian mediolateral axis, while WNT11-2 patterns the posterior midline. Moreover, we perform an extensive phylogenetic analysis on the Smed-wnt genes using a method that combines and integrates both sequence and structural alignments, enabling us to place all nine genes into Wnt subfamilies for the first time.

Gurley, Kyle A.; Elliott, Sarah A.; Simakov, Oleg; Schmidt, Heiko A.; Holstein, Thomas W.; Sanchez Alvarado, Alejandro

2010-01-01

113

PRMT5 and the role of symmetrical dimethylarginine in chromatoid bodies of planarian stem cells.  

PubMed

Planarian flatworms contain a population of adult stem cells (neoblasts) that proliferate and generate cells of all tissues during growth, regeneration and tissue homeostasis. A characteristic feature of neoblasts is the presence of chromatoid bodies, large cytoplasmic ribonucleoprotein (RNP) granules morphologically similar to structures present in the germline of many organisms. This study aims to reveal the function, and identify additional components, of planarian chromatoid bodies. We uncover the presence of symmetrical dimethylarginine (sDMA) on chromatoid body components and identify the ortholog of protein arginine methyltransferase PRMT5 as the enzyme responsible for sDMA modification in these proteins. RNA interference-mediated depletion of planarian PRMT5 results in defects in homeostasis and regeneration, reduced animal size, reduced number of neoblasts, fewer chromatoid bodies and increased levels of transposon and repetitive-element transcripts. Our results suggest that PIWI family member SMEDWI-3 is one sDMA-containing chromatoid body protein for which methylation depends on PRMT5. Additionally, we discover an RNA localized to chromatoid bodies, germinal histone H4. Our results reveal new components of chromatoid bodies and their function in planarian stem cells, and also support emerging studies indicative of sDMA function in stabilization of RNP granules and the Piwi-interacting RNA pathway. PMID:22318224

Rouhana, Labib; Vieira, Ana P; Roberts-Galbraith, Rachel H; Newmark, Phillip A

2012-02-08

114

Stem cell protection mechanisms in planarians: the role of some heat shock genes.  

PubMed

Planarians contain a large population of stem cells, named neoblasts, and use these for continuous turnover of all cell types. In addition, thanks to the amazing flexibility of these cells, planarians respond well to the effects of stressful situations, for example activating regeneration after trauma. How neoblasts respond to stress and support continuous proliferation, maintaining long-term stability, is still an open question. Heat shock proteins (HSPs) are a complex protein family with key roles in maintaining protein homeostasis, as well as in apoptosis and growth-related processes. We recently characterized some planarian homologs of hsp genes that are highly expressed in mammalian stem cells, and observed that some of them are critical for neoblast survival/maintenance. The results of these studies support the notion that some HSPs play crucial roles in the modulation of pathways regulating stem cell activity, regeneration and tissue repair. In this review we compare the evidence available for planarian hsp genes and focus on questions emerging from these results. PMID:22451000

Isolani, Maria-Emilia; Conte, Maria; Deri, Paolo; Batistoni, Renata

2012-01-01

115

In situ hybridization protocol for enhanced detection of gene expression in the planarian Schmidtea mediterranea  

PubMed Central

Background The freshwater planarian Schmidtea mediterranea has emerged as a powerful model for studies of regenerative, stem cell, and germ cell biology. Whole-mount in situ hybridization (WISH) and whole-mount fluorescent in situ hybridization (FISH) are critical methods for determining gene expression patterns in planarians. While expression patterns for a number of genes have been elucidated using established protocols, determining the expression patterns for particularly low-abundance transcripts remains a challenge. Results We show here that a short bleaching step in formamide dramatically enhances signal intensity of WISH and FISH. To further improve signal sensitivity we optimized blocking conditions for multiple anti-hapten antibodies, developed a copper sulfate quenching step that virtually eliminates autofluorescence, and enhanced signal intensity through iterative rounds of tyramide signal amplification. For FISH on regenerating planarians, we employed a heat-induced antigen retrieval step that provides a better balance between permeabilization of mature tissues and preservation of regenerating tissues. We also show that azide most effectively quenches peroxidase activity between rounds of development for multicolor FISH experiments. Finally, we apply these modifications to elucidate the expression patterns of a few low-abundance transcripts. Conclusion The modifications we present here provide significant improvements in signal intensity and signal sensitivity for WISH and FISH in planarians. Additionally, these modifications might be of widespread utility for whole-mount FISH in other model organisms.

2013-01-01

116

Memory and obesity affect the population dynamics of asexual freshwater planarians.  

PubMed

Asexual reproduction in multicellular organisms is a complex biophysical process that is not yet well understood quantitatively. Here, we report a detailed population study for the asexual freshwater planarian Schmidtea mediterranea, which can reproduce via transverse fission due to a large stem cell contingent. Our long-term observations of isolated non-interacting planarian populations reveal that the characteristic fission waiting time distributions for head and tail fragments differ significantly from each other. The stochastic fission dynamics of tail fragments exhibits non-negligible memory effects, implying that an accurate mathematical description of future data should be based on non-Markovian tree models. By comparing the effective growth of non-interacting planarian populations with those of self-interacting populations, we are able to quantify the influence of interactions between flatworms and physical conditions on the population growth. A surprising result is the non-monotonic relationship between effective population growth rate and nutrient supply: planarians exhibit a tendency to become 'obese' if the feeding frequency exceeds a critical level, resulting in a decreased reproduction activity. This suggests that these flatworms, which possess many genes homologous to those of humans, could become a new model system for studying dietary effects on reproduction and regeneration in multicellular organisms. PMID:21263170

Dunkel, Jörn; Talbot, Jared; Schötz, Eva-Maria

2011-01-24

117

PRMT5 and the role of symmetrical dimethylarginine in chromatoid bodies of planarian stem cells  

PubMed Central

Planarian flatworms contain a population of adult stem cells (neoblasts) that proliferate and generate cells of all tissues during growth, regeneration and tissue homeostasis. A characteristic feature of neoblasts is the presence of chromatoid bodies, large cytoplasmic ribonucleoprotein (RNP) granules morphologically similar to structures present in the germline of many organisms. This study aims to reveal the function, and identify additional components, of planarian chromatoid bodies. We uncover the presence of symmetrical dimethylarginine (sDMA) on chromatoid body components and identify the ortholog of protein arginine methyltransferase PRMT5 as the enzyme responsible for sDMA modification in these proteins. RNA interference-mediated depletion of planarian PRMT5 results in defects in homeostasis and regeneration, reduced animal size, reduced number of neoblasts, fewer chromatoid bodies and increased levels of transposon and repetitive-element transcripts. Our results suggest that PIWI family member SMEDWI-3 is one sDMA-containing chromatoid body protein for which methylation depends on PRMT5. Additionally, we discover an RNA localized to chromatoid bodies, germinal histone H4. Our results reveal new components of chromatoid bodies and their function in planarian stem cells, and also support emerging studies indicative of sDMA function in stabilization of RNP granules and the Piwi-interacting RNA pathway.

Rouhana, Labib; Vieira, Ana P.; Roberts-Galbraith, Rachel H.; Newmark, Phillip A.

2012-01-01

118

Withdrawal-like behavior in planarians is dependent on drug exposure duration.  

PubMed

Planarians display a concentration-related reduction in locomotor activity following their spontaneous withdrawal from opioids, cannabinoids, stimulants and benzodiazepines. This suggests that planarians display a withdrawal-like behavior that can be quantified as a reduction in locomotor activity. Because withdrawal-like behavior in previous studies has been quantified only following the cessation of a 60-min drug exposure, it is unclear whether the withdrawal response varies with drug exposure duration. Therefore, the goal of this study is to determine if the duration of drug exposure (0, 5, 15, 30, 45, 60min and 24h) to three different drugs - methamphetamine, cocaine and caffeine - affects the magnitude of withdrawal-like behavior (i.e., reduced locomotor activity) in planarians. Experiments revealed that methamphetamine (10microM) produced significant withdrawal-like behavior regardless of the exposure time (P<0.05). An exposure time of only 5min resulted in a significant reduction in locomotor activity. The peak effect, although occurring following a 24-h exposure, was only slightly greater than that caused by a 30-min exposure. For cocaine (10microM), a longer exposure time (15min) was required for the manifestation of significant withdrawal-like behavior. The peak cocaine effect was observed following a 24-h exposure. Caffeine (10microM) did not produce significant changes in locomotor activity during withdrawal or alter locomotor activity during acute exposure. The present results suggest that the magnitude of withdrawal-like behavior in planarians is dependent on both the duration and type of drug exposure, and that planarians do not display withdrawal to caffeine. PMID:18511196

Sacavage, Steve; Patel, Hiren; Zielinski, Mike; Acker, Jeneane; Phillips, Austin G; Raffa, Robert B; Rawls, Scott M

2008-04-30

119

Complete genome sequence of a novel extrachromosomal virus-like element identified in planarian Girardia tigrina  

PubMed Central

Background Freshwater planarians are widely used as models for investigation of pattern formation and studies on genetic variation in populations. Despite extensive information on the biology and genetics of planaria, the occurrence and distribution of viruses in these animals remains an unexplored area of research. Results Using a combination of Suppression Subtractive Hybridization (SSH) and Mirror Orientation Selection (MOS), we compared the genomes of two strains of freshwater planarian, Girardia tigrina. The novel extrachromosomal DNA-containing virus-like element denoted PEVE (Planarian Extrachromosomal Virus-like Element) was identified in one planarian strain. The PEVE genome (about 7.5 kb) consists of two unique regions (Ul and Us) flanked by inverted repeats. Sequence analyses reveal that PEVE comprises two helicase-like sequences in the genome, of which the first is a homolog of a circoviral replication initiator protein (Rep), and the second is similar to the papillomavirus E1 helicase domain. PEVE genome exists in at least two variant forms with different arrangements of single-stranded and double-stranded DNA stretches that correspond to the Us and Ul regions. Using PCR analysis and whole-mount in situ hybridization, we characterized PEVE distribution and expression in the planarian body. Conclusions PEVE is the first viral element identified in free-living flatworms. This element differs from all known viruses and viral elements, and comprises two potential helicases that are homologous to proteins from distant viral phyla. PEVE is unevenly distributed in the worm body, and is detected in specific parenchyma cells.

Rebrikov, Denis V; Bulina, Maria E; Bogdanova, Ekaterina A; Vagner, Loura L; Lukyanov, Sergey A

2002-01-01

120

Decreased neoblast progeny and increased cell death during starvation-induced planarian degrowth.  

PubMed

The development of a complex multicellular organism requires a careful coordination of growth, cell division, cell differentiation and cell death. All these processes must be under intricate and coordinated control, as they have to be integrated across all tissues. Freshwater planarians are especially plastic, in that they constantly replace somatic tissues from a pool of adult somatic stem cells and continuously undergo growth and degrowth as adult animals in response to nutrient availability. During these processes they appear to maintain perfect scale of tissues and organs. These life history traits make them an ideal model system to study growth and degrowth. We have studied the unique planarian process of degrowth. When food is not available, planarians are able to degrow to a minimum size, without any signs of adverse physiological outcomes. For example they maintain full regenerative capacity. Our current knowledge of how this is regulated at the molecular and cellular level is very limited. Planarian degrowth has been reported to result from a decrease in cell number rather than a decrease in cell size. Thus one obvious explanation for degrowth would be a decrease in stem cell proliferation. However evidence in the literature suggests this is not the case. We show that planarians maintain normal basal mitotic rates during degrowth but that the number of stem cell progeny decreases during starvation and degrowth. These observations are reversed upon feeding, indicating that they are dependent on nutritional status. An increase in cell death is also observed during degrowth, which is not rapidly reversed upon feeding. We conclude that degrowth is a result of cell death decreasing cell numbers and that the dynamics of neoblast self-renewal and differentiation adapt to nutrient conditions to allow maintenance of the neoblast population during the period of starvation. PMID:22252539

González-Estévez, Cristina; Felix, Daniel A; Rodríguez-Esteban, Gustavo; Aboobaker, A Aziz

2012-01-01

121

Inhibitory Smads and bone morphogenetic protein (BMP) modulate anterior photoreceptor cell number during planarian eye regeneration.  

PubMed

Planarians represent an excellent model to study the processes of body axis and organ re-specification during regeneration. Previous studies have revealed a conserved role for the bone morphogenetic protein (BMP) pathway and its intracellular mediators Smad1/5/8 and Smad4 in planarian dorsoventral (DV) axis re-establishment. In an attempt to gain further insight into the role of this signalling pathway in planarians, we have isolated and functionally characte-rized the inhibitory Smads (I-Smads) in Schmidtea mediterranea. Two I-Smad homologues have been identified: Smed-smad6/7-1 and Smed-smad6/7-2. Expression of smad6/7-1 was detected in the parenchyma, while smad6/7-2 was found to be ex-pressed in the central nervous system and the eyes. Neither single smad6/7-1 and smad6/7-2 nor double smad6/7-1,-2 silencing gave rise to any apparent disruption of the DV axis. However, both regenerating and intact smad6/7-2 (RNAi) planarians showed defects in eye morphogenesis and displayed small, rounded eyes that lacked the anterior subpopulation of photoreceptor cells. The number of pigment cells was also reduced in these animals at later stages of regeneration. In contrast, after low doses of Smed-bmp(RNAi), planarians regenerated larger eyes in which the anterior subpopulation of photoreceptor cells was expanded. Our results suggest that Smed-smad6/7-2 and Smed-bmp control the re-specification and maintenance of anterior photoreceptor cell number in S. mediterranea. PMID:22451003

González-Sastre, Alejandro; Molina, Ma Dolores; Saló, Emili

2012-01-01

122

Epigenetic regulation of planarian stem cells by the SET1/MLL family of histone methyltransferases.  

PubMed

Chromatin regulation is a fundamental mechanism underlying stem cell pluripotency, differentiation, and the establishment of cell type-specific gene expression profiles. To examine the role of chromatin regulation in stem cells in vivo, we study regeneration in the freshwater planarian Schmidtea mediterranea. These animals possess a high concentration of pluripotent stem cells, which are capable of restoring any damaged or lost tissues after injury or amputation. Here, we identify the S. mediterranea homologs of the SET1/MLL family of histone methyltransferases and COMPASS and COMPASS-like complex proteins and investigate their role in stem cell function during regeneration. We identified six S. mediterranea homologs of the SET1/MLL family (set1, mll1/2, trr-1, trr-2, mll5-1 and mll5-2), characterized their patterns of expression in the animal, and examined their function by RNAi. All members of this family are expressed in the stem cell population and differentiated tissues. We show that set1, mll1/2, trr-1, and mll5-2 are required for regeneration and that set1, trr-1 and mll5-2 play roles in the regulation of mitosis. Most notably, knockdown of the planarian set1 homolog leads to stem cell depletion. A subset of planarian homologs of COMPASS and COMPASS-like complex proteins are also expressed in stem cells and implicated in regeneration, but the knockdown phenotypes suggest that some complex members also function in other aspects of planarian biology. This work characterizes the function of the SET1/MLL family in the context of planarian regeneration and provides insight into the role of these enzymes in adult stem cell regulation in vivo. PMID:23235145

Hubert, Amy; Henderson, Jordana M; Ross, Kelly G; Cowles, Martis W; Torres, Jessica; Zayas, Ricardo M

2012-12-12

123

Pharmacological and Functional Genetic Assays to Manipulate Regeneration of the Planarian Dugesia japonica  

PubMed Central

Free-living planarian flatworms have a long history of experimental usage owing to their remarkable regenerative abilities1. Small fragments excised from these animals reform the original body plan following regeneration of missing body structures. For example if a 'trunk' fragment is cut from an intact worm, a new 'head' will regenerate anteriorly and a 'tail' will regenerate posteriorly restoring the original 'head-to-tail' polarity of body structures prior to amputation (Figure 1A). Regeneration is driven by planarian stem cells, known as 'neoblasts' which differentiate into ~30 different cell types during normal body homeostasis and enforced tissue regeneration. This regenerative process is robust and easy to demonstrate. Owing to the dedication of several pioneering labs, many tools and functional genetic methods have now been optimized for this model system. Consequently, considerable recent progress has been made in understanding and manipulating the molecular events underpinning planarian developmental plasticity2-9. The planarian model system will be of interest to a broad range of scientists. For neuroscientists, the model affords the opportunity to study the regeneration of an entire nervous system, rather than simply the regrowth/repair of single nerve cell process that typically are the focus of study in many established models. Planarians express a plethora of neurotransmitters10, represent an important system for studying evolution of the central nervous system11, 12 and have behavioral screening potential13, 14. Regenerative outcomes are amenable to manipulation by pharmacological and genetic apparoaches. For example, drugs can be screened for effects on regeneration simply by placing body fragments in drug-containing solutions at different time points after amputation. The role of individual genes can be studied using knockdown methods (in vivo RNAi), which can be achieved either through cycles of microinjection or by feeding bacterially-expressed dsRNA constructs8, 9, 15. Both approaches can produce visually striking phenotypes at high penetrance- for example, regeneration of bipolar animals16-21. To facilitate adoption of this model and implementation of such methods, we showcase in this video article protocols for pharmacological and genetic assays (in vivo RNAi by feeding) using the planarian Dugesia japonica.

Chan, John D.; Marchant, Jonathan S.

2011-01-01

124

Neurobiology of the basal platyhelminth Macrostomum lignano : map and digital 3D model of the juvenile brain neuropile  

Microsoft Academic Search

We have analyzed brain structure in Macrostomum lignano, a representative of the basal platyhelminth taxon Macrostomida. Using confocal microscopy and digital 3D modeling software\\u000a on specimens labeled with general markers for neurons (tyrTub), muscles (phalloidin), and nuclei (Sytox), an atlas and digital\\u000a model of the juvenile Macrostomum brain was generated. The brain forms a ganglion with a central neuropile surrounded

Joshua Morris; Albert Cardona; Maria Del Mar De Miguel-Bonet; Volker Hartenstein

2007-01-01

125

Planarians as a model to assess in vivo the role of matrix metalloproteinase genes during homeostasis and regeneration.  

PubMed

Matrix metalloproteinases (MMPs) are major executors of extracellular matrix remodeling and, consequently, play key roles in the response of cells to their microenvironment. The experimentally accessible stem cell population and the robust regenerative capabilities of planarians offer an ideal model to study how modulation of the proteolytic system in the extracellular environment affects cell behavior in vivo. Genome-wide identification of Schmidtea mediterranea MMPs reveals that planarians possess four mmp-like genes. Two of them (mmp1 and mmp2) are strongly expressed in a subset of secretory cells and encode putative matrilysins. The other genes (mt-mmpA and mt-mmpB) are widely expressed in postmitotic cells and appear structurally related to membrane-type MMPs. These genes are conserved in the planarian Dugesia japonica. Here we explore the role of the planarian mmp genes by RNA interference (RNAi) during tissue homeostasis and regeneration. Our analyses identify essential functions for two of them. Following inhibition of mmp1 planarians display dramatic disruption of tissues architecture and significant decrease in cell death. These results suggest that mmp1 controls tissue turnover, modulating survival of postmitotic cells. Unexpectedly, the ability to regenerate is unaffected by mmp1(RNAi). Silencing of mt-mmpA alters tissue integrity and delays blastema growth, without affecting proliferation of stem cells. Our data support the possibility that the activity of this protease modulates cell migration and regulates anoikis, with a consequent pivotal role in tissue homeostasis and regeneration. Our data provide evidence of the involvement of specific MMPs in tissue homeostasis and regeneration and demonstrate that the behavior of planarian stem cells is critically dependent on the microenvironment surrounding these cells. Studying MMPs function in the planarian model provides evidence on how individual proteases work in vivo in adult tissues. These results have high potential to generate significant information for development of regenerative and anti cancer therapies. PMID:23405188

Isolani, Maria Emilia; Abril, Josep F; Saló, Emili; Deri, Paolo; Bianucci, Anna Maria; Batistoni, Renata

2013-02-06

126

The More the Merrier?. Entropy and Statistics of Asexual Reproduction in Freshwater Planarians  

NASA Astrophysics Data System (ADS)

The trade-off between traits in life-history strategies has been widely studied for sexual and parthenogenetic organisms, but relatively little is known about the reproduction strategies of asexual animals. Here, we investigate clonal reproduction in the freshwater planarian Schmidtea mediterranea, an important model organism for regeneration and stem cell research. We find that these flatworms adopt a randomized reproduction strategy that comprises both asymmetric binary fission and fragmentation (generation of multiple offspring during a reproduction cycle). Fragmentation in planarians has primarily been regarded as an abnormal behavior in the past; using a large-scale experimental approach, we now show that about one third of the reproduction events in S. mediterranea are fragmentations, implying that fragmentation is part of their normal reproductive behavior. Our analysis further suggests that certain characteristic aspects of the reproduction statistics can be explained in terms of a maximum relative entropy principle.

Quinodoz, Sofia; Thomas, Michael A.; Dunkel, Jörn; Schötz, Eva-Maria

2011-04-01

127

Planarian activity differences when maintained in water pre-treated with magnetic fields: a nonlinear effect.  

PubMed

There have been multiple claims that exposing water to a static magnetic field affects its properties which influence living systems. To test this hypothesis, planarian subsequent to dissection were maintained in spring water that had been previously exposed for only one day to one of three (16, 160, or 1,600 G) intensity static magnetic fields or to a reference condition. Although there was no significant difference in regeneration rates over the subsequent seven-day period, there was a statistically significant nonlinear effect for planarian mobility and diffusion rates. Both mobility rates and diffusion velocity of a liquid within the water that had been exposed to the 16 G field was about twice that for water exposed to the other intensities. These results imply that nonlinear biophysical effects may emerge under specific conditions of intensity ranges for particular volumes of water. PMID:22047458

Gang, Noa; Persinger, Michael A

2011-12-01

128

Planarian Hh signaling regulates regeneration polarity and links Hh pathway evolution to cilia  

PubMed Central

The Hedgehog (Hh) signaling pathway plays multiple essential roles during metazoan development, homeostasis, and disease. Although core protein components are highly conserved, the variations in Hh signal transduction mechanisms exhibited by existing model systems (Drosophila, fish, and mammals) are difficult to understand. We characterize the Hh pathway in planarians. Hh signaling is essential for establishing the Anterior/Posterior axis during regeneration by modulating wnt expression. Moreover, RNAi methods to reduce signal transduction proteins Cos2/Kif27/Kif7, Fused, or Iguana do not result in detectable Hh signaling defects; however, these proteins are essential for planarian ciliogenesis. Our study expands the understanding of Hh signaling in the animal kingdom and suggests an ancestral mechanistic link between Hh signaling and the function of cilia.

Rink, Jochen C.; Gurley, Kyle A.; Elliott, Sarah A.; Alvarado, Alejandro Sanchez

2010-01-01

129

Planarian Hh signaling regulates regeneration polarity and links Hh pathway evolution to cilia.  

PubMed

The Hedgehog (Hh) signaling pathway plays multiple essential roles during metazoan development, homeostasis, and disease. Although core protein components are highly conserved, the variations in Hh signal transduction mechanisms exhibited by existing model systems (Drosophila, fish, and mammals) are difficult to understand. We characterized the Hh pathway in planarians. Hh signaling is essential for establishing the anterior/posterior axis during regeneration by modulating wnt expression. Moreover, RNA interference methods to reduce signal transduction proteins Cos2/Kif27/Kif7, Fused, or Iguana do not result in detectable Hh signaling defects; however, these proteins are essential for planarian ciliogenesis. Our study expands the understanding of Hh signaling in the animal kingdom and suggests an ancestral mechanistic link between Hh signaling and the function of cilia. PMID:19933103

Rink, Jochen C; Gurley, Kyle A; Elliott, Sarah A; Sánchez Alvarado, Alejandro

2009-10-22

130

Change in ATP and ADP content of fasting and feeding planarians (Polycelis nigra).  

PubMed

The ATP and ADP content of planarians subjected to starvation for two weeks followed by feeding for the same period was investigated. The ATP and ADP content during fasting increased and then, after feeding, returned to normal. The ATP/ADP ratio varied in the same way, which is consistent with the view that the adenylic nucleotide pool is implicated in the regulation of the energy metabolism of the organism. PMID:551828

Grudnitskii, V A; Dontsova, G V

131

Short communication Amphetamine-induced increase in planarian locomotor activity and block by UV light  

Microsoft Academic Search

The dopamine D2-receptor antagonist sulpiride decreases spontaneous locomotor velocity of planarians (pLMV) in an enantiomeric- selective and dose-dependent manner and is significantly attenuated by UV light (254 and 366 nm). We now report that amphetamine (10 AM) produced the opposite effect and was also reversed by UV light. These findings strengthen the hypothesis that the effects of dopaminergic ligands and

Robert B. Raffa; Andrea F. Martley

132

The planarian P2X homolog in the regulation of asexual reproduction.  

PubMed

The growth in size of freshwater planarians in response to nutrient intake is limited by the eventual separation of tail and body fragments in a process called fission. The resulting tail fragment regenerates the entire body as an artificially amputated tail fragment would do, and the body fragment regenerates a tail, resulting in two whole planarians. This regenerative ability is supported by pluripotent somatic stem cells, called neoblasts, which are distributed throughout almost the entire body of the planarian. Neoblasts are the only planarian cells with the ability to continuously proliferate and give rise to all types of cells during regeneration, asexual reproduction, homeostasis, and growth. In order to investigate the molecular characteristics of neoblasts, we conducted an extensive search for neoblast-specific genes using the High Coverage Expression Profiling (HiCEP) method, and tested the function of the resulting candidates by RNAi. Disruption of the expression of one candidate gene, DjP2X-A (Dugesia japonica membrane protein P2X homologue), resulted in a unique phenotype. DjP2X-A RNAi leads to an increase of fission events upon feeding. We confirmed by immunohistochemistry that DjP2X-A is a membrane protein, and elucidated its role in regulating neoblast proliferation, thereby explaining its unique phenotype. We found that DjP2X-A decreases the burst of neoblast proliferation that normally occurs after feeding. We also found that DjP2X-A is required for normal proliferation in starved animals. We propose that DjP2X-A modulates stem cell proliferation in response to the nutritional condition. PMID:22451005

Sakurai, Toshihide; Lee, Hayoung; Kashima, Makoto; Saito, Yumi; Hayashi, Tetsutaro; Kudome-Takamatsu, Tomomi; Nishimura, Osamu; Agata, Kiyokazu; Shibata, Norito

2012-01-01

133

Genome-Wide Analyses Reveal a Role for Peptide Hormones in Planarian Germline Development  

PubMed Central

Bioactive peptides (i.e., neuropeptides or peptide hormones) represent the largest class of cell-cell signaling molecules in metazoans and are potent regulators of neural and physiological function. In vertebrates, peptide hormones play an integral role in endocrine signaling between the brain and the gonads that controls reproductive development, yet few of these molecules have been shown to influence reproductive development in invertebrates. Here, we define a role for peptide hormones in controlling reproductive physiology of the model flatworm, the planarian Schmidtea mediterranea. Based on our observation that defective neuropeptide processing results in defects in reproductive system development, we employed peptidomic and functional genomic approaches to characterize the planarian peptide hormone complement, identifying 51 prohormone genes and validating 142 peptides biochemically. Comprehensive in situ hybridization analyses of prohormone gene expression revealed the unanticipated complexity of the flatworm nervous system and identified a prohormone specifically expressed in the nervous system of sexually reproducing planarians. We show that this member of the neuropeptide Y superfamily is required for the maintenance of mature reproductive organs and differentiated germ cells in the testes. Additionally, comparative analyses of our biochemically validated prohormones with the genomes of the parasitic flatworms Schistosoma mansoni and Schistosoma japonicum identified new schistosome prohormones and validated half of all predicted peptide-encoding genes in these parasites. These studies describe the peptide hormone complement of a flatworm on a genome-wide scale and reveal a previously uncharacterized role for peptide hormones in flatworm reproduction. Furthermore, they suggest new opportunities for using planarians as free-living models for understanding the reproductive biology of flatworm parasites.

Collins, James J.; Hou, Xiaowen; Romanova, Elena V.; Lambrus, Bramwell G.; Miller, Claire M.; Saberi, Amir; Sweedler, Jonathan V.; Newmark, Phillip A.

2010-01-01

134

The Dr-nanos gene is essential for germ cell specification in the planarian Dugesia ryukyuensis.  

PubMed

Homologs of nanos are required for the formation and maintenance of germline stem cell (GSC) systems and for gametogenesis in many metazoans. Planarians can change their reproductive mode seasonally, alternating between asexual and sexual reproduction; they develop and maintain their somatic stem cells (SSCs) and GCSs from pluripotent stem cells known as neoblasts. We isolated a nanos homolog, Dr-nanos, from the expressed sequence tags (ESTs) of the sexualized form of Dugesia ryukyuensis. We examined the expression of Dr-nanos in asexual and sexualized planarians by in situ hybridization and analyzed its function using RNA interference (RNAi) together with a planarian sexualization assay. A nanos homolog, Dr-nanos, was identified in the planarian D. ryukyuensis. Dr-nanos expression was observed in the ovarian primordial cells of the asexual worms. This expression increased in proportion to sexualization and was localized in the early germline cells of the ovaries and testes. In X-ray-irradiated worms, the expression of Dr-nanos decreased to a large extent, indicating that Dr-nanos is expressed in some subpopulations of stem cells, especially in GSCs. During the sexualization process, worms in which Dr-nanos was knocked down by RNAi exhibited decreased numbers of oogonia in the ovaries and failed to develop testes, whereas the somatic sexual organs were not affected. We conclude that Dr-nanos is essential for the development of germ cells in the ovaries and testes and may have a function in the early stages of germ cell specification, but not in the development of somatic sexual organs. PMID:22451004

Nakagawa, Haruka; Ishizu, Hirotsugu; Chinone, Ayako; Kobayashi, Kazuya; Matsumoto, Midori

2012-01-01

135

Planarian regeneration involves distinct stem cell responses to wounds and tissue absence  

PubMed Central

Regeneration requires signaling from a wound site for detection of the wound, and a mechanism that determines the nature of the injury to specify the appropriate regenerative response. Wound signals and tissue responses to wounds that elicit regeneration remain poorly understood. Planarians are able to regenerate from essentially any type of injury and present a novel system for the study of wound responses in regeneration initiation. Newly developed molecular and cellular tools now enable study of regeneration initiation using the planarian Schmidtea mediterranea. Planarian regeneration requires adult stem cells called neoblasts and amputation triggers two peaks in neoblast mitoses early in regeneration. We demonstrate that the first mitotic peak is a body-wide response to any injury and that a second, local, neoblast response is induced only when injury results in missing tissue. This second response was characterized by recruitment of neoblasts to wounds, even in areas that lack neoblasts in the intact animal. Subsequently, these neoblasts were induced to divide and differentiate near the wound, leading to formation of new tissue. We conclude that there exist two functionally distinct signaling phases of the stem cell wound response that distinguish between simple injury and situations that require the regeneration of missing tissue.

Wenemoser, Danielle; Reddien, Peter W.

2010-01-01

136

Loss of DNA Mismatch Repair Imparts a Selective Advantage in Planarian Adult Stem Cells  

PubMed Central

Lynch syndrome (LS) leads to an increased risk of early-onset colorectal and other types of cancer and is caused by germline mutations in DNA mismatch repair (MMR) genes. Loss of MMR function results in a mutator phenotype that likely underlies its role in tumorigenesis. However, loss of MMR also results in the elimination of a DNA damage-induced checkpoint/apoptosis activation barrier that may allow damaged cells to grow unchecked. A fundamental question is whether loss of MMR provides pre-cancerous stem cells an immediate selective advantage in addition to establishing a mutator phenotype. To test this hypothesis in an in vivo system, we utilized the planarian Schmidtea mediterranea which contains a significant population of identifiable adult stem cells. We identified a planarian homolog of human MSH2, a MMR gene which is mutated in 38% of LS cases. The planarian Smed-msh2 is expressed in stem cells and some progeny. We depleted Smed-msh2 mRNA levels by RNA-interference and found a striking survival advantage in these animals treated with a cytotoxic DNA alkylating agent compared to control animals. We demonstrated that this tolerance to DNA damage is due to the survival of mitotically active, MMR-deficient stem cells. Our results suggest that loss of MMR provides an in vivo survival advantage to the stem cell population in the presence of DNA damage that may have implications for tumorigenesis.

Hollenbach, Jessica P.; Resch, Alissa M.; Palakodeti, Dasaradhi; Graveley, Brenton R.; Heinen, Christopher D.

2011-01-01

137

The use of planarians as in vivo animal model to study laser biomodulation effects  

NASA Astrophysics Data System (ADS)

A variety of effects is attributed to the photo stimulation of tissues, such as improved healing of ulcers, analgesic and anti-inflammatory effects, stimulation of the proliferation of cells of different origins and stimulation of bone repair. Some investigations that make qualitative evaluations, like wound healing and evaluation of pain and edema, can be conducted in human subjects. However, deeper investigations on the mechanisms of action of the light stimulus and other quantitative works that requires biopsies or destructive analysis has to be carried out in animal models or in cell cultures. In this work, we propose the use of planarians as a model to study laser-tissue interaction. Contrasting with cell cultures and unicellular organisms, planarians are among the simplest organism having tissue layers, central nerve system, digestive and excretory system that might have been platforms for the evolution of the complex and highly organized tissues and organs found in higher organisms. For the present study, 685 nm laser radiation was employed. Planarians were cut transversally, in a plane posterior to the auricles. The body fragments were left to regenerate and the proliferation dynamics of stem cells was studied by using histological analysis. Maximum cell count was obtained for the laser treated group at the 4th experimental day. At that experimental time, we also had the largest difference between the irradiated and the non-irradiated control group. We concluded that the studied flatworm could be an interesting animal model for in vivo studies of laser-tissue interactions.

Munin, Egberto; Garcia, Neila Maria Rocha; Braz, Allison Gustavo; de Souza, Sandra Cristina; Alves, Leandro Procópio; Salgado, Miguel Angel Castillo; Pilla, Viviane

2007-03-01

138

A comparative transcriptomic analysis reveals conserved features of stem cell pluripotency in planarians and mammals.  

PubMed

Many long-lived species of animals require the function of adult stem cells throughout their lives. However, the transcriptomes of stem cells in invertebrates and vertebrates have not been compared, and consequently, ancestral regulatory circuits that control stem cell populations remain poorly defined. In this study, we have used data from high-throughput RNA sequencing to compare the transcriptomes of pluripotent adult stem cells from planarians with the transcriptomes of human and mouse pluripotent embryonic stem cells. From a stringently defined set of 4,432 orthologs shared between planarians, mice and humans, we identified 123 conserved genes that are ?5-fold differentially expressed in stem cells from all three species. Guided by this gene set, we used RNAi screening in adult planarians to discover novel stem cell regulators, which we found to affect the stem cell-associated functions of tissue homeostasis, regeneration, and stem cell maintenance. Examples of genes that disrupted these processes included the orthologs of TBL3, PSD12, TTC27, and RACK1. From these analyses, we concluded that by comparing stem cell transcriptomes from diverse species, it is possible to uncover conserved factors that function in stem cell biology. These results provide insights into which genes comprised the ancestral circuitry underlying the control of stem cell self-renewal and pluripotency. PMID:22696458

Labbé, Roselyne M; Irimia, Manuel; Currie, Ko W; Lin, Alexander; Zhu, Shu Jun; Brown, David D R; Ross, Eric J; Voisin, Veronique; Bader, Gary D; Blencowe, Benjamin J; Pearson, Bret J

2012-08-01

139

The maintenance and regeneration of the planarian excretory system are regulated by EGFR signaling.  

PubMed

The maintenance of organs and their regeneration in case of injury are crucial to the survival of all animals. High rates of tissue turnover and nearly unlimited regenerative capabilities make planarian flatworms an ideal system with which to investigate these important processes, yet little is known about the cell biology and anatomy of their organs. Here we focus on the planarian excretory system, which consists of internal protonephridial tubules. We find that these assemble into complex branching patterns with a stereotyped succession of cell types along their length. Organ regeneration is likely to originate from a precursor structure arising in the blastema, which undergoes extensive branching morphogenesis. In an RNAi screen of signaling molecules, we identified an EGF receptor (Smed-EGFR-5) as a crucial regulator of branching morphogenesis and maintenance. Overall, our characterization of the planarian protonephridial system establishes a new paradigm for regenerative organogenesis and provides a platform for exploring its functional and evolutionary homologies with vertebrate excretory systems. PMID:21828097

Rink, Jochen C; Vu, Hanh Thi-Kim; Sánchez Alvarado, Alejandro

2011-09-01

140

Existence of two sexual races in the planarian species switching between asexual and sexual reproduction.  

PubMed

In certain planarian species that are able to switch between asexual and sexual reproduction, determining whether a sexual has the ability to switch to the asexual state is problematic, which renders the definition of sexuals controversial. We experimentally show the existence of two sexual races, acquired and innate, in the planarian Dugesia ryukyuensis. Acquired sexuals used in this study were experimentally switched from asexuals. Inbreeding of acquired sexuals produced both innate sexuals and asexuals, but inbreeding of innate sexuals produced innate sexuals only and no asexuals. Acquired sexuals, but not innate sexuals, were forced to become asexuals by ablation and regeneration (asexual induction). This suggests that acquired sexuals somehow retain asexual potential, while innate sexuals do not. We also found that acquired sexuals have the potential to develop hyperplastic and supernumerary ovaries, while innate sexuals do not. In this regard, acquired sexuals were more prolific than innate sexuals. The differences between acquired and innate sexuals will provide a structure for examining the mechanism underlying asexual and sexual reproduction in planarians. PMID:22468837

Kobayashi, Kazuya; Maezawa, Takanobu; Nakagawa, Haruka; Hoshi, Motonori

2012-04-01

141

Gap junction proteins: master regulators of the planarian stem cell response to tissue maintenance and injury.  

PubMed

Gap junction (GJ) proteins are crucial mediators of cell-cell communication during embryogenesis, tissue regeneration and disease. GJ proteins form plasma membrane channels that facilitate passage of small molecules across cells and modulate signaling pathways and cellular behavior in different tissues. These properties have been conserved throughout evolution, and in most invertebrates GJ proteins are known as innexins. Despite their critical relevance for physiology and disease, the mechanisms by which GJ proteins modulate cell behavior are poorly understood. This review summarizes findings from recent work that uses planarian flatworms as a paradigm to analyze GJ proteins in the complexity of the whole organism. The planarian model allows access to a large pool of adult somatic stem cells (known as neoblasts) that support physiological cell turnover and tissue regeneration. Innexin proteins are present in planarians and play a fundamental role in controlling neoblast behavior. We discuss the possibility that GJ proteins participate as cellular sensors that inform neoblasts about local and systemic physiological demands. We believe that functional analyses of GJ proteins will bring a complementary perspective to studies that focus on the temporal expression of genes. Finally, integrating functional studies along with molecular genetics and epigenetic approaches would expand our understanding of cellular regulation in vivo and greatly enhance the possibilities for rationally modulating stem cell behavior in their natural environment. This article is part of a Special Issue entitled: The communicating junctions, roles and dysfunctions. PMID:22450236

Peiris, T Harshani; Oviedo, Néstor J

2012-03-16

142

The in vivo effect of chelidonine on the stem cell system of planarians.  

PubMed

The presence of adult pluripotent stem cells and the amazing regenerative capabilities make planarian flatworms an extraordinary experimental model to assess in vivo the effects of substances of both natural and synthetic origin on stem cell dynamics. This study focuses on the effects of chelidonine, an alkaloid obtained from Chelidonium majus. The expression levels of molecular markers specific for stem or differentiated cells were compared in chelidonine-treated and control planarians. The use of these markers demonstrates that chelidonine produces in vivo a significant anti-proliferative effect on planarian stem cells in a dose-dependent fashion. In response to chelidonine treatment mitotic abnormalities were also observed and the number of cells able to proceed to anaphase/telophase appeared significantly reduced with respect to the controls. Our results support the possibility that chelidonine acts on cell cycle progression by inhibition of tubulin polymerization. These studies provide a basis for preclinical evaluation in vivo of the effects of chelidonine on physiologically proliferating stem cells. PMID:22503932

Isolani, Maria Emilia; Pietra, Daniele; Balestrini, Linda; Borghini, Alice; Deri, Paolo; Imbriani, Marcello; Bianucci, Anna Maria; Batistoni, Renata

2012-04-03

143

TORC1 is required to balance cell proliferation and cell death in planarians.  

PubMed

Multicellular organisms are equipped with cellular mechanisms that enable them to replace differentiated cells lost to normal physiological turnover, injury, and for some such as planarians, even amputation. This process of tissue homeostasis is generally mediated by adult stem cells (ASCs), tissue-specific stem cells responsible for maintaining anatomical form and function. To do so, ASCs must modulate the balance between cell proliferation, i.e. in response to nutrients, and that of cell death, i.e. in response to starvation or injury. But how these two antagonistic processes are coordinated remains unclear. Here, we explore the role of the core components of the TOR pathway during planarian tissue homeostasis and regeneration and identified an essential function for TORC1 in these two processes. RNAi-mediated silencing of TOR in intact animals resulted in a significant increase in cell death, whereas stem cell proliferation and stem cell maintenance were unaffected. Amputated animals failed to increase stem cell proliferation after wounding and displayed defects in tissue remodeling. Together, our findings suggest two distinct roles for TORC1 in planarians. TORC1 is required to modulate the balance between cell proliferation and cell death during normal cell turnover and in response to nutrients. In addition, it is required to initiate appropriate stem cell proliferation during regeneration and for proper tissue remodeling to occur to maintain scale and proportion. PMID:22445864

Tu, Kimberly C; Pearson, Bret J; Sánchez Alvarado, Alejandro

2012-03-16

144

Restoration of anterior regeneration in a planarian with limited regenerative ability.  

PubMed

Variability of regenerative potential among animals has long perplexed biologists. On the basis of their exceptional regenerative abilities, planarians have become important models for understanding the molecular basis of regeneration. However, planarian species with limited regenerative abilities are also found. Despite the importance of understanding the differences between closely related, regenerating and non-regenerating organisms, few studies have focused on the evolutionary loss of regeneration, and the molecular mechanisms leading to such regenerative loss remain obscure. Here we examine Procotyla fluviatilis, a planarian with restricted ability to replace missing tissues, using next-generation sequencing to define the gene expression programs active in regeneration-permissive and regeneration-deficient tissues. We found that Wnt signalling is aberrantly activated in regeneration-deficient tissues. Notably, downregulation of canonical Wnt signalling in regeneration-deficient regions restores regenerative abilities: blastemas form and new heads regenerate in tissues that normally never regenerate. This work reveals that manipulating a single signalling pathway can reverse the evolutionary loss of regenerative potential. PMID:23883929

Sikes, James M; Newmark, Phillip A

2013-07-24

145

Identification of small non-coding RNAs in the planarian Dugesia japonica via deep sequencing.  

PubMed

Freshwater planarian flatworm possesses an extraordinary ability to regenerate lost body parts after amputation; it is perfect organism model in regeneration and stem cell biology. Recently, small RNAs have been an increasing concern and studied in many aspects, including regeneration and stem cell biology, among others. In the current study, the large-scale cloning and sequencing of sRNAs from the intact and regenerative planarian Dugesia japonica are reported. Sequence analysis shows that sRNAs between 18nt and 40nt are mainly microRNAs and piRNAs. In addition, 209 conserved miRNAs and 12 novel miRNAs are identified. Especially, a better screening target method, negative-correlation relationship of miRNAs and mRNA, is adopted to improve target prediction accuracy. Similar to miRNAs, a diverse population of piRNAs and changes in the two samples are also listed. The present study is the first to report on the important role of sRNAs during planarian Dugesia japonica regeneration. PMID:22425900

Qin, Yun-Fei; Zhao, Jin-Mei; Bao, Zhen-Xia; Zhu, Zhao-Yu; Mai, Jia; Huang, Yi-Bo; Li, Jian-Biao; Chen, Ge; Lu, Ping; Chen, San-Jun; Su, Lin-Lin; Fang, Hui-Min; Lu, Ji-Ke; Zhang, Yi-Zhe; Zhang, Shou-Tao

2012-03-08

146

Differential expression of microRNA patterns in planarian normal and regenerative tissues.  

PubMed

MicroRNAs (miRNAs) are ~22-nt small non-coding RNAs that regulate the expression of specific target genes in many eukaryotes. miRNAs have been shown to play important roles in stem cell maintenance, cell fate determination, and differentiation. Planarians are capable of regenerating entire body plans from tiny fragments; this regenerative capacity is facilitated by a population of pluripotent stem cells known as neoblasts. Planarians have been a classic model system for the study of many aspects of stem cell biology. However, very limited knowledge on miRNA involved in this regulatory mechanism exists. This study profiles the expression of miRNAs in the normal and regenerative tissues of planarians using miRCURY LNA array technology. Thirteen miRNAs showed significant differences in expression between these two tissues. To further confirm our results, we examined the expression of two miRNAs by qRT-PCR. Results show that some known miRNAs may play key roles in the regulatory mechanisms of regeneration. Our findings can be utilized in future research on miRNA function. PMID:21713409

Tian, Qing-Nan; Bao, Zhen-Xia; Lu, Ping; Qin, Yun-Fei; Chen, San-Jun; Liang, Feng; Mai, Jia; Zhao, Jin-Mei; Zhu, Zhao-Yu; Zhang, Yi-Zhe; Zhang, Shou-Tao

2011-06-29

147

Stem cell-based growth, regeneration, and remodeling of the planarian intestine  

PubMed Central

Although some animals are capable of regenerating organs, the mechanisms by which this is achieved are poorly understood. In planarians, pluripotent somatic stem cells called neoblasts supply new cells for growth, replenish tissues in response to cellular turnover, and regenerate tissues after injury. For most tissues and organs, however, the spatiotemporal dynamics of stem cell differentiation and the fate of tissue that existed prior to injury have not been characterized systematically. Utilizing in vivo imaging and bromodeoxyuridine pulse-chase experiments, we have analyzed growth and regeneration of the planarian intestine, the organ responsible for digestion and nutrient distribution. During growth, we observe that new gut branches are added along the entire anteroposterior axis. We find that new enterocytes differentiate throughout the intestine rather than in specific growth zones, suggesting that branching morphogenesis is achieved primarily by remodeling of differentiated intestinal tissues. During regeneration, we also demonstrate a previously unappreciated degree of intestinal remodeling, in which pre-existing posterior gut tissue contributes extensively to the newly formed anterior gut, and vice versa. By contrast to growing animals, differentiation of new intestinal cells occurs at preferential locations, including within newly generated tissue (the blastema), and along pre-existing intestinal branches undergoing remodeling. Our results indicate that growth and regeneration of the planarian intestine are achieved by coordinated differentiation of stem cells and the remodeling of pre-existing tissues. Elucidation of the mechanisms by which these processes are integrated will be critical for understanding organogenesis in a post-embryonic context.

Forsthoefel, David J.; Park, Amanda E.; Newmark, Phillip A.

2011-01-01

148

TORC1 is required to balance cell proliferation and cell death in planarians  

PubMed Central

Multicellular organisms are equipped with cellular mechanisms that enable them to replace differentiated cells lost to normal physiological turnover, injury, and for some such as planarians, even amputation. This process of tissue homeostasis is generally mediated by adult stem cells (ASCs), tissue-specific stem cells responsible for maintaining anatomical form and function. To do so, ASCs must modulate the balance between cell proliferation, i.e. in response to nutrients, and that of cell death, i.e. in response to starvation or injury. But how these two antagonistic processes are coordinated remains unclear. Here, we explore the role of the core components of the TOR pathway during planarian tissue homeostasis and regeneration and identified an essential function for TORC1 in these two processes. RNAi-mediated silencing of TOR in intact animals resulted in a significant increase in cell death, whereas stem cell proliferation and stem cell maintenance were unaffected. Amputated animals failed to increase stem cell proliferation after wounding and displayed defects in tissue remodeling. Together, our findings suggest two distinct roles for TORC1 in planarians. TORC1 is required to modulate the balance between cell proliferation and cell death during normal cell turnover and in response to nutrients. In addition, it is required to initiate appropriate stem cell proliferation during regeneration and for proper tissue remodeling to occur to maintain scale and proportion.

Tu, Kimberly C.; Pearson, Bret J.; Alvarado, Alejandro Sanchez

2012-01-01

149

An automated training paradigm reveals long-term memory in planarians and its persistence through head regeneration.  

PubMed

Planarian flatworms are a popular system for research into the molecular mechanisms that enable these complex organisms to regenerate their entire body, including the brain. Classical data suggest that they may also be capable of long-term memory. Thus, the planarian system may offer the unique opportunity to study brain regeneration and memory in the same animal. To establish a system for the investigation of the dynamics of memory in a regenerating brain, we developed a computerized training and testing paradigm that avoided the many issues that confounded previous, manual attempts to train planarians. We then used this new system to train flatworms in an environmental familiarization protocol. We show that worms exhibit environmental familiarization, and that this memory persists for at least 14 days - long enough for the brain to regenerate. We further show that trained, decapitated planarians exhibit evidence of memory retrieval in a savings paradigm after regenerating a new head. Our work establishes a foundation for objective, high-throughput assays in this molecularly tractable model system that will shed light on the fundamental interface between body patterning and stored memories. We propose planarians as key emerging model species for mechanistic investigations of the encoding of specific memories in biological tissues. Moreover, this system is lik ely to have important implications for the biomedicine of stem-cell-derived treatments of degenerative brain disorders in human adults. PMID:23821717

Shomrat, Tal; Levin, Michael

2013-07-02

150

Regionalization of DNA and protein synthesis in developing stages of the parasitic platyhelminth Echinococcus granulosus.  

PubMed

Echinococcus granulosus is a parasitic platyhelminth, which causes cystid hydatid disease, a major zoonosis involving canids as definitive hosts, and both human and herbivorous domestic animals as intermediate hosts. The disease is caused in intermediate hosts by hydatid cysts, formed upon ingestion of E. granulosus eggs excreted by canids. Protoscoleces, the developmental forms of the parasite infective to canids, are formed in the germinal cellular layer of hydatid cysts. We have found that protoscoleces develop from patches of proliferating cells present in the germinal layer of the hydatid cyst, while most of the other cells of the germinal layer are in a resting state. Further, patches of proliferating cells form buds, which elongate and develop a separate population of cycling cells. In these elongated buds, cell differentiation leads to the main structures of the protoscolex. Protein synthesis is very active among cells of early buds and coincides with their proliferating activity. By contrast, protein synthesis presents a much lower activity in the resting cells of the germinal layer surrounding the growing protoscoleces. In elongated buds at different stages of development, protein synthesis is found mainly close to cellular territories in which cell differentiation occurs. In free infective protoscoleces, cells in DNA synthesis are concentrated in the body of the larva while protein synthesis occurs in the entire larva. This is the first description of the regionalization of DNA and protein synthesis in developing stages of E. granulosus. PMID:14505346

Galindo, M; Paredes, R; Marchant, C; Mińo, V; Galanti, N

2003-10-01

151

Cellular organization and appearance of differentiated structures in developing stages of the parasitic platyhelminth Echinococcus granulosus.  

PubMed

Echinococcus granulosus is the causative agent of hydatidosis, a major zoonoses that affects humans and herbivorous domestic animals. The disease is caused by the pressure exerted on viscera by hydatid cysts that are formed upon ingestion of E. granulosus eggs excreted by canine. Protoscoleces, larval forms infective to canine, develop asynchronously and clonally from the germinal layer (GL) of hydatid cysts. In this report, we describe the cellular organization and the appearance of differentiated structures both in nascent buds and developed protoscoleces attached to the GL. Early protoscolex morphogenesis is a highly complex and dynamic process starting from the constitution of a foramen in the early bud, around which nuclei are distributed mainly at the lateral and apical regions. Similarly, distribution of nuclei in mature protoscoleces is not homogenous but underlies three cellular territories: the suckers, the rostellar pad, and the body, that surrounds the foramen. Several nuclei are associated to calcareous corpuscles (Cc), differentiated structures that are absent in the earlier bud stages. The number of nuclei is similar from the grown, elongated bud stage to the mature protoscolex attached to the GL, strongly suggesting that there is no significant cellular proliferation during final protoscolex development. The amount of DNA per nucleus is in the same range to the one described for most other platyhelminthes. Our results point to a sequential series of events involving cell proliferation, spatial cell organization, and differentiation, starting in early buds at the GL of fertile hydatid cysts leading to mature protoscoleces infective to canine. PMID:15526286

Martínez, Claudio; Paredes, R; Stock, R P; Saralegui, A; Andreu, M; Cabezón, C; Ehrlich, R; Galanti, N

2005-02-01

152

A Dual Platform Approach to Transcript Discovery for the Planarian Schmidtea Mediterranea to Establish RNAseq for Stem Cell and Regeneration Biology  

PubMed Central

The use of planarians as a model system is expanding and the mechanisms that control planarian regeneration are being elucidated. The planarian Schmidtea mediterranea in particular has become a species of choice. Currently the planarian research community has access to this whole genome sequencing project and over 70,000 expressed sequence tags. However, the establishment of massively parallel sequencing technologies has provided the opportunity to define genetic content, and in particular transcriptomes, in unprecedented detail. Here we apply this approach to the planarian model system. We have sequenced, mapped and assembled 581,365 long and 507,719,814 short reads from RNA of intact and mixed stages of the first 7 days of planarian regeneration. We used an iterative mapping approach to identify and define de novo splice sites with short reads and increase confidence in our transcript predictions. We more than double the number of transcripts currently defined by publicly available ESTs, resulting in a collection of 25,053 transcripts described by combining platforms. We also demonstrate the utility of this collection for an RNAseq approach to identify potential transcripts that are enriched in neoblast stem cells and their progeny by comparing transcriptome wide expression levels between irradiated and intact planarians. Our experiments have defined an extensive planarian transcriptome that can be used as a template for RNAseq and can also help to annotate the S. mediterranea genome. We anticipate that suites of other 'omic approaches will also be facilitated by building on this comprehensive data set including RNAseq across many planarian regenerative stages, scenarios, tissues and phenotypes generated by RNAi.

Wilson, Ray; Evans, Deborah; Jowett, Jamie; Hall, Amy; Lemay, Virginie; Lam, Sabrina; Aboobaker, A. Aziz

2010-01-01

153

[New occurrences of metacercariae of Austrodiplostomum compactum (Lutz, 1928) (Platyhelminthes: Digenea) eye flukes of fish from the Paraná Basin].  

PubMed

Austrodiplostomum compactum (Platyhelminthes, Digenea) eye flukes of several species of fishes. The presence of this parasite, in extreme cases, can cause swelling of the eyelids, displacement of the retina, opacity of the crystalline lens and blindness or even death. The present study it registers new occurrences of this metacercariae infecting the eyes of four new hosts of fish, Serrasalmus maculatus collected in the Rosana reservoir in the Paranapanema river and Hypostomus regani, Schizodon borellii and Auchenipterus osteomystax collected in the the Upper Paraná River floodplain. PMID:19245765

Yamada, Fábio Hideki; Moreira, Luis Henrique De A; Ceschini, Tiago L; Takemoto, Ricardo Massato; Pavanelli, Gilberto Cezar

154

Drpiwi-1 is essential for germline cell formation during sexualization of the planarian Dugesia ryukyuensis.  

PubMed

A piwi homolog is required for the regulation of stem cells, formation and maintenance of germline stem cells, and gametogenesis in many metazoans. Planarians can change their reproductive mode seasonally, both asexually and sexually, and develop and maintain germ cells and sexual organs. They have many pluripotent stem cells (neoblasts) that can differentiate into both somatic and germline stem cells. Thus, we searched for a piwi subfamily in the planarian Dugesia ryukyuensis. Four piwi homologs, identified as Drpiwi-1, -2, -3, and -4, were expressed in sexually reproductive worms. We then selectively destroyed the neoblasts by irradiating the worms with X-rays. In such worms, Drpiwi-1, -2, and -3 were not expressed at all, whereas Drpiwi-4 was expressed to the same degree as that in non-irradiated controls, indicating that Drpiwi-1, -2, and -3, but not Drpiwi-4, are expressed in neoblasts. During the regeneration process, Drpiwi-2(RNAi) and -3(RNAi) worms failed to regenerate after ablation, but Drpiwi-1 and -4(RNAi) worms regenerated. During the sexualizing process, Drpiwi-1(RNAi) worms failed to develop ovaries and testes, but somatic sexual organs were unaffected. Germ cell development was normal in Drpiwi-4(RNAi) worms. Therefore, Drpiwi-2 and -3 may be related to the regulation of neoblasts important for maintaining homeostasis, and Drpiwi-1 is essential for the development of germ cells but not somatic sexual organs. DrPiwi-1 is localized in the cytoplasm of stem cells and germline cells and may be involved in regulating some gene expression. We suggest that planarian Piwi controls germline formation via RNA silencing mechanisms. PMID:22024321

Nakagawa, Haruka; Ishizu, Hirotsugu; Hasegawa, Reiko; Kobayashi, Kazuya; Matsumoto, Midori

2011-10-15

155

Amphetamine-induced increase in planarian locomotor activity and block by UV light.  

PubMed

The dopamine D2-receptor antagonist sulpiride decreases spontaneous locomotor velocity of planarians (pLMV) in an enantiomeric-selective and dose-dependent manner and is significantly attenuated by UV light (254 and 366 nm). We now report that amphetamine (10 microM) produced the opposite effect and was also reversed by UV light. These findings strengthen the hypothesis that the effects of dopaminergic ligands and UV light on pLMV relate to interaction with neurotransmitter transduction process(es). PMID:15621023

Raffa, Robert B; Martley, Andrea F

2005-01-01

156

Phylogenetic diversity of microturbellarians in Japanese rice paddy fields, with special attention to the genus Stenostomum  

Microsoft Academic Search

Free-living fresh water Platyhelminthes except for the order Tricladida (planaria) are collectively called microturbellaria, most species of which are less than a few millimeters in length. The ecology and the fauna of microturbellarians in rice fields has not been clarified in detail since Okugawa (1932) reported the morphological and ecological characters of all microturbellarian species that had been observed in

Masatsugu Yamazaki; Susumu Asakawa; Jun Murase; Makoto Kimura

2012-01-01

157

Planarians maintain a constant ratio of different cell types during changes in body size by using the stem cell system.  

PubMed

Planarians change in body size depending upon whether they are in feeding or starving conditions. To investigate how planarians regulate this flexible system, the numbers of total cells and specific cell types were counted and compared among worms 2 mm to 9 mm in body length. The total cell number increased linearly with increasing body length, but the ratio of cell numbers between the head and the trunk portion was constant (1:3). Interestingly, counting the numbers of specific neurons in the eye and brain after immunostaining using cell type-specific antibodies revealed that the ratio between different neuron types was constant regardless of the brain and body size. These results suggest that planarians can maintain proportionality while changing their body size by maintaining a constant ratio of different cell types. To understand this system and reveal how planarians restore the original ratio during eye and brain regeneration, the numbers of specialized cells were Investigated during regeneration. The results further substantiate the existence of some form of "counting mechanism" that has the ability to regulate both the absolute and relative numbers of different cell types in complex organs such as the brain during cell turnover, starvation, and regeneration. PMID:19968467

Takeda, Hiroyuki; Nishimura, Kaneyasu; Agata, Kiyokazu

2009-12-01

158

Size Matters!. Birth Size and a Size-Independent Stochastic Term Determine Asexual Reproduction Dynamics in Freshwater Planarians  

NASA Astrophysics Data System (ADS)

Asexual reproduction by division in higher organisms is rare, because a prerequisite is the ability to regenerate an entire organism from a piece of the original body. Freshwater planarians are one of the few animals that can reproduce this way, but little is known about the regulation of their reproduction cycles or strategies. We have previously shown that a planarian's reproduction strategy is randomized to include fragmentations, producing multiple offspring, as well as binary fissions, and can be partially explained by a maximum relative entropy principle. In this study we attempt to decompose the factors controlling their reproduction cycle. Based on recent studies on the cell cycle of budding yeast, which suggest that molecular noise in gene expression and cell size at birth together control cell cycle variability, we investigated whether the variability in planarian reproduction waiting times could be similarly regulated. We find that such a model can indeed explain the observed distribution of waiting times between birth and next reproductive event, suggesting that birth size and a stochastic noise term govern the reproduction dynamics of asexual planarians.

Thomas, Michael A.; Quinodoz, Sofia; Schötz, Eva-Maria

2012-09-01

159

Genetic regulators of a pluripotent adult stem cell system in planarians identified by RNAi and clonal analysis.  

PubMed

Pluripotency is a central, well-studied feature of embryonic development, but the role of pluripotent cell regulation in somatic tissue regeneration remains poorly understood. In planarians, regeneration of entire animals from tissue fragments is promoted by the activity of adult pluripotent stem cells (cNeoblasts). We utilized transcriptional profiling to identify planarian genes expressed in adult proliferating, regenerative cells (neoblasts). We also developed quantitative clonal analysis methods for expansion and differentiation of cNeoblast descendants that, together with RNAi, revealed gene roles in stem cell biology. Genes encoding two zinc finger proteins, Vasa, a LIM domain protein, Sox and Jun-like transcription factors, two candidate RNA-binding proteins, a Setd8-like protein, and PRC2 (Polycomb) were required for proliferative expansion and/or differentiation of cNeoblast-derived clones. These findings suggest that planarian stem cells utilize molecular mechanisms found in germ cells and other pluripotent cell types and identify genetic regulators of the planarian stem cell system. PMID:22385657

Wagner, Daniel E; Ho, Jaclyn J; Reddien, Peter W

2012-03-01

160

Genetic regulators of a pluripotent adult stem cell system in planarians identified by RNAi and clonal analysis  

PubMed Central

Summary Pluripotency is a central, well-studied feature of embryonic development, but the role of pluripotent cell regulation in somatic tissue regeneration remains poorly understood. In planarians, regeneration of entire animals from tissue fragments is promoted by the activity of adult pluripotent stem cells (cNeoblasts). We utilized transcriptional profiling to identify planarian genes expressed in adult proliferating, regenerative cells (neoblasts). We also developed quantitative clonal analysis methods for expansion and differentiation of cNeoblast descendants that, together with RNAi, revealed gene roles in stem cell biology. Genes encoding two zinc finger proteins, Vasa, a LIM domain protein, Sox and Jun-like transcription factors, two candidate RNA-binding proteins, a Setd8-like protein, and PRC2 (Polycomb) were required for proliferative expansion and/or differentiation of cNeoblast-derived clones. These findings suggest that planarian stem cells utilize molecular mechanisms found in germ cells and other pluripotent cell types, and identify novel genetic regulators of the planarian stem cell system.

Wagner, Daniel E.; Ho, Jaclyn J.

2012-01-01

161

SMG-1 and mTORC1 Act Antagonistically to Regulate Response to Injury and Growth in Planarians  

PubMed Central

Planarian flatworms are able to both regenerate their whole bodies and continuously adapt their size to nutrient status. Tight control of stem cell proliferation and differentiation during these processes is the key feature of planarian biology. Here we show that the planarian homolog of the phosphoinositide 3-kinase-related kinase (PIKK) family member SMG-1 and mTOR complex 1 components are required for this tight control. Loss of smg-1 results in a hyper-responsiveness to injury and growth and the formation of regenerative blastemas that remain undifferentiated and that lead to lethal ectopic outgrowths. Invasive stem cell hyper-proliferation, hyperplasia, hypertrophy, and differentiation defects are hallmarks of this uncontrolled growth. These data imply a previously unappreciated and novel physiological function for this PIKK family member. In contrast we found that planarian members of the mTOR complex 1, tor and raptor, are required for the initial response to injury and blastema formation. Double smg-1 RNAi experiments with tor or raptor show that abnormal growth requires mTOR signalling. We also found that the macrolide rapamycin, a natural compound inhibitor of mTORC1, is able to increase the survival rate of smg-1 RNAi animals by decreasing cell proliferation. Our findings support a model where Smg-1 acts as a novel regulator of both the response to injury and growth control mechanisms. Our data suggest the possibility that this may be by suppressing mTOR signalling. Characterisation of both the planarian mTORC1 signalling components and another PIKK family member as key regulators of regeneration and growth will influence future work on regeneration, growth control, and the development of anti-cancer therapies that target mTOR signalling.

Gonzalez-Estevez, Cristina; Felix, Daniel A.; Smith, Matthew D.; Paps, Jordi; Morley, Simon J.; James, Victoria; Sharp, Tyson V.; Aboobaker, A. Aziz

2012-01-01

162

SMG-1 and mTORC1 act antagonistically to regulate response to injury and growth in planarians.  

PubMed

Planarian flatworms are able to both regenerate their whole bodies and continuously adapt their size to nutrient status. Tight control of stem cell proliferation and differentiation during these processes is the key feature of planarian biology. Here we show that the planarian homolog of the phosphoinositide 3-kinase-related kinase (PIKK) family member SMG-1 and mTOR complex 1 components are required for this tight control. Loss of smg-1 results in a hyper-responsiveness to injury and growth and the formation of regenerative blastemas that remain undifferentiated and that lead to lethal ectopic outgrowths. Invasive stem cell hyper-proliferation, hyperplasia, hypertrophy, and differentiation defects are hallmarks of this uncontrolled growth. These data imply a previously unappreciated and novel physiological function for this PIKK family member. In contrast we found that planarian members of the mTOR complex 1, tor and raptor, are required for the initial response to injury and blastema formation. Double smg-1 RNAi experiments with tor or raptor show that abnormal growth requires mTOR signalling. We also found that the macrolide rapamycin, a natural compound inhibitor of mTORC1, is able to increase the survival rate of smg-1 RNAi animals by decreasing cell proliferation. Our findings support a model where Smg-1 acts as a novel regulator of both the response to injury and growth control mechanisms. Our data suggest the possibility that this may be by suppressing mTOR signalling. Characterisation of both the planarian mTORC1 signalling components and another PIKK family member as key regulators of regeneration and growth will influence future work on regeneration, growth control, and the development of anti-cancer therapies that target mTOR signalling. PMID:22479207

González-Estévez, Cristina; Felix, Daniel A; Smith, Matthew D; Paps, Jordi; Morley, Simon J; James, Victoria; Sharp, Tyson V; Aboobaker, A Aziz

2012-03-29

163

Defining the molecular profile of planarian pluripotent stem cells using a combinatorial RNA-seq, RNA interference and irradiation approach  

PubMed Central

Background Planarian stem cells, or neoblasts, drive the almost unlimited regeneration capacities of freshwater planarians. Neoblasts are traditionally described by their morphological features and by the fact that they are the only proliferative cell type in asexual planarians. Therefore, they can be specifically eliminated by irradiation. Irradiation, however, is likely to induce transcriptome-wide changes in gene expression that are not associated with neoblast ablation. This has affected the accurate description of their specific transcriptomic profile. Results We introduce the use of Smed-histone-2B RNA interference (RNAi) for genetic ablation of neoblast cells in Schmidtea mediterranea as an alternative to irradiation. We characterize the rapid, neoblast-specific phenotype induced by Smed-histone-2B RNAi, resulting in neoblast ablation. We compare and triangulate RNA-seq data after using both irradiation and Smed-histone-2B RNAi over a time course as means of neoblast ablation. Our analyses show that Smed-histone-2B RNAi eliminates neoblast gene expression with high specificity and discrimination from gene expression in other cellular compartments. We compile a high confidence list of genes downregulated by both irradiation and Smed-histone-2B RNAi and validate their expression in neoblast cells. Lastly, we analyze the overall expression profile of neoblast cells. Conclusions Our list of neoblast genes parallels their morphological features and is highly enriched for nuclear components, chromatin remodeling factors, RNA splicing factors, RNA granule components and the machinery of cell division. Our data reveal that the regulation of planarian stem cells relies on posttranscriptional regulatory mechanisms and suggest that planarians are an ideal model for this understudied aspect of stem cell biology.

2012-01-01

164

Proteomic Profiling of the Planarian Schmidtea mediterranea and Its Mucous Reveals Similarities with Human Secretions and Those Predicted for Parasitic Flatworms*  

PubMed Central

The freshwater planarian Schmidtea mediterranea has been used in research for over 100 years, and is an emerging stem cell model because of its capability of regenerating large portions of missing body parts. Exteriorly, planarians are covered in mucous secretions of unknown composition, implicated in locomotion, predation, innate immunity, and substrate adhesion. Although the planarian genome has been sequenced, it remains mostly unannotated, challenging both genomic and proteomic analyses. The goal of the current study was to annotate the proteome of the whole planarian and its mucous fraction. The S. mediterranea proteome was analyzed via mass spectrometry by using multidimensional protein identification technology with whole-worm tryptic digests. By using a proteogenomics approach, MS data were searched against an in silico translated planarian transcript database, and by using the Swiss-Prot BLAST algorithm to identify proteins similar to planarian queries. A total of 1604 proteins were identified. The mucous subproteome was defined through analysis of a mucous trail fraction and an extract obtained by treating whole worms with the mucolytic agent N-acetylcysteine. Gene Ontology analysis confirmed that the mucous fractions were enriched with secreted proteins. The S. mediterranea proteome is highly similar to that predicted for the trematode Schistosoma mansoni associated with intestinal schistosomiasis, with the mucous subproteome particularly highly conserved. Remarkably, orthologs of 119 planarian mucous proteins are present in human mucosal secretions and tear fluid. We suggest planarians have potential to be a model system for the characterization of mucous protein function and relevant to parasitic flatworm infections and diseases underlined by mucous aberrancies, such as cystic fibrosis, asthma, and other lung diseases.

Bocchinfuso, Donald G.; Taylor, Paul; Ross, Eric; Ignatchenko, Alex; Ignatchenko, Vladimir; Kislinger, Thomas; Pearson, Bret J.; Moran, Michael F.

2012-01-01

165

Morphogenesis defects are associated with abnormal nervous system regeneration following roboA RNAi in planarians.  

PubMed

The process by which the proper pattern is restored to newly formed tissues during metazoan regeneration remains an open question. Here, we provide evidence that the nervous system plays a role in regulating morphogenesis during anterior regeneration in the planarian Schmidtea mediterranea. RNA interference (RNAi) knockdown of a planarian ortholog of the axon-guidance receptor roundabout (robo) leads to unexpected phenotypes during anterior regeneration, including the development of a supernumerary pharynx (the feeding organ of the animal) and the production of ectopic, dorsal outgrowths with cephalic identity. We show that Smed-roboA RNAi knockdown disrupts nervous system structure during cephalic regeneration: the newly regenerated brain and ventral nerve cords do not re-establish proper connections. These neural defects precede, and are correlated with, the development of ectopic structures. We propose that, in the absence of proper connectivity between the cephalic ganglia and the ventral nerve cords, neurally derived signals promote the differentiation of pharyngeal and cephalic structures. Together with previous studies on regeneration in annelids and amphibians, these results suggest a conserved role of the nervous system in pattern formation during blastema-based regeneration. PMID:17251262

Cebriŕ, Francesc; Newmark, Phillip A

2007-01-24

166

TOR signaling regulates planarian stem cells and controls localized and organismal growth.  

PubMed

Target of Rapamycin (TOR) controls an evolutionarily conserved signaling pathway that modulates cellular growth and division by sensing levels of nutrients, energy and stress. As such, TOR signaling is a crucial component of tissues and organs that translates systemic signals into cellular behavior. The ubiquitous nature of TOR signaling, together with the difficulty of analyzing tissue during cellular turnover and repair, have limited our understanding of how this kinase operates throughout the body. Here, we use the planarian model system to address TOR regulation at the organismal level. The planarian TOR homolog (Smed-TOR) is ubiquitously expressed, including stem cells (neoblasts) and differentiated tissues. Inhibition of TOR with RNA interference severely restricts cell proliferation, allowing the study of neoblasts with restricted proliferative capacity during regeneration and systemic cell turnover. Strikingly, TOR signaling is required for neoblast response to amputation and localized growth (blastema). However, in the absence of TOR signaling, regeneration takes place only within differentiated tissues. In addition, TOR is essential for maintaining the balance between cell division and cell death, and its dysfunction leads to tissue degeneration and lack of organismal growth in the presence of nutrients. Finally, TOR function is likely to be mediated through TOR Complex 1 as its disruption recapitulates signs of the TOR phenotype. Our data reveal novel roles for TOR signaling in controlling adult stem cells at a systemic level and suggest a new paradigm for studying TOR function during physiological turnover and regeneration. PMID:22427692

Peiris, T Harshani; Weckerle, Frank; Ozamoto, Elyse; Ramirez, Daniel; Davidian, Devon; García-Ojeda, Marcos E; Oviedo, Néstor J

2012-03-16

167

Identification of neoblast- and regeneration-specific miRNAs in the planarian Schmidtea mediterranea.  

PubMed

In recent years, the planarian Schmidtea mediterranea has emerged as a tractable model system to study stem cell biology and regeneration. MicroRNAs are small RNA species that control gene expression by modulating translational repression and mRNA stability and have been implicated in the regulation of various cellular processes. Though recent studies have identified several miRNAs in S. mediterranea, their expression in neoblast subpopulations and during regeneration has not been examined. Here, we identify several miRNAs whose expression is enriched in different neoblast subpopulations and in regenerating tissue at different time points in S. mediterranea. Some of these miRNAs were enriched within 3 h post-amputation and may, therefore, play a role in wound healing and/or neoblast migration. Our results also revealed miRNAs, such as sme-miR-2d-3p and the sme-miR-124 family, whose expression is enriched in the cephalic ganglia, are also expressed in the brain primordium during CNS regeneration. These results provide new insight into the potential biological functions of miRNAs in neoblasts and regeneration in planarians. PMID:23974438

Sasidharan, Vidyanand; Lu, Yi-Chien; Bansal, Dhiru; Dasari, Pranavi; Poduval, Deepak; Seshasayee, Aswin; Resch, Alissa M; Graveley, Brenton R; Palakodeti, Dasaradhi

2013-08-23

168

PBX/extradenticle is required to re-establish axial structures and polarity during planarian regeneration.  

PubMed

Recent advances in a number of systems suggest many genes involved in orchestrating regeneration are redeployed from similar processes in development, with others being novel to the regeneration process in particular lineages. Of particular importance will be understanding the architecture of regenerative genetic regulatory networks and whether they are conserved across broad phylogenetic distances. Here, we describe the role of the conserved TALE class protein PBX/Extradenticle in planarians, a representative member of the Lophotrocozoa. PBX/Extradenticle proteins play central roles in both embryonic and post-embryonic developmental patterning in both vertebrates and insects, and we demonstrate a broad requirement during planarian regeneration. We observe that Smed-pbx has pleiotropic functions during regeneration, with a primary role in patterning the anterior-posterior (AP) axis and AP polarity. Smed-pbx is required for expression of polarity determinants notum and wnt1 and for correct patterning of the structures polarized along the AP axis, such as the brain, pharynx and gut. Overall, our data suggest that Smed-pbx functions as a central integrator of positional information to drive patterning of regeneration along the body axis. PMID:23318635

Blassberg, Robert A; Felix, Daniel A; Tejada-Romero, Belen; Aboobaker, A Aziz

2013-01-14

169

PBX/extradenticle is required to re-establish axial structures and polarity during planarian regeneration  

PubMed Central

Recent advances in a number of systems suggest many genes involved in orchestrating regeneration are redeployed from similar processes in development, with others being novel to the regeneration process in particular lineages. Of particular importance will be understanding the architecture of regenerative genetic regulatory networks and whether they are conserved across broad phylogenetic distances. Here, we describe the role of the conserved TALE class protein PBX/Extradenticle in planarians, a representative member of the Lophotrocozoa. PBX/Extradenticle proteins play central roles in both embryonic and post-embryonic developmental patterning in both vertebrates and insects, and we demonstrate a broad requirement during planarian regeneration. We observe that Smed-pbx has pleiotropic functions during regeneration, with a primary role in patterning the anterior-posterior (AP) axis and AP polarity. Smed-pbx is required for expression of polarity determinants notum and wnt1 and for correct patterning of the structures polarized along the AP axis, such as the brain, pharynx and gut. Overall, our data suggest that Smed-pbx functions as a central integrator of positional information to drive patterning of regeneration along the body axis.

Blassberg, Robert A.; Felix, Daniel A.; Tejada-Romero, Belen; Aboobaker, A. Aziz

2013-01-01

170

The planarian neoblast: the rambling history of its origin and some current black boxes.  

PubMed

First described by Randolph in 1897, the nature and main features of planarian neoblasts have a long rambling history. While their morphologically undifferentiated features have long been recognized, their origin and actual role during regeneration have been highly debated. Here I summarize the main stages of this rambling history: 1) undifferentiated, wandering cells of uncertain origin with a main, albeit undefined, role in regeneration (1890-1940s); 2) quiescent, undifferentiated cells whose main function is to build the blastema during regeneration, an idea which culminated in the 'neoblast theory' of the French School (1940-1960); 3) neoblasts as temporal, undifferentiated cells arising by dedifferentiation from differentiated cells (the 'cell dedifferentiation theory'; 1960-1980s); 4) a new paradigm, starting in the late 1970s-early 1980s, that brought together the role of neoblasts as the main cell for regeneration, with its more important role as somatic stem cells for the daily wear and tear of tissues and as the source of germ cells; and 5) more recent developments that culminate in the report of rescuing lethally irradiated planarians by injection of single neoblasts, which makes of neoblasts an unrivaled toti-, pluripotent somatic stem cell system in the Animal Kingdom. I finally discuss some "black boxes" regarding neoblasts which still baffle us, namely their phylogenetic and ontogenetic origins, their role in body size control, how their pool is regulated during growth and degrowth, the logic of their proliferative control, and some 'old' long-sought missing tools. PMID:22252540

Baguńŕ, Jaume

2012-01-01

171

An RbAp48-like gene regulates adult stem cells in planarians.  

PubMed

Retinoblastoma-associated proteins 46 and 48 (RbAp46 and RbAp48) are factors that are components of different chromatin-modelling complexes, such as polycomb repressive complex 2, the activity of which is related to epigenetic gene regulation in stem cells. To date, no direct findings are available on the in vivo role of RbAp48 in stem-cell biology. We recently identified DjRbAp48 - a planarian (Dugesia japonica) homologue of human RBAP48 - expression of which is restricted to the neoblasts, the adult stem cells of planarians. In vivo silencing of DjRbAp48 induces lethality and inability to regenerate, even though neoblasts proliferate and accumulate after wounding. Despite a partial reduction in neoblast number, we were always able to detect a significant number of these cells in DjRbAp48 RNAi animals. Parallel to the decrease in neoblasts, a reduction in the number of differentiated cells and the presence of apoptotic-like neoblasts were detectable in RNAi animals. These findings suggest that DjRbAp48 is not involved in neoblast maintenance, but rather in the regulation of differentiation of stem-cell progeny. We discuss our data, taking into account the possibility that DjRbAp48 might control the expression of genes necessary for cell differentiation by influencing chromatin architecture. PMID:20124416

Bonuccelli, Lucia; Rossi, Leonardo; Lena, Annalisa; Scarcelli, Vittoria; Rainaldi, Giuseppe; Evangelista, Monica; Iacopetti, Paola; Gremigni, Vittorio; Salvetti, Alessandra

2010-02-02

172

Long-range Neural and Gap Junction Protein-mediated Cues Control Polarity During Planarian Regeneration  

PubMed Central

SUMMARY Having the ability to coordinate the behavior of stem cells to induce regeneration of specific large-scale structures would have far reaching consequences in the treatment of degenerative diseases, acute injury, and aging. Thus, identifying and learning to manipulate the sequential steps that determine the fate of new tissue within the overall morphogenetic program of the organism is fundamental. We identified novel early signals, mediated by the central nervous system and 3 innexin proteins, which determine the fate and axial polarity of regenerated tissue in planarians. Modulation of gap junction-dependent and neural signals specifically induces ectopic anterior regeneration blastemas in posterior and lateral wounds. These ectopic anterior blastemas differentiate new brains that establish permanent primary axes re-established during subsequent rounds of unperturbed regeneration. These data reveal powerful novel controls of pattern formation and suggest a constructive model linking nervous inputs and polarity determination in early stages of regeneration.

Oviedo, Nestor J.; Morokuma, Junji; Walentek, Peter; Kema, Ido P.; Gu, Man Bock; Ahn, Joo-Myung; Hwang, Jung Shan; Gojobori, Takashi; Levin, Michael

2010-01-01

173

Molecular analysis of stem cells and their descendents during cell turnover and regeneration in the planarian Schmidtea mediterranea  

PubMed Central

In adult planarians the replacement of cells lost to physiological turnover or injury is sustained by the proliferation and differentiation of stem cells known as neoblasts. Neoblast lineage relationships and the molecular changes that take place during differentiation into the appropriate cell types are poorly understood. Here we report the identification and characterization of a cohort of genes specifically expressed in neoblasts and their descendents. We find that genes with severely downregulated expression after irradiation molecularly define at least three discrete subpopulations of cells. Simultaneous BrdU labeling and in situ hybridization experiments in intact and regenerating animals indicate that these cell subpopulations are related by lineage. Our data demonstrate not only the ability to measure and study the in vivo population dynamics of adult stem cells during tissue homeostasis and regeneration, but also the utility of studies in planarians to broadly inform stem cell biology in adult organisms.

Eisenhoffer, George T.; Kang, Hara; Alvarado, Alejandro Sanchez

2008-01-01

174

Evaluation of endogenous reference genes for analysis of gene expression with real-time RT-PCR during planarian regeneration.  

PubMed

It is important that endogenous reference genes for real-time RT-PCR be empirically evaluated for stability in different cell types, developmental stages, and/or sample treatment. To select the most stable endogenous reference genes during planarian regeneration, three housekeeping genes, 18S rRNA, ACTB and DjEF2, were identified and established expression levels by real-time RT-PCR. The data were analyzed by GeNorm and NormFinder software. Expression levels of the Djsix-1 gene were studied in parallel with ACTB and DjEF2 both or each and 18S rRNA as reference during regeneration. The results showed that ACTB was the most stable expressed reference gene in the planarian regeneration. PMID:21161407

Yuwen, Yan-Qing; Dong, Zi-Mei; Wang, Qing-Hua; Sun, Xiao-Juan; Shi, Chang-Ying; Chen, Guang-Wen

2010-12-14

175

Global irradiation effects, stem cell genes and rare transcripts in the planarian transcriptome.  

PubMed

Stem cells are the closest relatives of the totipotent primordial cell, which is able to spawn millions of daughter cells and hundreds of cell types in multicellular organisms. Stem cells are involved in tissue homeostasis and regeneration, and may play a major role in cancer development. Among animals, planarians host a model stem cell type, called the neoblast, which essentially confers immortality. Gaining insights into the global transcriptional landscape of these exceptional cells takes an unprecedented turn with the advent of Next Generation Sequencing methods. Two Digital Gene Expression transcriptomes of Schmidtea mediterranea planarians, with or without neoblasts lost through irradiation, were produced and analyzed. Twenty one bp NlaIII tags were mapped to transcripts in the Schmidtea and Dugesia taxids. Differential representation of tags in normal versus irradiated animals reflects differential gene expression. Canonical and non-canonical tags were included in the analysis, and comparative studies with human orthologs were conducted. Transcripts fell into 3 categories: invariant (including housekeeping genes), absent in irradiated animals (potential neoblast-specific genes, IRDOWN) and induced in irradiated animals (potential cellular stress response, IRUP). Different mRNA variants and gene family members were recovered. In the IR-DOWN class, almost all of the neoblast-specific genes previously described were found. In irradiated animals, a larger number of genes were induced rather than lost. A significant fraction of IRUP genes behaved as if transcript versions of different lengths were produced. Several novel potential neoblast-specific genes have been identified that varied in relative abundance, including highly conserved as well as novel proteins without predicted orthologs. Evidence for a large body of antisense transcripts, for example regulated antisense for the Smed-piwil1 gene, and evidence for RNA shortening in irradiated animals is presented. Novel neoblast-specific candidates include a peroxiredoxin protein that appears to be preferentially expressed in human embryonic stem cells. PMID:22450998

Galloni, Mireille

2012-01-01

176

Comprehensive gene expression analyses in pluripotent stem cells of a planarian, Dugesia japonica.  

PubMed

The neoblasts are the only somatic stem cells in planarians possessing pluripotency, and can give rise to all types of cells, including germline cells. Recently, accumulated knowledge about the transcriptome and expression dynamics of various pluripotent somatic stem cells has provided important opportunities to understand not only fundamental mechanisms of pluripotency, but also stemness across species at the molecular level. The neoblasts can easily be eliminated by radiation. Also, by using fluorescence activated cell sorting (FACS), we can purify and collect many neoblasts, enabling identification of neoblast-related genes by comparison of the gene expression level among intact and X-ray-irradiated animals, and purified neoblasts. In order to find such genes, here we employed the high coverage expression profiling (HiCEP) method, which enables us to observe and compare genome-wide gene expression levels between different samples without advance sequence information, in the planarian D. japonica as a model organism of pluripotent stem cell research. We compared expression levels of ~17,000 peaks corresponding to independent genes among different samples, and obtained 102 peaks as candidates. Expression analysis of genes identified from those peaks by in situ hybridization revealed that at least 42 genes were expressed in the neoblasts and in neoblast-related cells that had a different distribution pattern in the body than neoblasts. Also, single-cell PCR analysis of those genes revealed heterogeneous expression of some genes in the neoblast population. Thus, using multidimensional gene expression analyses, we were able to obtain a valuable data set of neoblast-related genes and their expression patterns. PMID:22450997

Shibata, Norito; Hayashi, Tetsutaro; Fukumura, Ryutaro; Fujii, Junsuke; Kudome-Takamatsu, Tomomi; Nishimura, Osamu; Sano, Syozo; Son, Fuyan; Suzuki, Nobuko; Araki, Ryoko; Abe, Masumi; Agata, Kiyokazu

2012-01-01

177

Specific features of the planarian Dugesia tigrina regeneration and mollusk Helix albescens nociception under weak electromagnetic shielding  

NASA Astrophysics Data System (ADS)

It has been demonstrated that weak electromagnetic shielding stimulates regeneration in the planarian Dugesia tigrina, the stimulating intensity being dependent on both the initial state of the animals, which is determined by season, and their functional asymmetry. As has been shown, the effect of a weak electromagnetic field induces phasic changes in the nociceptive sensitivity of the mollusk Helix albescens: an increase in the sensitivity to a thermal stimulus is replaced by the development of the hypalgesic effect.

Temur'yants, N. A.; Demtsun, N. A.; Kostyuk, A. S.; Yarmolyuk, N. S.

2012-12-01

178

SpolvlgA is a DDX3/PL10-related DEAD-box RNA helicase expressed in blastomeres and embryonic cells in planarian embryonic development.  

PubMed

Planarian flatworms have an impressive regenerative power. Although their embryonic development is still poorly studied and is highly derived it still displays some simple characteristics. We have identified SpolvlgA, a Schmidtea polychroa homolog of the DDX3/PL10 DEAD-box RNA helicase DjvlgA from the planarian species Dugesia japonica. This gene has been previously described as being expressed in planarian adult stem cells (neoblasts), as well as the germ line. Here we present the expression pattern of SpolvlgA in developing embryos of S. polychroa and show that it is expressed from the first cleavage rounds in blastomere cells and blastomere-derived embryonic cells. These cells are undifferentiated cells that engage in a massive wave of differentiation during stage 5 of development. SpolvlgA expression highlights this wave of differentiation, where nearly all previous structures are substituted by blastomere-derived embryonic cells. In late stages of development SpolvlgA is expressed in most proliferating and differentiating cells. Thus, SpolvlgA is a gene expressed in planarian embryos from the first stages of development and a good marker for the zygote-derived cell lineage in these embryos. Expression in adult worms is also monitored and is found in the planarian germ line, where it is showed to be expressed in spermatogonia, spermatocytes and differentiating spermatids. PMID:19159016

Solana, Jordi; Romero, Rafael

2009-01-06

179

Complete Sequence of the mitochondrial genome of the tapeworm Hymenolepis diminuta: Gene arrangements indicate that platyhelminths are eutrochozoans  

SciTech Connect

Using ''long-PCR'' we have amplified in overlapping fragments the complete mitochondrial genome of the tapeworm Hymenolepis diminuta (Platyhelminthes: Cestoda) and determined its 13,900 nucleotide sequence. The gene content is the same as that typically found for animal mitochondrial DNA (mtDNA) except that atp8 appears to be lacking, a condition found previously for several other animals. Despite the small size of this mtDNA, there are two large non-coding regions, one of which contains 13 repeats of a 31 nucleotide sequence and a potential stem-loop structure of 25 base pairs with an 11-member loop. Large potential secondary structures are identified also for the non-coding regions of two other cestode mtDNAs. Comparison of the mitochondrial gene arrangement of H. diminuta with those previously published supports a phylogenetic position of flatworms as members of the Eutrochozoa, rather than being basal to either a clade of protostomes or a clade of coelomates.

von Nickisch-Rosenegk, Markus; Brown, Wesley M.; Boore, Jeffrey L.

2001-01-01

180

Evidence against gastrointestinal pseudoparasitism by the land planarian, Bipalium kewense Moseley 1878.  

PubMed

The possibility of gastrointestinal pseudoparasitism by the free-living land planarian, Bipalium kewense, was tested by feeding and survival experiments. The intestinal tracts of three dogs were negative for B. kewense after the individual dogs were fed eight, three, and four large worms and autopsied at 12 h, 3 h, and 45 min, respectively. Survival of the worms in Warburg flasks, under N2, AT 27 C, was 2 h or less.. In Gas Pak jars (CO2 + H2) at 37 C, survival was less than 60 min. Aerobically, at 37 C, survival varied from 45 to 60 min. Attra-tion of the worms to stool material was examined by placing planaria inside square whose boundries were constructed of fecal smears. Bipalium kewense exposed to canine feces showed strong avoidance reactions. Urea, a nitrogen end product of these worms, was also shown to be a negative stimulus for B. kewense. Failure to establish even short term passage in the digestive tract, lethality of 37 C and anaerobic environments, and sensitivity to feces makes gastrointestinal pseudoparasitism unlikely in these organisms. PMID:833002

Daly, J J; Matthews, H M; Farris, H E

1977-01-01

181

An insulin-like peptide regulates size and adult stem cells in planarians.  

PubMed

Animal growth depends on nutritional intake during development. In many animals, nutritional status is uncoupled from moderation of adult stature after adult size is achieved. However, some long-lived animals continue to regulate adult size and fertility in a nutrition-dependent manner. For example, the regenerating flatworm Schmidtea mediterranea becomes smaller, or degrows, during periods of starvation. These animals provide an opportunity to readily observe adult stem cell population dynamics in response to nutritional cues. We explored the role of insulin signaling in S. mediterranea. We disrupted insulin signaling via RNA interference and showed that animals, despite eating, degrew similarly to starved animals. Utilizing in situ hybridization and immunofluorescence, we assessed cellular changes in proliferative populations including the planarian adult stem cell population (neoblasts) and the germline. Both impaired insulin signaling and nutritional deprivation correlated with decreased neoblast proliferation. Additionally, insulin signaling played a role in supporting spermatogenesis that was distinct from the effects of starvation. In sum, we have demonstrated that insulin signaling is responsible for regulation of adult animal size and tissue homeostasis in an organism with plastic adult size. Importantly, insulin signaling continued to affect stem cell and germline populations in a mature organism. Furthermore, we have shown that adult organisms can differentially regulate specific cell populations as a result of environmental challenges. PMID:22252538

Miller, Claire M; Newmark, Phillip A

2012-01-01

182

Production of asexual and sexual offspring in the triploid sexual planarian Dugesia ryukyuensis.  

PubMed

Certain freshwater planarians reproduce asexually as well as sexually, and their chromosomal ploidies include polyploidy, aneuploidy and mixoploidy. Previously, we successfully performed an experiment in which a clonal population produced by asexual reproduction of the Dugesia ryukyuensis (OH strain) switched to the sexual mode of reproduction. Worms of this strain are triploid with a pericentric inversion on Chromosome 4. The worms were switched to sexual reproduction after being fed with sexually mature Bdellocephala brunnea, which is a sexually reproducing species. The resulting sexualized OH strain produced cocoons filled with several eggs. Two putative factors, Mendelian factor(s) and chromosomal control(s), have been proposed as determining the reproductive mode. The present study demonstrated that inbreeding of the resultant sexualized worms produced the following four types of offspring through sexual reproduction: diploid asexual worms, triploid asexual worms, diploid sexual worms and triploid sexual worms. The chromosomal mutation on Chromosome 4 was inherited by these offspring independent of their reproductive mode. These results provide two important pieces of information: (i) the putative genetic factor was not necessarily inherited in a Mendelian fashion; and (ii) the reproductive mode is not regulated by chromosomal changes such as polyploidy or chromosomal mutations. This suggests that asexuality in D. ryukyuensis is regulated by an unknown factor(s) other than a Mendelian factor or a chromosomal control. PMID:21392298

Kobayashi, Kazuya; Arioka, Sachiko; Hoshi, Motonori; Matsumoto, Midori

2009-09-01

183

Amputation induces stem cell mobilization to sites of injury during planarian regeneration.  

PubMed

How adult stem cell populations are recruited for tissue renewal and repair is a fundamental question of biology. Mobilization of stem cells out of their niches followed by correct migration and differentiation at a site of tissue turnover or injury are important requirements for proper tissue maintenance and regeneration. However, we understand little about the mechanisms that control this process, possibly because the best studied vertebrate adult stem cell systems are not readily amenable to in vivo observation. Furthermore, few clear examples of the recruitment of fully potent stem cells, compared with limited progenitors, are known. Here, we show that planarian stem cells directionally migrate to amputation sites during regeneration. We also show that during tissue homeostasis they are stationary. Our study not only uncovers the existence of specific recruitment mechanisms elicited by amputation, but also sets the stage for the systematic characterization of evolutionarily conserved stem cell regulatory processes likely to inform stem cell function and dysfunction in higher organisms, including humans. PMID:22899852

Guedelhoefer, Otto C; Sánchez Alvarado, Alejandro

2012-08-16

184

Gene expression of pluripotency determinants is conserved between mammalian and planarian stem cells.  

PubMed

Freshwater planaria possess extreme regeneration capabilities mediated by abundant, pluripotent stem cells (neoblasts) in adult animals. Although planaria emerged as an attractive in vivo model system for stem cell biology, gene expression in neoblasts has not been profiled comprehensively and it is unknown how molecular mechanisms for pluripotency in neoblasts relate to those in mammalian embryonic stem cells (ESCs). We purified neoblasts and quantified mRNA and protein expression by sequencing and shotgun proteomics. We identified ?4000 genes specifically expressed in neoblasts, including all ?30 known neoblast markers. Genes important for pluripotency in ESCs, including regulators as well as targets of OCT4, were well conserved and upregulated in neoblasts. We found conserved expression of epigenetic regulators and demonstrated their requirement for planarian regeneration by knockdown experiments. Post-transcriptional regulatory genes characteristic for germ cells were also enriched in neoblasts, suggesting the existence of a common ancestral state of germ cells and ESCs. We conclude that molecular determinants of pluripotency are conserved throughout evolution and that planaria are an informative model system for human stem cell biology. PMID:22543868

Onal, Pinar; Grün, Dominic; Adamidi, Catherine; Rybak, Agnieszka; Solana, Jordi; Mastrobuoni, Guido; Wang, Yongbo; Rahn, Hans-Peter; Chen, Wei; Kempa, Stefan; Ziebold, Ulrike; Rajewsky, Nikolaus

2012-04-27

185

Gene expression of pluripotency determinants is conserved between mammalian and planarian stem cells  

PubMed Central

Freshwater planaria possess extreme regeneration capabilities mediated by abundant, pluripotent stem cells (neoblasts) in adult animals. Although planaria emerged as an attractive in vivo model system for stem cell biology, gene expression in neoblasts has not been profiled comprehensively and it is unknown how molecular mechanisms for pluripotency in neoblasts relate to those in mammalian embryonic stem cells (ESCs). We purified neoblasts and quantified mRNA and protein expression by sequencing and shotgun proteomics. We identified ?4000 genes specifically expressed in neoblasts, including all ?30 known neoblast markers. Genes important for pluripotency in ESCs, including regulators as well as targets of OCT4, were well conserved and upregulated in neoblasts. We found conserved expression of epigenetic regulators and demonstrated their requirement for planarian regeneration by knockdown experiments. Post-transcriptional regulatory genes characteristic for germ cells were also enriched in neoblasts, suggesting the existence of a common ancestral state of germ cells and ESCs. We conclude that molecular determinants of pluripotency are conserved throughout evolution and that planaria are an informative model system for human stem cell biology.

Onal, Pinar; Grun, Dominic; Adamidi, Catherine; Rybak, Agnieszka; Solana, Jordi; Mastrobuoni, Guido; Wang, Yongbo; Rahn, Hans-Peter; Chen, Wei; Kempa, Stefan; Ziebold, Ulrike; Rajewsky, Nikolaus

2012-01-01

186

Characterization of novel genes expressed specifically in the sexual organs of the planarian Dugesia ryukyuensis.  

PubMed

The planarian Dugesia ryukyuensis reproduces both asexually (fissiparous) and sexually (oviparous) and can switch from the asexual mode to the sexual mode. By feeding with mature Bdellocephala brunnea oviparous worms, the fissiparous worms, which do not possess sexual organs, can be converted to fully sexualized worms in a process termed sexualization. As sexualization proceeds, the sexual organs are formed uniformly and five stages (stages 15) of the process have been identified histologically. In order to clarify the sexualization process, we attempted to isolate the genes expressed specifically at stage 5 by the differential display method. We isolated five genes expressed in the testis and two genes expressed in the yolk gland, which is an organ specific to sexualized worms. By BLAST search, one of the testis-specific genes was coded as testis-specific alpha-tubulin and two yolk gland-specific genes are similar to ribose-phosphate pyrophosphokinase I and F-box/SPRY-domain protein 1. Drs1, Drs2 and Drs3 were expressed in spermatocytes and spermatids from the early stage of spermatogenesis and Drs4 and Drs5 were expressed in spermatogonia, spermatocytes and spermatids. These genes are useful markers for elucidating the sexualization process. PMID:17554688

Hase, Sumitaka; Kashiwagi, Emiko; Kobayashi, Kazuya; Hoshi, Motonori; Matsumoto, Midori

2007-01-01

187

Combining life-history and toxicokinetic parameters to interpret differences in sensitivity to cadmium between populations of Polycelis tenuis (Platyhelminthes).  

PubMed

Five populations of the planarian Polycelis tenuis, collected from four locations in a metal contaminated stream and from one location in an unpolluted tributary stream, were compared for their sensitivity to cadmium by measuring survival, reproduction, and body size changes during 28 days of exposure via the water. Survival data and bioconcentration of cadmium measured at the end of the experiment enabled the use of a kinetic-based toxicity model, estimating the lethal body concentration, the uptake rate constant, the elimination rate constant, and the ultimate LC(50) as estimated toxicokinetic parameters. Accurate body size selection and a one-month acclimatization period to standardized laboratory conditions were applied to enhance the comparability between populations. An increased elimination rate constant and a greater ultimate LC(50) were estimated for one of the populations, located downstream from an ancient lead-zinc mine. The cadmium concentration causing an effect on reproduction varied considerably (a factor of 10) among populations but did not differ significantly between populations. For the number of reproducing individuals as a function of cadmium concentration, EC(50) and steepness of slope of the concentration/effect relationship tended to be correlated positively (P<0.10), indicating that selection in the field may have occurred. Cadmium was found to enhance shrinkage of P. tenuis under conditions of food limitation. PMID:10499984

Indeherberg, M B; van Straalen, N M; Schockaert, E R

1999-09-01

188

A LIM-homeobox gene is required for differentiation of Wnt-expressing cells at the posterior end of the planarian body.  

PubMed

Planarians have high regenerative ability, which is dependent on pluripotent adult somatic stem cells called neoblasts. Recently, canonical Wnt/?-catenin signaling was shown to be required for posterior specification, and Hedgehog signaling was shown to control anterior-posterior polarity via activation of the Djwnt1/P-1 gene at the posterior end of planarians. Thus, various signaling molecules play an important role in planarian stem cell regulation. However, the molecular mechanisms directly involved in stem cell differentiation have remained unclear. Here, we demonstrate that one of the planarian LIM-homeobox genes, Djislet, is required for the differentiation of Djwnt1/P-1-expressing cells from stem cells at the posterior end. RNA interference (RNAi)-treated planarians of Djislet [Djislet(RNAi)] show a tail-less phenotype. Thus, we speculated that Djislet might be involved in activation of the Wnt signaling pathway in the posterior blastema. When we carefully examined the expression pattern of Djwnt1/P-1 by quantitative real-time PCR during posterior regeneration, we found two phases of Djwnt1/P-1 expression: the first phase was detected in the differentiated cells in the old tissue in the early stage of regeneration and then a second phase was observed in the cells derived from stem cells in the posterior blastema. Interestingly, Djislet is expressed in stem cell-derived DjPiwiA- and Djwnt1/P-1-expressing cells, and Djislet(RNAi) only perturbed the second phase. Thus, we propose that Djislet might act to trigger the differentiation of cells expressing Djwnt1/P-1 from stem cells. PMID:21828095

Hayashi, Tetsutaro; Motoishi, Minako; Yazawa, Shigenobu; Itomi, Kazu; Tanegashima, Chiharu; Nishimura, Osamu; Agata, Kiyokazu; Tarui, Hiroshi

2011-09-01

189

Stem cells from innate sexual but not acquired sexual planarians have the capability to form a sexual individual.  

PubMed

Planarian species may harbor as many as three populations with different reproductive strategies. Animals from innate asexual (AS) and innate sexual (InS) populations reproduce only by fission and cross-fertilization, respectively, whereas the third population switches seasonally between the two reproductive modes. AS worms can be experimentally sexualized by feeding them with minced InS worms; we termed the resulting animals "acquired sexual" (AqS) worms. Both AqS and InS worms exhibit sexualizing activity when used as feed, suggesting that they maintain their sexual state via endogenous sexualizing substances, although the mechanisms underlying determination of reproductive strategy and sexual switching in these metazoans remain enigmatic. Therefore, we compared the endogenous sexualizing activity of InS worms and AqS worms. First, we amputated mature worms and assessed if they could re-enter a sexual state. Regenerants of InS worms, but not AqS worms, were only sexual, indicating that sexual state regulation comprises two steps: (1) autonomous initiation of sexualizing substance production and (2) maintenance of the sexual state by continuous production of sexualizing substances. Next, InS neoblasts were characterized by transplantation, finding that they successfully engrafted, proliferated, and replaced all recipient cells. Under such conditions, the AS recipients of InS worm neoblasts, but not those of AqS worms, became sexual. These results clearly show that there is a neoblast-autonomous determination of reproductive strategy in planarians. PMID:22968921

Nodono, Hanae; Ishino, Yugo; Hoshi, Motonori; Matsumoto, Midori

2012-09-25

190

Evolution of the major lineages of tapeworms (Platyhelminthes: Cestoidea) inferred from 18S ribosomal DNA and elongation factor-1alpha.  

PubMed

The interrelationships of the tapeworms (Platyhelminthes: Cestoidea) were inferred by analysis of complete gene sequences (approximately 2,200 bp) of 18S small subunit ribosomal DNA (18S) and partial gene sequences (approximately 900 bp) of elongation factor-1alpha (Ef-1alpha). New collections were made of 23 species representing each of the 14 currently recognized orders of tapeworms, including the Amphilinidea, Gyrocotylidea, and the 12 orders of the Eucestoda. Sequences were determined directly from polymerase chain reaction (PCR) products by either manual or automated methods. Nucleotide sequences of platyhelminth species outside of the Cestoidea were obtained for rooting the resulting trees. The 18S sequences were aligned with reference to the secondary structural features of the gene and the Ef-1alpha sequences were aligned with reference to their corresponding amino acid residues. Significant length variation among taxa was observed in the V2, V4, and V7 variable regions of the 18S gene. Such positions where sequences could not be aligned confidently were excluded from the analyses. Third codon positions of the Ef-1alpha gene were inferred to be saturated at an ordinal level of comparison. In addition, a short (approximately 35 bp) intron region of the Ef-1alpha gene was found to be shared only among the eucestode taxa, with the exception of Spathebothrium simplex (Spathebothriidea), which lacked the intron. Complete alignments showing structural features of the genes and sites excluded from analysis are provided as appendices. The sequence data were partitioned into 7 data sets in order to examine the effects of analyses on different subsets of the data. Analyses were conducted on the 2 genes independently, different codon positions of Ef-1alpha, amino acid sequences of Ef-1alpha, and combinations thereof. All subsets of the data were analyzed under the criterion of maximum parsimony as well as minimum evolution using both maximum-likelihood estimated, and LogDet-transformed distances. Results varied among the different data partitions and methods of analysis. Nodes with strong character support, however, were consistently recovered, and a general pattern of evolution was observed. Monophyly of the Cestoidea (Amphilinidea + Gyrocotylidea + Eucestoda) and Eucestoda and the traditionally accepted positions of the Amphilinidea and Gyrocotylidea as sister lineages to the Eucestoda were supported. Within the Eucestoda, the Spathebothriidea was found to be the sister of all other eucestodes. The remaining orders generally formed a diphyletic pattern of evolution consisting of separate difossate and tetrafossate lineages. This pattern was not universally observed among the analyses, primarily because the trypanorhynch and diphyllidean taxa showed instability in their phylogenetic position. Additional relationships that showed high levels of nodal support included a sister relationship between the Pseudophyllidea and Haplobothriidea and a clade uniting the Cyclophyllidea, Nippotaeniidea, and Tetrabothriidea. The Tetraphyllidea, as currently defined, was found to be paraphyletic without the inclusion of the orders Proteocephalidea and, possibly, Lecanicephalidea. Ordinal status of a monophyletic Litobothriidea, currently classified within the Tetraphyllidea, was found to be supported from a phylogenetic perspective. PMID:10647048

Olson, P D; Caira, J N

1999-12-01

191

Modeling planarian regeneration: a primer for reverse-engineering the worm.  

PubMed

A mechanistic understanding of robust self-assembly and repair capabilities of complex systems would have enormous implications for basic evolutionary developmental biology as well as for transformative applications in regenerative biomedicine and the engineering of highly fault-tolerant cybernetic systems. Molecular biologists are working to identify the pathways underlying the remarkable regenerative abilities of model species that perfectly regenerate limbs, brains, and other complex body parts. However, a profound disconnect remains between the deluge of high-resolution genetic and protein data on pathways required for regeneration, and the desired spatial, algorithmic models that show how self-monitoring and growth control arise from the synthesis of cellular activities. This barrier to progress in the understanding of morphogenetic controls may be breached by powerful techniques from the computational sciences-using non-traditional modeling approaches to reverse-engineer systems such as planaria: flatworms with a complex bodyplan and nervous system that are able to regenerate any body part after traumatic injury. Currently, the involvement of experts from outside of molecular genetics is hampered by the specialist literature of molecular developmental biology: impactful collaborations across such different fields require that review literature be available that presents the key functional capabilities of important biological model systems while abstracting away from the often irrelevant and confusing details of specific genes and proteins. To facilitate modeling efforts by computer scientists, physicists, engineers, and mathematicians, we present a different kind of review of planarian regeneration. Focusing on the main patterning properties of this system, we review what is known about the signal exchanges that occur during regenerative repair in planaria and the cellular mechanisms that are thought to underlie them. By establishing an engineering-like style for reviews of the molecular developmental biology of biomedically important model systems, significant fresh insights and quantitative computational models will be developed by new collaborations between biology and the information sciences. PMID:22570595

Lobo, Daniel; Beane, Wendy S; Levin, Michael

2012-04-26

192

Weak extremely-low-frequency magnetic field-induced regeneration anomalies in the planarian, Dugesia tigrina  

SciTech Connect

The authors recently reported that cephalic regeneration in the planarian Dugesia tigrina was significantly delayed in populations exposed continuously to combined parallel DC and AC magnetic fields. This effect was consistent with hypotheses suggesting an underlying resonance phenomenon. The authors report here, in a parallel series of investigations on the same model system, that the incidence of regeneration anomalies presenting as tumor-like protuberances also increases significantly (P < .001) in association with exposure to weak 60 Hz magnetic fields, with peak intensities ranging between 1.0 and 80.0 {micro}T. These anomalies often culminate in the complete disaggregation of the organism. Similar to regeneration rate effects, the incidence of regeneration anomalies is specifically dependent upon the planaria possessing a fixed orientation with respect to the applied magnetic field vectors. However, unlike the regeneration rate effects, the AC magnetic field alone, in the absence of any measurable DC field, is capable of producing these anomalies. Moreover, the incidence of regeneration anomalies follows a clear dose-response relationship as a function of AC magnetic field intensity, with the threshold for induced electric field intensity estimated at 5 {micro} V/m. The addition of either 51.1 or 78.4 {micro}T DC magnetic fields, applied in parallel combination with the AC field, enhances the appearance of anomalies relative to the 60 Hz AC field alone, but only at certain AC field intensities. Thus, whereas the previous study of regeneration rate effects appeared to involve exclusively resonance interactions, the regeneration anomalies reported here appear to result primarily from Faraday induction coupling.

Jenrow, K.A.; Smith, C.H.; Liboff, A.R. [Oakland Univ., Rochester, MI (United States). Dept. of Physics

1996-12-31

193

The Mystery of the Vanished Citations: James McConnell's Forgotten 1960s Quest for Planarian Learning, a Biochemical Engram, and Celebrity  

Microsoft Academic Search

During the 1960s, at a time of skepticism about the possibility of invertebrate learning, James McConnell and other researchers attracted to the glamour created by McConnell for planarian learning established invertebrate learning with a Pavlovian conditioning paradigm and a wide variety of control groups and procedures that are still used today in work on the biochemistry of learning and memory.

Mark Rilling

1996-01-01

194

Flumazenil-sensitive dose-related physical dependence in planarians produced by two benzodiazepine and one non-benzodiazepine benzodiazepine receptor agonists  

PubMed Central

Two benzodiazepine (midazolam and clorazepate) and one non-benzodiazepine (zolpidem) benzodiazepine-receptor agonists produced dose-related physical dependence, as evidenced by abstinence-induced decrease in planarian locomotor velocity (pLMV) when drug-exposed planarians were placed into drug-free water, but not when they were placed into drug-containing water (i.e., an abstinence-induced withdrawal, since the effect was only obtained in the removal of dug and not in the continued presence of drug). We have previously shown that the decrease in pLMV is associated with specific and transient withdrawal signs. In the present study, the selective benzodiazepine receptor antagonist flumazenil significantly antagonized (P < 0.05), by co-application, the ability of each agonist to produce the withdrawal. These results: (1) suggest that benzodiazepine receptor agonists, for two different chemical categories, produce dose-related physical dependence manifested as abstinence-induced withdrawal in this simple and convenient model, and (2) in the absence of cloning or radioligand binding literature, suggest a possible specific interaction site (receptor?) for these compounds in planarians.

Raffa, Robert B.; Cavallo, Federica; Capasso, Anna

2009-01-01

195

Transcription factors lhx1/5-1 and pitx are required for the maintenance and regeneration of serotonergic neurons in planarians.  

PubMed

In contrast to most adult organisms, freshwater planarians can regenerate any injured body part, including their entire nervous system. This allows for the analysis of genes required for both the maintenance and regeneration of specific neural subtypes. In addition, the loss of specific neural subtypes may uncover previously unknown behavioral roles for that neural population in the context of the adult animal. Here we show that two homeodomain transcription factor homologs, Smed-lhx1/5-1 and Smed-pitx, are required for the maintenance and regeneration of serotonergic neurons in planarians. When either lhx1/5-1 or pitx was knocked down by RNA interference, the expression of multiple canonical markers for serotonergic neurons was lost. Surprisingly, the loss of serotonergic function uncovered a role for these neurons in the coordination of motile cilia on the ventral epidermis of planarians that are required for their nonmuscular gliding locomotion. Finally, we show that in addition to its requirement in serotonergic neurons, Smed-pitx is required for proper midline patterning during regeneration, when it is required for the expression of the midline-organizing molecules Smed-slit in the anterior and Smed-wnt1 in the posterior. PMID:23903188

Currie, Ko W; Pearson, Bret J

2013-07-31

196

Platyhelminths as paleogeographical indicators  

Microsoft Academic Search

Turbellarians do not feature as examples in the present discussions on the theory and method of analytical biogeography. It is argued, however, that turbellarian distributional records form good examples of large-scale biogeographic patterns resulting from continental breakup. Some turbellarian taxa also indicate biogeographic links across the Pacific Ocean, which can be visualized readily by means of track construction. Amphi-pacific organismal

Ronald Sluys

1995-01-01

197

Toxicity of selenium (Na2SeO3) and mercury (HgCl2) on the planarian Dugesia gonocephala.  

PubMed

The toxicity of selenium (Na2SeO3) and mercury (HgCl2) was determined by using a freshwater planarian which is particularly sensitive to pollution, and belongs to a fissiparous breed of Dugesia gonocephala. The mortality and fissiparity frequency of the subjects were studied. They were exposed to intense treatments (48 hours) or for medium to long periods of time (21 days) to either the single compounds or a combination of both, and were fed or fasting. The lethal effect of sodium selenite is correlated to the food intake, whereas the toxicity of mercurous chloride is probably the result of a fixative effect which does not depend on feeding. The 21-day treatment with the first compound has a non-negligible lethal effect which is probably due to an accumulation phenomenon. At doses where an antioxidant effect prevails, fissiparity is stimulated. On the other hand, the second compound reduces reproduction frequency to half the base values. Compared to the Paracentrotus lividus, the Dugesia gonocephala offers various advantages concerning toxicological experiments; besides being easier to handle in the laboratory, it is available all year round and is not subject to seasonal cycles. It is also more susceptible to the toxic effect of mercury, which is a common and highly toxic pollutant, than the sea urchin. PMID:2616901

Congiu, A M; Casu, S; Ugazio, G

1989-10-01

198

Substitution saturation and nuclear paralogs of commonly employed phylogenetic markers in the Caryophyllidea, an unusual group of non-segmented tapeworms (Platyhelminthes).  

PubMed

Caryophyllidean cestodes (Platyhelminthes) represent an unusual group of tapeworms lacking serially repeated body parts that potentially diverged from the common ancestor of the Eucestoda prior to the evolution of segmentation. Here we evaluate the utility of two nuclear and two mitochondrial molecular markers (ssrDNA and lsrDNA, nad3 and cox1) for use in circumscribing generic boundaries and estimating interrelationships in the group. We show that these commonly employed markers do not contain sufficient signal to infer well-supported phylogenetic estimates due to substitution saturation. Moreover, we detected multiple trnK+nad3+trnS+trnW+cox1 haplotypes within individuals, indicating a history of gene exchange between the mitochondrial and nuclear genomes. The presence of such nuclear paralogs (i.e. numts), to our knowledge described here in cestodes for the first time, together with the results of phylogenetic, saturation and split-decomposition analyses all suggest that finding informative markers for estimating caryophyllidean evolution is unusually problematic in comparison to other major lineages of tapeworms. PMID:22366732

Brabec, Jan; Scholz, Tomáš; Králová-Hromadová, Ivica; Bazsalovicsová, Eva; Olson, Peter D

2012-02-16

199

Platyhelminth mitochondrial DNA: Evidence for early evolutionary origin of a tRNA ser AGN that contains a dihydrouridine arm replacement loop, and of serine-specifying AGA and AGG codons  

Microsoft Academic Search

Summary  The nucleotide sequence of a segment of the mitochondrial DNA (mtDNA) molecule of the liver flukeFasciola hepatica (phylum Platyhelminthes, class Trematoda) has been determined, within which have been identified the genes for tRNAala, tRNAasp, respiratory chain NADH dehydrogenase subunit I (ND1), tRNAasn, tRNApro, tRNAile, tRNAlys, ND3, tRNAserAGN, tRNAtrp, and cytochromec oxidase subunit I (COI). The 11 genes are arranged in

James R. Garey; David R. Wolstenholme

1989-01-01

200

Expression patterns of Xenopus FGF receptor-like 1/nou-darake in early Xenopus development resemble those of planarian nou-darake and Xenopus FGF8.  

PubMed

Fibroblast growth factors (FGFs) mediate many cell-to-cell signaling events during early development. Nou-darake (ndk), a gene encoding an FGF receptor (FGFR)-like molecule, was found to be highly and specifically expressed in the head region of the planarian Dugesia japonica, and its functional analyses provided strong molecular evidence for the existence of a brain-inducing circuit based on the FGF signaling pathway. To analyze the role of ndk during vertebrate development, we isolated the Xenopus ortholog of ndk, the vertebrate FGFR-like 1 gene (XFGFRL1). Expression of XFGFRL1/Xndk was first detected in the anterior region at the late gastrula stage and dramatically increased at the early neurula stage in an overall anterior mesendodermal region, including the prechordal plate, paraxial mesoderm, anterior endoderm, and archenteron roof. This anterior expression pattern resembles that of ndk in planarians, suggesting that the expression of FGFRL1/ndk is conserved in evolution between these two distantly diverged organisms. During the tail bud stages, XFGFRL1/Xndk expression was detected in multiple regions, including the forebrain, eyes, midbrain-hindbrain boundary, otic vesicles, visceral arches, and somites. In many of these regions, XFGFRL1/Xndk was coexpressed with XFGF8, indicating that XFGFRL1/Xndk is a member of the XFGF8 synexpression group, which includes sprouty, sef, and isthmin. PMID:15254904

Hayashi, Shuichi; Itoh, Mari; Taira, Sumiko; Agata, Kiyokazu; Taira, Masanori

2004-08-01

201

The PIWI proteins SMEDWI-2 and SMEDWI-3 are required for stem cell function and piRNA expression in planarians  

PubMed Central

PIWI proteins are expressed in germ cells in a wide variety of metazoans, where they participate in the synthesis and function of a novel class of small RNAs called PIWI associated RNAs (piRNAs). One function of piRNAs is to preserve the integrity of the germline genome by silencing transposons, though they also participate in epigenetic and post-transcriptional gene regulation. In the planarian Schmidtea mediterranea, the PIWI proteins SMEDWI-1 and SMEDWI-2 are expressed in neoblasts and SMEDWI-2 is required for regeneration and homeostasis. Here, we identify a diverse population of ?32-nucleotide small RNAs that strongly resemble vertebrate and insect piRNAs and map to hundreds of thousands of loci in the S. mediterranea genome. The expression of these RNAs occurs predominantly in neoblasts and is not restricted to the germline. RNAi knockdown of either SMEDWI-2 or a newly identified PIWI protein, SMEDWI-3, impairs regeneration and homeostasis and decreases the levels of both piRNAs and neoblasts. Therefore, SMEDWI-2 and SMEDWI-3 are required for piRNA expression, regeneration, and neoblast function in S. mediterranea.

Palakodeti, Dasaradhi; Smielewska, Magda; Lu, Yi-Chien; Yeo, Gene W.; Graveley, Brenton R.

2008-01-01

202

Developmental diversity in free-living flatworms  

PubMed Central

Flatworm embryology has attracted attention since the early beginnings of comparative evolutionary biology. Considered for a long time the most basal bilaterians, the Platyhelminthes (excluding Acoelomorpha) are now robustly placed within the Spiralia. Despite having lost their relevance to explain the transition from radially to bilaterally symmetrical animals, the study of flatworm embryology is still of great importance to understand the diversification of bilaterians and of developmental mechanisms. Flatworms are acoelomate organisms generally with a simple centralized nervous system, a blind gut, and lacking a circulatory organ, a skeleton and a respiratory system other than the epidermis. Regeneration and asexual reproduction, based on a totipotent neoblast stem cell system, are broadly present among different groups of flatworms. While some more basally branching groups - such as polyclad flatworms - retain the ancestral quartet spiral cleavage pattern, most flatworms have significantly diverged from this pattern and exhibit unique strategies to specify the common adult body plan. Most free-living flatworms (i.e. Platyhelminthes excluding the parasitic Neodermata) are directly developing, whereas in polyclads, also indirect developers with an intermediate free-living larval stage and subsequent metamorphosis are found. A comparative study of developmental diversity may help understanding major questions in evolutionary biology, such as the evolution of cleavage patterns, gastrulation and axial specification, the evolution of larval types, and the diversification and specialization of organ systems. In this review, we present a thorough overview of the embryonic development of the different groups of free-living (turbellarian) platyhelminths, including the Catenulida, Macrostomorpha, Polycladida, Lecithoepitheliata, Proseriata, Bothrioplanida, Rhabdocoela, Fecampiida, Prolecithophora and Tricladida, and discuss their main features under a consensus phylogeny of the phylum.

2012-01-01

203

Stem cells and the Planarian Schmidtea mediterranea  

Microsoft Academic Search

In recent years, stem cells have been heralded as potential therapeutic agents to address a large number of degenerative diseases. Yet, in order to rationally utilize these cells as effective therapeutic agents, and\\/or improve treatment of stem-cell-associated malignancies such as leukemias and carcinomas, a better understanding of the basic biological properties of stem cells needs to be acquired. A major

Alejandro Sánchez Alvarado

2007-01-01

204

Stem cell systems and regeneration in planaria.  

PubMed

Planarians are members of the Platyhelminthes (flatworms). These animals have evolved a remarkable stem cell system. A single pluripotent adult stem cell type ("neoblast") gives rise to the entire range of cell types and organs in the planarian body plan, including a brain, digestive-, excretory-, sensory- and reproductive systems. Neoblasts are abundantly present throughout the mesenchyme and divide continuously. The resulting stream of progenitors and turnover of differentiated cells drive the rapid self-renewal of the entire animal within a matter of weeks. Planarians grow and literally de-grow ("shrink") by the food supply-dependent adjustment of organismal turnover rates, scaling body plan proportions over as much as a 50-fold size range. Their dynamic body architecture further allows astonishing regenerative abilities, including the regeneration of complete and perfectly proportioned animals even from tiny tissue remnants. Planarians as an experimental system, therefore, provide unique opportunities for addressing a spectrum of current problems in stem cell research, including the evolutionary conservation of pluripotency, the dynamic organization of differentiation lineages and the mechanisms underlying organismal stem cell homeostasis. The first part of this review focuses on the molecular biology of neoblasts as pluripotent stem cells. The second part examines the fascinating mechanistic and conceptual challenges posed by a stem cell system that epitomizes a universal design principle of biological systems: the dynamic steady state. PMID:23138344

Rink, Jochen C

2012-11-09

205

Ultrastructural aspects of the germovitellarium of two prorhynchids (Platyhelminthes, Lecithoepitheliata)  

Microsoft Academic Search

The female gonad of two fresh-water prorhynchids, Geocentrophora baltica and Prorhynchus stagnalis, has been investigated by means of conventional electron microscopy and cytochemical techniques. Both species have an unpaired germovitellarium located under the gut; accessory cells surround the germovitellarium of G. baltica. The germovitellarium consists of a restricted germinative area where early differentiating oocytes and vitellocytes are randomly associated, and

ALESSANDRA FALLENI

1997-01-01

206

Hox genes from the tapeworm Taenia asiatica (Platyhelminthes: Cestoda).  

PubMed

Hox genes are important in forming the anterior-posterior body axis pattern in the early developmental stage of animals. The conserved nature of the genomic organization of Hox genes is well known in diverse metazoans. To understand the Hox gene architecture in human-infecting Taenia tapeworms, we conducted a genomic survey of the Hox gene using degenerative polymerase chain reaction primers in Taenia asiatica. Six Hox gene orthologs from 276 clones were identified. Comparative analysis revealed that T. asiatica has six Hox orthologs, including two lab/Hox1, two Hox3, one Dfd/Hox4, and one Lox2/Lox4. The results suggest that Taenia Hox genes may have undergone independent gene duplication in two Hox paralogs. The failure to detect Post1/2 orthologs in T. asiatica may suggest that sequence divergence or the secondary loss of the posterior genes has occurred in the lineage leading to the cestode and trematode. PMID:17265186

Kim, Kyu-Heon; Lee, Yong Seok; Jeon, Hyeong-Kyu; Park, Joong-Ki; Kim, Chang-Bae; Eom, Keeseon S

2007-04-01

207

Ca(2+) channels and praziquantel: A view from the free world.  

PubMed

Targeting the cellular Ca(2+) channels and pumps that underpin parasite Ca(2+) homeostasis may realize novel antihelmintic agents. Indeed, the antischistosomal drug praziquantel (PZQ) is a key clinical agent that has been proposed to work in this manner. Heterologous expression data has implicated an action of PZQ on voltage-operated Ca(2+) channels, although the relevant in vivo target of this drug has remained undefined over three decades of clinical use. The purpose of this review is to bring new perspective to this issue by discussing the potential utility of free-living planarian flatworms for providing new insight into the mechanism of PZQ action. First, we discuss in vivo functional genetic data from the planarian system that broadly supports the molecular data collected in heterologous systems and the 'Ca(2+) hypothesis' of PZQ action. On the basis of these similarities we highlight our current knowledge of platyhelminth voltage operated Ca(2+) channels, their unique molecular pharmacology and the downstream functional PZQ interactome engaged by dysregulation of Ca(2+) influx that has potential to yield novel antischistosomal targets. Overall the broad dataset underscores a common theme of PZQ-evoked disruptions of Ca(2+) homeostasis in trematodes, cestodes and turbellarians, and showcases the utility of the planarian model for deriving insight into drug action and targets in parasitic flatworms. PMID:23246536

Chan, John D; Zarowiecki, Magdalena; Marchant, Jonathan S

2012-12-16

208

Redescripción y algunos aspectos ecológicos de Girardia tigrina, G. cameliae y G. paramensis (Dugesiidae, Tricladida) en Antioquia, Colombia Redescription and some ecological aspects of Girardia tigrina, G. cameliae y G. paramensis (Dugesiidae, Tricladida) in Antioquia, Colombia  

Microsoft Academic Search

Palabras clave: taxonomía, ecología, Turbellaria, planarias, Neotropico. Abstract. The Turbellaria comprises 2 orders: Catenulida and Rhabditophora. In the latter, the suborder Paludicola contains the families Dendrocoelidae, Dugesiidae and Planariidae. In this study, we found Girardia cameliae, Girardia paramensis and Giardia tigrina (Dugesiidae) in 22 aquatic systems using manual capture in river ponds zones of central and southeastern Antioquia. In 3

Mauricio A. Muńoz; Imelda Vélez

2007-01-01

209

The cell biology of schistosomes: a window on the evolution of the early metazoa.  

PubMed

This review of schistosome cell biology has a dual purpose; its intent is to alert two separate research communities to the activities of the other. Schistosomes are by far and away the best-characterised platyhelminths, due to their medical and economic importance, but seem to be almost totally ignored by researchers on the free-living lower metazoans. Equally, in their enthusiasm for the parasitic way of life, schistosome researchers seldom pay attention to the work on free-living animals that could inform their molecular investigations. The publication of transcriptomes and/or genomes for Schistosoma mansoni and Schistosoma japonicum, the sponge Archimedon, the cnidarians Nematostella and Hydra and the planarian Schmidtea provide the raw material for comparisons. Apart from interrogation of the databases for molecular similarities, there have been differences in technical approach to these lower metazoans; widespread application of whole mount in situ hybridisation to Schmidtea contrasts with the application of targeted proteomics to schistosomes. Using schistosome cell biology as the template, the key topics of cell adhesion, development, signalling pathways, nerve and muscle, and epithelia, are reviewed, where possible interspersing comparisons with the sponge, cnidarian and planarian data. The biggest jump in the evolution of cellular capabilities appears to be in the transition from a diploblast to triploblast level of organisation associated with development of a mobile and plastic body form. PMID:21976269

Wilson, R Alan

2012-07-01

210

Insight into the role of cetaceans in the life cycle of the tetraphyllideans (Platyhelminthes: Cestoda)  

Microsoft Academic Search

Four types of tetraphyllidean larvae infect cetaceans worldwide: two plerocercoids differing in size, ‘small’ (SP) and ‘large’ (LP), and two merocercoids referred to as Phyllobothrium delphini and Monorygma grimaldii. The latter merocercoid larvae parasitize marine mammals exclusively and exhibit a specialised cystic structure. Adult stages are unknown for any of the larvae and thus the role of cetaceans in the

F. J. Aznar; C. Agustí; D. T. J. Littlewood; J. A. Raga; P. D. Olson

2007-01-01

211

Dynamics of the platyhelminth fauna of wood ducks in relation to food habits and reproductive state.  

PubMed

The dynamics of the intestinal helminth fauna of wood ducks (Aix sponsa) and the relationship between changes in food habits and helminth populations during the bird's reproductive cycle were studied in southeast Missouri. A total of 11 species of helminths, comprised of four species of trematodes and seven species of cestodes, were recovered from the digestive tracts of 155 wood ducks. All species except one were found in both sexes. Significant differences were found in parasite numbers by season (spring vs. fall), sex, and stage of the reproductive cycle. Helminth infection in both sexes was higher in the spring than during fall courtship; however, the magnitude of the seasonal difference was considerably greater in females. Females contained significantly more helminths than males during spring, but no sex-related differences were detected in the fall. Seasonal changes and sex-related differences in parasite numbers corresponded closely with the consumption of invertebrates that serve as intermediate hosts. The average number of parasites in females was similar during fall courtship and pre-egg-laying, suggesting that little or no new infection occurred during winter. The most intensive infections were found in laying females and were attributed to hyperphagia and increased invertebrate consumption during egg production. A large decrease in the parasite numbers in females between incubation and fall courtship indicated that most of the helminths acquired by laying females were lost during summer; therefore, the long-term effects of the increased infection are probably negligible. PMID:6854477

Drobney, R D; Train, C T; Fredrickson, L H

1983-04-01

212

First report of Gyrodactylus spp. (Platyhelminthes: Monogenea) in the western Mediterranean Sea: molecular and morphological descriptions.  

PubMed

The Gyrodactylus spp. fauna on species of gobies, Pomatoschistus, Gobiusculus, and Knipowitschia (Gobiidae: Teleostei), from the western Mediterranean and Adriatic seas is strikingly similar to that found in the Baltic Sea and eastern Atlantic Ocean, both in morphology and in internal transcribed spacer (ITS) rRNA. The fauna consisted of Gyrodactylus branchialis, G. ostendicus, G. gondae, G. rugiensis, G. rugiensoides, and G. arcuatus. No new species have been found. A morphometric comparison between G. branchialis from the Mediterranean and Adriatic seas and its type locality in Ostend (Belgium) showed significant differences in ventral bar and marginal hook features. The morphometric variation was lower in G. rugiensis, whereas no significant differences were found in G. ostendicus. Gyrodactylus branchialis and G. ostendicus collected on P. microps were slightly different in the ITS rDNA (-0.6%) compared with specimens on the closely related P. marmoratus, probably reflecting ongoing speciation. A hybrid zone was identified in the Vaccarčs lagoon complex (France) where both host species are sympatric. There was no clear geographic or host-related pattern in the variation found in the ITS2 rDNA in G. arcuatus sampled from P. microps, G. flavescens, Pungitius pungitius, K. panizzae, and its original host Gasterosteus aculeatus (2 polymorphic sites). Of all studied species, only G. arcuatus, G. rugiensis, and G. rugiensoides showed minor intraspecific variation in the ITS rDNA. Hence, the physical separation by a shoreline of more than 10,000 km is hardly reflected in the parasite ITS rDNA. PMID:16995382

Huyse, T; Pampoulie, C; Audenaert, V; Volckaert, F A M

2006-08-01

213

A molecular approach for the identification of meiofaunal turbellarians (Platyhelminthes, Turbellaria)  

Microsoft Academic Search

In the absence of reliable morphological characters, or in conjunction with morphology-based identifications, meiofaunal turbellarians may also be identified using the nucleotide sequence of a portion of the large subunit of the ribosomal RNA (26\\/28S rRNA). A 284 base pair-long region of the 26\\/28S rRNA has been identified by isolating genomic DNA from ten species of turbellarians belonging to four

M. K. Litvaitis; G. Nunn; W. K. Thomas; T. D. Kocher

1994-01-01

214

Intracytoplasmic ciliary elements in epidermal cells of Syndesmis echinorum and Paravortex cardii (Platyhelminthes, Dalyellioida).  

PubMed

Epidermal cells of Syndesmis echinorum and Paravortex cardii contain many intracytoplasmic ciliary components: clusters of centrioles disorganized and incomplete short axonemes composed of loosely organized microtubules of irregular lengths, fully formed axonemes though some with fewer than nine doublets, and ciliary rootlets. Furthermore, conspicuous dense granules are found in solitary groups in the cytoplasm. Clusters of dense granules are also closely associated with Golgi complexes and developing axonemal microtubules. Since the dense granules decrease in number as the axonemes increase, it is likely that the granules are involved in the formation of axonemal microtubules. Ciliary elements are especially abundant in epidermal cells of Paravortex cardii embryos, some of them resembling those previously described by several authors in differentiating ciliated cells engaged in centriologenesis and ciliogenesis. Attention has been focused on the relative proportion and position of these elements, as well as the different morphology and several assembling states that they exhibit in epidermal cells of adult S. echinorum and adults and embryos of P. cardii. A functional interpretation of some of the findings is given, which allows us to suggest a sequence of ciliogenetic events that occur in epidermal cells of both species. PMID:1518068

Cifrian, B; García-Corrales, P; Martínez-Alos, S

1992-08-01

215

Bucephalidae (Platyhelminthes: Digenea) of Plectropomus (Serranidae: Epinephelinae) in the tropical Pacific.  

PubMed

We examined four species of Plectropomus Oken, 1817 (Serranidae: Epinephelinae), Plectropomus areolatus (Rüppell), Plectropomus laevis (Lacepčde), Plectropomus leopardus (Lacepčde) and Plectropomus maculatus (Bloch) from sites off Heron Island and Lizard Island on the Great Barrier Reef, Australia (GBR), and the Gambier Islands, French Polynesia. Three new species of Neidhartia Nagaty, 1937, five new species of Prosorhynchus Odhner, 1905, and one previously described species, Prosorhynchus freitasi Nagaty, 1937, are characterised. The three species of Neidhartia, Neidhartia haywardi n. sp., Neidhartia plectropomi n. sp. and Neidhartia tyleri n. sp. are readily distinguishable morphologically. Two of the six species of Prosorhynchus (Prosorhynchus lesteri n. sp. and Prosorhynchus wrightae n. sp.) are easily distinguished from their other congeners by morphology but the other four species (P. freitasi, Prosorhynchus heronensis n. sp., Prosorhynchus munozae n. sp. and Prosorhynchus plectropomi n. sp.) are generally similar in morphology and were only distinguished initially by comparing their ITS2 rRNA sequences. Three additional taxa, one from the GBR and two from French Polynesia, were recognised as distinct on the basis that their ITS2 rRNA sequences differed from those of the new taxa described here; these species remain unnamed for the present. Inter-specific divergence observed within these genera in the ITS2 rRNA ranged from 10 to 42 base pairs (4-16 %) for species of Neidhartia and 2-57 base pairs (3-21 %) for species of Prosorhynchus. Inter-generic divergences were 42-55 base pairs (17-21 %). No intraspecific variation in the ITS2 rRNA region was observed for any of the six species for which multiple sequence replicates were obtained. Phylogenetic analysis of 12 operational taxa from Plectropomus together with sequences of three other species from epinepheline serranids demonstrated that Neidhartia and Prosorhynchus were reciprocally monophyletic with the exception that P. wrightae n. sp. fell either within or basal to the Neidhartia species. The richness of bucephalids in species of Plectropomus appears to be exceptional within the Serranidae relative to that observed in other serranid genera in the tropical Indo-West Pacific. PMID:23728730

Bott, Nathan J; Miller, Terrence L; Cribb, Thomas H

2013-06-01

216

Phylogeny of the Polystomatidae (Platyhelminthes, Monogenea), with particular reference to Polystoma integerrimum  

Microsoft Academic Search

Polystome phylogeny is examined, with emphasis on the dimorphism of the frog parasite Polystoma integerrimum, which exists in a fully differentiated and a neotenic form, and the evolutionary development of exclusively neotenic genera. Protopolystoma, which infects the aquatic toad Xenopus, has essentially the same morphology as the neotenic (branchial) adult of P. integerrimum and is interpreted as a neotenic genus;

J. B. Williams

1995-01-01

217

Phylogenetic relationships within the polyopisthocotylean monogeneans (Platyhelminthes) inferred from partial 28S rDNA sequencesq  

Microsoft Academic Search

Recent studies based on molecular data (18S rDNA and partial 28S rDNA) and morphology did not resolve a terminal polytomy within the Polyopisthocotylea. Here, we have used sequences from the full domain D2 of the 28S rDNA for 24 species (18 new sequences) with three phylogenetic methods, maximum parsimony, neighbour-joining and maximum likelihood, to infer the relationships among the Polyopistho-

Richard Jovelin; Jean-Lou Justine

218

Mystery tubes coiled around deep-water tropical gorgonians: fecampiid cocoons (Platyhelminthes: Fecampiida) resembling Solenogastres (Mollusca)  

Microsoft Academic Search

During the examination of a large suite of tropical deep-water molluscs, a number of Solenogastres were found, some of them typically curled around gorgonian stems. A subsequent closer examination of the Solenogastres revealed another type of object also curled around the gor- gonians, which strongly resembled Solenogastres but lacked their external features. These objects proved to be cocoons with egg

Claudia Handl; Philippe Bouchet

2007-01-01

219

Ultrastructural and cytochemical studies of the female gonad of Prorhynchus sp. (Platyhelminthes, Lecithoepitheliata)  

Microsoft Academic Search

The female gonad of Prorhynchus is heterocellular (neoophoran organization) and consists of an unpaired, elongate germovitellarium enveloped by a finely granular extracellular lamina. It is composed of a posterior germinative area where early oocytes are randomly associated with differentiating vitellocytes and a growth area with follicular organization. In each follicle a single oocyte is surrounded by a layer of vitellocytes.

Alessandra Falleni; Paolo Lucchesi; Vittorio Gremigni

1995-01-01

220

A new species of Gieysztoria (Platyhelminthes; Rhabdocoela; Dalyelliidae) from a freshwater lake in Queensland, Australia  

Microsoft Academic Search

A new species of Dalyelliidae, Gieysztoria queenslandica, is described from a freshwater lake in Brisbane, Queensland, Australia. Gieysztoria queenslandica n. sp. is a member of the Aequales group of Gieysztoria and differs from its congeners by possession of an S-shaped ovary, a Y-shaped oviduct leading to a separate receptaculum seminis, and the shape and size of the male copulatory organ.

RICK HOCHBERG; LESTER R. G. CANNON

221

A multienzyme network functions in intestinal protein digestion by a platyhelminth parasite.  

PubMed

Proteases frequently function not only as individual enzymes but also in cascades or networks. A notable evolutionary switch occurred in one such protease network that is involved in protein digestion in the intestine. In vertebrates, this is largely the work of trypsin family serine proteases, whereas in invertebrates, cysteine proteases of the papain family and aspartic proteases assume the role. Utilizing a combination of protease class-specific inhibitors and RNA interference, we deconvoluted such a network of major endopeptidases functioning in invertebrate intestinal protein digestion, using the parasitic helminth, Schistosoma mansoni as an experimental model. We show that initial degradation of host blood proteins is ordered, occasionally redundant, and substrate-specific. Although inhibition of parasite cathepsin D had a greater effect on primary cleavage of hemoglobin, inhibition of cathepsin B predominated in albumin degradation. Nevertheless, in both cases, inhibitor combinations were synergistic. An asparaginyl endopeptidase (legumain) also synergized with cathepsin B and L in protein digestion, either by zymogen activation or facilitating substrate cleavage. This protease network operates optimally in acidic pH compartments either in the gut lumen or in vacuoles of the intestinal lining cells. Defining the role of each of these major enzymes now provides a clearer understanding of the function of a complex protease network that is conserved throughout invertebrate evolution. It also provides insights into which of these proteases are logical targets for development of chemotherapy for schistosomiasis, a major global health problem. PMID:17028179

Delcroix, Melaine; Sajid, Mohammed; Caffrey, Conor R; Lim, Kee-C; Dvorák, Jan; Hsieh, Ivy; Bahgat, Mahmoud; Dissous, Colette; McKerrow, James H

2006-10-06

222

Spermatozoa of tapeworms (Platyhelminthes, Eucestoda): advances in ultrastructural and phylogenetic studies.  

PubMed

New data on spermiogenesis and the ultrastructure of spermatozoa of 'true' tapeworms (Eucestoda) are summarized. Since 2001, more than 50 species belonging to most orders of the Eucestoda have been studied or reinvestigated, particularly members of the Caryophyllidea, Spathebothriidea, Diphyllobothriidea, Bothriocephalidea, Trypanorhyncha, Tetraphyllidea, Proteocephalidea, and Cyclophyllidea. A new classification of spermatozoa of eucestodes into seven basic types is proposed and a key to their identification is given. For the first time, a phylogenetic tree inferred from spermatological characters is provided. New information obtained in the last decade has made it possible to fill numerous gaps in the character data matrix, enabling us to carry out a more reliable analysis of the evolution of ultrastructural characters of sperm and spermiogenesis in eucestodes. The tree is broadly congruent with those based on morphological and molecular data, indicating that convergent evolution of sperm characters in cestodes may not be as common as in other invertebrate taxa. The main gaps in the current knowledge of spermatological characters are mapped and topics for future research are outlined, with special emphasis on those characters that might provide additional information about the evolution of tapeworms and their spermatozoa. Future studies should be focused on representatives of those major groups (families and orders) in which molecular data indicate paraphyly or polyphyly (e.g. 'Tetraphyllidea' and Trypanorhyncha) and on those that have a key phylogenetic position among eucestodes (e.g. Diphyllidea, 'Tetraphyllidea', Lecanicephalidea, Nippotaeniidea). PMID:20015312

Levron, Céline; Miquel, Jordi; Oros, Mikulás; Scholz, Tomás

2010-12-15

223

Insight into the role of cetaceans in the life cycle of the tetraphyllideans (Platyhelminthes: Cestoda).  

PubMed

Four types of tetraphyllidean larvae infect cetaceans worldwide: two plerocercoids differing in size, 'small' (SP) and 'large' (LP), and two merocercoids referred to as Phyllobothrium delphini and Monorygma grimaldii. The latter merocercoid larvae parasitize marine mammals exclusively and exhibit a specialised cystic structure. Adult stages are unknown for any of the larvae and thus the role of cetaceans in the life cycle of these species has been a long-standing problem. The SP and LP forms are thought to be earlier stages of P. delphini and M. grimaldii that are presumed to infect large pelagic sharks that feed on cetaceans. A molecular analysis of the D2 variable region of the large subunit ribosomal DNA gene based on several individuals of each larval type collected from three Mediterranean species of cetaceans showed consistent and unique molecular signatures for each type regardless of host species or site of infection. The degree of divergence suggested that LP, P. delphini and M. grimaldii larvae may represent separate species, whereas SP may be conspecific with M. grimaldii. In all host species, individuals of SP accumulated in the gut areas in which the lymphoid tissue was especially developed. We suggest therefore that these larvae use the lymphatic system to migrate to the abdominal peritoneum and mesenteries where they develop into forms recognizable as M. grimaldii. The plerocercoid stage of P. delphini remains unknown. In a partial phylogenetic tree of the Tetraphyllidea, all larvae formed a clade that included a representative of the genus Clistobothrium, some species of which parasitize sharks such as the great white which is known to feed on cetaceans. A bibliographic examination of tetraphyllidean infections in marine mammals indicated that these larvae are acquired mostly offshore. In summary, the evidence suggests that cetaceans play a significant role in the life cycle of these larvae. In addition, it seems clear that cetaceans act as natural intermediate hosts for P. delphini and M. grimaldii, as within these hosts they undergo development from the plerocercoid stage to the merocercoid stage. Because tetraphyllidean species use fish, cephalopods and other marine invertebrates as intermediate hosts, the inclusion of cetaceans in the life cycle would have facilitated their transmission to apex predators such as the large, lamnid sharks. The biological significance of infections of LP in cetaceans is unclear, but infections do not seem to be accidental as such larvae show high prevalence and abundance as well as a high degree of site specificity, particularly in the anal crypts and bile ducts. PMID:17161403

Aznar, F J; Agustí, C; Littlewood, D T J; Raga, J A; Olson, P D

2006-11-27

224

Twelve new species of dactylogyrids (Platyhelminthes, Monogenea) from West African barbels (Teleostei, Cyprinidae), with some biogeographical  

Microsoft Academic Search

Fifteen species of dactylogyrid monogeneans, belong to Dactylogyms and Dogielius were observed in seven different African species of Barbus and Varicorhiizus (Teleostei, Cyprinidae). The barbels examined in West Africa were: Barbus occideritalis Boulenger, 1911, known in the large Sahel-Sudan rivers and in Gabon; B. waldroni Nordman, 1935, B. petitjearii Daget. 1962, B. sacratus Daget, 1963, B. parawaldroni Lévęque, Thys van

Jean-Françoi Guégan; Alain Lambert

1990-01-01

225

Sequence and secondary structure variation in the Gyrodactylus (Platyhelminthes: Monogenea) ribosomal RNA gene array.  

PubMed

Nucleotide sequences were determined for the rRNA internal transcribed spacers 1 and 2 (ITS1 and 2) and the 5' terminus of the large subunit rRNA in selected Gyrodactylus species. Examination of primary sequence variation and secondary structure models in ITS2 and variable region V4 of the small subunit rRNA revealed that structure was largely conserved despite significant variation in sequence. ITS1 sequences were highly variable, and models of structure were unreliable but, despite this, show some resemblance to structures predicted in Digenea. ITS2 models demonstrated binding of the 3' end of 5.8S rRNA to the 5' end of the large subunit rRNA and enabled the termini of these genes to be defined with greater confidence than previously. The structure model shown here may prove useful in future phylogenetic analyses. PMID:10864256

Cunningham, C O; Aliesky, H; Collins, C M

2000-06-01

226

Wnt gene loss in flatworms.  

PubMed

Wnt genes encode secreted glycoproteins that act in cell-cell signalling to regulate a wide array of developmental processes, ranging from cellular differentiation to axial patterning. Discovery that canonical Wnt/?-catenin signalling is responsible for regulating head/tail specification in planarian regeneration has recently highlighted their importance in flatworm (phylum Platyhelminthes) development, but examination of their roles in the complex development of the diverse parasitic groups has yet to be conducted. Here, we characterise Wnt genes in the model tapeworm Hymenolepis microstoma and mine genomic resources of free-living and parasitic species for the presence of Wnts and downstream signalling components. We identify orthologs through a combination of BLAST and phylogenetic analyses, showing that flatworms have a highly reduced and dispersed complement that includes orthologs of only five subfamilies (Wnt1, Wnt2, Wnt4, Wnt5 and Wnt11) and fewer paralogs in parasitic flatworms (5-6) than in planarians (9). All major signalling components are identified, including antagonists and receptors, and key binding domains are intact, indicating that the canonical (Wnt/?-catenin) and non-canonical (planar cell polarity and Wnt/Ca(2+)) pathways are functional. RNA-Seq data show expression of all Hymenolepis Wnts and most downstream components in adults and larvae with the notable exceptions of wnt1, expressed only in adults, and wnt2 expressed only in larvae. The distribution of Wnt subfamilies in animals corroborates the idea that the last common ancestor of the Cnidaria and Bilateria possessed all contemporary Wnts and highlights the extent of gene loss in flatworms. PMID:21892738

Riddiford, Nick; Olson, Peter D

2011-09-03

227

The effects of starvation on the planarian worm Polycelis tenuis Iijima.  

PubMed

Employing a combination of microscopical, biochemical and autoradiographic techniques, the primary effects of starvation on adult polycelis tenuis have been studied. Over a five week period of starvation there is on average a 32% decrease in the size of the organism. This decrease is contributed to by a reduction in mitosis and an increase in cell shrinkage autolysis and death. During starvation (following a sharp rise in RNA synthesis) there is a distinct sequence of events; four peaks of acid phosphatase activity can be resolved. The first is associated with the immediate response of the gastrodermis to feeding; the second (after 6 to 7 days) with increased autophagy and dedifferentiation in the gland cells and with muscle lysis of cells. The third peak (after 14 to 15 days) is contributed to largely by the lysis of cells in the gut and the fourth peak (after 25 to 26 days) is caused by an extensive lysis of the reproductive system. Fine structural changes involving increased intracellular vacuolation, autophagy, crinophagy, atrophy of muscle, increased intercellular space and loss of basement membrane matrix have been related to changes in enzyme pattern. Nerve cells appear unchanged throughout the first five weeks of starvation. Pigment and gland cells loose their characteristic granules, dedifferentiate and become morphologically similar to the undifferentiatied neoblasts. Dedifferentiation and the mechanisms involved in the survival of starvation are discussed. PMID:954048

Bowen, E D; Ryder, T A; Dark, C

1976-06-14

228

Multicellularity, stem cells, and the neoblasts of the planarian Schmidtea mediterranea  

Microsoft Academic Search

All multicellular organisms depend on stem cells for their survival and perpetuation. Their central role in reproductive, embryonic, and post- embryonic processes, combined with their wide phylogenetic distribution in both the plant and animal kingdoms intimates that the emergence of stem cells may have been a prerequisite in the evolution of multicellular organisms. We present an evolutionary perspective on stem

Alejandro Sanchez Alvarado; Hara Kang

2005-01-01

229

Double-stranded RNA specifically disrupts gene expression during planarian regeneration  

Microsoft Academic Search

Metazoan regeneration is one of the least understood fundamental problems of biology. The lack of progress in understanding this phenomenon at the molecular level has been due to the poor regenerative abilities of the genetic organisms used for developmental studies, as well as the difficulties encountered with molecular and genetic ma- nipulations of the commonly studied vertebrate models (the urodele

ALEJANDRO SANCHEZ ALVARADO; PHILLIP A. NEWMARK

1999-01-01

230

Weak extremely-low-frequency magnetic fields and regeneration in the planarian Dugesia tigrina  

SciTech Connect

Extremely-low-frequency (ELF), low-intensity magnetic fields have been shown to influence cell signaling processes in a variety of systems, both in vivo and in vitro. Similar effects have been demonstrated for nervous system development and neurite outgrowth. The authors report that regeneration in planaria, which incorporates many of these processes, is also affected by ELF magnetic fields. The rate of cephalic regeneration, reflected by the mean regeneration time (MRT), for planaria populations regenerating under continuous exposure to combined DC (78.4 {mu}T) and AC (60.0 Hz at 10.0 {mu}T{sub peak}) magnetic fields applied in parallel was found to be significantly delayed (P {much_lt} 0.001) by 48 {+-} 1 h relative to two different types of control populations (MRT {minus}140 {+-} 12 h). One control population was exposed to only the AC component of this field combination, while the other experienced only the ambient geomagnetic field. All measurements were conducted in a low-gradient, low-noise magnetics laboratory under well-maintained temperature conditions. This delay in regeneration was shown to be dependent on the planaria having a fixed orientation with respect to the magnetic field vectors. Results also indicate that this orientation-dependent transduction process does not result from Faraday induction but is consistent with a Ca{sup 2+} cyclotron resonance mechanism. Data interpretation also permits the tentative conclusion that the effect results from an inhibition of events at an early stage in the regeneration process before the onset of proliferation and differentiation.

Jenrow, K.A.; Smith, C.H.; Liboff, A.R. [Oakland Univ., Rochester, MI (United States). Dept. of Physics

1995-06-01

231

Acute toxic responses of the freshwater planarian, Dugesia dorotocephala, to methylmercury  

SciTech Connect

Toxic responses of planaria to various aquatic habitat concentrations of methylmercury chloride (MMC) were investigated. One hundred percent lethality occurred within 5 h in 2 ppM MMC, 24 h in 1 ppM MMC, and 5 days in 0.5 ppM MMC. No deaths occurred in 0.2 ppM MMC over a 10 day period, however, non-lethal toxic responses were observed. Varying degrees of head resorption, progressing caudally from the snout were observed. With continuing exposure, partial head regeneration and recovery toward more normal appearance occurred by 10 days. Teratogenic effects were observed in surgical decapitation experiments. Head regeneration was retarded in 0.1 and 0.2 ppM MMC. Malformations, visible lesions, or gross behavioral abnormalities were produced by 2 week exposure of planaria to concentrations of 20 ppB MMC or lower. (RJC)

Best, J.B.; Morita, M.; Ragin, J.; Best, J. Jr.

1981-07-01

232

RESPONSES OF THE PLANARIAN, DUGESIA, AND THE PROTOZOAN, PARAMECIUM, TO VERY WEAK HORIZONTAL MAGNETIC FIELDS  

Microsoft Academic Search

The question of whether living things are sensitive to terrestrial magnetism has undoubtedly fleeted through the minds of innumerable persons since this geophysical factor first became known. But neither the naturalist, observing the behavior of organisms in the field during their continuing responses to the myriads of more obvious physical factors, nor the experimental biologist, casually testing the response of

FRANK A. BROWN

233

Identification of the Boudicca and Sinbad retrotransposons in the genome of the human blood fluke Schistosoma haematobium.  

PubMed

Schistosomes have a comparatively large genome, estimated for Schistosoma mansoni to be about 270 megabase pairs (haploid genome). Recent findings have shown that mobile genetic elements constitute significant proportions of the genomes of S. mansoni and S. japonicum. Much less information is available on the genome of the third major human schistosome, S. haematobium. In order to investigate the possible evolutionary origins of the S. mansoni long terminal repeat retrotransposons Boudicca and Sinbad, several genomes were searched by Southern blot for the presence of these retrotransposons. These included three species of schistosomes, S. mansoni, S. japonicum, and S. haematobium, and three related platyhelminth genomes, the liver flukes Fasciola hepatica and Fascioloides magna and the planarian, Dugesia dorotocephala. In addition, Homo sapiens and three snail host genomes, Biomphalaria glabrata, Oncomelania hupensis, and Bulinus truncatus, were examined for possible indications of a horizontal origin for these retrotransposons. Southern hybridization analysis indicated that both Boudicca and Sinbad were present in the genome of S. haematobium. Furthermore, low stringency Southern hybridization analyses suggested that a Boudicca-like retrotransposon was present in the genome of B. truncatus, the snail host of S. haematobium. PMID:17072464

Copeland, Claudia S; Lewis, Fred A; Brindley, Paul J

2006-08-01

234

Mitochondrial pathway of apoptosis is ancestral in metazoans  

PubMed Central

The mitochondrial pathway of apoptosis is the major mechanism of physiological cell death in vertebrates. In this pathway, proapoptotic members of the Bcl-2 family cause mitochondrial outer membrane permeabilization (MOMP), allowing the release of cytochrome c, which interacts with Apaf-1 to trigger caspase activation and apoptosis. Despite conservation of Bcl-2, Apaf-1, and caspases in invertebrate phyla, the existence of the mitochondrial pathway in any invertebrate is, at best, controversial. Here we show that apoptosis in a lophotrochozoan, planaria (phylum Platyhelminthes), is associated with MOMP and that cytochrome c triggers caspase activation in cytosolic extracts from these animals. Further, planarian Bcl-2 family proteins can induce and/or regulate cell death in yeast and can replace Bcl-2 proteins in mammalian cells to regulate MOMP. These results suggest that the mitochondrial pathway of apoptosis in animals predates the emergence of the vertebrates but was lost in some lineages (e.g., nematodes). In further support of this hypothesis, we surveyed the ability of cytochrome c to trigger caspase activation in cytosolic extracts from a variety of organisms and found this effect in cytosolic extracts from invertebrate deuterostomes (phylum Echinodermata).

Bender, Cheryl E.; Fitzgerald, Patrick; Tait, Stephen W. G.; Llambi, Fabien; McStay, Gavin P.; Tupper, Douglas O.; Pellettieri, Jason; Alvarado, Alejandro Sanchez; Salvesen, Guy S.; Green, Douglas R.

2012-01-01

235

CALORIFIC VALUES IN THE PHYLUM PLATYHELMINTHES: THE RELATIONSHIP BETWEEN POTENTIAL ENERGY, MODE OF LIFE AND THE EVOLUTION OF ENTOPARASITISM  

Microsoft Academic Search

An analysis of the calorific values (kcal\\/g) of seventeen species of animals from six phyla has shown that they have a skewed distribution with a modal fre quency at or near the lower range limit (Slobodkin and Richman, 1%1 ) . This was regarded as support for the hypothesis that natural selection generally favors production of the maximum number of

P. CALOW; J. B. JENNINGS

236

Development of mitochondrial loop-mediated isothermal amplification for detection of the small liver fluke Opisthorchis viverrini (Opisthorchiidae; Trematoda; Platyhelminthes).  

PubMed

Mitochondrial DNA sequences offer major advantages over the more usual nuclear targets for loop-mediated isothermal amplification approaches (mito-LAMP) because multiple copies occur in every cell. Four LAMP primers [F3, FIP(F1c+F2), BIP(B1c+B2), and B3] were designed based on the mitochondrial nad1 sequence of Opisthorchis viverrini and used for a highly specific assay (mito-OvLAMP) to distinguish DNA of O. viverrini from that of another opisthorchiid (Clonorchis sinensis) and other trematodes (Haplorchis pumilio, Haplorchis taichui, Fasciola hepatica, and Fasciola gigantica). Conventional PCR was applied using F3/B3 primer pairs to verify the specificity of the primers for O. viverrini DNA templates. All LAMP-positive samples could be detected with the naked eye in sunlight, by gel electrophoresis (stained with ethidium bromide), and by addition of SYBR green I to the product in sunlight or under UV light. Only DNA from O. viverrini yielded amplification products by LAMP (and by PCR verification), and the LAMP limit of detection was as little as 100 fg (10(-4) ng DNA), indicating that this assay is 10 to 100 times more sensitive than PCR. Field testing was done using representative egg and metacercarial samples collected from localities where the fluke is endemic. With the advantages of simplicity, rapidity, sensitivity, and cost effectiveness, mito-OvLAMP is a good tool for molecular detection and epidemiology studies in regions or countries where O. viverrini is endemic, which can lead to more effective control of opisthorchiasis and trematodiasis. PMID:22322346

Le, Thanh Hoa; Nguyen, Nga Thi Bich; Truong, Nam Hai; De, Nguyen Van

2012-02-08

237

Cryptostylochus hullensis sp. nov. (Polycladida, Acotylea, Platyhelminthes): A possible case of transoceanic dispersal on a ship's hull  

NASA Astrophysics Data System (ADS)

In July 1993, the car carrier “Faust” entered Bremerhaven after a voyage from the North-American Atlantic coast to Europe. In a dockyard, five living specimens of the order Polycladida were collected from the hull of the ship. This could be a possible case of trans-atlantic dispersal of plathelminths living as fouling organisms of ships. The specimens found represent a new species of the genus Cryptostylochus Faubel, 1983, Cryptostylochus hullensis sp. nov.

Faubel, A.; Gollasch, S.

1996-12-01

238

A comprehensive molecular phylogeny of dalytyphloplanida (platyhelminthes: rhabdocoela) reveals multiple escapes from the marine environment and origins of symbiotic relationships.  

PubMed

In this study we elaborate the phylogeny of Dalytyphloplanida based on complete 18S rDNA (156 sequences) and partial 28S rDNA (125 sequences), using a Maximum Likelihood and a Bayesian Inference approach, in order to investigate the origin of a limnic or limnoterrestrial and of a symbiotic lifestyle in this large group of rhabditophoran flatworms. The results of our phylogenetic analyses and ancestral state reconstructions indicate that dalytyphloplanids have their origin in the marine environment and that there was one highly successful invasion of the freshwater environment, leading to a large radiation of limnic and limnoterrestrial dalytyphloplanids. This monophyletic freshwater clade, Limnotyphloplanida, comprises the taxa Dalyelliidae, Temnocephalida, and most Typhloplanidae. Temnocephalida can be considered ectosymbiotic Dalyelliidae as they are embedded within this group. Secondary returns to brackish water and marine environments occurred relatively frequently in several dalyeliid and typhloplanid taxa. Our phylogenies also show that, apart from the Limnotyphloplanida, there have been only few independent invasions of the limnic environment, and apparently these were not followed by spectacular speciation events. The distinct phylogenetic positions of the symbiotic taxa also suggest multiple origins of commensal and parasitic life strategies within Dalytyphloplanida. The previously established higher-level dalytyphloplanid clades are confirmed in our topologies, but many of the traditional families are not monophyletic. Alternative hypothesis testing constraining the monophyly of these families in the topologies and using the approximately unbiased test, also statistically rejects their monophyly. PMID:23536894

Van Steenkiste, Niels; Tessens, Bart; Willems, Wim; Backeljau, Thierry; Jondelius, Ulf; Artois, Tom

2013-03-25

239

Massive parasitism by Gussevia tucunarense (Platyhelminthes: Monogenea: Dactylogyridae) in fingerlings of bujurqui-tucunare cultured in the Peruvian Amazon.  

PubMed

A high infestation of the monogenean Gussevia tucunarense in a cultivation of bujurqui-tucunare was reported. The prevalence was 100%. The mean intensity and abundance of the parasite was 164.4 of parasites per individual. This is the first report of a high infestation by G. tucunarense in C. semifasciatus cultured from the Peruvian Amazon. PMID:23666660

Mathews, Patrick D; Mertins, Omar; Mathews, John P D; Orbe, Rosa I

2013-05-11

240

BIOLOGY OF NEOCHILDA FUSCA N. GEN., N. SP. FROM THE NORTHEASTERN COAST OF THE UNITED STATES (PLATYHELMINTHES : TURBELLARIA)  

Microsoft Academic Search

A dark brown acoel has been collected for a number of years in bottom mud samples from Great Harbor and Buzzards Bay near Woods Hole, Massachusetts for use in class and experimental work. In the summer of 1965 I found a large population of the same species living intertidally in the salt marsh bordering Barn stable Harbor near West Dennis,

LOUISE BUSH

1975-01-01

241

A new and alien species of ``oyster leech'' (Platyhelminthes, Polycladida, Stylochidae) from the brackish North Sea Canal, The Netherlands  

NASA Astrophysics Data System (ADS)

A new species of polyclad flatworm, Imogine necopinata Sluys, sp. nov., is described from a brackish habitat in The Netherlands. Taxonomic affinities with Asian species and the ecology of the animals suggest that the species is an introduced, exotic component of the Dutch fauna. The new species belongs to a group of worms with species that are known to predate on oysters.

Sluys, Ronald; Faubel, Anno; Rajagopal, Sanjeevi; Velde, Gerard Van Der

2005-11-01

242

Development of Mitochondrial Loop-Mediated Isothermal Amplification for Detection of the Small Liver Fluke Opisthorchis viverrini (Opisthorchiidae; Trematoda; Platyhelminthes)  

PubMed Central

Mitochondrial DNA sequences offer major advantages over the more usual nuclear targets for loop-mediated isothermal amplification approaches (mito-LAMP) because multiple copies occur in every cell. Four LAMP primers [F3, FIP(F1c+F2), BIP(B1c+B2), and B3] were designed based on the mitochondrial nad1 sequence of Opisthorchis viverrini and used for a highly specific assay (mito-OvLAMP) to distinguish DNA of O. viverrini from that of another opisthorchiid (Clonorchis sinensis) and other trematodes (Haplorchis pumilio, Haplorchis taichui, Fasciola hepatica, and Fasciola gigantica). Conventional PCR was applied using F3/B3 primer pairs to verify the specificity of the primers for O. viverrini DNA templates. All LAMP-positive samples could be detected with the naked eye in sunlight, by gel electrophoresis (stained with ethidium bromide), and by addition of SYBR green I to the product in sunlight or under UV light. Only DNA from O. viverrini yielded amplification products by LAMP (and by PCR verification), and the LAMP limit of detection was as little as 100 fg (10?4 ng DNA), indicating that this assay is 10 to 100 times more sensitive than PCR. Field testing was done using representative egg and metacercarial samples collected from localities where the fluke is endemic. With the advantages of simplicity, rapidity, sensitivity, and cost effectiveness, mito-OvLAMP is a good tool for molecular detection and epidemiology studies in regions or countries where O. viverrini is endemic, which can lead to more effective control of opisthorchiasis and trematodiasis.

Nguyen, Nga Thi Bich; Truong, Nam Hai; De, Nguyen Van

2012-01-01

243

Phylogenetic relationships of the Gorgoderidae (Platyhelminthes: Trematoda), including the proposal of a new subfamily (Degeneriinae n. subfam.).  

PubMed

Phylogenetic analyses of a range of gorgoderid trematodes based on ITS2 and partial 28S rDNA data lead us to propose the Degeneriinae n. subfam. for the genus Degeneria in recognition of its phylogenetic isolation and distinctive morphology and biology. The current concepts of the subfamilies Anaporrhutinae and Gorgoderinae were supported. Within the Gorgoderinae, the large genus Phyllodistomum is shown to be paraphyletic relative to Pseudophyllodistomum and Xystretrum. Notably, the clade of marine Phyllodistomum does not form a clade with the other marine genus, Xystretrum. Distinct clades within the Gorgoderinae correspond variously to identity of first intermediate host, form of cercaria and their marine or freshwater habitat. We are not yet in a position to propose separate genera for these clades. PMID:23760874

Cutmore, Scott C; Miller, Terrence L; Curran, Stephen S; Bennett, Michael B; Cribb, Thomas H

2013-06-13

244

Genetic differentiation among populations of Minona ileanae (Platyhelminthes: Proseriata) from the Red Sea and the Suez Canal  

Microsoft Academic Search

The phenomenon of Lessepsian migration has stimulated the interest of biologists ever since the opening of the Suez Canal in 1869, concerning, in particular, the possible effects of migrants on Mediterranean autochthonous communities. So far, most attention has been devoted to macrofaunal taxa – yet, the nature of the sandy shores of the Canal may constitute an ideal habitat for

Tiziana Lai; Marco Curini-Galletti; Marco Casu

2008-01-01

245

Effects of Microphallus papillorobustus (Platyhelminthes: Trematoda) on serotonergic immunoreactivity and neuronal architecture in the brain of Gammarus insensibilis (Crustacea: Amphipoda).  

PubMed Central

The larval flatworm Microphallus papillorobustus encysts in the protocerebrum of its intermediate host, Gammarus insensibilis, and changes the gammarid's responses to mechanical and photic stimuli. The resulting aberrant escape behaviour renders infected gammarids more susceptible to predation by birds, the definitive hosts of the parasite. We used immunocytochemical methods to explore the mechanisms underlying these subtle behavioural modifications. Whole mounts of gammarid brains were labelled with fluorescent anti-serotonin and anti-synapsin antibodies and viewed using confocal microscopy. Two types of change were observed in infected brains: the intensity of the serotonergic label was altered in specific regions of the brain, and the architecture of some serotonergic tracts and neurons was affected. A morphometric analysis of the distribution of the label showed that serotonergic immunoreactivity was decreased significantly (by 62%) in the optic neuropils, but not in the olfactory lobes, in the presence of the parasite. In addition, the optic tracts and the tritocerebral giant neurons were stunted in parasitized individuals. Published evidence demonstrates changes in serotonin levels in hosts ranging from crustaceans to mammals infected by parasites as diverse as protozoans and helminths. The present study suggests that the degeneration of discrete sets of serotonergic neurons might underlie the serotonergic imbalance and thus contribute to host manipulation.

Helluy, S; Thomas, F

2003-01-01

246

Morphological and molecular characterisation of Gyrodactylus salmonis (Platyhelminthes, Monogenea) isolates collected in Mexico from rainbow trout (Oncorhynchus mykiss Walbaum).  

PubMed

Gyrodactylus salmonis (Yin et Sproston, 1948) isolates collected from feral rainbow trout, Oncorhynchus mykiss (Walbaum) in Veracruz, southeastern Mexico are described. Morphological and molecular variation of these isolates to G. salmonis collected in Canada and the U.S.A. is characterised. Morphologically, the marginal hook sickles of Mexican isolates of G. salmonis closely resemble those of Canadian specimens - their shaft and hook regions align closely with one another; only features of the sickle base and a prominent bridge to the toe permit their separation. The 18S sequence determined from the Mexican specimens was identical to two variable regions of SSU rDNA obtained from a Canadian population of G. salmonis. Internal transcribed spacer (ITS) regions (spanning ITS1, 5.8S and ITS2) of Mexican isolates of G. salmonis are identical to ITS sequences of an American population of G. salmonis and to Gyrodactylus salvelini Kuusela, Zi?tara et Lumme, 2008 from Finland. Analyses of the ribosomal RNA gene of Mexican isolates of G. salmonis show 98-99% similarity to those of Gyrodactylus gobiensis Gläser, 1974, Gyrodactylus salaris Malmberg, 1957, and Gyrodactylus rutilensis Gläser, 1974. Mexican and American isolates of G. salmonis are 98% identical, as assessed by sequencing the mitochondrial cox1 gene. Oncorhynchus mykiss is one of the most widely-dispersed fish species in the world and has been shown to be an important vector for parasite/disease transmission. Considering that Mexican isolates of G. salmonis were collected well outside the native distribution range of all salmonid fish, we discuss the possibility that the parasites were translocated with their host through the aquacultural trade. In addition, this study includes a morphological review of Gyrodactylus species collected from rainbow trout and from other salmonid fish of the genus Oncorhynchus which occur throughout North America. PMID:22169222

Rubio-Godoy, Miguel; Paladini, Giuseppe; Freeman, Mark A; García-Vásquez, Adriana; Shinn, Andrew P

2011-11-07

247

Light and electron microscopy observations of embryogenesis and egg development in the human liver fluke, Opisthorchis viverrini (Platyhelminthes, Digenea).  

PubMed

Eggs of most species digenean flukes hatch in the external environment to liberate larvae that seek and penetrate a snail intermediate host. Those of the human liver flukes, Opisthorchis viverrini, hatch within the gastrointestinal canal of their snail hosts. While adult parasites are primarily responsible for the pathology in cases of human opisthorchiasis, their eggs also contribute by inducing granulomata and in serving as nidi for gallstone formation. In view of the peculiar biology of O. viverrini eggs and their contribution to pathology, we investigated embryogenesis in this species by light and transmission electron microscopy. Egg development was traced from earliest stages of coalescence in the ootype until full embryonation in the distal region of the uterus. Fully mature eggs were generally impermeable to resin and could not be examined by conventional electron microscopy methods. However, the use of high-pressure freezing and freeze-substitution fixation of previously fixed eggs enabled the internal structure of mature eggs, particularly the subshell envelopes, to be elucidated. Fertilization occurs in the ootype, and the large zygote is seen therein with a single spermatozoon wrapped around its plasma membrane. As the zygote begins to divide, the spent vitellocytes are pushed to the periphery of the eggs, where they progressively degrade. The early eggshell is formed in the ootype by coalescing eggshell precursor material released by approximately six vitelline cells. The early eggs have a thinner eggshell and are larger than, but lack the characteristic shape of, mature eggs. Characteristic shell ornamentation, the "muskmelon" appearance of eggs, appears after eggshell polymerization in the ootype. Pores are not present in the shell of O. viverrini eggs. The inner and outer envelopes are poorly formed in this species, with the outer envelope evident beneath the eggshell at the opercular pole of the mature egg. The miracidium has a conical anterior end that lacks the distinctive lamellar appearance of the terebratorium of other digeneans, such as the schistosomes. The miracidium is richly glandular, containing an apical gland in the anterior end, large cephalic gland, and posterior secretory glands. Each gland contains a secretory product with different structure. The paucity of vitelline cells associating with eggs, the reduced size of eggs, and reduced complexity of the extraembryonic envelopes are interpreted as adaptations to the peculiar hatching biology of the miracidia. PMID:21786067

Khampoosa, Panita; Jones, Malcolm K; Lovas, Erica M; Srisawangwong, Tuanchai; Laha, Thewarach; Piratae, Supawadee; Thammasiri, Chalida; Suwannatrai, Apiporn; Sripanidkulchai, Bungorn; Eursitthichai, Veerachai; Tesana, Smarn

2011-07-23

248

Light and electron microscopy observations of embryogenesis and egg development in the human liver fluke, Opisthorchis viverrini (Platyhelminthes, Digenea)  

Microsoft Academic Search

Eggs of most species digenean flukes hatch in the external environment to liberate larvae that seek and penetrate a snail\\u000a intermediate host. Those of the human liver flukes, Opisthorchis viverrini, hatch within the gastrointestinal canal of their snail hosts. While adult parasites are primarily responsible for the pathology\\u000a in cases of human opisthorchiasis, their eggs also contribute by inducing granulomata

Panita Khampoosa; Malcolm K. Jones; Erica M. Lovas; Tuanchai Srisawangwong; Thewarach Laha; Supawadee Piratae; Chalida Thammasiri; Apiporn Suwannatrai; Bungorn Sripanidkulchai; Veerachai Eursitthichai; Smarn Tesana

249

Two new species of Temnocephala (Platyhelminthes, Temnocephalida) from the South American snake-necked turtle Hydromedusa tectifera (Testudines, Chelidae).  

PubMed

Temnocephala brevicornis Monticelli, 1889 is the only species of the genus Temnocephala Blanchard, 1849 reported from chelonians to date. During a survey of the species of Temnocephala extant in southern Uruguay, two new species were found on the chelonian Hydromedusa tectifera Cope, 1869. They are described here as Temnocephala pereirai n. sp. and Temnocephala cuocoloi n. sp. Both resemble T. brevicornis, but differ in the morphometry of the penial stylet, and in qualitative details of the reproductive complex. Temnocephala pereirai n. sp. differs from T. brevicornis by having a massive, cylindrical sphincter in the distal portion of the vagina, and a seminal vesicle that opens into the subpolar to equatorial portion of the contractile vesicle. In addition, the penial stylet in Temnocephala pereirai n. sp. is large in relation to body size, straight and more slender, having the distal portion of its shaft slightly sinuous, and a smaller introvert equipped with about 16 distal crowns of smaller spines. Temnocephala cuocoloi n. sp. is most similar to T. brevicornis, but differs by having a smaller, curved penial stylet that has a smaller introvert in relation to stylet size, with about 10 distal crowns of smaller spines. A key to the species of the Temnocephala from chelonians is provided. This study supports the validity of the following characters previously proposed for the taxonomy of the genus Temnocephala: the shape of the sphincters in the female reproductive system, the shape of the penial stylet, and the number, size, and position of spines in the introvert. PMID:21110724

Volonterio, Odile

2010-12-01

250

Evidence for host-specific clades of tetraphyllidean tapeworms (Platyhelminthes: Eucestoda) revealed by analysis of 18S ssrDNA.  

PubMed

Sequence data from the V4 and V7-V9 variable regions of the 18S small subunit ribosomal DNA (ssrDNA) gene were used to examine relationships among 26 tetraphyllidean and two lecanicephalidean taxa. Newly collected specimens of 21 of the tetraphyllidean species were used to generate ssrDNA sequences that were combined with sequences previously available, including those of two diphyllidean taxa used for outgroup rooting. The sequences were aligned by eye according to secondary structural motifs of the conserved core of the molecule. Of the 1520 sites in the alignment, 874 (58%) were excluded from analysis due to alignment gaps and lack of positional homology as inferred by manual inspection. Genetic variability of the ssrDNA gene regions compared was greater than would be expected, based on the present taxonomy of the ingroup species, and the genetic divergences among tetraphyllidean 'families' and genera were comparable to that among tapeworm orders. Phylogenetic hypotheses were generated by the methods of maximum parsimony and maximum likelihood (GTR + I + Gamma nucleotide substitution model). Four most parsimonious trees resulted from analysis by maximum parsimony. Strict consensus of the four trees supported the monophyly of the Tetraphyllidea, with the lecanicephalidean taxa forming a sister lineage. Among the tetraphyllidean taxa included in the analysis were three major clades: a basal clade including species of the phyllobothriid genera Anthocephalum, Echeneibothrium, Rhinebothrium, Rhodobothrium and Spongiobothrium; a clade uniting the phyllobothriids of the genus Duplicibothrium with the dioecotaeniid genus Dioecotaenia; and a larger sister clade to the Duplicibothrium + Dioecotaenia clade that included the phyllobothriid genera Caulohothrium, Ceratobothrium, Clistobothrium, Paraoryigmatobothrium and Prosobothrium, the litobothriid genus Litobothrium and the onchobothriid genera Acanthobothrium, Calliobothrium, Phoreiobothrium and Platybothrium. Maximum likelihood analysis resulted in a topology that was congruent where nodes were strongly supported by parsimony analysis, but differed in the relative positions of the well-supported clades. In addition,maximum likelihood analysis grouped the lecanicephalidean taxa among the tetraphyllidean taxa, indicating paraphyly of the order Tetraphyllidea as currently defined. Relationships suggested by both methods of analysis reflected common host associations of the taxa better than their current classification, suggesting that coevolution has had a significant role in the evolution of the group. PMID:10579434

Olson, P D; Ruhnke, T R; Sanney, J; Hudson, T

1999-09-01

251

Added resolution among ordinal level relationships of tapeworms (Platyhelminthes: Cestoda) with complete small and large subunit nuclear ribosomal RNA genes.  

PubMed

The addition of large subunit ribosomal DNA (lsrDNA) to small subunit ribosomal DNA (ssrDNA) has been shown to add resolution to phylogenies at various taxonomic levels for a diversity of phyla. We added nearly complete lsrDNA (4057-4593bp) sequences to ssrDNA (1940-2228bp) for 26 ingroup and 3 outgroup taxa in an attempt to provide an improved ordinal phylogeny for the Cestoda. Ten lsrDNA and seven ssrDNA sequences were generated from new taxa and 13 existing partial lsrDNA sequences were sequenced to completion. The majority of phylogenetic signal in the combined analysis came from lsrDNA (69.6% of parsimonious informative sites, as opposed to 30.4% obtained from ssrDNA), resulting in almost identical topologies for lsrDNA and lsr+ssrDNA (pairwise symmetric distance=6) in model-based analyses. Topology testing found trees based on partial lsrDNA (domains D1-D3)+ssrDNA and complete lsr+ssrDNA to differ significantly; the addition of lsrDNA domains D4-D12 had a significant effect on topology. Overall nodal support was greatest in the combined analysis and weakest for ssrDNA only. Our molecular phylogenies differed significantly from those based on morphology alone. Acetabulate lineages form a monophyletic group, with the Tetraphyllidea being paraphyletic. Support for the combined data was high for the following topology: (Litobothriidea (Lecanicephalidea (Rhinebothrium/Rhodobothrium (Clistobothrium (Pachybothrium(Acanthobothrium Proteocephalidea) (Mesocestoididae, Nippotaeniidea, Cyclophyllidea, Tetrabothriidea)))))); all genus names refer to tetraphyllidean lineages. Although the interrelationships among the four most derived taxa remain uncertain, overall ambiguity of the acetabulate interrelationships was reduced. The Pseudophyllidea were recovered as polyphyletic, with support for a sister-group relationship between Diphyllobothriidae and Haplobothriidea. The monophyly of the Trypanorhyncha was recovered for the first time based on molecular data. The positions of the Trypanorhyncha, Diphyllidea and "Bothriocephaliidea" in relation to other orders remains ambiguous. Higher congruence was found between trees based on model-based phylogenetic methods than with those constructed under the parsimony criterion. Although some uncertainties remain, the addition of lsrDNA D4-D12 has provided an overall more resolved and better supported cestode phylogeny, which further promotes the utility of complete lsrDNA as phylogenetic marker where ssrDNA alone proves inadequate. PMID:17485227

Waeschenbach, Andrea; Webster, Bonnie L; Bray, Rodney A; Littlewood, D T J

2007-04-03

252

Adding resolution to ordinal level relationships of tapeworms (Platyhelminthes: Cestoda) with large fragments of mtDNA.  

PubMed

The construction of a stable phylogeny for the Cestoda, indicating the interrelationships of recognised orders and other major lineages, has proceeded iteratively since the group first received attention from phylogenetic systematists. Molecular analyses using nuclear ribosomal RNA gene fragments from the small (ssrDNA) and large (lsrDNA) subunits have been used to test competing evolutionary scenarios based on morphological data but could not arbitrate between some key conflicting hypotheses. To the ribosomal data, we have added a contiguous fragment of mitochondrial (mt) genome data (mtDNA) of partial nad1-trnN-trnP-trnI-trnK-nad3-trnS-trnW-cox1-trnT-rrnL-trnC-partial rrnS, spanning 4034-4447 bp, where new data for this region were generated for 18 species. Bayesian analysis of mtDNA and rDNA as nucleotides, and where appropriate as amino acids, demonstrated that these two classes of genes provide complementary signal across the phylogeny. In all analyses, except when using mt amino acids only, the Gyrocotylidea is sister group to all other Cestoda (Nephroposticophora), and Amphilinidea forms the sister group to the Eucestoda. However, an earliest-diverging position of Amphilinidea is strongly supported in the mt amino acid analysis. Amphilinidea exhibit a unique tRNA arrangement (nad1-trnI-trnL2-trnP-trnK-trnV-trnA-trnN-nad3), whereas Gyrocotylidea shares that of the derived lineages, providing additional evidence of the uniqueness of amphilinid genes and genomes. The addition of mtDNA to the rDNA genes supported the Caryophyllidea as the sister group to (Spathebothriidea+remaining Eucestoda), a hypothesis consistently supported by morphology. This relationship suggests a history of step-wise evolutionary transitions from simple monozoic, unsegmented tapeworms to the more familiar polyzoic, externally segmented (strobilate) forms. All our data partitions recovered Haplobothriidea as the sister group to Diphyllobothriidae. The sister-group relationship between Diphyllidea and Trypanorhyncha, as previously established using rDNA, is not supported by the mt data, although it is supported by the combined mt and rDNA analysis. With regards to the more derived taxa, in all except the mt amino acid analysis, the following topology is supported: (Bothriocephalidea (Litobothriidea (Lecanicephalidea (Rhinebothriidea (Tetraphyllidea, (Acanthobothrium, Proteocephalidea), (Nippotaeniidea, Mesocestoididae, Tetrabothriidea, Cyclophyllidea)))))), where the Tetraphyllidea are paraphyletic. Evidence from the mt data provides strong (nucleotides) to moderate (amino acids) support for Tetraphyllidea forming a group to the inclusion of Proteocephalidea, with the latter consistently forming the sister group to Acanthobothrium. The interrelationships among Nippotaeniidea, Mesocestoididae, Tetrabothriidea and Cyclophyllidea remain ambiguous and require further systematic attention. Mitochondrial and nuclear rDNA data provide conflicting signal for certain parts of the cestode tree. In some cases mt data offer results in line with morphological evidence, such as the interrelationships of the early divergent lineages. Also, Tetraphyllidea, although remaining paraphyletic with the inclusion of the Proteocephalidea, does not include the most derived cestodes; a result which has consistently been obtained with rDNA. PMID:22406529

Waeschenbach, Andrea; Webster, B L; Littlewood, D T J

2012-03-03

253

Twelve new species of dactylogyrids (Platyhelminthes, Monogenea) from West African barbels (Teleostei, Cyprinidae), with some biogeographical implications  

Microsoft Academic Search

Fifteen species of dactylogyrid monogeneans, belong to Dactylogyrus and Dogielius were observed in seven different African species of Barbus and Varicorhinus (Teleostei, Cyprinidae). The barbels examined in West Africa were: Barbus occidentalis Boulenger, 1911, known in the large Sahel-Sudan rivers and in Gabon: B. waldroni Nordman, 1935, B. petitjeani Daget, 1962, B. sacratus Daget, 1963, B. parawaldroni Lévęque, Thys van

Jean-François Guégan; Alain Lambert

1990-01-01

254

[Histological and cytological study of neoblasts and fixed parenchymal cells in Dendrocoelum lacteum planarians deprived of their anterior region].  

PubMed

On amputating either at the root of the pharynx or between the mouth and the genital pore, the posterior parts which are thereby isolated do not regenerate. However they are capable of surviving for ever a year, without feeding. The wounded region is occupied by neoblasts which degenerate within a few days after an initial secretory activity. Their degeneration is of autophagic type. The abortive blastems disappear only slowly, because the degeneration of their neoblasts is partly compensated by the continual immigration of new regeneration cells which come from the posterior region. Several months after amputation, under the effect of prolonged starvation, neoblasts and fixed parenchymal cells begin to degenerate. PMID:1217901

Stéphan-Dubois, F; Bautz, A

255

Ameliorating Effect of Chloride on Nitrite Toxicity to Freshwater Invertebrates with Different Physiology: a Comparative Study Between Amphipods and Planarians  

Microsoft Academic Search

High nitrite concentrations in freshwater ecosystems may cause toxicity to aquatic animals. These living organisms can take\\u000a nitrite up from water through their chloride cells, subsequently suffering oxidation of their respiratory pigments (hemoglobin,\\u000a hemocyanin). Because NO2? and Cl? ions compete for the same active transport site, elevated chloride concentrations in the aquatic environment have the potential\\u000a of reducing nitrite toxicity.

A. Alonso; J. A. Camargo

2008-01-01

256

Intermediate filaments in muscle and epithelial cells of nematodes  

PubMed Central

Current concepts of the developmentally controlled multigene family of intermediate filament (IF) proteins expect the origin of their complexity in evolutionary precursors preceding all vertebrate classes. Among invertebrates, however, firm ultrastructural as well as molecular documentation of IFs is restricted to some giant axons and to epithelia of a few molluscs and annelids. As Ascaris lumbricoides is easily dissected into clean tissues, IF expression in this large nematode was analyzed by electron microscopic and biochemical procedures and a monoclonal antibody reacting with all mammalian IF proteins. We document for the first time the presence of IFs in muscle cells of an invertebrate. They occur in three muscle types (irregular striated pharynx muscle, obliquely striated body muscle, uterus smooth muscle). IFs are also found in the epithelia studied (syncytial epidermis, intestine, ovary, testis). Immunoblots on muscles, pharynx, intestine, uterus, and epidermis identify a pair of polypeptides (with apparent molecular masses of 71 and 63 kD) as IF constituents. In vitro reconstitution of filaments was obtained with the proteins purified from body muscle. In the small nematode Caenorhabditis elegans IF proteins are so far found only in the massive desmosome-anchored tonofilament bundles which traverse a special epithelial cell type, the marginal cells of the pharynx. We speculate that IFs may occur in most but perhaps not all invertebrates and that they may not occur in all cells in large amounts. As electron micrographs of the epidermis of a planarian--a member of the Platyhelminthes--reveal IFs, the evolutionary origin of this cytoplasmic structure can be expected either among the lowest metazoa or already in some unicellular eukaryotes.

1986-01-01

257

Ultrastructural study of the spermatozoon of the digenean Enodiotrema reductum Looss, 1901 (Platyhelminthes, Plagiorchioidea, Plagiorchiidae), parasite of the green turtle Chelonia mydas (Linnaeus, 1758) in Senegal.  

PubMed

This study describes the ultrastructural organisation of the spermatozoon of a digenean Enodiotrema reductum (Pligiorchiida: Plagiorchiidae) from the green turtle Chelonia mydas (Linnaeus, 1758). This is the first report of E. reductum from Senegal. The mature spermatozoon of E. reductum is filiform and exhibits two axonemes of the 9?+?"1" pattern of the Trepaxonemata, a nucleus, parallel cortical microtubules, an extramembranar ornamentation associated with spine-like bodies and granules of glycogen, among other ultrastructural features. The spermatozoon of E. reductum is distinguished by the presence of a moniliform mitochondrion composed of a bulge associated with a long cord and of a central cytoplasmic expansion. This work represents the first utrastructural study of any representative of the large family Plagiorchiidae. Our results are compared with previously published data on spermatozoa of other digenean taxa. PMID:22488200

Ndiaye, Papa Ibnou; Quilichini, Yann; Sčne, Aminata; Tkach, Vasyl V; Bâ, Cheikh Tidiane; Marchand, Bernard

2012-04-11

258

Characterization of a UDP-N-acetyl-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase with an unusual lectin domain from the platyhelminth parasite Echinococcus granulosus  

PubMed Central

As part of a general project aimed at elucidating the initiation of mucin-type O-glycosylation in helminth parasites, we have characterized a novel ppGalNAc-T (UDP-N-acetyl-D-galactosamine:polypeptide N-acetylgalactosaminyltransferase) from the cestode Echinococcus granulosus (Eg-ppGalNAc-T1). A full-length cDNA was isolated from a library of the tissue-dwelling larval stage of the parasite, and found to code for a 654-amino-acid protein containing all the structural features of ppGalNAc-Ts. Functional characterization of a recombinant protein lacking the transmembrane domain showed maximal activity at 28 °C, in the range 6.5–7.5 pH units and in the presence of Cu2+. In addition, it transferred GalNAc to a broad range of substrate peptides, derived from human mucins and O-glycosylated parasite proteins, including acceptors containing only serine or only threonine residues. Interestingly, the C-terminal region of Eg-ppGalNAc-T1 bears a highly unusual lectin domain, considerably longer than the one from other members of the family, and including only one of the three ricin B repeats generally present in ppGalNAc-Ts. Furthermore, a search for conserved domains within the protein C-terminus identified a fragment showing similarity to a recently defined domain, specialized in the binding of organic phosphates (CYTH). The role of the lectin domain in the determination of the substrate specificity of these enzymes suggests that Eg-ppGalNAc-T1 would be involved in the glycosylation of a special type of substrate. Analysis of the tissue distribution by in situ hybridization and immunohistochemistry revealed that this transferase is expressed in the hydatid cyst wall and the subtegumental region of larval worms. Therefore it could participate in the biosynthesis of O-glycosylated parasite proteins exposed at the interface between E. granulosus and its hosts.

2004-01-01

259

Development and evaluation of a single-step duplex PCR for simultaneous detection of Fasciola hepatica and Fasciola gigantica (family Fasciolidae, class Trematoda, phylum Platyhelminthes).  

PubMed

A single-step multiplex PCR (here referred to as a duplex PCR) has been developed for simultaneous detection and diagnosis of Fasciola hepatica and F. gigantica. These species overlap in distribution in many countries of North and East Africa and Central and Southeast Asia and are similar in egg morphology, making identification from fecal samples difficult. Based on a comparative alignment of mitochondrial DNA (mtDNA) spanning the region of cox1-trnT-rrnL, two species-specific forward primers were designed, FHF (for F. hepatica) and FGF (for F. gigantica), and a single reverse primer, FHGR (common for both species). Conventional PCR followed by sequencing was applied using species-specific primer pairs to verify the specificity of primers and the identity of Fasciola DNA templates. Duplex PCR (using three primers) was used for testing with the DNA extracted from adult worms, miracidia, and eggs, producing amplicons of 1,031 bp for F. hepatica and 615 bp for F. gigantica. The duplex PCR failed to amplify from DNA of other common liver and intestinal trematodes, including two opisthorchiids, three heterophyids, an echinostomid, another fasciolid, and a taeniid cestode. The sensitivity assay showed that the duplex PCR limit of detection for each Fasciola species was between 0.012 ng and 0.006 ng DNA. Evaluation using DNA templates from 32 Fasciola samples (28 adults and 4 eggs) and from 25 field-collected stools of ruminants and humans revealed specific bands of the correct size and the presence of Fasciola species. This novel mtDNA duplex PCR is a sensitive and fast tool for accurate identification of Fasciola species in areas of distributional and zonal overlap. PMID:22692744

Le, Thanh Hoa; Nguyen, Khue Thi; Nguyen, Nga Thi Bich; Doan, Huong Thi Thanh; Le, Xuyen Thi Kim; Hoang, Chau Thi Minh; De, Nguyen Van

2012-06-12

260

Development and Evaluation of a Single-Step Duplex PCR for Simultaneous Detection of Fasciola hepatica and Fasciola gigantica (Family Fasciolidae, Class Trematoda, Phylum Platyhelminthes)  

PubMed Central

A single-step multiplex PCR (here referred to as a duplex PCR) has been developed for simultaneous detection and diagnosis of Fasciola hepatica and F. gigantica. These species overlap in distribution in many countries of North and East Africa and Central and Southeast Asia and are similar in egg morphology, making identification from fecal samples difficult. Based on a comparative alignment of mitochondrial DNA (mtDNA) spanning the region of cox1-trnT-rrnL, two species-specific forward primers were designed, FHF (for F. hepatica) and FGF (for F. gigantica), and a single reverse primer, FHGR (common for both species). Conventional PCR followed by sequencing was applied using species-specific primer pairs to verify the specificity of primers and the identity of Fasciola DNA templates. Duplex PCR (using three primers) was used for testing with the DNA extracted from adult worms, miracidia, and eggs, producing amplicons of 1,031 bp for F. hepatica and 615 bp for F. gigantica. The duplex PCR failed to amplify from DNA of other common liver and intestinal trematodes, including two opisthorchiids, three heterophyids, an echinostomid, another fasciolid, and a taeniid cestode. The sensitivity assay showed that the duplex PCR limit of detection for each Fasciola species was between 0.012 ng and 0.006 ng DNA. Evaluation using DNA templates from 32 Fasciola samples (28 adults and 4 eggs) and from 25 field-collected stools of ruminants and humans revealed specific bands of the correct size and the presence of Fasciola species. This novel mtDNA duplex PCR is a sensitive and fast tool for accurate identification of Fasciola species in areas of distributional and zonal overlap.

Nguyen, Khue Thi; Nguyen, Nga Thi Bich; Doan, Huong Thi Thanh; Le, Xuyen Thi Kim; Hoang, Chau Thi Minh; De, Nguyen Van

2012-01-01

261

Speciation and host–parasite relationships in the parasite genus Gyrodactylus (Monogenea, Platyhelminthes) infecting gobies of the genus Pomatoschistus (Gobiidae, Teleostei)  

Microsoft Academic Search

Using species-level phylogenies, the speciation mode of Gyrodactylus species infecting a single host genus was evaluated. Eighteen Gyrodactylus species were collected from gobies of the genus Pomatoschistus and sympatric fish species across the distribution range of the hosts. The V4 region of the ssrRNA and the internal transcribed spacers encompassing the 5.8S rRNA gene were sequenced; by including published sequences

Tine Huyse; Vanessa Audenaert; Filip A. M. Volckaert

2003-01-01

262

Factors affecting the distribution and abundance of the commensal Temnocephala iheringi (Platyhelminthes: Temnocephalidae) among the southernmost populations of the apple snail Pomacea canaliculata (Mollusca: Ampullariidae)  

Microsoft Academic Search

Temnocephala iheringi is the most common temnocephalan inhabiting the mantle cavity of the apple snail Pomacea canaliculata, a freshwater neotropical gastropod that has become a serious rice pest in Southeastern Asia. T. iheringi has been recorded from Mato Grosso (Brazil) to water bodies associated with the Río de la Plata river (Argentina). During an extensive survey in the southern limit

Pablo R. Martín; Alejandra L. Estebenet; Silvana Burela

2005-01-01

263

A serological study of the diet of British, lake-dwelling Glossiphonia complanata (L.) (Hirudinea: Glossiphoniidae)  

Microsoft Academic Search

Monthly or two-monthly samples of G. complanata were taken from the stony littoral of five unproductive lakes in N. Wales and seven productive lakes in Cheshire and Shropshire, England, over two years. Satisfactory antisera were produced against ten potential prey groups, viz. Tricladida, Oligochaeta, Mollusca, Cladocera, Asellus, Amphipoda, Chironomidae, Ephemeroptera, Plecoptera and Trichoptera. These were used in the precipitin test

Johnstone O. Young

1981-01-01

264

Prey preference and gregarious attacks by the invasive flatworm Platydemus manokwari  

Microsoft Academic Search

The flatworm Platydemus manokwari (Tricladida: Rhynchodemidae) preys on various species of land snail, and its introduction to areas outside of its native\\u000a range is thought to have caused the extinction of native land snails on several Pacific islands. Platydemus manokwari occurs in areas where land snails have been absent since its invasion, suggesting that the flatworm can prey on animals

Shinji Sugiura

2010-01-01

265

Adaptations to life in the sulfide system: a comparison of oxygen detoxifying enzymes in thiobiotic and oxybiotic meiofauna (and freshwater planarians)  

Microsoft Academic Search

Although thiobiotic meiofauna live under anoxia, they have higher levels of the oxygen-detoxifying enzymes catalase and superoxide dismutase (SOD) than oxybiotic species. Microoxyphilic oxybios have lower enzyme activities than other oxybiotic species. All meiofauna have lower activities than oxybiotic macrofauna. Catalase and SOD activities increased in direct proportion to sulfide tolerance in thiobios and to ambient oxygen concentration in oxybios.

A. C. Morrill; E. N. Powell; R. R. Bidigare; J. M. Shick

1988-01-01

266

The Turbellaria of some Friesland lakes with incidental records of Gasteropoda and Hirudinea  

Microsoft Academic Search

INTRODUCTION During the week 7th to 12th June, 1971 opportunity was taken of accompanying members of the I.B.P. team based at the Tjeukemeer, the Netherlands, on a boat journey through the Friesland lakes. The main aim of the author was to collect Turbellaria (Tricladida and Microturbellaria) from the vegetation and stones in the littoral zone of 16 lakes (see Table

J. O. Young

1972-01-01

267

Characterization of the stem cell system of the acoel Isodiametra pulchra  

Microsoft Academic Search

BACKGROUND: Tissue plasticity and a substantial regeneration capacity based on stem cells are the hallmark of several invertebrate groups such as sponges, cnidarians and Platyhelminthes. Traditionally, Acoela were seen as an early branching clade within the Platyhelminthes, but became recently positioned at the base of the Bilateria. However, little is known on how the stem cell system in this new

Katrien De Mulder; Georg Kuales; Daniela Pfister; Maxime Willems; Bernhard Egger; Willi Salvenmoser; Marlene Thaler; Anne-Kathrin Gorny; Martina Hrouda; Gaëtan Borgonie; Peter Ladurner

2009-01-01

268

Dactylogyrids (Platyhelminthes: Monogenoidea) parasitizing butterfly fishes (Teleostei: Chaetodontidae) from the coral reefs of Palau, Moorea, Wallis, New Caledonia, and Australia: species of Euryhaliotrematoides n. gen. and Aliatrema n. gen.  

PubMed

Seven species of Euryhaliotrematoides n. gen. and 1 species of Aliatrema n. gen. (Monogenoidea; Dactylogyridae) are described and reported from the gills of 15 species of butterfly fishes (Chaetodontidae) from the coral reefs of Moorea (French Polynesia), Wallis (Wallis and Futuna), Heron and Lizard (Australia), Palau (Micronesia), and New Caledonia: Aliatrema cribbi n. sp. from Chaetodon auriga, Chaetodon lunula, Chaetodon trifasciatus, Chaetodon ulietensis, Chaetodon vagabundus, Forcipiger flavisissimus, and Heniochus chrysostomus; Euryhaliotrematoides annulocirrus n. comb. from C. auriga, C. lunula, and C. vagabundus; Euryhaliotrematoides aspistis n. sp. from C. auriga, Chaetodon citrinellus, C. lunula, Chaetodon reticulatus, C. ulietensis, and C. vagabundus; Euryhaliotrematoides berenguelae n. sp. from C. citrinellus, Chaetodon ornatissimus, and F. flavisissimus; Euryhaliotrematoides grandis n. comb. from C. auriga, C. citrinellus, Chaetodon ephippium, Chaetodon kleinii, Chaetodon lineolatus, C. lunula, C. ornatissimus, C. trifasciatus, C. vagabundus, and H. chrysostomus; Euryhaliotrematoides microphallus n. comb. from C. auriga, C. citrinellus, C. ephippium, C. kleinii, C. lunula, C. ornatissimus, C. reticulatus, Chaetodon trifascialis, C. trifasciatus, C. vagabundus, F. flavisissimus, and H. chrysostomus; Euryhaliotrematoides pirulum n. sp. from C. auriga, C. citrinellus, C. lunula, C. trifasciatus, and C. vagabundus; and Euryhaliotrematoides triangulovagina n. comb. from C. auriga, C. citrinellus, C. kleinii, C. lunula, C. ornatissimus, C. vagabundus, F. flavisissimus, H. chrysostomus, and Hemitaurichthys polylepis. All reports of previously described species are new locality records. With exceptions of E. grandis and E. annulocirrus on C. auriga and C. lunula and E. triangulovagina and E. microphallus on C. auriga, all reports are new host records. Haliotrema hainanensis and H. affinis are considered junior subjective synonyms of E. triangulovagina and E. annulocirrus, respectively. Aliatrema n. gen. is characterized by marine dactylogyrids with tandem gonads (germarium pretesticular), haptoral hooks with upright acute thumbs, a coiled copulatory organ with counterclockwise rings and funnel-shaped base but lacking an accessory piece, and a dextral vaginal pore. Euryhaliotrematoides n. gen. is characterized by marine dactylogyrids having tandem gonads (germarium pretesticular), haptoral hooks with upright acute thumbs, a coiled copulatory organ with counterclockwise rings and funnel-shaped base, a vas deferens looping the left intestinal cecum, and a dextral vaginal pore. PMID:15165056

Plaisance, Laetitia; Kritsky, Delane C

2004-04-01

269

An unusual blood sequestering tapeworm ( Sanguilevator yearsleyi n. gen., n. sp.) from Borneo with description of Cathetocephalus resendezi n. sp. from Mexico and molecular support for the recognition of the order Cathetocephalidea (Platyhelminthes: Eucestoda)  

Microsoft Academic Search

Sanguilevator yearsleyi n. gen., n. sp. and Cathetocephalus resendezi n. sp. are described from the Broadfin shark, Lamiopsis temmincki, in Malaysian Borneo and Carcharhinus leucas in Mexico, respectively. The new genus is unusual in its possession of internal chambers and channels in its scolex that appear to house extensive quantities of host white and red blood cells, respectively. Histology reveals

J. N. Caira; J. Mega; T. R. Ruhnke

2005-01-01

270

SmedGD: the Schmidtea mediterranea genome database  

Microsoft Academic Search

The planarian Schmidtea mediterranea is rapidly emerging as a model organism for the study of regeneration, tissue homeostasis and stem cell biology. The recent sequencing, assembly and annotation of its genome are expected to further buoy the biomedical importance of this organism. In order to make the extensive data associated with the genome sequence accessible to the biomedical and planarian

Sofia M. C. Robb; Eric Ross; Alejandro Sánchez Alvarado

2008-01-01

271

The Worm's Turn.  

ERIC Educational Resources Information Center

|Discusses experiments involving classical conditioning of planarians. Suitable for advanced high school students or college-level independent study, flatworms are trained to curl up under a bright light. Then the planarians to are subjected to controlled reproduction experiments to determine whether the learned behavior is inherited by their…

Miller, John C., Jr.

1983-01-01

272

Parasitic Infections of Man and Animals in Hawaii.  

National Technical Information Service (NTIS)

Contents: Classification of internal parasites of man and animals in Hawaii(Protozoa, Nemathelminthes (Roundworms), Platyhelminthes (Tapeworms and Flukes)); Life cycles of parasites; Routes of infection of internal parasites; Mollusks of parasitological i...

J. E. Alicata

1964-01-01

273

[The determination of the species classification of Baikal planarian cocoons found in the stomach of the black grayling (Thymallus arcticus baicalensis) by a comparative analysis of the nucleotide sequences of the ribosomal RNA gene].  

PubMed

Comparative analysis of nucleotide sequences of gene 18S of ribosome RNA was carried out. The results show that the genetic sequences of the given locus could be used as a molecular marker to identify the species of planaria irrespective of ontogenetic stage. The articles deals with problem of specific determination of cocoons of Baikal planaria from the stomach of Baikal black grayling using comparative analysis of nucleotide sequences of ribosome RNA fragments with known sequences determined earlier for Baikal planaria. The cocoons belong to two species of Rimacephalus. The authors discuss also the importance of feeding relationships of planaria and benthophage fish to investigate the biotic factors that influence the evolution of Baikal planaria. PMID:10520297

Kuznedelov, K D; Dziuba, E V

274

Phylogenetic position of Nemertea derived from phylogenomic data.  

PubMed

Nemertea and Platyhelminthes have traditionally been grouped together because they possess a so-called acoelomate organization, but lateral vessels and rhynchocoel of nemerteans have been regarded as coelomic cavities. Additionally, both taxa show spiral cleavage patterns prompting the placement of Nemertea as sister to coelomate Protostomia, that is, either to Neotrochozoa (Mollusca and Annelida) or to Teloblastica (Neotrochozoa plus Arthropoda). Some workers maintain a sister group relationship of Nemertea and Platyhelminthes as Parenchymia because of an assumed homology of Götte's and Müller's larvae of polyclad Platyhelminthes and the pilidium larvae of heteronemerteans. So far, molecular data were only able to significantly reject a sister group relationship to Teloblastica. Although phylogenomic data are available for Platyhelminthes, Annelida, Mollusca, and Arthropoda, they are lacking for Nemertea. Herein, we present the first analysis specifically addressing nemertean phylogenetic position using phylogenomic data. More specifically, we collected expressed sequence tag data from Lineus viridis (O.F. Müller, 1774) and combined it with available data to produce a data set of 9,377 amino acid positions from 60 ribosomal proteins. Maximum likelihood analyses and Bayesian inferences place Nemertea in a clade together with Annelida and Mollusca. Furthermore, hypothesis testing significantly rejected a sister group relationship to either Platyhelminthes or Teloblastica. The Coelomata hypothesis, which groups coelomate taxa together to the exclusion of acoelomate and pseudocoelomate taxa, is not congruent with our results. Thus, the supposed acoelomate organization evolved independently in Nemertea and Platyhelminthes. In Nemertea, evolution of acoely is most likely due to a secondary reduction of the coelom as it is found in certain species of Mollusca and Annelida. Though looking very similar, the Götte's and Müller's larvae of polyclad Platyhelminthes are not homologous to the pilidium larvae of heteronemerteans. Finally, the convergent evolution of segmentation in Annelida and Arthropoda is further substantiated. PMID:18222945

Struck, Torsten H; Fisse, Frauke

2008-01-24

275

Innate sexuality determines the mechanisms of telomere maintenance.  

PubMed

Recently, telomere length has been shown to be differentially regulated in asexually and sexually reproducing planarians. In addition, it was found that asexual worms maintain telomere length somatically during reproduction by fission or when regeneration is induced by amputation, whereas sexual worms only achieve telomere elongation through sexual reproduction. We have established an experimental bioassay system to induce switching from asexual to sexual reproduction in planarians, that is, sexualization. In this study, the relationship between the reproductive mode and telomere maintenance was investigated using innate asexually reproducing worms, innate sexually reproducing worms, and experimentally sexualized worms. Here, we show that innate asexual planarians maintain telomere length during cell division and that innate sexual planarians exhibit telomere shortening. However, experimental sexualized worms maintain telomere length during cell division. These results indicate that innate sexuality is linked to the mechanism of telomere maintenance. PMID:23319366

Tasaka, Kenta; Yokoyama, Naoki; Nodono, Hanae; Hoshi, Motonori; Matsumoto, Midori

2013-01-01

276

Small RNA pathways in Schmidtea mediterranea.  

PubMed

Planarians are bilaterally symmetrical fresh water organisms capable of regenerating body parts from small fragments following bodily injury. Planarians possess a specialized population of pluripotent cells called neoblasts, which are responsible for their unique regenerative ability. The study of planarian stem cell biology and regeneration has traditionally focused on the transcription factors and proteins that regulate signal transduction pathways. New evidence shows that small RNA molecules are important players in stem cell function and regeneration, yet little is known about the exact nature of their regulatory roles during the regenerative process. In this review, we discuss biogenesis of microRNAs and piwiRNAs and their functional role in key developmental pathways in vertebrates and invertebrates with an emphasis on recent studies on planarian small RNA pathways. PMID:22450996

Resch, Alissa M; Palakodeti, Dasaradhi

2012-01-01

277

SmedGD: the Schmidtea mediterranea genome database.  

PubMed

The planarian Schmidtea mediterranea is rapidly emerging as a model organism for the study of regeneration, tissue homeostasis and stem cell biology. The recent sequencing, assembly and annotation of its genome are expected to further buoy the biomedical importance of this organism. In order to make the extensive data associated with the genome sequence accessible to the biomedical and planarian communities, we have created the Schmidtea mediterranea Genome Database (SmedGD). SmedGD integrates in a single web-accessible portal all available data associated with the planarian genome, including predicted and annotated genes, ESTs, protein homologies, gene expression patterns and RNAi phenotypes. Moreover, SmedGD was designed using tools provided by the Generic Model Organism Database (GMOD) project, thus making its data structure compatible with other model organism databases. Because of the unique phylogenetic position of planarians, SmedGD (http://smedgd.neuro.utah.edu) will prove useful not only to the planarian research community, but also to those engaged in developmental and evolutionary biology, comparative genomics, stem cell research and regeneration. PMID:17881371

Robb, Sofia M C; Ross, Eric; Sánchez Alvarado, Alejandro

2007-09-18

278

Regeneration of neuronal cell types in Schmidtea mediterranea: an immunohistochemical and expression study.  

PubMed

Freshwater planarians are unique in their ability to regenerate a complete Central Nervous System (CNS) from almost any small piece of their body in just a few days. The planarian CNS contains a pair of anterior cephalic ganglia lying on top of two ventral nerve cords that extend along the length of the animal. Studies of planarian CNS regeneration have generally used pan-neural markers, which provide only a general overview of the process. Nevertheless, some reports have started to characterize the genes that are required for this process. In this study, to obtain a more detailed description of planarian neural regeneration, we monitored the regeneration of neuronal populations specifically labelled with antibodies against serotonin, allatostatin, neuropeptide F, GYRFamide and FMRFamide. We also characterized the regeneration of dopaminergic and octopaminergic cell populations by in situ hybridization. Finally, we characterized the expression pattern of a set of receptors for neurotransmitters, neuropeptides and hormones that are suggested to play a role in the regeneration process itself. Together, these data provide a more detailed description of the cellular events occurring during anterior and posterior CNS regeneration in planarians and provide the foundations for future mechanistic studies into the regeneration process in this important model system. PMID:22451002

Fraguas, Susanna; Barberán, Sara; Ibarra, Begońa; Stöger, Linda; Cebriŕ, Francesc

2012-01-01

279

Toward next-generation sequencing of mitochondrial genomes — Focus on parasitic worms of animals and biotechnological implications  

Microsoft Academic Search

Helminths (worms) include parasitic nematodes (roundworms) and platyhelminths (flatworms). These worms are abundant, and many of them are of agricultural, aquacultural, veterinary and medical importance and cause substantial socioeconomic losses worldwide. The genetic characterization of parasitic nematodes using advanced molecular tools is central to the diagnosis of infections and the control of parasitism. The accurate analysis of genetic variation also

Aaron R. Jex; D. Timothy J. Littlewood; Robin B. Gasser

2010-01-01

280

Parasitic incidence in a cyprinid fish Labeo rohita (Ham.) at river Song in Doon valley (Uttarakhand)  

Microsoft Academic Search

On examining 144 specimens of Labeo rohita (Ham.) for parasites occurrence; as many as 34 fish were found positive to harbour ciliophorans (Trichodina sp. and Ichthyophthirius multifilis Fouquet, 1876), monogeneans (Haplocleidus vachi Tripathi, 1959 and Dactylogyrus glossogobii Jain, 1960), trematode (Allocreadium mahaseri Pandey, 1939), nematode (Camallanus (Zeylanema) anabantis Pearse, 1933) and acanthocephalan (Sachalinorhynchus sp). Highest prevalence was shown by platyhelminthes

Jayti Upadhyay; R. K. Jauhari; N. Pemola Devi

281

Identification of Echinococcus granulosus microRNAs and their expression in different life cycle stages and parasite genotypes  

Microsoft Academic Search

The aetiological agent of cystic hydatid disease, the platyhelminth parasite Echinococcus granulosus, undergoes a series of metamorphic events during its complex life cycle. One of its developmental stages, the protoscolex, shows a remarkable degree of heterogeneous morphogenesis, being able to develop either into the vesicular or strobilar direction. Another level of complexity is added by the existence of genotypes or

M. Cucher; L. Prada; G. Mourglia-Ettlin; S. Dematteis; F. Camicia; S. Asurmendi; M. Rosenzvit

2011-01-01

282

Thyroid hormone receptor orthologues from invertebrate species with emphasis on Schistosoma mansoni  

Microsoft Academic Search

BACKGROUND:: Thyroid hormone receptors (TRs) function as molecular switches in response to thyroid hormone to regulate gene transcription. TRs were previously believed to be present only in chordates. RESULTS:: We isolated two TR genes from the Schistosoma mansoni and identified TR orthologues from other invertebrates: the platyhelminths, S. japonium and Schmidtea mediterranea, the mollusc, Lottia gigantean and the arthropod Daphnia

Wenjie Wu; Edward G Niles; Philip T LoVerde

2007-01-01

283

Asexual reproduction and the turbellarian archetype  

Microsoft Academic Search

The turbellarian archetype is widely believed to have been a hermaphrodite lacking asexual reproduction, and such asexual reproduction as is now seen in the Turbellaria (as paratomy and architomy) is generally assumed to have arisen secondarily several times independently. Asexual reproduction clearly prevails among the most primitive metazoans such as the placozoans, sponges, and radiates, however, and if the Platyhelminthes

Reinhard M. Rieger

1986-01-01

284

Back in time: a new systematic proposal for the Bilateria  

Microsoft Academic Search

Conventional wisdom suggests that bilateral organisms arose from ancestors that were radially, rather than bilaterally, symmetrical and, therefore, had a single body axis and no mesoderm. The two main hypotheses on how this transformation took place consider either a simple organism akin to the planula larva of extant cnidarians or the acoel Platyhelminthes (planuloid-acoeloid theory), or a rather complex organism

Pere Martinez; Jordi Paps; Marta Riutort

2009-01-01

285

Elucidating the transcriptome of Fasciola hepatica — A key to fundamental and biotechnological discoveries for a neglected parasite  

Microsoft Academic Search

Liver flukes of animals are parasitic flatworms (Platyhelminthes: Digenea) of major socioeconomic importance in many countries. Key representatives, such as Fasciola hepatica and F. gigantica, cause “liver fluke disease” (= fascioliasis), which is of major animal health significance worldwide. In particular, F. hepatica is a leading cause of production losses to the livestock (mainly sheep and cattle) and meat industries

Neil D. Young; Ross S. Hall; Aaron R. Jex; Cinzia Cantacessi; Robin B. Gasser

2010-01-01

286

Temnocephalan symbionts of the freshwater crayfish Cherax quadricarinatus from northern Australia  

Microsoft Academic Search

Four species of turbellarian temnocephalan symbionts (Platyhelminthes: Temnocephalida) are reported for the first time from the external surfaces of Cherax quadricarinatus, a freshwater crayfish from northern Australia. Three of these species — Temnocephala rouxii Merton, 1913, Notodactylus handschini (Baer, 1945), and Diceratocephala boschmai Baer, 1953 — are known previously from related crayfish in New Guinea. The newly discovered fourth species,

Lester R. G. Cannon

1991-01-01

287

A New Family of Receptor Tyrosine Kinases with a Venus Flytrap Binding Domain in Insects and Other Invertebrates Activated by Aminoacids  

Microsoft Academic Search

BackgroundTyrosine kinase receptors (RTKs) comprise a large family of membrane receptors that regulate various cellular processes in cell biology of diverse organisms. We previously described an atypical RTK in the platyhelminth parasite Schistosoma mansoni, composed of an extracellular Venus flytrap module (VFT) linked through a single transmembrane domain to an intracellular tyrosine kinase domain similar to that of the insulin

Arnaud Ahier; Philippe Rondard; Nadčge Gouignard; Naji Khayath; Siluo Huang; Jacques Trolet; Daniel J. Donoghue; Monique Gauthier; Jean-Philippe Pin; Colette Dissous; Gordon Langsley

2009-01-01

288

A New Family of Receptor Tyrosine Kinases with a Venus Flytrap Binding Domain in Insects and Other Invertebrates Activated by Aminoacids  

Microsoft Academic Search

Background: Tyrosine kinase receptors (RTKs) comprise a large family of membrane receptors that regulate various cellular processes in cell biology of diverse organisms. We previously described an atypical RTK in the platyhelminth parasite Schistosoma mansoni, composed of an extracellular Venus flytrap module (VFT) linked through a single transmembrane domain to an intracellular tyrosine kinase domain similar to that of the

Arnaud Ahier; Philippe Rondard; Nadčge Gouignard; Naji Khayath; Siluo Huang; Jacques Trolet; Daniel J. Donoghue; Monique Gauthier; Jean-Philippe Pin; Colette Dissous

2009-01-01

289

Effect of Gamma Irradiation on Different Stages of Fasciola hepatica.  

National Technical Information Service (NTIS)

Fascioliasis or hepatic distomatosis is one of the most widespread liver diseases throughout the world. It is caused by a parasitic worm of the class Trematoda of the phylum Platyhelminthes. This flatworm is commonly known as liver-fluke in the United Sta...

J. L. Torres J. Chiriboga

1976-01-01

290

Anthelmintic efficacy of Flemingia vestita: genistein-induced effect on the activity of nitric oxide synthase and nitric oxide in the trematode parasite, Fasciolopsis buski  

Microsoft Academic Search

The root-tuber peel of Flemingia vestita, an indigenous leguminous plant of Meghalaya (Northeast India), has usage in local traditional medicine as curative against worm infections. The peel and its active component, genistein, have been shown to cause flaccid paralysis, deformity of tegumental architecture and alterations in the activity of several enzymes in platyhelminth parasites. To investigate further the mode of

Pradip K Kar; Veena Tandon; Nirmalendu Saha

2002-01-01

291

Mitochondrial genetic code in cestodes  

Microsoft Academic Search

The flatworm mitochondrial genetic code, which has been used for all species of the Platyhelminthes, is mainly characterized by AUA codon for isoleucine, AAA codon for asparagine and UAA codon for tyrosine. In eight species of cestodes (Echinococcus multilocularis, Echinococcus granulosus, Taenia solium, Taenia saginata, Taenia hydatigena, Taenia crassiceps, Hymenolepis nana and Mesocestoides corti), the cytochrome c oxidase subunit I

Minoru Nakao; Yasuhito Sako; Noriko Yokoyama; Masahito Fukunaga; Akira Ito

2000-01-01

292

Using parasites to infer host population history: a new rationale for parasite conservation  

Microsoft Academic Search

Only one of the 5000 extant louse species (Phthiraptera) and no species of flea (Siphonaptera), parasitic helminth (Platyhelminthes), parasitic nematode (Nemata), mite, or tick (Acari) is listed as threatened by the IUCN, despite impassioned pleas for parasite conservation beginning more than a decade ago. Although they should be conserved for their own sake, past arguments, highlighting the intrinsic and utilitarian

Noah Kerness Whiteman; Patricia G. Parker

2005-01-01

293

To Be or Not to Be a Flatworm: The Acoel Controversy  

PubMed Central

Since first described, acoels were considered members of the flatworms (Platyhelminthes). However, no clear synapomorphies among the three large flatworm taxa - the Catenulida, the Acoelomorpha and the Rhabditophora - have been characterized to date. Molecular phylogenies, on the other hand, commonly positioned acoels separate from other flatworms. Accordingly, our own multi-locus phylogenetic analysis using 43 genes and 23 animal species places the acoel flatworm Isodiametra pulchra at the base of all Bilateria, distant from other flatworms. By contrast, novel data on the distribution and proliferation of stem cells and the specific mode of epidermal replacement constitute a strong synapomorphy for the Acoela plus the major group of flatworms, the Rhabditophora. The expression of a piwi-like gene not only in gonadal, but also in adult somatic stem cells is another unique feature among bilaterians. These two independent stem-cell-related characters put the Acoela into the Platyhelminthes-Lophotrochozoa clade and account for the most parsimonious evolutionary explanation of epidermal cell renewal in the Bilateria. Most available multigene analyses produce conflicting results regarding the position of the acoels in the tree of life. Given these phylogenomic conflicts and the contradiction of developmental and morphological data with phylogenomic results, the monophyly of the phylum Platyhelminthes and the position of the Acoela remain unresolved. By these data, both the inclusion of Acoela within Platyhelminthes, and their separation from flatworms as basal bilaterians are well-supported alternatives.

Arendt, Detlev; Borgonie, Gaetan; Funayama, Noriko; Gschwentner, Robert; Hartenstein, Volker; Hobmayer, Bert; Hooge, Matthew; Hrouda, Martina; Ishida, Sachiko; Kobayashi, Chiyoko; Kuales, Georg; Nishimura, Osamu; Pfister, Daniela; Rieger, Reinhard; Salvenmoser, Willi; Smith, Julian; Technau, Ulrich; Tyler, Seth; Agata, Kiyokazu; Salzburger, Walter; Ladurner, Peter

2009-01-01

294

A limnological reconnaissance of the Falkland Islands; with particular reference to the waterfleas (Arthropoda: Anomopoda)  

Microsoft Academic Search

Forty?eight freshwater bodies on the Falkland Islands, including 33 lakes and pools, and 12 rivers and streams, were sampled for freshwater invertebrates. This study yielded 129 species of invertebrates (79 Rotifera, 34 Arthropoda, six Platyhelminthes, three Gastrotricha, two Nematoda, two Annelida, two Mollusca, and one Tardigrada) plus two fish species bringing the known Falkland Islands freshwater fauna to more than

Herbert J. G. Dartnall; Werner Hollwedel

2007-01-01

295

Variation in worm assemblages associated with Pomacea canaliculata (Caenogastropoda, Ampullariidae) in sites near the Río de la Plata estuary, Argentina  

Microsoft Academic Search

Pomacea canaliculata is a common gastropod in freshwater habitats from Central and North- ern Argentina, extending northwards into the Amazon basin. Several Platyhelminthes have been reported associated to P. canaliculata, sharing an intimate relationship with this gastropod host. The objectives of this study were to describe the symbiotic species assemblages associated to P. canaliculata in the study area, and to

C. DAMBORENEA; F. BRUSA; A. PAOLA

296

Suppression of cathepsin B expression in Schistosoma mansoni by RNA interference  

Microsoft Academic Search

In this paper, we used the genetic manipulation technique known as RNA-interference to suppress the expression of a target, cathepsin B, gene in the platyhelminth parasite, Schistosoma mansoni. Parasites were cultured for 6 days in the presence of double stranded RNA derived from the cathepsin B cDNA sequence or from two control sequences. Relative to the controls, the cathepsin B

Patrick J Skelly; Akram Da'dara; Donald A Harn

2003-01-01

297

Specialized progenitors and regeneration.  

PubMed

Planarians are flatworms capable of regenerating all body parts. Planarian regeneration requires neoblasts, a population of dividing cells that has been studied for over a century. Neoblast progeny generate new cells of blastemas, which are the regenerative outgrowths at wounds. If the neoblasts comprise a uniform population of cells during regeneration (e.g. they are all uncommitted and pluripotent), then specialization of new cell types should occur in multipotent, non-dividing neoblast progeny cells. By contrast, recent data indicate that some neoblasts express lineage-specific transcription factors during regeneration and in uninjured animals. These observations raise the possibility that an important early step in planarian regeneration is the specialization of neoblasts to produce specified rather than naďve blastema cells. PMID:23404104

Reddien, Peter W

2013-03-01

298

Specialized progenitors and regeneration  

PubMed Central

Planarians are flatworms capable of regenerating all body parts. Planarian regeneration requires neoblasts, a population of dividing cells that has been studied for over a century. Neoblast progeny generate new cells of blastemas, which are the regenerative outgrowths at wounds. If the neoblasts comprise a uniform population of cells during regeneration (e.g. they are all uncommitted and pluripotent), then specialization of new cell types should occur in multipotent, non-dividing neoblast progeny cells. By contrast, recent data indicate that some neoblasts express lineage-specific transcription factors during regeneration and in uninjured animals. These observations raise the possibility that an important early step in planarian regeneration is the specialization of neoblasts to produce specified rather than naďve blastema cells.

Reddien, Peter W.

2013-01-01

299

Deciphering the molecular machinery of stem cells: a look at the neoblast gene expression profile  

PubMed Central

Background Mammalian stem cells are difficult to access experimentally; model systems that can regenerate offer an alternative way to characterize stem cell related genes. Planarian regeneration depends on adult pluripotent stem cells - the neoblasts. These cells can be selectively destroyed using X-rays, enabling comparison of organisms lacking stem cells with wild-type worms. Results Using a genomic approach we produced an oligonucleotide microarray chip (the Dj600 chip), which was designed using selected planarian gene sequences. Using this chip, we compared planarians treated with high doses of X-rays (which eliminates all neoblasts) with wild-type worms, which led to identification of a set of putatively neoblast-restricted genes. Most of these genes are involved in chromatin modeling and RNA metabolism, suggesting that epigenetic modifications and post-transcriptional regulation are pivotal in neoblast regulation. Comparing planarians treated with low doses of X-rays (after which some radiotolerant neoblasts re-populate the planarian body) with specimens irradiated with high doses and unirradiated control worms, we identified a group of genes that were upregulated as a consequence of low-dose X-ray treatment. Most of these genes encode proteins that are known to regulate the balance between death and survival of the cell; our results thus suggest that genetic programs that control neoblast cytoprotection, proliferation, and migration are activated by low-dose X-rays. Conclusion The broad differentiation potential of planarian neoblasts is unparalleled by any adult stem cells in the animal kingdom. In addition to our validation of the Dj600 chip as a valuable platform, our work contributes to elucidating the molecular mechanisms that regulate the self-renewal and differentiation of neoblasts.

Rossi, Leonardo; Salvetti, Alessandra; Marincola, Francesco M; Lena, Annalisa; Deri, Paolo; Mannini, Linda; Batistoni, Renata; Wang, Ena; Gremigni, Vittorio

2007-01-01

300

The cellular basis for animal regeneration  

PubMed Central

The ability of animals to regenerate missing parts is a dramatic and poorly understood aspect of biology. The sources of new cells for these regenerative phenomena have been sought for decades. Recent advances involving cell fate tracking in complex tissues have shed new light on the cellular underpinnings of regeneration in Hydra, planarians, zebrafish, Xenopus, and Axolotl. Planarians accomplish regeneration with use of adult pluripotent stem cells, whereas several vertebrates utilize a collection of lineage-restricted progenitors from different tissues. Together, an array of cellular strategies—from pluripotent stem cells to tissue-specific stem cells and dedifferentiation—are utilized for regeneration.

Tanaka, Elly; Reddien, Peter W.

2011-01-01

301

Regeneration: The origin of cancer or a possible cure?  

Microsoft Academic Search

A better understanding of the forces controlling cell growth will be essential for developing effective therapies in regenerative medicine and cancer. Historically, the literature has linked cancer and tissue regeneration—proposing regeneration as both the source of cancer and a method to inhibit tumorigenesis. This review discusses two powerful regeneration models, the vertebrate urodele amphibians and invertebrate planarians, in light of

Néstor J. Oviedo; Wendy S. Beane

2009-01-01

302

Combining Classical and Molecular Approaches Elaborates on the Complexity of Mechanisms Underpinning Anterior Regeneration  

Microsoft Academic Search

The current model of planarian anterior regeneration evokes the establishment of low levels of Wnt signalling at anterior wounds, promoting anterior polarity and subsequent elaboration of anterior fate through the action of the TALE class homeodomain PREP. The classical observation that decapitations positioned anteriorly will regenerate heads more rapidly than posteriorly positioned decapitations was among the first to lead to

Deborah J. Evans; Suthira Owlarn; Belen Tejada Romero; Chen Chen; A. Aziz Aboobaker

2011-01-01

303

Inhibition of Regeneration of Planaria by Midlethal Exposures to X Rays.  

National Technical Information Service (NTIS)

The paper reports the results of studies on the radiosensitivity of the planarian worm, Dugesis dorotocephala, and on the effect of X-rays on the capacity to regenerate after decapitation. The LD50/60 of intact worms is 1303 R (95% confidence interval 119...

D. S. Nachtwey K. Kendall

1968-01-01

304

Temporally-Patterned Magnetic Fields Induce Complete Fragmentation in Planaria  

PubMed Central

A tandem sequence composed of weak temporally-patterned magnetic fields was discovered that produced 100% dissolution of planarian in their home environment. After five consecutive days of 6.5 hr exposure to a frequency-modulated magnetic field (0.1 to 2 µT), immediately followed by an additional 6.5 hr exposure on the fifth day, to another complex field (0.5 to 5 µT) with exponentially increasing spectral power 100% of planarian dissolved within 24 hr. Reversal of the sequence of the fields or presentation of only one pattern for the same duration did not produce this effect. Direct video evidence showed expansion (by visual estimation ?twice normal volume) of the planarian following the first field pattern followed by size reduction (estimated ?1/2 of normal volume) and death upon activation of the second pattern. The contortions displayed by the planarian during the last field exposure suggest effects on contractile proteins and alterations in the cell membrane’s permeability to water.

Murugan, Nirosha J.; Karbowski, Lukasz M.; Lafrenie, Robert M.; Persinger, Michael A.

2013-01-01

305

Early embryogenesis of planaria: a cryptic larva feeding on maternal resources  

Microsoft Academic Search

The early planarian embryo presents a complete ciliated epidermis and a pharynx and feeds on maternal yolk cells. In this paper, we report on all the elements involved in the formation of such an autonomous embryo, which we name cryptic larva. First, we provide a description of the spherical and fusiform yolk cells and their relationship with the blastomeres, from

Albert Cardona; Volker Hartenstein; Rafael Romero

2006-01-01

306

Teratological research using in vitro systems. V. Nonmammalian model systems.  

PubMed Central

In this review of alternative tests to whole-animal rodent studies, the use of sub-mammalian and sub-vertebrate systems is investigated. The history, methodology, known limitations, end points, dose response, and requirements of virus, hydra, planarian, cricket, fish, amphibia, Drosophila, and chicken embryo systems are discussed.

Collins, T F

1987-01-01

307

Smed454 dataset: unravelling the transcriptome of Schmidtea mediterranea  

PubMed Central

Background Freshwater planarians are an attractive model for regeneration and stem cell research and have become a promising tool in the field of regenerative medicine. With the availability of a sequenced planarian genome, the recent application of modern genetic and high-throughput tools has resulted in revitalized interest in these animals, long known for their amazing regenerative capabilities, which enable them to regrow even a new head after decapitation. However, a detailed description of the planarian transcriptome is essential for future investigation into regenerative processes using planarians as a model system. Results In order to complement and improve existing gene annotations, we used a 454 pyrosequencing approach to analyze the transcriptome of the planarian species Schmidtea mediterranea Altogether, 598,435 454-sequencing reads, with an average length of 327 bp, were assembled together with the ~10,000 sequences of the S. mediterranea UniGene set using different similarity cutoffs. The assembly was then mapped onto the current genome data. Remarkably, our Smed454 dataset contains more than 3 million novel transcribed nucleotides sequenced for the first time. A descriptive analysis of planarian splice sites was conducted on those Smed454 contigs that mapped univocally to the current genome assembly. Sequence analysis allowed us to identify genes encoding putative proteins with defined structural properties, such as transmembrane domains. Moreover, we annotated the Smed454 dataset using Gene Ontology, and identified putative homologues of several gene families that may play a key role during regeneration, such as neurotransmitter and hormone receptors, homeobox-containing genes, and genes related to eye function. Conclusions We report the first planarian transcript dataset, Smed454, as an open resource tool that can be accessed via a web interface. Smed454 contains significant novel sequence information about most expressed genes of S. mediterranea. Analysis of the annotated data promises to contribute to identification of gene families poorly characterized at a functional level. The Smed454 transcriptome data will assist in the molecular characterization of S. mediterranea as a model organism, which will be useful to a broad scientific community.

2010-01-01

308

The urbilaterian brain revisited: novel insights into old questions from new flatworm clades.  

PubMed

Flatworms are classically considered to represent the simplest organizational form of all living bilaterians with a true central nervous system. Based on their simple body plans, all flatworms have been traditionally grouped together in a single phylum at the base of the bilaterians. Current molecular phylogenomic studies now split the flatworms into two widely separated clades, the acoelomorph flatworms and the platyhelminth flatworms, such that the last common ancestor of both clades corresponds to the urbilaterian ancestor of all bilaterian animals. Remarkably, recent comparative neuroanatomical analyses of acoelomorphs and platyhelminths show that both of these flatworm groups have complex anterior brains with surprisingly similar basic neuroarchitectures. Taken together, these findings imply that fundamental neuroanatomical features of the brain in the two separate flatworm groups are likely to be primitive and derived from the urbilaterian brain. PMID:23143292

Bailly, Xavier; Reichert, Heinrich; Hartenstein, Volker

2012-11-10

309

Gnathostomulida—An Enigmatic Metazoan Phylum from both Morphological and Molecular Perspectives  

Microsoft Academic Search

On the basis of few and contentious morphological characters Gnathostomulids have been thought to be the sister-group of either the Platyhelminthes or the Syndermata (Rotifera+Acanthocephala). We provide a full 18S rDNA sequence for a species ofGnathostomulaand attempt to resolve its position among the Metazoa, on the basis of molecular evidence. Sixty sequences, representing 30 nominal phyla and including new entoproct

D. Timothy J. Littlewood; Maximilian J. Telford; Karen A. Clough; Klaus Rohde

1998-01-01

310

DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates.  

PubMed

We describe "universal" DNA primers for polymerase chain reaction (PCR) amplification of a 710-bp fragment of the mitochondrial cytochrome c oxidase subunit I gene (COI) from 11 invertebrate phyla: Echinodermata, Mollusca, Annelida, Pogonophora, Arthropoda, Nemertinea, Echiura, Sipuncula, Platyhelminthes, Tardigrada, and Coelenterata, as well as the putative phylum Vestimentifera. Preliminary comparisons revealed that these COI primers generate informative sequences for phylogenetic analyses at the species and higher taxonomic levels. PMID:7881515

Folmer, O; Black, M; Hoeh, W; Lutz, R; Vrijenhoek, R

1994-10-01

311

Phylogenetic Position of Phylum Nemertini, Inferred from 18s rRNA Sequences: Molecular Data as a Test of Morphological Character Homology  

Microsoft Academic Search

Partial 18s rRNA sequence of the nemertine Cerebratulus lacteus was obtained and compared with those of coelomate metazoans and acoelomate platyhelminths to test whether nemertines share a most recent common ancestor with the platy- helminths, as traditionally has been implied, or whether nemertines lie within a protostome coelomate clade, as suggested by more recent morphological analyses. Maximum-parsimony analysis supports the

J. McClintock Turbeville; Katharine G. Field

1992-01-01

312

Genetic characterisation of Fasciola samples from different host species and geographical localities revealed the existence of F. hepatica and F. gigantica in Niger  

Microsoft Academic Search

In the present study, 16 samples representing Fasciola (Platyhelminthes: Trematoda: Digenea) from sheep and cattle from seven geographical locations in Niger were characterized\\u000a genetically by sequences of the first (ITS-1) and second (ITS-2) internal transcribed spacers (ITS) of nuclear ribosomal DNA\\u000a (rDNA). The ITS rDNA was amplified from individual liver flukes by polymerase chain reaction (PCR), and the amplicons were

H. Ali; L. Ai; H. Q. Song; S. Ali; R. Q. Lin; B. Seyni; G. Issa; X. Q. Zhu

2008-01-01

313

Characterization of Fasciola samples from different host species and geographical localities in Spain by sequences of internal transcribed spacers of rDNA  

Microsoft Academic Search

In the present study, 25 samples representing Fasciola (Platyhelminthes: Trematoda: Digenea) from nine host species and 19 geographical locations in Spain were characterized genetically\\u000a by sequences of the first (ITS-1) and second (ITS-2) internal transcribed spacers (ITS) of nuclear ribosomal DNA (rDNA). The\\u000a ITS rDNA was amplified from individual liver flukes by polymerase chain reaction (PCR), and the amplicons were

S. Alasaad; C. Q. Huang; Q. Y. Li; J. E. Granados; C. García-Romero; J. M. Pérez; X. Q. Zhu

2007-01-01

314

Genetic characterization of Fasciola hepatica from Tunisia and Algeria based on mitochondrial and nuclear DNA sequences  

Microsoft Academic Search

Fasciolosis caused by Fasciola spp. (Platyhelminthes: Trematoda: Digenea) is considered the most important helminth infection of ruminants in tropical countries,\\u000a causing considerable socioeconomic problems. In the present study, samples identified morphologically as Fasciola hepatica from sheep and cattle from different geographical locations of Tunisia and Algeria were genetically characterised by sequences\\u000a of the first (ITS-1), the 5.8S and second (ITS-2)

Sarra Farjallah; Daria Sanna; Nabil Amor; Benakriche Ben Mehel; Maria Cristina Piras; Paolo Merella; Marco Casu; Marco Curini-Galletti; Khaled Said; Giovanni Garippa

2009-01-01

315

Molecular Phylogeny of Metazoan Intermediate Filament Proteins  

Microsoft Academic Search

.   We have cloned cytoplasmic intermediate filament (IF) proteins from a large number of invertebrate phyla using cDNA probes,\\u000a the monoclonal antibody IFA, peptide sequence information, and various RT-PCR procedures. Novel IF protein sequences reported\\u000a here include the urochordata and nine protostomic phyla, i.e., Annelida, Brachiopoda, Chaetognatha, Echiura, Nematomorpha,\\u000a Nemertea, Platyhelminthes, Phoronida, and Sipuncula. Taken together with the wealth of

Andreas Erber; Dieter Riemer; Marc Bovenschulte; Klaus Weber

1998-01-01

316

Neuroschistosomiasis  

Microsoft Academic Search

Schistosomiasis (bilharzia) is a neglected tropical disease caused by digenetic trematode platyhelminths of the genus Schistosoma. Neuroschistosomiasis is one of the most severe clinical outcomes associated with schistosome infection. Neurological complications\\u000a early during the course of infection are thought to occur through in situ egg deposition following aberrant migration of adult\\u000a worms to the brain or spinal cord. The presence

Allen G. RossDonald; Donald P. McManus; Jeremy Farrar; Richard J. Hunstman; Darren J. Gray; Yue-Sheng Li

317

A phylogenetic analysis of myosin heavy chain type II sequences corroborates that Acoela and Nemertodermatida are basal bilaterians  

PubMed Central

Bilateria are currently subdivided into three superclades: Deuterostomia, Ecdysozoa, and Lophotrochozoa. Within this new taxonomic frame, acoelomate Platyhelminthes, for a long time held to be basal bilaterians, are now considered spiralian lophotrochozoans. However, recent 18S rDNA [small subunit (SSU)] analyses have shown Platyhelminthes to be polyphyletic with two of its orders, the Acoela and the Nemertodermatida, as the earliest extant bilaterians. To corroborate such position and avoid the criticisms of saturation and long-branch effects thrown on the SSU molecule, we have searched for independent molecular data bearing good phylogenetic information at deep evolutionary nodes. Here we report a phylogenetic analysis of DNA sequences from the myosin heavy chain type II (myosin II) gene from a large set of metazoans, including acoels and nemertodermatids. Our study demonstrates, both for the myosin II data set alone and for a combined SSU + myosin II data set, that Platyhelminthes are polyphyletic and that acoels and nemertodermatids are the extant earliest bilaterians. Hence, the common bilaterian ancestor was not, as currently held, large and complex but small, simple, and likely with direct development. This scenario has far-reaching implications for understanding the evolution of major body plans and for perceptions of the Cambrian evolutionary explosion.

Ruiz-Trillo, I.; Paps, J.; Loukota, M.; Ribera, C.; Jondelius, U.; Baguna, J.; Riutort, M.

2002-01-01

318

Combined effect of invertebrate predation and sublethal pesticide exposure on the behavior and survival of Asellus aquaticus (Crustacea; Isopoda).  

PubMed

Invertebrate communities of lentic habitats comprise, amongst others, the crustacean Asellus aquaticus (Isopoda) and the turbellarian Dendrocoelum lacteum (Tricladida). Because D. lacteum preferentially preys on A. aquaticus, contaminants introduced into the aquatic environment may affect this predator-prey interaction, finally influencing the performance of the predator. However, no studies investigating implications of organic pollutants on this food web subsystem currently exist. Hence, the present study assessed short-term implications of pesticides with different modes of action, namely, the triazole fungicide tebuconazole and the pyrethroid insecticide lambda-cyhalothrin, during a 72 h trial. The experiments for tebuconazole showed a statistically significant decrease in predatory success of D. lacteum. Lambda-cyhalothrin, in contrast, increased predation success by 40%, which is, however, not statistically significant. Both the decrease and the increase in predation seemed to be primarily driven by an altered activity of the prey A. aquaticus. This may be hypothesized because any shift in the prey's activity influenced its probability to stick to mucus, a viscous substance released by D. lacteum, or to encounter the predator directly. PMID:22223070

Bundschuh, Mirco; Appeltauer, Andreas; Dabrunz, André; Schulz, Ralf

2012-01-06

319

Effects and molecular mechanisms of the biological action of weak and extremely weak magnetic fields  

Microsoft Academic Search

A number of effects of weak combined (static and alternating) magnetic fields with an alternating component of tens and hundreds\\u000a nT at a collinear static field of 42 ?T, which is equivalent to the geomagnetic field, have been found: activation of fission\\u000a and regeneration of planarians Dugesia tigrina, inhibition of the growth of the Ehrlich ascites carcinoma in mice, stimulation

V. V. Novikov; V. O. Ponomarev; G. V. Novikov; V. V. Kuvichkin; E. V. Yablokova; E. E. Fesenko

2010-01-01

320

Crystal structure of the N-terminal RecA-like domain of a DEAD-box RNA helicase, the Dugesia japonica vasa-like gene B protein  

Microsoft Academic Search

The Dugesia japonica vasa-like gene B (DjVLGB) protein is a DEAD-box RNA helicase of a planarian, which is well known for its strong regenerative capacity. DjVLGB shares sequence similarity to the Drosophila germ-line-specific DEAD-box RNA helicase Vasa, and even higher similarity to its paralogue, mouse PL10. In this study, we solved the crystal structure of the DjVLGB N-terminal RecA-like domain.

Kazuki Kurimoto; Yutaka Muto; Naomi Obayashi; Takaho Terada; Mikako Shirouzu; Takashi Yabuki; Masaaki Aoki; Eiko Seki; Takayoshi Matsuda; Takanori Kigawa; Hiromi Okumura; Akiko Tanaka; Norito Shibata; Maki Kashikawa; Kiyokazu Agata; Shigeyuki Yokoyama

2005-01-01

321

Towards a bioinformatics of patterning: a computational approach to understanding regulative morphogenesis  

PubMed Central

Summary The mechanisms underlying the regenerative abilities of certain model species are of central importance to the basic understanding of pattern formation. Complex organisms such as planaria and salamanders exhibit an exceptional capacity to regenerate complete body regions and organs from amputated pieces. However, despite the outstanding bottom-up efforts of molecular biologists and bioinformatics focused at the level of gene sequence, no comprehensive mechanistic model exists that can account for more than one or two aspects of regeneration. The development of computational approaches that help scientists identify constructive models of pattern regulation is held back by the lack of both flexible morphological representations and a repository for the experimental procedures and their results (altered pattern formation). No formal representation or computational tools exist to efficiently store, search, or mine the available knowledge from regenerative experiments, inhibiting fundamental insights from this huge dataset. To overcome these problems, we present here a new class of ontology to encode formally and unambiguously a very wide range of possible morphologies, manipulations, and experiments. This formalism will pave the way for top-down approaches for the discovery of comprehensive models of regeneration. We chose the planarian regeneration dataset to illustrate a proof-of-principle of this novel bioinformatics of shape; we developed a software tool to facilitate the formalization and mining of the planarian experimental knowledge, and cured a database containing all of the experiments from the principal publications on planarian regeneration. These resources are freely available for the regeneration community and will readily assist researchers in identifying specific functional data in planarian experiments. More importantly, these applications illustrate the presented framework for formalizing knowledge about functional perturbations of morphogenesis, which is widely applicable to numerous model systems beyond regenerating planaria, and can be extended to many aspects of functional developmental, regenerative, and evolutionary biology.

Lobo, Daniel; Malone, Taylor J.; Levin, Michael

2013-01-01

322

Energy-dependent UV light-induced disruption of (?)sulpiride antagonism of dopamine  

Microsoft Academic Search

The dopamine D2 receptor antagonist sulpiride decreases the spontaneous locomotor activity of Planaria in an enantiomeric-selective and dose-dependent manner. We now report that (?)sulpiride (0.1 ?M)-induced decrease of planarian locomotor activity is significantly (P<0.05) attenuated by low-energy (366 nm) ultraviolet (UV) light and to a greater extent by high-energy (254 nm) UV light. The phenomenon offers a novel approach for

Robert B Raffa; Joseph M Valdez; Lauren J Holland; Robert J Schulingkamp

2000-01-01

323

The adaptive evolution divergence of triosephosphate isomerases between parasitic and free-living flatworms and the discovery of a potential universal target against flatworm parasites.  

PubMed

Triosephosphate isomerase (TIM) is an important drug target or vaccine candidate for pathogenetic organisms such as schistosomes. Parasitic and free-living flatworms shared their last common ancestor but diverged from each other for adapting to parasitic and free-living lives afterwards, respectively. Therefore, adaptive evolution divergence must have occurred between them. Here, for the first time, TIMs were identified from three free-living planarian flatworms, namely Dugesia japonica, Dugesia ryukyuensis, and Schmidtea mediterranea. When these were compared with parasitic flatworms and other organisms, the following results were obtained: (1) planarian TIM genes each contain only one intron, while parasitic flatworm genes each contain other four introns, which are usually present in common metazoans, suggesting planarian-specific intron loss must have occurred; (2) planarian TIM protein sequences are more similar to those of vertebrates rather than to their parasitic relatives or other invertebrates. This implies that relatively rapid evolution occurred in parasitic flatworm TIMs; (3) All the investigated parasitic flatworm TIMs contain a unique tripeptide insert (SXD/E), which may imply its insertion importance to the adaptation of parasitic life. Moreover, our homology modeling results showed the insert region was largely surface-exposed and predicted to be of a B cell epitope location. Finally, the insert is located within one of the three regions previously suggested to be promising immunogenic epitopes in Schistosoma mansoni TIM. Therefore, this unique insert might be significant to developing new effective vaccines or specific drugs against all parasitic flatworm diseases such as schistosomiasis and taeniosis/cysticercosis. PMID:21246382

Chen, Bing; Wen, Jian-Fan

2011-01-19

324

Towards a bioinformatics of patterning: a computational approach to understanding regulative morphogenesis.  

PubMed

The mechanisms underlying the regenerative abilities of certain model species are of central importance to the basic understanding of pattern formation. Complex organisms such as planaria and salamanders exhibit an exceptional capacity to regenerate complete body regions and organs from amputated pieces. However, despite the outstanding bottom-up efforts of molecular biologists and bioinformatics focused at the level of gene sequence, no comprehensive mechanistic model exists that can account for more than one or two aspects of regeneration. The development of computational approaches that help scientists identify constructive models of pattern regulation is held back by the lack of both flexible morphological representations and a repository for the experimental procedures and their results (altered pattern formation). No formal representation or computational tools exist to efficiently store, search, or mine the available knowledge from regenerative experiments, inhibiting fundamental insights from this huge dataset. To overcome these problems, we present here a new class of ontology to encode formally and unambiguously a very wide range of possible morphologies, manipulations, and experiments. This formalism will pave the way for top-down approaches for the discovery of comprehensive models of regeneration. We chose the planarian regeneration dataset to illustrate a proof-of-principle of this novel bioinformatics of shape; we developed a software tool to facilitate the formalization and mining of the planarian experimental knowledge, and cured a database containing all of the experiments from the principal publications on planarian regeneration. These resources are freely available for the regeneration community and will readily assist researchers in identifying specific functional data in planarian experiments. More importantly, these applications illustrate the presented framework for formalizing knowledge about functional perturbations of morphogenesis, which is widely applicable to numerous model systems beyond regenerating planaria, and can be extended to many aspects of functional developmental, regenerative, and evolutionary biology. PMID:23429669

Lobo, Daniel; Malone, Taylor J; Levin, Michael

2012-11-26

325

Telomere maintenance and telomerase activity are differentially regulated in asexual and sexual worms  

PubMed Central

In most sexually reproducing animals, replication and maintenance of telomeres occurs in the germ line and during early development in embryogenesis through the use of telomerase. Somatic cells generally do not maintain telomere sequences, and these cells become senescent in adults as telomeres shorten to a critical length. Some animals reproduce clonally and must therefore require adult somatic mechanisms for maintaining their chromosome ends. Here we study the telomere biology of planarian flatworms with apparently limitless regenerative capacity fueled by a population of highly proliferative adult stem cells. We show that somatic telomere maintenance is different in asexual and sexual animals. Asexual animals maintain telomere length somatically during reproduction by fission or when regeneration is induced by amputation, whereas sexual animals only achieve telomere elongation through sexual reproduction. We demonstrate that this difference is reflected in the expression and alternate splicing of the protein subunit of the telomerase enzyme. Asexual adult planarian stem cells appear to maintain telomere length over evolutionary timescales without passage through a germ-line stage. The adaptations we observe demonstrate indefinite somatic telomerase activity in proliferating stem cells during regeneration or reproduction by fission, and establish planarians as a pertinent model for studying telomere structure, function, and maintenance.

Tan, Thomas C. J.; Rahman, Ruman; Jaber-Hijazi, Farah; Felix, Daniel A.; Chen, Chen; Louis, Edward J.; Aboobaker, Aziz

2012-01-01

326

Inhibition of planar cell polarity extends neural growth during regeneration, homeostasis, and development.  

PubMed

The ability to stop producing or replacing cells at the appropriate time is essential, as uncontrolled growth can lead to loss of function and even cancer. Tightly regulated mechanisms coordinate the growth of stem cell progeny with the patterning needs of the host organism. Despite the importance of proper termination during regeneration, cell turnover, and embryonic development, very little is known about how tissues determine when patterning is complete during these processes. Using planarian flatworms, we show that the planar cell polarity (PCP) pathway is required to stop the growth of neural tissue. Although traditionally studied as regulators of tissue polarity, we found that loss of the PCP genes Vangl2, DAAM1, and ROCK by RNA interference (individually or together) resulted in supernumerary eyes and excess optical neurons in intact planarians, while regenerating planarians had continued hyperplasia throughout the nervous system long after controls ceased new growth. This failure to terminate growth suggests that neural tissues use PCP as a readout of patterning, highlighting a potential role for intact PCP as a signal to stem and progenitor cells to halt neuronal growth when patterning is finished. Moreover, we found this mechanism to be conserved in vertebrates. Loss of Vangl2 during normal development, as well as during Xenopus tadpole tail regeneration, also leads to the production of excess neural tissue. This evolutionarily conserved function of PCP represents a tractable new approach for controlling the growth of nerves. PMID:22339734

Beane, Wendy S; Tseng, Ai-Sun; Morokuma, Junji; Lemire, Joan M; Levin, Michael

2012-03-23

327

Telomere maintenance and telomerase activity are differentially regulated in asexual and sexual worms.  

PubMed

In most sexually reproducing animals, replication and maintenance of telomeres occurs in the germ line and during early development in embryogenesis through the use of telomerase. Somatic cells generally do not maintain telomere sequences, and these cells become senescent in adults as telomeres shorten to a critical length. Some animals reproduce clonally and must therefore require adult somatic mechanisms for maintaining their chromosome ends. Here we study the telomere biology of planarian flatworms with apparently limitless regenerative capacity fueled by a population of highly proliferative adult stem cells. We show that somatic telomere maintenance is different in asexual and sexual animals. Asexual animals maintain telomere length somatically during reproduction by fission or when regeneration is induced by amputation, whereas sexual animals only achieve telomere elongation through sexual reproduction. We demonstrate that this difference is reflected in the expression and alternate splicing of the protein subunit of the telomerase enzyme. Asexual adult planarian stem cells appear to maintain telomere length over evolutionary timescales without passage through a germ-line stage. The adaptations we observe demonstrate indefinite somatic telomerase activity in proliferating stem cells during regeneration or reproduction by fission, and establish planarians as a pertinent model for studying telomere structure, function, and maintenance. PMID:22371573

Tan, Thomas C J; Rahman, Ruman; Jaber-Hijazi, Farah; Felix, Daniel A; Chen, Chen; Louis, Edward J; Aboobaker, Aziz

2012-02-27

328

Thyroid hormone receptor orthologues from invertebrate species with emphasis on Schistosoma mansoni  

PubMed Central

Background: Thyroid hormone receptors (TRs) function as molecular switches in response to thyroid hormone to regulate gene transcription. TRs were previously believed to be present only in chordates. Results: We isolated two TR genes from the Schistosoma mansoni and identified TR orthologues from other invertebrates: the platyhelminths, S. japonium and Schmidtea mediterranea, the mollusc, Lottia gigantean and the arthropod Daphnia pulex. Phylogenetic analysis of the DNA binding domain and/or ligand binding domain shows that invertebrate and vertebrate TRs cluster together, TRs from the vertebrates and from the jawless vertebrate (lamprey) clustered within separate subgroups, Platyhelminth TRs cluster outside of the vertebrate TR subgroups and that the schistosome TRs and S. mediterranea TRs clustered within separate subgroups. Alignment of the C-terminus of the A/B domain revealed a conserved TR-specific motif, termed TR 'N-terminus signature sequence', with a consensus sequence of (G/P)YIPSY(M/L)XXXGPE(D/E)X. Heterodimer formation between S. mansoni TRs and SmRXR1 suggests that the invertebrate TR protein gained the ability to form a heterodimer with RXR. ESMA analysis showed that SmTR? could bind to a conserved DNA core motif as a monomer or homodimer. Conclusion: Vertebrate TR genes originated from a common ancestor of the Bilateria. TR genes underwent duplication independently in the Protostomia and Deuterostomia. The duplication of TRs in deuterostomes occurred after the split of jawless and jawed vertebrates. In protostomes, TR genes underwent duplication in Platyhelminths, occurring independently in trematode and turbellarian lineages. Using S. mansoni TRs as an example, invertebrate TRs exhibited the ability to form a dimer with RXR prior to the emergence of the vertebrate TRs and were able to bind to vertebrate TR core DNA elements as a monomer or homodimer.

Wu, Wenjie; Niles, Edward G; LoVerde, Philip T

2007-01-01

329

Phylogenetic analysis reveals wide distribution of globin X  

PubMed Central

The vertebrate globin gene repertoire consists of seven members that differ in terms of structure, function and phyletic distribution. While hemoglobin, myoglobin, cytoglobin, and neuroglobin are present in almost all gnathostomes examined so far, other globin genes, like globin X, are much more restricted in their phyletic distribution. Till today, globin X has only been found in teleost fish and Xenopus. Here, we report that globin X is also present in the genomes of the sea lamprey, ghost shark and reptiles. Moreover, the identification of orthologs of globin X in crustacean, insects, platyhelminthes, and hemichordates confirms its ancient origin.

2011-01-01

330

The defensive function of trichocysts in Paramecium tetraurelia against metazoan predators compared with the chemical defense of two species of toxin-containing ciliates.  

PubMed

The time-honored assumption about the defensive function of trichocysts in Paramecium against predators was recently verified experimentally against different species of unicellular predators. In the present study, we examined the defensive function of trichocysts against three metazoan predators, Cephalodella sp. (Rotifera), Eucypris sp. (Arthropoda), and Stenostomum sphagnetorum (Platyhelminthes). The results confirmed the defensive function of trichocysts against two of these metazoan predators (Cephalodella sp. and Eucypris sp.), while they seem ineffective against S. sphagnetorum. We also compared the defensive efficiency of the trichocysts of P. tetraurelia with that of toxin-containing extrusomes of two ciliates. PMID:23537235

Buonanno, Federico; Harumoto, Terue; Ortenzi, Claudio

2013-04-01

331

Changes in mitochondrial genetic codes as phylogenetic characters: Two examples from the flatworms  

PubMed Central

Shared molecular genetic characteristics other than DNA and protein sequences can provide excellent sources of phylogenetic information, particularly if they are complex and rare and are consequently unlikely to have arisen by chance convergence. We have used two such characters, arising from changes in mitochondrial genetic code, to define a clade within the Platyhelminthes (flatworms), the Rhabditophora. We have sampled 10 distinct classes within the Rhabditophora and find that all have the codon AAA coding for the amino acid Asn rather than the usual Lys and AUA for Ile rather than the usual Met. We find no evidence to support claims that the codon UAA codes for Tyr in the Platyhelminthes rather than the standard stop codon. The Rhabditophora are a very diverse group comprising the majority of the free-living turbellarian taxa and the parasitic Neodermata. In contrast, three other classes of turbellarian flatworm, the Acoela, Nemertodermatida, and Catenulida, have the standard invertebrate assignments for these codons and so are convincingly excluded from the rhabditophoran clade. We have developed a rapid computerized method for analyzing genetic codes and demonstrate the wide phylogenetic distribution of the standard invertebrate code as well as confirming already known metazoan deviations from it (ascidian, vertebrate, echinoderm/hemichordate).

Telford, Maximilian J.; Herniou, Elisabeth A.; Russell, Robert B.; Littlewood, D. Timothy J.

2000-01-01

332

sine oculis in basal Metazoa.  

PubMed

We report the recovery of homologs of Six1/2/sine oculis (so), a homeodomain-containing member of the Six-gene family, from a diverse set of basal Metazoa, including representatives of the poriferan classes Demospongia, Calcarea and Hexactinellida, the cnidarian classes Hydrozoa, Scyphozoa and Anthozoa, as well as a ctenophore. so sequences were also recovered from a platyhelminth, an echiurid and two bivalve molluscs, members of the super-phyletic group Lophotrochozoa. In the case of the platyhelminth, multiple distinct so sequences were recovered, as well as a member of the related group Six4/5/D-Six4. Extended sequences of the so gene were recovered from the demosponge, Haliclona sp., and the scyphozoan Aurelia aurita via PCR, and 3' RACE. The affinities of all recovered sequences were assessed using a parsimony analysis based on both nucleic and amino acid sequence and using successive character weighting. Our results indicate that so is highly conserved across the animal kingdom. Preliminary expression data for Aurelia reveal that transcripts of the so homolog are present in the manubrium as well as in the rhopalia, which contain the statocyst and eyes, in the free-swimming ephyra and juvenile stages of these jellyfish. PMID:15221378

Bebenek, Ilona G; Gates, Ruth D; Morris, Joshua; Hartenstein, Volker; Jacobs, David K

2004-06-25

333

Phylogenetic position of phylum Nemertini, inferred from 18S rRNA sequences: molecular data as a test of morphological character homology.  

PubMed

Partial 18S rRNA sequence of the nemertine Cerebratulus lacteus was obtained and compared with those of coelomate metazoans and acoelomate platyhelminths to test whether nemertines share a most recent common ancestor with the platyhelminths, as traditionally has been implied, or whether nemertines lie within a protostome coelomate clade, as suggested by more recent morphological analyses. Maximum-parsimony analysis supports the inclusion of the nemertine within a protostome-coelomate clade that falls within a more inclusive coelomate clade. Bootstrap analysis indicates strong support for a monophyletic Coelomata composed of a deuterostome and protostome-coelomate clade. Support for a monophyletic protostome Coelomata is weak. Inference by distance analysis is consistent with that of maximum parsimony. Analysis of down-weighted paired sites by maximum parsimony reveals variation in topology only within the protostome-coelomate clade. The relationships among the protostome coelomates cannot be reliably inferred from the partial sequences, suggesting that coelomate protostomes diversified rapidly. Results with evolutionary parsimony are consistent with the inclusion of the nemertine in a coelomate clade. The molecular inference corroborates recent morphological character analyses that reveal no synapomorphies of nemertines and flatworms but instead suggest that the circulatory system and rhynchocoel of nemertines are homologous to coelomic cavities of protostome coelomates, thus supporting the corresponding hypothesis that nemertines belong within a protostome-coelomate clade. The sequence data provide an independent test of morphological character homology. PMID:1560760

Turbeville, J M; Field, K G; Raff, R A

1992-03-01

334

Lipid biosynthesis in the marine flatworm Convoluta roscoffensis and its algal symbiont Platymonas convoluta.  

PubMed

As a part of an investigations on the lipid metabolism in Platyhelminthes, the acoel Convoluta roscoffensis, which harbors the green alga Platymonas convoluta as a symbiont, was studied. Isotopic tracer experiments established that the acoel lacks the ability to synthesize de novo long-chain saturated and unsaturated fatty acids and depends on its algal symbiont for these compounds. The acoel's fatty acid composition closely resembles that of the alga but differs from those of other animals; the acoel's polyunsaturated fatty acids are of the plant type (omega 3 family) rather than of the animal type (omega 6 family). The acoel also lacks the ability to synthesize sterols de novo. It contains 24-methylenecholesterol synthesized by the algae and, in addition, significant amounts of cholesterol, which is probably a host modification product of the algal sterol. With fatty acids provided by the symbiont, the acoel has the ability to synthesize its own complex lipids. The acoel contains relatively large amounts of triglyceride, phosphatidylcholine, and ethanolamine plasmalogen. These compounds are either not present at all or present only in minute amounts in the symbiotic alga. Since acoels belong to the most primitive forms of the present-day flatworms, the observed metabolic defects in this organism suggest that mechanisms for the biosynthesis of fatty acids and sterols were lost early during the evolution of the Platyhelminthes, and that this phenomenon is widespread within the phylum. PMID:465514

Meyer, H; Provasoli, L; Meyer, F

1979-06-21

335

Identification of novel glutathione transferases in Echinococcus granulosus. An evolutionary perspective.  

PubMed

Glutathione transferase enzymes (GSTs) constitute a major detoxification system in helminth parasites and have been related to the modulation of host immune response mechanisms. At least three different GSTs classes have been described in Platyhelminthes: Mu, Sigma and Omega. Mining the genome of Echinococcus multilocularis and the ESTs databases of Taenia solium and E. granulosus identified two new GSTs from the cestode E. granulosus, named EgGST2 and EgGST3. It also revealed that the Omega class of GSTs was absent from the Taenidae family. EgGST2 and EgGST3 are actively expressed in the parasite. In order to know the origin of these new GSTs, in silico analyses were performed. While EgGST2 is classified as belonging to the Sigma class, the data obtained for EgGST3 allowed a less clear interpretation. The study of the evolutionary relatedness based on the C-terminal domain sequence, gene structure conservation and three-dimensional structure predictions, suggests that EgGST3 is derived from the Platyhelminthes' Sigma-class cluster. Interestingly, the N-terminal domain displays some characteristic Omega-class residues, including a Cys residue that is likely to be involved in the catalytic mechanism. We discuss different evolutionary scenarios that could explain the observed patterns. PMID:22659461

Iriarte, Andrés; Arbildi, Paula; La-Rocca, Silvana; Musto, Héctor; Fernández, Verónica

2012-05-31

336

Variation in worm assemblages associated with Pomacea canaliculata (Caenogastropoda, Ampullariidae) in sites near the Río de la Plata estuary, Argentina.  

PubMed

Pomacea canaliculata is a common gastropod in freshwater habitats from Central and Northern Argentina, extending northwards into the Amazon basin. Several Platyhelminthes have been reported associated to P. canaliculata, sharing an intimate relationship with this gastropod host. The objectives of this study were to describe the symbiotic species assemblages associated to P. canaliculata in the study area, and to disclose differences among them. Samples were taken in three typical small streams and one artificial lentic lagoon, all connected with the Rio de la Plata estuary. The 81.53% were infested with different symbiotic (sensu lato) species. Among the Platyhelminthes, the commensal Temnocephala iheringi Haswell, 1893 was highly prevalent in all samples, always in the mantle cavity. Four trematode taxa were recognized: (a) metacercariae of Echinostoma parcespinosum Lutz, 1924 in the mantle cavity and sporocysts in the digestive gland; (b) metacercariae of Dietziella egregia (Dietz, 1909) in the pericardial cavity; (c) unidentified xiphidiocercariae and (d) unidentified sporocysts and furcocercariae in the digestive gland. Nematode larvae and oligochaetes were found in two localities in the mantle cavity. Among the Annelida, Helobdella ampullariae Ringuelet, 1945 was found in the mantle cavity and lung of snails only from one locality. Our results show that although some of the symbionts are present in all localities, others are restricted to some particular ones, whether in their absolute numbers or in their relative abundance. Thus, each hosting population at the studied localities may be defined by the particular combination of symbionts that bears. PMID:17375466

Damborenea, C; Brusa, E; Paola, A

2006-12-01

337

Energy-dependent UV light-induced disruption of (-)sulpiride antagonism of dopamine.  

PubMed

The dopamine D2 receptor antagonist sulpiride decreases the spontaneous locomotor activity of Planaria in an enantiomeric-selective and dose-dependent manner. We now report that (-)sulpiride (0.1 microM)-induced decrease of planarian locomotor activity is significantly (P<0.05) attenuated by low-energy (366 nm) ultraviolet (UV) light and to a greater extent by high-energy (254 nm) UV light. The phenomenon offers a novel approach for studying dopamine D2 receptor transduction processes in a simple in vivo model. PMID:11040357

Raffa, R B; Valdez, J M; Holland, L J; Schulingkamp, R J

2000-10-20

338

dlx and sp6-9 Control Optic Cup Regeneration in a Prototypic Eye  

PubMed Central

Optic cups are a structural feature of diverse eyes, from simple pit eyes to camera eyes of vertebrates and cephalopods. We used the planarian prototypic eye as a model to study the genetic control of optic cup formation and regeneration. We identified two genes encoding transcription factors, sp6-9 and dlx, that were expressed in the eye specifically in the optic cup and not the photoreceptor neurons. RNAi of these genes prevented formation of visible optic cups during regeneration. Planarian regeneration requires an adult proliferative cell population with stem cell-like properties called the neoblasts. We found that optic cup formation occurred only after migration of progressively differentiating progenitor cells from the neoblast population. The eye regeneration defect caused by dlx and sp6-9 RNAi can be explained by a failure to generate these early optic cup progenitors. Dlx and Sp6-9 genes function as a module during the development of diverse animal appendages, including vertebrate and insect limbs. Our work reveals a novel function for this gene pair in the development of a fundamental eye component, and it utilizes these genes to demonstrate a mechanism for total organ regeneration in which extensive cell movement separates new cell specification from organ morphogenesis.

Lapan, Sylvain W.; Reddien, Peter W.

2011-01-01

339

Conservation status of Chinese species: (2) Invertebrates.  

PubMed

A total of 2441 invertebrate species were evaluated using the IUCN Red List Criteria and Regional Guidelines. Approximately 30 experts were involved in this project, which covered a wide range of species, including jellyfish, corals, planarians, snails, mollusks, bivalves, decapods, benthic crustaceans, arachnids (spiders, scorpions), butterflies, moths, beetles, sea cucumbers, sea urchins, sea stars, acorn worms and lancelets. In general, invertebrate species in China were found to be severely threatened, with 0.9% being critically endangered, 13.44% endangered and 20.63% vulnerable. All species of hermatypic corals and planarians are threatened. More than 80% of evaluated species face serious threat due to habitat destruction by coral collection, logging, non-woody vegetation collection, timber plantations, non-timber plantations, extraction and/or livestock. Other threats are intrinsic factors, harvesting by humans, alien invasive species and pollution. The main intrinsic factors contributing to the high levels of threat are limited dispersal and restricted range. No conservation measures have been taken for 70% of the threatened invertebrates evaluated. Existing conservation measures include: strengthening of national and international legislation (Convention on International Trade in Endangered Species of Wild Fauna and Flora), increasing public awareness, studying population trends/monitoring, and establishment of protected areas. The major conservation measure employed is strengthening of policies. Relative to the situation worldwide (2006 IUCN Red List), there is little information available about invertebrate extinctions in China. PMID:21396022

Xie, Yan; Wang, Sung

2007-06-01

340

Increased mobility and stem-cell proliferation rate in Dugesia tigrina induced by 880nm light emitting diode.  

PubMed

The therapeutic effects elicited by photobiostimulation in the near infrared range may be associated with increased proliferation rate of particular cell-types. The present study utilized commercial light emitting diodes to investigate the effects of low-level near-infrared radiation on the proliferation rate of stem cells in amputated planarian. Whole and amputated animals were exposed to either ambient diurnal lighting, darkness, white light, red light, or near-infrared (880 nm) light. Irradiation was consistent for the duration of the experiments and was provided using commercial 5mm light emitting diodes (?1.0 mW/m(2) in power density and ?0.01 J/cm(2) in radiant exposure). Compared to other groups amputated planarian exposed to near-infrared displayed increased mobility by the 3rd day of exposure (F((4,26))=4.31, p<0.04, ?(2)=41%). Higher densities of stem cells were measured in these worms 84 h post injury (F((4,72))=4.78, p<0.01, ?(2)=21%). These findings suggest that non-coherent light sources with power-densities about 1000 times lower than contemporary low-power laser settings remain effective in generating photobiostimulation effects and warrants further investigation on stem-cell proliferation induced by near-infrared light emitting diodes. PMID:21146998

Wu, Hsia-Pai Patrick; Persinger, Michael A

2010-11-24

341

Tissue absence initiates regeneration through Follistatin-mediated inhibition of Activin signaling.  

PubMed

Regeneration is widespread, but mechanisms that activate regeneration remain mysterious. Planarians are capable of whole-body regeneration and mount distinct molecular responses to wounds that result in tissue absence and those that do not. A major question is how these distinct responses are activated. We describe a follistatin homolog (Smed-follistatin) required for planarian regeneration. Smed-follistatin inhibition blocks responses to tissue absence but does not prevent normal tissue turnover. Two activin homologs (Smed-activin-1 and Smed-activin-2) are required for the Smed-follistatin phenotype. Finally, Smed-follistatin is wound-induced and expressed at higher levels following injuries that cause tissue absence. These data suggest that Smed-follistatin inhibits Smed-Activin proteins to trigger regeneration specifically following injuries involving tissue absence and identify a mechanism critical for regeneration initiation, a process important across the animal kingdom. DOI:http://dx.doi.org/10.7554/eLife.00247.001. PMID:24040508

Gavińo, Michael A; Wenemoser, Danielle; Wang, Irving E; Reddien, Peter W

2013-09-10

342

Heterochromatin protein 1 promotes self-renewal and triggers regenerative proliferation in adult stem cells.  

PubMed

Adult stem cells (ASCs) capable of self-renewal and differentiation confer the potential of tissues to regenerate damaged parts. Epigenetic regulation is essential for driving cell fate decisions by rapidly and reversibly modulating gene expression programs. However, it remains unclear how epigenetic factors elicit ASC-driven regeneration. In this paper, we report that an RNA interference screen against 205 chromatin regulators identified 12 proteins essential for ASC function and regeneration in planarians. Surprisingly, the HP1-like protein SMED-HP1-1 (HP1-1) specifically marked self-renewing, pluripotent ASCs, and HP1-1 depletion abrogated self-renewal and promoted differentiation. Upon injury, HP1-1 expression increased and elicited increased ASC expression of Mcm5 through functional association with the FACT (facilitates chromatin transcription) complex, which consequently triggered proliferation of ASCs and initiated blastema formation. Our observations uncover an epigenetic network underlying ASC regulation in planarians and reveal that an HP1 protein is a key chromatin factor controlling stem cell function. These results provide important insights into how epigenetic mechanisms orchestrate stem cell responses during tissue regeneration. PMID:23629965

Zeng, An; Li, Yong-Qin; Wang, Chen; Han, Xiao-Shuai; Li, Ge; Wang, Jian-Yong; Li, Dang-Sheng; Qin, Yong-Wen; Shi, Yufang; Brewer, Gary; Jing, Qing

2013-04-29

343

Heterochromatin protein 1 promotes self-renewal and triggers regenerative proliferation in adult stem cells  

PubMed Central

Adult stem cells (ASCs) capable of self-renewal and differentiation confer the potential of tissues to regenerate damaged parts. Epigenetic regulation is essential for driving cell fate decisions by rapidly and reversibly modulating gene expression programs. However, it remains unclear how epigenetic factors elicit ASC-driven regeneration. In this paper, we report that an RNA interference screen against 205 chromatin regulators identified 12 proteins essential for ASC function and regeneration in planarians. Surprisingly, the HP1-like protein SMED–HP1-1 (HP1-1) specifically marked self-renewing, pluripotent ASCs, and HP1-1 depletion abrogated self-renewal and promoted differentiation. Upon injury, HP1-1 expression increased and elicited increased ASC expression of Mcm5 through functional association with the FACT (facilitates chromatin transcription) complex, which consequently triggered proliferation of ASCs and initiated blastema formation. Our observations uncover an epigenetic network underlying ASC regulation in planarians and reveal that an HP1 protein is a key chromatin factor controlling stem cell function. These results provide important insights into how epigenetic mechanisms orchestrate stem cell responses during tissue regeneration.

Zeng, An; Li, Yong-Qin; Wang, Chen; Han, Xiao-Shuai; Li, Ge; Wang, Jian-Yong; Li, Dang-Sheng; Qin, Yong-Wen; Shi, Yufang; Brewer, Gary

2013-01-01

344

Mini review on chemotherapy of taeniasis and cysticercosis due to Taenia solium in Asia, and a case report with 20 tapeworms in China.  

PubMed

A 43-year-old Tibetan woman living in northwest Sichuan, China, confirmed to be a taeniasis carrier of Taenia solium was treated with pumpkin seeds combined with Areca nut extract in October 2009. All 20 tapeworms except one without scolex were expelled under good conditions. She was free of secondary cysticercosis within one year follow up. Although the first choice for treatment of taeniasis is still praziquantel, it may often cause serious side effect on asymptomatic cysticercosis cases to suddenly become symptomatic within a half day of the treatment. Therefore, the problems in treatment of taeniasis and/or cysticercosis in Asia are briefly overviewed, since other platyhelminthic diseases including schistosomiasis, opisthorchiasis etc. are more common and praziquantel is strongly recommended for mass treatment of these trematodiases with no idea on the co-infection with eggs of T. solium which cause asymptomatic cysticercosis. PMID:23959481

Ito, A; Li, T; Chen, X; Long, C; Yanagida, T; Nakao, M; Sako, Y; Okamoto, M; Wu, Y; Raoul, F; Giraudoux, P; Craig, P S

2013-06-01

345

New approaches for understanding mechanisms of drug resistance in schistosomes.  

PubMed

Schistosomes are parasitic flatworms that cause schistosomiasis, a neglected tropical disease that affects hundreds of millions worldwide. Treatment and control of schistosomiasis relies almost entirely on the single drug praziquantel (PZQ), making the prospect of emerging drug resistance particularly worrisome. This review will survey reports of PZQ (and other drug) resistance in schistosomes and other platyhelminths, and explore mechanisms by which drug resistance might develop. Newer genomic and post-genomic strategies that offer the promise of better understanding of how drug resistance might arise in these organisms will be discussed. These approaches could also lead to insights into the mode of action of these drugs and potentially provide markers for monitoring the emergence of resistance. PMID:23552512

Greenberg, Robert M

2013-04-03

346

Lophotrochozoan mitochondrial genomes  

SciTech Connect

Progress in both molecular techniques and phylogeneticmethods has challenged many of the interpretations of traditionaltaxonomy. One example is in the recognition of the animal superphylumLophotrochozoa (annelids, mollusks, echiurans, platyhelminthes,brachiopods, and other phyla), although the relationships within thisgroup and the inclusion of some phyla remain uncertain. While much ofthis progress in phylogenetic reconstruction has been based on comparingsingle gene sequences, we are beginning to see the potential of comparinglarge-scale features of genomes, such as the relative order of genes.Even though tremendous progress is being made on the sequencedetermination of whole nuclear genomes, the dataset of choice forgenome-level characters for many animals across a broad taxonomic rangeremains mitochondrial genomes. We review here what is known aboutmitochondrial genomes of the lophotrochozoans and discuss the promisethat this dataset will enable insight into theirrelationships.

Valles, Yvonne; Boore, Jeffrey L.

2005-10-01

347

New approaches for understanding mechanisms of drug resistance in schistosomes  

PubMed Central

SUMMARY Schistosomes are parasitic flatworms that cause schistosomiasis, a neglected tropical disease that affects hundreds of millions worldwide. Treatment and control of schistosomiasis relies almost entirely on the single drug praziquantel (PZQ), making the prospect of emerging drug resistance particularly worrisome. This review will survey reports of PZQ (and other drug) resistance in schistosomes and other platyhelminths, and explore mechanisms by which drug resistance might develop. Newer genomic and post-genomic strategies that offer the promise of better understanding of how drug resistance might arise in these organisms will be discussed. These approaches could also lead to insights into the mode of action of these drugs and potentially provide markers for monitoring the emergence of resistance.

GREENBERG, ROBERT M.

2013-01-01

348

Producing parasitic helminth reference and draft genomes at the Wellcome Trust Sanger Institute.  

PubMed

The Wellcome Trust Sanger Institute (WTSI) is producing de novo reference quality genomes for parasitic helminth species from platyhelminth tapeworms (cestodes), flukes (trematodes) and roundworms (nematodes) primarily using second-generation (Illumina and 454) sequencing technologies. The reference genomes will be followed with draft coverage from a number of related strains or species. Comparing species- or strain-specific differences will help to unravel the genomic basis for differences in the organism's biology and ultimately contribute towards identifying potential novel targets for vaccine therapies. Second-generation sequencing technologies are revolutionizing parasite genomics. This article reviews the impact that sequencing technologies has had on genomics and how it has shaped the parasitic helminth genome sequencing initiative at WTSI. PMID:21707658

Holroyd, N; Sanchez-Flores, A

349

Phylogenetic study of the arginine-vasotocin/arginine-vasopressin-like immunoreactive system in invertebrates.  

PubMed

1. A phylogenetic study of arg-vasotocin (AVT)/arg-vasopressin (AVP)-like immunoreactive cells was performed by the PAP method in the central nervous system of invertebrates. 2. The immunoreactivity was detected in the nerve cells of Hydra magnipapillata of the Coelenterata; Neanthes japonica and Pheretima communissima of the Annelida; Pomacea canaliculata, Aplysia kurodai, Oncidium verrucosum, Bradybaena similaris, Achatina fulica, Limax marginatus and Meretrix lamarckii of the Mollusca; Gnorimosphaeroma rayi, Hemigrapsus sanguineus, Gryllus bimaculatus and Baratha brassicae of the Arthropoda; Asterina pectinifera of the Echinodermata; and Halocynthia roretzi of the Protochordata. 3. No immunoreactivity was detected in Bipalium sp. of the Platyhelminthes, or in Procambarus clarkii and Helice tridens of the Arthropoda. 4. From these results, it appears that AVT/AVP is a phylogenetically ancient peptide which is present in a wide variety of invertebrates. 5. The actions of AVT/AVP and its presence in invertebrates are discussed. PMID:2907440

Mizuno, J; Takeda, N

1988-01-01

350

Phylogenetic study of the oxytocin-like immunoreactive system in invertebrates.  

PubMed

1. A phylogenetic study of oxytocin (OXT)-like immunoreactive cells was performed by the PAP method in the central nervous system of invertebrates. 2. The immunoreactivity was detected in the nerve cells of Hydra magnipapillata of the Coelenterata; Neanthes japonica and Pheretima communissima of the Annelida; Oncidium verrucosum, Limax marginatus and Meretrix lamarckii of the Mollusca; and Baratha brassica of the Arthropoda. 3. No immunoreactive cells were found in Bipalium sp. of the Platyhelminthes; Pomacea canaliculata, Aplysia kurodai, Bradybaena similaris and Achatina fulica of the Mollusca; and Gnorimosphaeroma rayi, Procambarus clarkii, Hemigrapsus sanguineus, Helice tridens and Gryllus bimaculatus of the Arthropoda; Asterina pectinifera of the Echinodermata; and Halocynthia roretzi of the Protochordata. 4. These results demonstrate that an OXT-immunoreactive substance is widely present not only in vertebrates but also in invertebrates. 5. OXT seems to have been introduced into these invertebrates at an early stage of their phylogenetic history. PMID:2907439

Mizuno, J; Takeda, N

1988-01-01

351

Investigation of molluscan phylogeny on the basis of 18S rRNA sequences.  

PubMed

The 18S rRNA sequences of 12 molluscs, representing the extant classes Gastropoda, Bivalvia, Polyplacophora, Scaphopoda, and Caudofoveata, were determined and compared with selected known 18S rRNA sequences of Metazoa, including other Mollusca. These data do not provide support for a close relationship between Platyhelminthes (Turbellaria) and Mollusca, but rather suggest that the latter group belongs to a clade of eutrochozoan coelomates. The 18S rRNA data fail to recover molluscan, bivalve, or gastropod monophyly. However, the branching pattern of the eutrochozoan phyla and classes is unstable, probably due to the explosive Cambrian radiation during which these groups arose. Similarly, the 18S rRNA data do not provide a reliable signal for the molluscan interclass relationships. Nevertheless, we obtained strong preliminary support for phylogenetic inferences at more restricted taxonomic levels, such as the monophyly of Polyplacophora, Caenogastropoda, Euthyneura, Heterodonta, and Arcoida. PMID:8952075

Winnepenninckx, B; Backeljau, T; De Wachter, R

1996-12-01

352

Calcium channels of schistosomes: unresolved questions and unexpected answers  

PubMed Central

Parasitic flatworms of the genus Schistosoma are the causative agents of schistosomiasis, a highly prevalent, neglected tropical disease that causes significant morbidity in hundreds of millions of people worldwide. The current treatment of choice against schistosomiasis is praziquantel (PZQ), which is known to affect Ca2+ homeostasis in schistosomes, but which has an undefined molecular target and mode of action. PZQ is the only available antischistosomal drug in most parts of the world, making reports of PZQ resistance particularly troubling. Voltage-gated Ca2+ (Cav) channels have been proposed as possible targets for PZQ, and, given their central role in the neuromuscular system, may also serve as targets for new anthelmintic therapeutics. Indeed, ion channels constitute the majority of targets for current anthelmintics. Cav channel subunits from schistosomes and other platyhelminths have several unique properties that make them attractive as potential drug targets, and that could also provide insights into structure-function relationships in, and evolution of, Cav channels.

Salvador-Recatala, Vicenta; Greenberg, Robert M.

2011-01-01

353

Ultrastructural study on the spermiogenesis and spermatozoon of the metacercariae of Microphallus primas (Digenea), a parasite of Carcinus maenas.  

PubMed

The thread-like spermatozoon of the crab parasite Microphallus primas was studied by electron microscopy. A survey of the head region of the spermatozoon reveals three features hitherto unknown in Platyhelminthes spermatozoa. The first is the aberrant inclusion of the nucleus within one of the two axonemes, limited to the head end region. The second is the coexistence, in the same axoneme, of two patterns, 9 + 0 (doublets without dynein arms) and 9 + "1". The third is the presence of a layer of cortical microtubules running longitudinally from the zone where the nucleus goes from axoneme to the tail region (where the two flagella start). The sequence of events in spermatogenesis is similar to that described for most of the Digenea trematodes, and the spermiogenesis process conforms to a common plan in nearly the whole group. PMID:2310565

Castilho, F; Barandela, T

1990-02-01

354

Lophotrochozoan mitochondrial genomes.  

PubMed

Progress in both molecular techniques and phylogenetic methods has challenged many of the interpretations of traditional taxonomy. One example is in the recognition of the animal superphylum Lophotrochozoa (annelids, mollusks, echiurans, platyhelminthes, brachiopods, and other phyla), although the relationships within this group and the inclusion of some phyla remain uncertain. While much of this progress in phylogenetic reconstruction has been based on comparing single gene sequences, there are also higher order features of genomes, such as the relative order of genes, that have contributed, and this seems likely to be even more fruitful in the future. Even though tremendous progress is being made on the sequence determination of whole nuclear genomes, the dataset of choice for genome-level characters for many animals across a broad taxonomic range remains mitochondrial genomes. We review here what is known about mitochondrial genomes of the lophotrochozoans and how comparisons of some of these features may be useful in discerning the phylogeny of this group. PMID:21672765

Vallčs, Yvonne; Boore, Jeffrey L

2006-06-30

355

Early embryogenesis of planaria: a cryptic larva feeding on maternal resources.  

PubMed

The early planarian embryo presents a complete ciliated epidermis and a pharynx and feeds on maternal yolk cells. In this paper, we report on all the elements involved in the formation of such an autonomous embryo, which we name cryptic larva. First, we provide a description of the spherical and fusiform yolk cells and their relationship with the blastomeres, from the laying of the egg capsule up to their final fate in mid embryonic stages. Then, we describe the early cleavage and the subsequent development of the tissues of the cryptic larva, namely, the primary epidermis, the embryonic pharynx, and a new cell type, the star cells. Finally, we discuss the possibility that the cryptic larva either constitutes a vestigial larva or, more likely, is the evolutionary result of the competition between multiple embryos for the limited and shared maternal resources in the egg capsule. PMID:16932928

Cardona, Albert; Hartenstein, Volker; Romero, Rafael

2006-08-24

356

[Morphofunctional organization of reserve stem cells providing for asexual and sexual reproduction of invertebrates].  

PubMed

Published and original data indicating evolutionary conservation of the morphofunctional organization of reserve stem cells providing for asexual and sexual reproduction of invertebrates are reviewed. Stem cells were studied in representatives of five animal types: archeocytes in sponge Oscarella malakhovi (Porifera), large interstitial cells in colonial hydroid Obelia longissima (Cnidaria), neoblasts in an asexual race of planarian Girardia tigrina (Platyhelmintes), stem cells in colonial rhizocephalans Peltogasterella gracilis, Polyascus polygenea, and Thylacoplethus isaevae (Arthropoda), and colonial ascidian Botryllus tuberatus (Chordata). Stem cells in animals of such diverse taxa feature the presence of germinal granules, are positive for proliferating cell nuclear antigen, demonstrate alkaline phosphatase activity (at marker of embryonic stem cells and primary germ cells in vertebrates), and rhizocephalan stem cells express the vasa-like gene (such genes are expressed in germline cells of different metazoans). The self-renewing pool of stem cells is the cellular basis of the reproductive strategy including sexual and asexual reproduction. PMID:19405444

Isaeva, V V; Akhmadieva, A V; Aleksandriova, Ia N; Shukaliuk, A I

357

Disruption of a drug-induced choice behavior by UV light.  

PubMed

A considerable body of evidence suggests that UV light disrupts ligand binding in vitro. In vivo, UV light effects have been reported to disrupt simple behaviors such as spontaneous locomotor activity. However, there are no reports of UV light blocking a more complex drug-altered behavior. We now report that: (1) cocaine dose-relatedly reversed planarians' usual selection of dark over light (from 19.4+/-4.4% to 73.3+/-6.7%) (this effect was not due to an increase in locomotor activity, since cocaine only minimally increases locomotor activity at the highest dose tested); and (2) the cocaine-induced alteration of behavioral choice was attenuated significantly (P<0.05) by UV light (366 nm and 254 nm). The results demonstrate alteration of a relatively complex choice behavior by UV light. PMID:14557725

Raffa, R B; Dasrath, C S; Brown, D R

2003-11-01

358

Deep sequencing identifies regulated small RNAs in Dugesia japonica.  

PubMed

MicroRNAs (miRNAs), which are 18 ~ 24 nucleotides length, play important roles in regulating the expression of gene at the post-transcription level. Dugesia japonica is a branch of planarian organism. It is a model organism for studying the role of miRNAs in stem cell function. Next generation sequencing technology was used to identify the miRNAs of D. japonica. Bioinformatic analysis showed that 262 miRNA and miRNA* sequences were discovered, of which 102 miRNAs were the same as Schmidtea mediterranea and 160 miRNAs were related to other animals. There were 21 miRNAs expressed differentially after amputation. Results also revealed that some key miRNAs might play essential roles in the regeneration progress and some miRNAs might take part in the regulation progress of polarity regeneration in D. japonica. PMID:23314792

Xu, Zhenbiao; Chen, Maoshan; Ren, Zhonggan; Zhang, Nian; Xu, Hanmei; Liu, Xiao; Tian, Geng; Song, Linxia; Yang, Huanming

2013-01-12

359

A lack of commitment for over 500 million years: conserved animal stem cell pluripotency  

PubMed Central

EMBO J 31 12, 2755–2769 (2012); published online 04272012 Stem cells, both adult and germline, are the key cells underpinning animal evolution. Yet, surprisingly little is known about the evolution of their shared key feature: pluripotency. Now using genome-wide expression profiling of pluripotent planarian adult stem cells (pASCs), Önal et al (2012) present evidence for deep molecular conservation of pluripotency. They characterise the expression profile of pASCs and identify conserved expression profiles and functions for genes required for mammalian pluripotency. Their analyses suggest that molecular pluripotency mechanisms may be conserved, and tantalisingly that pluripotency in germ stem cells (GSCs) and somatic stem cells (SSCs) may have had shared common evolutionary origins.

Aboobaker, A Aziz; Kao, Damian

2012-01-01

360

SAPling: a Scan-Add-Print barcoding database system to label and track asexual organisms  

PubMed Central

SUMMARY We have developed a ‘Scan-Add-Print’ database system, SAPling, to track and monitor asexually reproducing organisms. Using barcodes to uniquely identify each animal, we can record information on the life of the individual in a computerized database containing its entire family tree. SAPling has enabled us to carry out large-scale population dynamics experiments with thousands of planarians and keep track of each individual. The database stores information such as family connections, birth date, division date and generation. We show that SAPling can be easily adapted to other asexually reproducing organisms and has a strong potential for use in large-scale and/or long-term population and senescence studies as well as studies of clonal diversity. The software is platform-independent, designed for reliability and ease of use, and provided open source from our webpage to allow project-specific customization.

Thomas, Michael A.; Schotz, Eva-Maria

2011-01-01

361

Opposing roles of voltage-gated Ca2+ channels in neuronal control of regenerative patterning  

PubMed Central

There is intense interest in developing methods to regulate proliferation and differentiation of stem cells into neuronal fates for the purposes of regenerative medicine. One way to do this is through in vivo pharmacological engineering using small molecules. However, a key challenge is identification of relevant signaling pathways and therein drugable targets to manipulate stem cell behaviour efficiently in vivo. Here, we use the planarian flatworm as a simple chemical-genetic screening model for nervous system regeneration to show that the isoquinoline drug praziquantel (PZQ) acts as a small molecule neurogenic to produce two-headed animals with integrated central nervous systems following regeneration. Characterization of the entire family of planarian voltage-operated Ca2+ channel alpha subunits (Cav?), followed by in vivo RNAi of specific Cav subunits revealed that PZQ subverted regeneration by activation of a specific voltage-gated Ca2+ channel isoform (Ca 1A). PZQ-evoked Ca2+ v entry via Cav1A served to inhibit neuronally-derived Hedgehog signals, as evidenced by data showing that RNAi of Cav1A prevented PZQ-evoked bipolarity, Ca2+ entry and decreases in wnt1 and wnt11-5 levels. Surprisingly the action of PZQ was opposed by Ca2+ influx through a closely related neuronal Cav isoform (Cav1B), establishing a novel interplay between specific Cav1 channel isoforms, Ca2+ entry and neuronal Hedgehog signaling. These data map PZQ efficacy to specific neuronal Cav complexes in vivo and underscore that both activators (Cav1A) and inhibitors of Ca2+ influx (Cav1B) can act as small molecule neurogenics in vivo on account of the unique coupling of Ca2+ channels to neuronally-derived polarity cues.

Zhang, Dan; Chan, John D.; Nogi, Taisaku; Marchant, Jonathan S.

2011-01-01

362

The TALE Class Homeobox Gene Smed-prep Defines the Anterior Compartment for Head Regeneration  

PubMed Central

Planaria continue to blossom as a model system for understanding all aspects of regeneration. They provide an opportunity to understand how the replacement of missing tissues from preexisting adult tissue is orchestrated at the molecular level. When amputated along any plane, planaria are capable of regenerating all missing tissue and rescaling all structures to the new size of the animal. Recently, rapid progress has been made in understanding the developmental pathways that control planarian regeneration. In particular Wnt/beta-catenin signaling is central in promoting posterior fates and inhibiting anterior identity. Currently the mechanisms that actively promote anterior identity remain unknown. Here, Smed-prep, encoding a TALE class homeodomain, is described as the first gene necessary for correct anterior fate and patterning during planarian regeneration. Smed-prep is expressed at high levels in the anterior portion of whole animals, and Smed-prep(RNAi) leads to loss of the whole brain during anterior regeneration, but not during lateral regeneration or homeostasis in intact worms. Expression of markers of different anterior fated cells are greatly reduced or lost in Smed-prep(RNAi) animals. We find that the ectopic anterior structures induced by abrogation of Wnt signaling also require Smed-prep to form. We use double knockdown experiments with the S. mediterranea ortholog of nou-darake (that when knocked down induces ectopic brain formation) to show that Smed-prep defines an anterior fated compartment within which stem cells are permitted to assume brain fate, but is not required directly for this differentiation process. Smed-prep is the first gene clearly implicated as being necessary for promoting anterior fate and the first homeobox gene implicated in establishing positional identity during regeneration. Together our results suggest that Smed-prep is required in stem cell progeny as they form the anterior regenerative blastema and is required for specifying anterior cell fates and correct patterning.

Felix, Daniel A.; Aboobaker, A. Aziz

2010-01-01

363

What Role Do Annelid Neoblasts Play? A Comparison of the Regeneration Patterns in a Neoblast-Bearing and a Neoblast-Lacking Enchytraeid Oligochaete  

PubMed Central

The term ‘neoblast’ was originally coined for a particular type of cell that had been observed during annelid regeneration, but is now used to describe the pluripotent/totipotent stem cells that are indispensable for planarian regeneration. Despite having the same name, however, planarian and annelid neoblasts are morphologically and functionally distinct, and many annelid species that lack neoblasts can nonetheless substantially regenerate. To further elucidate the functions of the annelid neoblasts, a comparison was made between the regeneration patterns of two enchytraeid oligochaetes, Enchytraeus japonensis and Enchytraeus buchholzi, which possess and lack neoblasts, respectively. In E. japonensis, which can reproduce asexually by fragmentation and subsequent regeneration, neoblasts are present in all segments except for the eight anterior-most segments including the seven head-specific segments, and all body fragments containing neoblasts can regenerate a complete head and a complete tail, irrespective of the region of the body from which they were originally derived. In E. japonensis, therefore, no antero-posterior gradient of regeneration ability exists in the trunk region. However, when amputation was carried out within the head region, where neoblasts are absent, the number of regenerated segments was found to be dependent on the level of amputation along the body axis. In E. buchholzi, which reproduces only sexually and lacks neoblasts in all segments, complete heads were never regenerated and incomplete (hypomeric) heads could be regenerated only from the anterior region of the body. Such an antero-posterior gradient of regeneration ability was observed for both the anterior and posterior regeneration in the whole body of E. buchholzi. These results indicate that the presence of neoblasts correlates with the absence of an antero-posterior gradient of regeneration ability along the body axis, and suggest that the annelid neoblasts are more essential for efficient asexual reproduction than for the regeneration of missing body parts.

Myohara, Maroko

2012-01-01

364

What role do annelid neoblasts play? A comparison of the regeneration patterns in a neoblast-bearing and a neoblast-lacking enchytraeid oligochaete.  

PubMed

The term 'neoblast' was originally coined for a particular type of cell that had been observed during annelid regeneration, but is now used to describe the pluripotent/totipotent stem cells that are indispensable for planarian regeneration. Despite having the same name, however, planarian and annelid neoblasts are morphologically and functionally distinct, and many annelid species that lack neoblasts can nonetheless substantially regenerate. To further elucidate the functions of the annelid neoblasts, a comparison was made between the regeneration patterns of two enchytraeid oligochaetes, Enchytraeus japonensis and Enchytraeus buchholzi, which possess and lack neoblasts, respectively. In E. japonensis, which can reproduce asexually by fragmentation and subsequent regeneration, neoblasts are present in all segments except for the eight anterior-most segments including the seven head-specific segments, and all body fragments containing neoblasts can regenerate a complete head and a complete tail, irrespective of the region of the body from which they were originally derived. In E. japonensis, therefore, no antero-posterior gradient of regeneration ability exists in the trunk region. However, when amputation was carried out within the head region, where neoblasts are absent, the number of regenerated segments was found to be dependent on the level of amputation along the body axis. In E. buchholzi, which reproduces only sexually and lacks neoblasts in all segments, complete heads were never regenerated and incomplete (hypomeric) heads could be regenerated only from the anterior region of the body. Such an antero-posterior gradient of regeneration ability was observed for both the anterior and posterior regeneration in the whole body of E. buchholzi. These results indicate that the presence of neoblasts correlates with the absence of an antero-posterior gradient of regeneration ability along the body axis, and suggest that the annelid neoblasts are more essential for efficient asexual reproduction than for the regeneration of missing body parts. PMID:22615975

Myohara, Maroko

2012-05-16

365

Opposing roles of voltage-gated Ca2+ channels in neuronal control of regenerative patterning.  

PubMed

There is intense interest in developing methods to regulate proliferation and differentiation of stem cells into neuronal fates for the purposes of regenerative medicine. One way to do this is through in vivo pharmacological engineering using small molecules. However, a key challenge is identification of relevant signaling pathways and therein druggable targets to manipulate stem cell behavior efficiently in vivo. Here, we use the planarian flatworm as a simple chemical-genetic screening model for nervous system regeneration to show that the isoquinoline drug praziquantel (PZQ) acts as a small molecule neurogenic to produce two-headed animals with integrated CNSs following regeneration. Characterization of the entire family of planarian voltage-operated Ca(2+) channel ? subunits (Ca(v)?), followed by in vivo RNAi of specific Ca(v) subunits, revealed that PZQ subverted regeneration by activation of a specific voltage-gated Ca(2+) channel isoform (Ca(v)1A). PZQ-evoked Ca(2+) entry via Ca(v)1A served to inhibit neuronally derived Hedgehog signals, as evidenced by data showing that RNAi of Ca(v)1A prevented PZQ-evoked bipolarity, Ca(2+) entry, and decreases in wnt1 and wnt11-5 levels. Surprisingly, the action of PZQ was opposed by Ca(2+) influx through a closely related neuronal Ca(v) isoform (Ca(v)1B), establishing a novel interplay between specific Ca(v)1 channel isoforms, Ca(2+) entry, and neuronal Hedgehog signaling. These data map PZQ efficacy to specific neuronal Ca(v) complexes in vivo and underscore that both activators (Ca(v)1A) and inhibitors (Ca(v)1B) of Ca(2+) influx can act as small molecule neurogenics in vivo on account of the unique coupling of Ca(2+) channels to neuronally derived polarity cues. PMID:22049441

Zhang, Dan; Chan, John D; Nogi, Taisaku; Marchant, Jonathan S

2011-11-01

366

Adult somatic stem cells in the human parasite, Schistosoma mansoni  

PubMed Central

Summary Schistosomiasis is among the most prevalent human parasitic diseases, affecting more than 200 million people worldwide1. The etiological agents of this disease are trematode flatworms (Schistosoma) that live and lay eggs within the vasculature of the host. These eggs lodge in host tissues, causing inflammatory responses that are the primary cause of morbidity. Because these parasites can live and reproduce within human hosts for decades2, elucidating the mechanisms that promote their longevity is of fundamental importance. Although adult pluripotent stem cells, called neoblasts, drive long-term homeostatic tissue maintenance in long-lived free-living flatworms3,4 (e.g., planarians), and neoblast-like cells have been described in some parasitic tapeworms5, little is known about whether similar cell types exist in any trematode species. Here, we describe a population of neoblast-like cells in the trematode Schistosoma mansoni. These cells resemble planarian neoblasts morphologically and share their ability to proliferate and differentiate into derivatives of multiple germ layers. Capitalizing on available genomic resources6,7 and RNAseq-based gene expression profiling, we find that these schistosome neoblast-like cells express a fibroblast growth factor receptor ortholog. Using RNA interference we demonstrate that this gene is required for the maintenance of these neoblast-like cells. Our observations suggest that adaptation of developmental strategies shared by free-living ancestors to modern-day schistosomes likely contributed to the success of these animals as long-lived obligate parasites. We expect that future studies deciphering the function of these neoblast-like cells will have important implications for understanding the biology of these devastating parasites.

Collins, James J.; Wang, Bo; Lambrus, Bramwell G.; Tharp, Marla; Iyer, Harini; Newmark, Phillip A.

2013-01-01

367

Modular evolution of glutathione peroxidase genes in association with different biochemical properties of their encoded proteins in invertebrate animals  

PubMed Central

Background Phospholipid hydroperoxide glutathione peroxidases (PHGPx), the most abundant isoforms of GPx families, interfere directly with hydroperoxidation of lipids. Biochemical properties of these proteins vary along with their donor organisms, which has complicated the phylogenetic classification of diverse PHGPx-like proteins. Despite efforts for comprehensive analyses, the evolutionary aspects of GPx genes in invertebrates remain largely unknown. Results We isolated GPx homologs via in silico screening of genomic and/or expressed sequence tag databases of eukaryotic organisms including protostomian species. Genes showing strong similarity to the mammalian PHGPx genes were commonly found in all genomes examined. GPx3- and GPx7-like genes were additionally detected from nematodes and platyhelminths, respectively. The overall distribution of the PHGPx-like proteins with different biochemical properties was biased across taxa; selenium- and glutathione (GSH)-dependent proteins were exclusively detected in platyhelminth and deuterostomian species, whereas selenium-independent and thioredoxin (Trx)-dependent enzymes were isolated in the other taxa. In comparison of genomic organization, the GSH-dependent PHGPx genes showed a conserved architectural pattern, while their Trx-dependent counterparts displayed complex exon-intron structures. A codon for the resolving Cys engaged in reductant binding was found to be substituted in a series of genes. Selection pressure to maintain the selenocysteine codon in GSH-dependent genes also appeared to be relaxed during their evolution. With the dichotomized fashion in genomic organizations, a highly polytomic topology of their phylogenetic trees implied that the GPx genes have multiple evolutionary intermediate forms. Conclusion Comparative analysis of invertebrate GPx genes provides informative evidence to support the modular pathways of GPx evolution, which have been accompanied with sporadic expansion/deletion and exon-intron remodeling. The differentiated enzymatic properties might be acquired by the evolutionary relaxation of selection pressure and/or biochemical adaptation to the acting environments. Our present study would be beneficial to get detailed insights into the complex GPx evolution, and to understand the molecular basis of the specialized physiological implications of this antioxidant system in their respective donor organisms.

Bae, Young-An; Cai, Guo-Bin; Kim, Seon-Hee; Zo, Young-Gun; Kong, Yoon

2009-01-01

368

Anti-schistosomal Intervention Targets Identified by Lifecycle Transcriptomic Analyses  

PubMed Central

Background Novel methods to identify anthelmintic drug and vaccine targets are urgently needed, especially for those parasite species currently being controlled by singular, often limited strategies. A clearer understanding of the transcriptional components underpinning helminth development will enable identification of exploitable molecules essential for successful parasite/host interactions. Towards this end, we present a combinatorial, bioinformatics-led approach, employing both statistical and network analyses of transcriptomic data, for identifying new immunoprophylactic and therapeutic lead targets to combat schistosomiasis. Methodology/Principal Findings Utilisation of a Schistosoma mansoni oligonucleotide DNA microarray consisting of 37,632 elements enabled gene expression profiling from 15 distinct parasite lifecycle stages, spanning three unique ecological niches. Statistical approaches of data analysis revealed differential expression of 973 gene products that minimally describe the three major characteristics of schistosome development: asexual processes within intermediate snail hosts, sexual maturation within definitive vertebrate hosts and sexual dimorphism amongst adult male and female worms. Furthermore, we identified a group of 338 constitutively expressed schistosome gene products (including 41 transcripts sharing no sequence similarity outside the Platyhelminthes), which are likely to be essential for schistosome lifecycle progression. While highly informative, statistics-led bioinformatics mining of the transcriptional dataset has limitations, including the inability to identify higher order relationships between differentially expressed transcripts and lifecycle stages. Network analysis, coupled to Gene Ontology enrichment investigations, facilitated a re-examination of the dataset and identified 387 clusters (containing 12,132 gene products) displaying novel examples of developmentally regulated classes (including 294 schistosomula and/or adult transcripts with no known sequence similarity outside the Platyhelminthes), which were undetectable by the statistical comparisons. Conclusions/Significance Collectively, statistical and network-based exploratory analyses of transcriptomic datasets have led to a thorough characterisation of schistosome development. Information obtained from these experiments highlighted key transcriptional programs associated with lifecycle progression and identified numerous anti-schistosomal candidate molecules including G-protein coupled receptors, tetraspanins, Dyp-type peroxidases, fucosyltransferases, leishmanolysins and the netrin/netrin receptor complex.

Fitzpatrick, Jennifer M.; Peak, Emily; Perally, Samirah; Chalmers, Iain W.; Barrett, John; Yoshino, Timothy P.; Ivens, Alasdair C.; Hoffmann, Karl F.

2009-01-01

369

Proteomic analysis of the Echinococcus granulosus metacestode during infection of its intermediate host.  

PubMed

Cystic hydatid disease (CHD) is caused by infection with the Echinococcus granulosus metacestode and affects both humans and livestock. In this work, we performed a proteomic analysis of the E. granulosus metacestode during infection of its intermediate bovine host. Parasite proteins were identified in different metacestode components (94 from protoscolex, 25 from germinal layer and 20 from hydatid cyst fluid), along with host proteins (58) that permeate into the hydatid cyst, providing new insights into host-parasite interplay. E. granulosus and platyhelminth EST data allowed successful identification of proteins potentially involved in downregulation of host defenses, highlighting possible evasion mechanisms adopted by the parasite to establish infection. Several intracellular proteins were found in hydatid cyst fluid, revealing a set of newly identified proteins that were previously thought to be inaccessible for inducing or modulating the host immune response. Host proteins identified in association with the hydatid cyst suggest that the parasite may bind/adsorb host molecules with nutritional and/or immune evasion purposes, masking surface antigens or inhibiting important effector molecules of host immunity, such as complement components and calgranulin. Overall, our results provide valuable information on parasite survival strategies in the adverse host environment and on the molecular mechanisms underpinning CHD immunopathology. PMID:20217864

Monteiro, Karina M; de Carvalho, Marcos O; Zaha, Arnaldo; Ferreira, Henrique B

2010-05-01

370

Seaweed extracts as a natural control against the monogenean ectoparasite, Neobenedenia sp., infecting farmed barramundi (Lates calcarifer).  

PubMed

Aqueous extracts from common tropical seaweeds were evaluated for their effect on the life cycle of the commercially important ectoparasite, Neobenedenia sp. (Platyhelminthes: Monogenea), through the survival of attached adult parasites, period of embryonic development, hatching success and oncomiracidia (larvae) infection success. There was no significant effect of any extract on the survival of adult parasites attached to fish hosts or infection success by oncomiracidia. However, the extracts of two seaweeds, Ulva sp. and Asparagopsis taxiformis, delayed embryonic development and inhibited egg hatching. The extract of A. taxiformis was most effective, inhibiting embryonic development of Neobenedenia sp. and reducing hatching success to 3% compared with 99% for the seawater control. Furthermore, of the 3% of eggs that hatched, time to first and last hatch was delayed (days 14 and 18) compared with the seawater control (days 5 and 7). Asparagopsis taxiformis shows the most potential for development as a natural treatment to manage monogenean infections in intensive aquaculture with the greatest impact at the embryo stage. PMID:23068914

Hutson, Kate S; Mata, Leonardo; Paul, Nicholas A; de Nys, Rocky

2012-10-12

371

HelmCoP: an online resource for helminth functional genomics and drug and vaccine targets prioritization.  

PubMed

A vast majority of the burden from neglected tropical diseases result from helminth infections (nematodes and platyhelminthes). Parasitic helminthes infect over 2 billion, exerting a high collective burden that rivals high-mortality conditions such as AIDS or malaria, and cause devastation to crops and livestock. The challenges to improve control of parasitic helminth infections are multi-fold and no single category of approaches will meet them all. New information such as helminth genomics, functional genomics and proteomics coupled with innovative bioinformatic approaches provide fundamental molecular information about these parasites, accelerating both basic research as well as development of effective diagnostics, vaccines and new drugs. To facilitate such studies we have developed an online resource, HelmCoP (Helminth Control and Prevention), built by integrating functional, structural and comparative genomic data from plant, animal and human helminthes, to enable researchers to develop strategies for drug, vaccine and pesticide prioritization, while also providing a useful comparative genomics platform. HelmCoP encompasses genomic data from several hosts, including model organisms, along with a comprehensive suite of structural and functional annotations, to assist in comparative analyses and to study host-parasite interactions. The HelmCoP interface, with a sophisticated query engine as a backbone, allows users to search for multi-factorial combinations of properties and serves readily accessible information that will assist in the identification of various genes of interest. HelmCoP is publicly available at: http://www.nematode.net/helmcop.html. PMID:21760913

Abubucker, Sahar; Martin, John; Taylor, Christina M; Mitreva, Makedonka

2011-07-08

372

The unique ultrastructure of the uterus of the Gyrocotylidea Poche, 1926 (Cestoda) and its phylogenetic implications.  

PubMed

The members of the order Gyrocotylidea are monozoic tapeworms and generally considered to be the most primitive group of the Cestoda in terms of the evolution of this platyhelminth class. As part of a series of ultrastructural studies on Gyrocotyle urna (Wagener, 1852), three regions of the uterus were distinguished. The proximal region of the uterus is characterised by underlying perikarya, the presence of septate junctions within the epithelial wall and two types of specialised outer coverings, lamellae and cilia. The middle, syncytial region of the uterus is covered by short lamellae and marked by a concentration of sunken glandular perikarya (uterine glands). Glandular spheroidal granules (c.0.45 microm in diameter) of moderately dense content and a fine fibrillar texture are liberated by migration through the luminal membrane. The epithelium of the sac-shaped, distal region of the uterine duct is interrupted by cytoplasmic processes of sunken epithelial bodies, covered with lamellae and contains septate junctions. A muscular sphincter surrounds the narrow, terminal region of the distal uterine duct. The ultrastructural pattern of the uterus of the Gyrocotylidea has important discriminating traits (i.e. the presence of sunken perikarya along its entire length, septate junctions within the uterine epithelial cytoplasm of the proximal and distal regions, and cilia on the surface of its proximal region) unique among the Neodermata and which may represent autapomorphic character states for the group. PMID:19731092

Poddubnaya, Larisa G; Kuchta, Roman; Levron, Céline; Gibson, David I; Scholz, Tomás

2009-09-04

373

The complete mitochondrial genome of the onychophoran Epiperipatus biolleyi reveals a unique transfer RNA set and provides further support for the ecdysozoa hypothesis.  

PubMed

Onychophora (velvet worms) play a crucial role in current discussions on position of arthropods. The ongoing Articulata/Ecdysozoa debate is in need of additional ground pattern characters for Panarthropoda (Arthropoda, Tardigrada, and Onychophora). Hence, Onychophora is an important outgroup taxon in resolving the relationships among arthropods, irrespective of whether morphological or molecular data are used. To date, there has been a noticeable lack of mitochondrial genome data from onychophorans. Here, we present the first complete mitochondrial genome sequence of an onychophoran, Epiperipatus biolleyi (Peripatidae), which shows several characteristic features. Specifically, the gene order is considerably different from that in other arthropods and other bilaterians. In addition, there is a lack of 9 tRNA genes usually present in bilaterian mitochondrial genomes. All these missing tRNAs have anticodon sequences corresponding to 4-fold degenerate codons, whereas the persisting 13 tRNAs all have anticodons pairing with 2-fold degenerate codons. Sequence-based phylogenetic analysis of the mitochondrial protein-coding genes provides a robust support for a clade consisting of Onychophora, Priapulida, and Arthropoda, which confirms the Ecdysozoa hypothesis. However, resolution of the internal ecdysozoan relationships suffers from a cluster of long-branching taxa (including Nematoda and Platyhelminthes) and a lack of data from Tardigrada and further nemathelminth taxa in addition to nematodes and priapulids. PMID:17934206

Podsiadlowski, Lars; Braband, Anke; Mayer, Georg

2007-10-13

374

Evolutionarily Ancient Association of the FoxJ1 Transcription Factor with the Motile Ciliogenic Program  

PubMed Central

It is generally believed that the last eukaryotic common ancestor (LECA) was a unicellular organism with motile cilia. In the vertebrates, the winged-helix transcription factor FoxJ1 functions as the master regulator of motile cilia biogenesis. Despite the antiquity of cilia, their highly conserved structure, and their mechanism of motility, the evolution of the transcriptional program controlling ciliogenesis has remained incompletely understood. In particular, it is presently not known how the generation of motile cilia is programmed outside of the vertebrates, and whether and to what extent the FoxJ1-dependent regulation is conserved. We have performed a survey of numerous eukaryotic genomes and discovered that genes homologous to foxJ1 are restricted only to organisms belonging to the unikont lineage. Using a mis-expression assay, we then obtained evidence of a conserved ability of FoxJ1 proteins from a number of diverse phyletic groups to activate the expression of a host of motile ciliary genes in zebrafish embryos. Conversely, we found that inactivation of a foxJ1 gene in Schmidtea mediterranea, a platyhelminth (flatworm) that utilizes motile cilia for locomotion, led to a profound disruption in the differentiation of motile cilia. Together, all of these findings provide the first evolutionary perspective into the transcriptional control of motile ciliogenesis and allow us to propose a conserved FoxJ1-regulated mechanism for motile cilia biogenesis back to the origin of the metazoans.

Ho, Hao Kee; Babu, Deepak; Eitel, Michael; Narasimhan, Vijayashankaranarayanan; Tiku, Varnesh; Westbrook, Jody; Schierwater, Bernd; Roy, Sudipto

2012-01-01

375

Progress in nemertean biology: development and phylogeny.  

PubMed

This paper reviews progress in developmental biology and phylogeny of the Nemertea, a common but poorly studied spiralian taxon of considerable ecological and evolutionary significance. Analyses of reproductive biology (including calcium dynamics during fertilization and oocyte maturation), larval morphology and development and developmental genetics have significantly extended our knowledge of spiralian developmental biology. Developmental genetics studies have in addition provided characters useful for reconstructing metazoan phylogeny. Reinvestigation of the cell lineage of Cerebratulus lacteus using fluorescent tracers revealed that endomesoderm forms from the 4d cell as in other spiralians and that ectomesoderm is derived from the 3a and 3b cells as in annelids, echiurans and molluscs. Studies examining blastomere specification show that cell fates are established precociously in direct developers and later in indirect developers. Morphological characters used to estimate the phylogenetic position of nemerteans are critically re-evaluated, and cladistic analyses of morphology reveal that conflicting hypotheses of nemertean relationships result because of different provisional homology statements. Analyses that include disputed homology statements (1, gliointerstitial cell system 2, coelomic circulatory system) suggest that nemerteans form the sister taxon to the coelomate spiralian taxa rather than the sister taxon to Platyhelminthes. Analyses of small subunit rRNA (18S rDNA) sequences alone or in combination with morphological characters support the inclusion of the nemerteans in a spiralian coelomate clade nested within a more inclusive lophotrochozoan clade. Ongoing evaluation of nemertean relationships with mitochondrial gene rearrangements and other molecular characters is discussed. PMID:21708766

Turbeville, J M

2002-07-01

376

Invasive species threat: parasite phylogenetics reveals patterns and processes of host-switching between non-native and native captive freshwater turtles.  

PubMed

One of the major threats to biodiversity involves biological invasions with direct consequences on the stability of ecosystems. In this context, the role of parasites is not negligible as it may enhance the success of invaders. The red-eared slider, Trachemys scripta elegans, has been globally considered among the worst invasive species. Since its introduction through the pet trade, T. s. elegans is now widespread and represents a threat for indigenous species. Because T. s. elegans coexists with Emys orbicularis and Mauremys leprosa in Europe, it has been suggested it may compete with the native turtle species and transmit pathogens. We examined parasite transfer from American captive to the two native species that co-exist in artificial pools of a Turtle Farm in France. As model parasite species we used platyhelminth worms of the family Polystomatidae (Monogenea) because polystomes have been described from American turtles in their native range. Phylogenetic relationships among polystomes parasitizing chelonian host species that are geographically widespread show patterns of diversification more complex than expected. Using DNA barcoding to identify species from adult and/or polystome eggs, several cases of host switching from exotic to indigenous individuals were illustrated, corroborating that parasite transmission is important when considering the pet trade and in reintroduction programmes to reinforce wild populations of indigenous species. PMID:21767431

Verneau, O; Palacios, C; Platt, T; Alday, M; Billard, E; Allienne, J-F; Basso, C; DU Preez, L H

2011-07-18

377

Presence of galactosylated core fucose on N-glycans in the planaria Dugesia japonica.  

PubMed

Planarial species are of especial interest to biologists due to the phenomenon of pluripotency and, in comparison to other developmental processes, it can be hypothesised that glycan-lectin interactions may play a role. In order to examine the N-glycans of one of these organisms, Dugesia japonica, peptide:N-glycosidase A was employed and the released glycans were subject to pyridylamination, HPLC and mass spectrometric analysis. A range of oligomannosidic glycans was observed with a trimethylated Man(5) GlcNAc(2) structure being the dominant species. Three glycans were also observed to contain deoxyhexose; in particular, a glycan with the composition Hex(4) HexNAc(2) Fuc(1) Me(2) was revealed by exoglycosidase digestion, in combination with MS/MS, to contain a galactosylated core ?1,6-fucose residue, whereas this core modification was found to be capped with a methylhexose residue in the case of a Hex(5) HexNAc(2) Fuc(1) Me(3) structure. This is the first report of these types of structures in a platyhelminth and indicates that the 'GalFuc' modification of N-glycans is not just restricted to molluscs and nematodes. PMID:21630384

Paschinger, Katharina; Razzazi-Fazeli, Ebrahim; Furukawa, Kiyoshi; Wilson, Iain B H

2011-06-01

378

Mixed Infections and Hybridisation in Monogenean Parasites  

PubMed Central

Theory predicts that sexual reproduction promotes disease invasion by increasing the evolutionary potential of the parasite, whereas asexual reproduction tends to enhance establishment success and population growth rate. Gyrodactylid monogeneans are ubiquitous ectoparasites of teleost fish, and the evolutionary success of the specious Gyrodactylus genus is thought to be partly due to their use of various modes of reproduction. Gyrodactylus turnbulli is a natural parasite of the guppy (Poecilia reticulata), a small, tropical fish used as a model for behavioural, ecological and evolutionary studies. Using experimental infections and a recently developed microsatellite marker, we conclusively show that monogenean parasites reproduce sexually. Conservatively, we estimate that sexual recombination occurs and that between 3.7–10.9% of the parasites in our experimental crosses are hybrid genotypes with ancestors from different laboratory strains of G. turnbulli. We also provide evidence of hybrid vigour and/or inter-strain competition, which appeared to lead to a higher maximum parasite load in mixed infections. Finally, we demonstrate inbreeding avoidance for the first time in platyhelminths which may influence the distribution of parasites within a host and their subsequent exposure to the host's localized immune response. Combined reproductive modes and inbreeding avoidance may explain the extreme evolutionary diversification success of parasites such as Gyrodactylus, where host-parasite coevolution is punctuated by relatively frequent host switching.

Schelkle, Bettina; Faria, Patricia J.; Johnson, Mireille B.; van Oosterhout, Cock; Cable, Joanne

2012-01-01

379

A New Family of Receptor Tyrosine Kinases with a Venus Flytrap Binding Domain in Insects and Other Invertebrates Activated by Aminoacids  

PubMed Central

Background Tyrosine kinase receptors (RTKs) comprise a large family of membrane receptors that regulate various cellular processes in cell biology of diverse organisms. We previously described an atypical RTK in the platyhelminth parasite Schistosoma mansoni, composed of an extracellular Venus flytrap module (VFT) linked through a single transmembrane domain to an intracellular tyrosine kinase domain similar to that of the insulin receptor. Methods and Findings Here we show that this receptor is a member of a new family of RTKs found in invertebrates, and particularly in insects. Sixteen new members of this family, named Venus Kinase Receptor (VKR), were identified in many insects. Structural and phylogenetic studies performed on VFT and TK domains showed that VKR sequences formed monophyletic groups, the VFT group being close to that of GABAB receptors and the TK one being close to that of insulin receptors. We show that a recombinant VKR is able to autophosphorylate on tyrosine residues, and report that it can be activated by L-arginine. This is in agreement with the high degree of conservation of the alpha amino acid binding residues found in many amino acid binding VFTs. The presence of high levels of vkr transcripts in larval forms and in female gonads indicates a putative function of VKR in reproduction and/or development. Conclusion The identification of RTKs specific for parasites and insect vectors raises new perspectives for the control of human parasitic and infectious diseases.

Ahier, Arnaud; Rondard, Philippe; Gouignard, Nadege; Khayath, Naji; Huang, Siluo; Trolet, Jacques; Donoghue, Daniel J.; Gauthier, Monique; Pin, Jean-Philippe; Dissous, Colette

2009-01-01

380

Lessons from parasitic flatworms about evolution and historical biogeography of their vertebrate hosts.  

PubMed

Cophylogenetic studies investigate the evolutionary trends within host-parasite associations. Examination of the different levels of fidelity between host and parasite phylogenies provides a powerful tool to inspect patterns and processes of parasite diversification over host evolution and geological times. Within the phylum Platyhelminthes, the monogeneans are mainly fish parasites. The Polystomatidae, however, are known from the sarcopterygian Australian lungfish and tetrapods such as amphibians, freshwater turtles, and the African hippopotamus. Cophylogenetic and biogeographic vicariance analyses, supplemented by molecular calibrations, showed that the Polystomatidae may track the evolutionary history of the first aquatic tetrapods in the Palaeozoic age. Evolutionary lines of the major polystome lineages would also be intimately related to the evolution of their hosts over hundreds of millions years. Since the Mesozoic, evolution of polystomes would have been shaped mainly by plate tectonics during the break-up of Gondwanaland and subsequent dispersal of ancestral neobatrachian host lineages. Therefore the Polystomatidae could serve as a novel model to improve cophylogenetic tools and to inspect a suite of questions about the evolution of vertebrate hosts. PMID:19281948

Verneau, Olivier; Du Preez, Louis; Badets, Mathieu

2008-11-29

381

Phylogeny and mitochondrial gene order variation in Lophotrochozoa in the light of new mitogenomic data from Nemertea  

PubMed Central

Background The new animal phylogeny established several taxa which were not identified by morphological analyses, most prominently the Ecdysozoa (arthropods, roundworms, priapulids and others) and Lophotrochozoa (molluscs, annelids, brachiopods and others). Lophotrochozoan interrelationships are under discussion, e.g. regarding the position of Nemertea (ribbon worms), which were discussed to be sister group to e.g. Mollusca, Brachiozoa or Platyhelminthes. Mitochondrial genomes contributed well with sequence data and gene order characters to the deep metazoan phylogeny debate. Results In this study we present the first complete mitochondrial genome record for a member of the Nemertea, Lineus viridis. Except two trnP and trnT, all genes are located on the same strand. While gene order is most similar to that of the brachiopod Terebratulina retusa, sequence based analyses of mitochondrial genes place nemerteans close to molluscs, phoronids and entoprocts without clear preference for one of these taxa as sister group. Conclusion Almost all recent analyses with large datasets show good support for a taxon comprising Annelida, Mollusca, Brachiopoda, Phoronida and Nemertea. But the relationships among these taxa vary between different studies. The analysis of gene order differences gives evidence for a multiple independent occurrence of a large inversion in the mitochondrial genome of Lophotrochozoa and a re-inversion of the same part in gastropods. We hypothesize that some regions of the genome have a higher chance for intramolecular recombination than others and gene order data have to be analysed carefully to detect convergent rearrangement events.

Podsiadlowski, Lars; Braband, Anke; Struck, Torsten H; von Dohren, Jorn; Bartolomaeus, Thomas

2009-01-01

382

Spiral cleavage and early embryology of a loxosomatid entoproct and the usefulness of spiralian apical cross patterns for phylogenetic inferences  

PubMed Central

Background Among the four major bilaterian clades, Deuterostomia, Acoelomorpha, Ecdysozoa, and Lophotrochozoa, the latter shows an astonishing diversity of bodyplans. While the largest lophotrochozoan assemblage, the Spiralia, which at least comprises Annelida, Mollusca, Entoprocta, Platyhelminthes, and Nemertea, show a spiral cleavage pattern, Ectoprocta, Brachiopoda and Phoronida (the Lophophorata) cleave radially. Despite a vast amount of recent molecular phylogenetic analyses, the interrelationships of lophotrochozoan phyla remain largely unresolved. Thereby, Entoprocta play a key role, because they have frequently been assigned to the Ectoprocta, despite their differently cleaving embryos. However, developmental data on entoprocts employing modern methods are virtually non-existent and the data available rely exclusively on sketch drawings, thus calling for thorough re-investigation. Results By applying fluorescence staining in combination with confocal microscopy and 3D-imaging techniques, we analyzed early embryonic development of a basal loxosomatid entoproct. We found that cleavage is asynchronous, equal, and spiral. An apical rosette, typical for most spiralian embryos, is formed. We also identified two cross-like cellular arrangements that bear similarities to both, a "molluscan-like" as well as an "annelid-like" cross, respectively. Conclusions A broad comparison of cleavage types and apical cross patterns across Lophotrochozoa shows high plasticity of these character sets and we therefore argue that these developmental traits should be treated and interpreted carefully when used for phylogenetic inferences.

2012-01-01

383

Combination of de novo assembly of massive sequencing reads with classical repeat prediction improves identification of repetitive sequences in Schistosoma mansoni.  

PubMed

The genome of the parasitic platyhelminth Schistosoma mansoni is composed of approximately 40% of repetitive sequences of which roughly 20% correspond to transposable elements. When the genome sequence became available, conventional repeat prediction programs were used to find these repeats, but only a fraction could be identified. To exhaustively characterize the repeats we applied a new massive sequencing based strategy: we re-sequenced the genome by next generation sequencing, aligned the sequencing reads to the genome and assembled all multiple-hit reads into contigs corresponding to the repetitive part of the genome. We present here, for the first time, this de novo repeat assembly strategy and we confirm that such assembly is feasible. We identified and annotated 4,143 new repeats in the S. mansoni genome. At least one third of the repeats are transcribed. This strategy allowed us also to identify 14 new microsatellite markers, which can be used for pedigree studies. Annotations and the combined (previously known and new) 5,420 repeat sequences (corresponding to 47% of the genome) are available for download (http://methdb.univ-perp.fr/downloads/). PMID:22381218

Lepesant, Julie M J; Roquis, David; Emans, Rémi; Cosseau, Céline; Arancibia, Nathalie; Mitta, Guillaume; Grunau, Christoph

2012-02-21

384

Statistical modelling and phylogenetic analysis of a deaminase domain.  

PubMed

Deamination reactions are catalyzed by a variety of enzymes including those involved in nucleoside/nucleotide metabolism and cytosine to uracil (C-->U) and adenosine to inosine (A-->I) mRNA editing. The active site of the deaminase (DM) domain in these enzymes contains a conserved histidine (or rarely cysteine), two cysteines and a glutamate proposed to act as a proton shuttle during deamination. Here, a statistical model, a hidden Markov model (HMM), of the DM domain has been created which identifies currently known DM domains and suggests new DM domains in viral, bacterial and eucaryotic proteins. However, no DM domains were identified in the currently predicted proteins from the archaeon Methanococcus jannaschii and possible causes for, and a potential means to ameliorate this situation are discussed. In some of the newly identified DM domains, the glutamate is changed to a residue that could not function as a proton shuttle and in one instance (Mus musculus spermatid protein TENR) the cysteines are also changed to lysine and serine. These may be non-competent DM domains able to bind but not act upon their substrate. Phylogenetic analysis using an HMM-generated alignment of DM domains reveals three branches with clear substructure in each branch. The results suggest DM domains that are candidates for yeast, platyhelminth, plant and mammalian C-->U and A-->I mRNA editing enzymes. Some bacterial and eucaryotic DM domains form distinct branches in the phylogenetic tree suggesting the existence of common, novel substrates. PMID:9541871

Mian, I S; Moser, M J; Holley, W R; Chatterjee, A

1998-01-01

385

HelmCoP: An Online Resource for Helminth Functional Genomics and Drug and Vaccine Targets Prioritization  

PubMed Central

A vast majority of the burden from neglected tropical diseases result from helminth infections (nematodes and platyhelminthes). Parasitic helminthes infect over 2 billion, exerting a high collective burden that rivals high-mortality conditions such as AIDS or malaria, and cause devastation to crops and livestock. The challenges to improve control of parasitic helminth infections are multi-fold and no single category of approaches will meet them all. New information such as helminth genomics, functional genomics and proteomics coupled with innovative bioinformatic approaches provide fundamental molecular information about these parasites, accelerating both basic research as well as development of effective diagnostics, vaccines and new drugs. To facilitate such studies we have developed an online resource, HelmCoP (Helminth Control and Prevention), built by integrating functional, structural and comparative genomic data from plant, animal and human helminthes, to enable researchers to develop strategies for drug, vaccine and pesticide prioritization, while also providing a useful comparative genomics platform. HelmCoP encompasses genomic data from several hosts, including model organisms, along with a comprehensive suite of structural and functional annotations, to assist in comparative analyses and to study host-parasite interactions. The HelmCoP interface, with a sophisticated query engine as a backbone, allows users to search for multi-factorial combinations of properties and serves readily accessible information that will assist in the identification of various genes of interest. HelmCoP is publicly available at: http://www.nematode.net/helmcop.html.

Taylor, Christina M.; Mitreva, Makedonka

2011-01-01

386

Polyethyleneimine (PEI) Mediated siRNA Gene Silencing in the Schistosoma mansoni Snail Host, Biomphalaria glabrata  

PubMed Central

An in vivo, non-invasive technique for gene silencing by RNA interference (RNAi) in the snail, Biomphalaria glabrata, has been developed using cationic polymer polyethyleneimine (PEI) mediated delivery of long double-stranded (ds) and small interfering (si) RNA. Cellular delivery was evaluated and optimized by using a ‘mock’ fluorescent siRNA. Subsequently, we used the method to suppress expression of Cathepsin B (CathB) with either the corresponding siRNA or dsRNA of this transcript. In addition, the knockdown of peroxiredoxin (Prx) at both RNA and protein levels was achieved with the PEI-mediated soaking method. B. glabrata is an important snail host for the transmission of the parasitic digenean platyhelminth, Schistosoma mansoni that causes schistosomiasis in the neotropics. Progress is being made to realize the genome sequence of the snail and to uncover gene expression profiles and cellular pathways that enable the snail to either prevent or sustain an infection. Using PEI complexes, a convenient soaking method has been developed, enabling functional gene knockdown studies with either dsRNA or siRNA. The protocol developed offers a first whole organism method for host-parasite gene function studies needed to identify key mechanisms required for parasite development in the snail host, which ultimately are needed as points for disrupting this parasite mediated disease.

Knight, Matty; Miller, Andre; Liu, Yijia; Scaria, Puthupparampil; Woodle, Martin; Ittiprasert, Wannaporn

2011-01-01

387

On the quantitative distribution and community structure of the meio and macrofaunal communities in the coastal area of the Central Adriatic Sea (Italy).  

PubMed

Many coastal areas have served as repositories of different anthropogenic and naturally induced organic material and nutrients. The major sources thereof are riverine inputs which strongly influence the spatial and temporal distribution of benthic communities. In this study, the benthic foraminiferal, meiofaunal, and macrofaunal colonies in front of three rivers in a poorly known, but environmentally valuable, area of the Central Adriatic Sea have been examined concurrently. The physico-chemical parameters of bottom water and sediment characteristics were determined in order to characterize both the sediment-water interface and the benthic environments. Although changes in the biota are neither univocal nor unidirectional, a moderate influence of riverine input on the different communities' components can be inferred. The most affected taxa are foraminifera and copepods and, to a lesser extent, meiofaunal polychaetes and platyhelminthes. These results are also tested by the ABC curves, which reveal that the macrofaunal communities closest to the river mouths are moderately disturbed. This integrated investigation documents, for the first time, how benthic communities can be used as an early warning indicator with which to monitor the health quality of a coastal ecosystem. PMID:21120690

Frontalini, Fabrizio; Semprucci, Federica; Coccioni, Rodolfo; Balsamo, Maria; Bittoni, Paolo; Covazzi-Harriague, Anabella

2010-12-01

388

Transcriptome Analysis Reveals Strain-Specific and Conserved Stemness Genes in Schmidtea mediterranea  

PubMed Central

The planarian Schmidtea mediterranea is a powerful model organism for studying stem cell biology due to its extraordinary regenerative ability mediated by neoblasts, a population of adult somatic stem cells. Elucidation of the S. mediterranea transcriptome and the dynamics of transcript expression will increase our understanding of the gene regulatory programs that regulate stem cell function and differentiation. Here, we have used RNA-Seq to characterize the S. mediterranea transcriptome in sexual and asexual animals and in purified neoblast and differentiated cell populations. Our analysis identified many uncharacterized genes, transcripts, and alternatively spliced isoforms that are differentially expressed in a strain or cell type-specific manner. Transcriptome profiling of purified neoblasts and differentiated cells identified neoblast-enriched transcripts, many of which likely play important roles in regeneration and stem cell function. Strikingly, many of the neoblast-enriched genes are orthologs of genes whose expression is enriched in human embryonic stem cells, suggesting that a core set of genes that regulate stem cell function are conserved across metazoan species.

Lu, Yi-Chien; Horowitz, Michael; Graveley, Brenton R.

2012-01-01

389

Automated analysis of behavior: a computer-controlled system for drug screening and the investigation of learning.  

PubMed

Efforts to understand cognition will be greatly facilitated by computerized systems that enable the automated analysis of animal behavior. A number of controversies in the invertebrate learning field have resulted from difficulties inherent in manual experiments. Driven by the necessity to overcome these problems during investigation of neural function in planarian flatworms and frog larvae, we designed and developed a prototype for an inexpensive, flexible system that enables automated control and analysis of behavior and learning. Applicable to a variety of small animals such as flatworms and zebrafish, this system allows automated analysis of innate behavior, as well as of learning and memory in a plethora of conditioning paradigms. We present here the schematics of a basic prototype, which overcomes experimenter effects and operator tedium, enabling a large number of animals to be analyzed with transparent on-line access to primary data. A scaled-up version of this technology represents an efficient methodology to screen pharmacological and genetic libraries for novel neuroactive reagents of basic and biomedical relevance. PMID:16779826

Hicks, Caitlin; Sorocco, Debra; Levin, Michael

2006-08-01

390

RNA interference can target pre-mRNA: consequences for gene expression in a Caenorhabditis elegans operon.  

PubMed Central

In nematodes, flies, trypanosomes, and planarians, introduction of double-stranded RNA results in sequence-specific inactivation of gene function, a process termed RNA interference (RNAi). We demonstrate that RNAi against the Caenorhabditis elegans gene lir-1, which is part of the lir-1/lin-26 operon, induced phenotypes very different from a newly isolated lir-1 null mutation. Specifically, lir-1(RNAi) induced embryonic lethality reminiscent of moderately strong lin-26 alleles, whereas the lir-1 null mutant was viable. We show that the lir-1(RNAi) phenotypes resulted from a severe loss of lin-26 gene expression. In addition, we found that RNAi directed against lir-1 or lin-26 introns induced similar phenotypes, so we conclude that lir-1(RNAi) targets the lir-1/lin-26 pre-mRNA. This provides direct evidence that RNA interference can prevent gene expression by targeting nuclear transcripts. Our results highlight that caution may be necessary when interpreting RNA interference without the benefit of mutant alleles.

Bosher, J M; Dufourcq, P; Sookhareea, S; Labouesse, M

1999-01-01

391

In vivo comparison of harmine efficacy against psychostimulants: preferential inhibition of the cocaine response through a glutamatergic mechanism  

PubMed Central

Harmine is a ?-carboline compound that targets glutamatergic, monoaminergic, and GABAergic pathways underlying drug addiction. We compared the efficacy of harmine against different psychoactive drugs using an invertebrate (planarian) assay designed to quantify ‘C-shape’ responses. Harmine itself (0.01 – 10 µM) did not produce C-shapes. However, when applied over the same concentration range, harmine significantly inhibited C-shapes elicited by cocaine, with a concentration of 0.1 µM producing almost 90% inhibition. Consistent with its putative actions, harmine produced a similar, though less efficacious, inhibition of C-shapes elicited by the substituted amphetamines methamphetamine and mephedrone (4-methylmethcathinone) but was much less effective against nicotine. When tested in the presence of the glutamate transporter inhibitor dihydrokainate (DHK) (0.1, 1 µM), harmine (0.1 µM) efficacy against cocaine-induced C-shapes was significantly reduced. Harmine also attenuated C-shapes elicited by N-methyl-D-aspartate (NMDA) and by glutamate itself. The present data suggest that harmine displays preferential efficacy against different addictive substances (cocaine > amphetamines > nicotine) and, at least for cocaine, is dependent on the glutamate system.

Owaisat, Suzan; Raffa, Robert B.; Rawls, Scott M.

2012-01-01

392

Invertebrate models for biomedical research, testing, and education.  

PubMed

Invertebrate animals have been used as medicinals for 4,000 years and have served as models for research and teaching since the late 1800s. Interest in invertebrate models has increased over the past several decades as the research community has responded to public concerns about the use of vertebrate animals in research. As a result, invertebrates are being evaluated and recognized as models for many diseases and conditions. Their use has led to discoveries in almost every area of biology and medicine--from embryonic development to aging processes. Species range from terrestrial invertebrates such as nematodes and insects to freshwater and marine life including planarians, crustaceans, molluscs, and many others. The most often used models are the fruit fly Drosophila melanogaster and the minuscule nematode Caenorhabditis elegans. Topics in this article are categorized by biologic system, process, or disease with discussion of associated invertebrate models. Sections on bioactive products discovered from invertebrates follow the models section, and the article concludes with uses of invertebrates in teaching. The models reviewed can serve as references for scientists, researchers, veterinarians, institutional animal care and use committees (IACUCs), and others interested in alternatives to vertebrate animals. PMID:21709307

Wilson-Sanders, Susan E

2011-01-01

393

Origin and Evolution of Dishevelled  

PubMed Central

Dishevelled (Dsh or Dvl) is an important signaling protein, playing a key role in Wnt signaling and relaying cellular information for several developmental pathways. Dsh is highly conserved among metazoans and has expanded into a multigene family in most bilaterian lineages, including vertebrates, planarians, and nematodes. These orthologs, where explored, are known to have considerable overlap in function, but evidence for functional specialization continues to mount. We performed a comparative analysis of Dsh across animals to explore protein architecture and identify conserved and divergent features that could provide insight into functional specialization with an emphasis on invertebrates, especially nematodes. We find evidence of dynamic evolution of Dsh, particularly among nematodes, with taxa varying in ortholog number from one to three. We identify a new domain specific to some nematode lineages and find an unexpected nuclear localization signal conserved in many Dsh orthologs. Our findings raise questions of protein evolution in general and provide clues as to how animals have dealt with the complex intricacies of having a protein, such as Dsh, act as a central messenger hub connected to many different and vitally important pathways. We discuss our findings in the context of functional specialization and bring many testable hypotheses to light.

Dillman, Adler R.; Minor, Paul J.; Sternberg, Paul W.

2013-01-01

394

Transcriptome analysis reveals strain-specific and conserved stemness genes in Schmidtea mediterranea.  

PubMed

The planarian Schmidtea mediterranea is a powerful model organism for studying stem cell biology due to its extraordinary regenerative ability mediated by neoblasts, a population of adult somatic stem cells. Elucidation of the S. mediterranea transcriptome and the dynamics of transcript expression will increase our understanding of the gene regulatory programs that regulate stem cell function and differentiation. Here, we have used RNA-Seq to characterize the S. mediterranea transcriptome in sexual and asexual animals and in purified neoblast and differentiated cell populations. Our analysis identified many uncharacterized genes, transcripts, and alternatively spliced isoforms that are differentially expressed in a strain or cell type-specific manner. Transcriptome profiling of purified neoblasts and differentiated cells identified neoblast-enriched transcripts, many of which likely play important roles in regeneration and stem cell function. Strikingly, many of the neoblast-enriched genes are orthologs of genes whose expression is enriched in human embryonic stem cells, suggesting that a core set of genes that regulate stem cell function are conserved across metazoan species. PMID:22496805

Resch, Alissa M; Palakodeti, Dasaradhi; Lu, Yi-Chien; Horowitz, Michael; Graveley, Brenton R

2012-04-04

395

A sex-specific transcription factor controls male identity in a simultaneous hermaphrodite.  

PubMed

Evolutionary transitions between hermaphroditic and dioecious reproductive states are found in many groups of animals. To understand such transitions, it is important to characterize diverse modes of sex determination utilized by metazoans. Currently, little is known about how simultaneous hermaphrodites specify and maintain male and female organs in a single individual. Here we show that a sex-specific gene, Smed-dmd-1 encoding a predicted doublesex/male-abnormal-3 (DM) domain transcription factor, is required for specification of male germ cells in a simultaneous hermaphrodite, the planarian Schmidtea mediterranea. dmd-1 has a male-specific role in the maintenance and regeneration of the testes and male accessory reproductive organs. In addition, a homologue of dmd-1 exhibits male-specific expression in Schistosoma mansoni, a derived, dioecious flatworm. These results demonstrate conservation of the role of DM domain genes in sexual development in lophotrochozoans and suggest one means by which modulation of sex-specific pathways can drive the transition from hermaphroditism to dioecy. PMID:23652002

Chong, Tracy; Collins, James J; Brubacher, John L; Zarkower, David; Newmark, Phillip A

2013-01-01

396

The identification and characteristics of Echinoparyphium rubrum (Cort. 1914) new comb. (Trematoda, Echinostomatidae) based on experimental evidence of the life cycle  

USGS Publications Warehouse

The life cycle of Echinoparyphium rubrum (Cort, 1914) comb. n. has been completed experimentally. All of the developmental stages egg, miracidium, sporocyst, mother and daughter rediae, cercaria, metacercaria, and adult were examined and described. The miracidia infected freshwater snails of the genus Physa , P. gyrina and P. occidentalis. Attempts to infect snails of the genera Lymnaea, L. auricularis, L. peregra, L. truncatula and Bulinus, B. truncatus failed. Cercariae infected various pulmonate and prosobranch freshwater snails, mussels, frogs, water turtles and planarians. The adults developed in the small intestine of birds and mammals. The identity and major characteristics of Echinoparyphium rubrum are discussed. Synonyms of E. rubrum are Cercaria rubra Cort, 1914; Cercaria biflexa Faust, 1917; Cercaria chisolenata Faust, 1918; Echinostoma callawayensis Barker et Noll, 1915; Echinostoma revolutum of Johnson (1920); Echinoparyphium elegans of Cannon (1938), of Bain and Trelfall (1977), of Mahoney and Trelfall (1977); and Echinoparyphium recurvatum of Jilek (1977), Harley (1972), Sankurathri and Holmes (1976). Comparisons are made between E. rubrum and its 43-collar-spined allies: E. flexum from North America, E. cinctum from Europe, E. dunni from Asia and E. elegans from Africa.

Kanev, I.; Sorensen, R.; Sterner, M.; Cole, R.; Fried, B.

1998-01-01

397

Detection of Rat Lungworm in Intermediate, Definitive, and Paratenic Hosts Obtained from Environmental Sources  

PubMed Central

Angiostrongylus cantonensis is the most common parasite causing human eosinophilic meningitis worldwide. The geographical distribution of this disease has changed dramatically in the last few decades. Various methods have been used to detect A. cantonensis in host animals around the world. A survey of mollusks collected on the island of Hawa‘i in 2005 using PCR showed an infection rate of 24–78% depending on the mollusk species. In this study, samples from intermediate, definitive, and paratenic hosts were analyzed to further determine the presence of A. cantonensis in the United States. All samples were from Hawa‘i, except for the apple snails (Pomacea maculata) that were collected in New Orleans, Louisiana. Angiostrongylus cantonensis was detected in the majority of species examined, including the apple snails from New Orleans and flatworms (planarians) from Hawa‘i. Among the mollusks examined, the semi-slug Parmarion martensi had the highest parasite load, with an average larval burden of 445 larvae in 25 mg of tissue, as estimated by real-time PCR. In contrast, slime excreted from these highly infected mollusks contained no or very little A. cantonensis DNA. Analysis of definitive hosts (Rattus spp.) showed discrepancies between morphological and PCR-based identification; 54% of the rats were positive based on morphology, while 100% of tissue samples from these animals were positive by real-time PCR. This indicates that necropsies of rodents could underestimate the infection rates in definitive hosts of A. cantonensis.

Bishop, Henry S; da Silva, Alexandre J

2013-01-01

398

Charles Manning Child (1869-1954): the past, present, and future of metabolic signaling.  

PubMed

Charles Manning Child's work focused on metabolic gradients and their influence on organismal development. Early in the 20th century, his work had considerable currency, but by the second half of the century he had become little more than a historical footnote. Yet today Child's ideas are once again topical. While there were issues of cause and effect that Child and his students were never able to address adequately, in hindsight the extent of his eclipse hardly seems warranted. In fact, the demise of Child's theories may have resulted from larger changes in the nature of biology in the early 20th century. Child frequently studied planarians, hydroids, and other animals that are capable of asexual, agametic reproduction, and his theories most clearly apply to such organisms. In contrast, Thomas Hunt Morgan, initially one of Child's competitors in studies of regeneration, later developed the field of transmission genetics based on fruit flies, which can only reproduce via gametes. Child's theories and model systems were largely casualties of the success of Morgan's mechanistic paradigm. Nevertheless, in modern biology metabolic gradients, recast in terms of redox signaling, have become central to understanding both normal and pathological development. PMID:16353198

Blackstone, Neil W

2006-01-15

399

A sex-specific transcription factor controls male identity in a simultaneous hermaphrodite  

PubMed Central

Evolutionary transitions between hermaphroditic and dioecious reproductive states are found in many groups of animals. To understand such transitions, it is important to characterize diverse modes of sex determination utilized by metazoans. Currently, little is known about how simultaneous hermaphrodites specify and maintain male and female organs in a single individual. Here we show that a sex-specific gene, Smed-dmd-1 encoding a predicted doublesex/male-abnormal-3 (DM) domain transcription factor, is required for specification of male germ cells in a simultaneous hermaphrodite, the planarian Schmidtea mediterranea. dmd-1 has a male-specific role in the maintenance and regeneration of the testes and male accessory reproductive organs. In addition, a homologue of dmd-1 exhibits male-specific expression in Schistosoma mansoni, a derived, dioecious flatworm. These results demonstrate conservation of the role of DM domain genes in sexual development in lophotrochozoans and suggest one means by which modulation of sex-specific pathways can drive the transition from hermaphroditism to dioecy.

Chong, Tracy; Collins, James J.; Brubacher, John L.; Zarkower, David; Newmark, Phillip A.