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Sample records for mediterranean ascidian ciona

  1. Delineating metamorphic pathways in the ascidian Ciona intestinalis.

    PubMed

    Nakayama-Ishimura, Akie; Chambon, Jean-phillippe; Horie, Takeo; Satoh, Nori; Sasakura, Yasunori

    2009-02-15

    In most ascidians, metamorphosis of tadpole-like swimming larvae is accompanied by dynamic changes in their shape to form sessile adults. The mechanisms underlying ascidian metamorphosis have been debated for a long time. Although recent molecular studies have revealed the presence of various molecules involving in this process, the basic mechanism of the metamorphic events is still unclear. For example, it has not been solved whether all metamorphic events are organized by the same single pathway or by multiple, independent pathways. In the present study, we approached this question using the ascidian Ciona intestinalis. When the papillae and preoral lobes of the larvae were cut off, the papillae-cut larvae initiated certain trunk metamorphic events such as the formation of an ampulla, body axis rotation and adult organ growth without other metamorphic events. This observation indicates that metamorphic events can be divided into at least two groups, events initiated in the papillae-cut larva and events not initiated in this larva. In addition to this observation, we have isolated a novel mutant, tail regression failed (trf), which shows similar phenotypes to those of papillae-cut larvae. The phenotypes of trf mutants are basically different from those of swimming juvenile mutants (Sasakura, Y., Nakashima, K., Awazu, S., Matsuoka, T., Nakayama, A., Azuma, J., Satoh, N., 2005. Transposon-mediated insertional mutagenesis revealed the functions of animal cellulose synthase in the ascidian Ciona intestinalis. Proc. Natl. Acad. Sci. U. S. A. 102, 15134-15139.), which also show abnormal metamorphosis. These findings suggest a model by which ascidian metamorphic events can be classified into four groups initiated by different pathways.

  2. SL RNA genes of the ascidian tunicates Ciona intestinalis and Ciona savignyi.

    PubMed

    Yeats, Brendan; Matsumoto, Jun; Mortimer, Sandra I; Shoguchi, Eiichi; Satoh, Nori; Hastings, Kenneth E M

    2010-02-01

    We characterized by bioinformatics the trans-spliced leader donor RNA (SL RNA) genes of two ascidians, Ciona intestinalis and Ciona savignyi. The Ciona intestinalis genome contains approximately 670 copies of the SL RNA gene, principally on a 264-bp tandemly repeated element. Fluorescent in-situ hybridization mapped most of the repeats to a single site on the short arm of chromosome 8. The Ciona intestinalis genome also contains approximately 100 copies of a >3.6-kb element that carries 1) an SL RNA-related sequence (possible a pseudogene) and 2) genes for the U6 snRNA and a histone-like protein. The Ciona savignyi genome contains two SL RNA gene classes having the same SL sequence as Ciona intestinalis but differing in the intron-like segments. These reside in similar but distinct repeat units of 575 bp ( approximately 410 copies) and 552 bp ( approximately 250 copies) that are arranged as separate tandem repeats. In neither Ciona species is the 5S RNA gene present within the SL RNA gene repeat unit. Although the number of SL RNA genes is similar, there is little sequence similarity between the intestinalis and savignyi repeat units, apart from the region encoding the SL RNA itself. This suggests that cis-regulatory elements involved in transcription and 3'-end processing are likely to be present within the transcribed region. The genomes of both Ciona species also include > 100 dispersed short elements containing the 16-nt SL sequence and up to 6 additional nucleotides of the SL RNA sequence.

  3. Toll-like Receptors of the Ascidian Ciona intestinalis

    PubMed Central

    Sasaki, Naoko; Ogasawara, Michio; Sekiguchi, Toshio; Kusumoto, Shoichi; Satake, Honoo

    2009-01-01

    Key transmembrane proteins in the innate immune system, Toll-like receptors (TLRs), have been suggested to occur in the genome of non-mammalian organisms including invertebrates. However, authentic invertebrate TLRs have been neither structurally nor functionally investigated. In this paper, we originally present the structures, localization, ligand recognition, activities, and inflammatory cytokine production of all TLRs of the ascidian Ciona intestinalis, designated as Ci-TLR1 and Ci-TLR2. The amino acid sequence of Ci-TLR1 and Ci-TLR2 were found to possess unique structural organization with moderate sequence similarity to functionally characterized vertebrate TLRs. ci-tlr1 and ci-tlr2 genes were expressed predominantly in the stomach and intestine as well as in hemocytes. Ci-TLR1 and Ci-TLR2 expressed in HEK293 cells, unlike vertebrate TLRs, were localized to both the plasma membrane and endosomes. Intriguingly, both Ci-TLR1 and Ci-TLR2 stimulate NF-κB induction in response to multiple pathogenic ligands such as double-stranded RNA, and bacterial cell wall components that are differentially recognized by respective vertebrate TLRs, revealing that Ci-TLRs recognize broader pathogen-associated molecular patterns than vertebrate TLRs. The Ci-TLR-stimulating pathogenic ligands also induced the expression of Ci-TNFα in the intestine and stomach where Ci-TLRs are expressed. These results provide evidence that the TLR-triggered innate immune systems are essentially conserved in ascidians, and that Ci-TLRs possess “hybrid” biological and immunological functions, compared with vertebrate TLRs. Moreover, it is presumed that chordate TLR ancestors also acquired the Ci-TLR-like multiple cellular localization and pathogen-associated molecular pattern recognition. PMID:19651780

  4. Primary genetic linkage maps of the ascidian, Ciona intestinalis.

    PubMed

    Kano, Shungo; Satoh, Nori; Sordino, Paolo

    2006-01-01

    For whole-genome analysis in a basal chordate (protochordate), we used F1 pseudo-testcross mapping strategy and amplified fragment length polymorphism (AFLP) markers to construct primary linkage maps of the ascidian tunicate Ciona intestinalis. Two genetic maps consisted of 14 linkage groups, in agreement with the haploid chromosome number, and contained 276 and 125 AFLP loci derived from crosses between British and Neapolitan individuals. The two maps covered 4218.9 and 2086.9 cM, respectively, with an average marker interval of 16.1 and 18.9 cM. We observed a high recombinant ratio, ranging from 25 to 49 kb/cM, which can explain the high degree of polymorphism in this species. Some AFLP markers were converted to sequence tagged sites (STSs) by sequence determination, in order to create anchor markers for the fragmental physical map. Our recombination tools provide basic knowledge of genetic status and whole genome organization, and genetic markers to assist positional cloning in C. intestinalis.

  5. The nervous system of the adult ascidian Ciona intestinalis Type A (Ciona robusta): Insights from transgenic animal models

    PubMed Central

    Osugi, Tomohiro; Sasakura, Yasunori

    2017-01-01

    The nervous system of ascidians is an excellent model system to provide insights into the evolutionary process of the chordate nervous system due to their phylogenetic positions as the sister group of vertebrates. However, the entire nervous system of adult ascidians has yet to be functionally and anatomically investigated. In this study, we have revealed the whole dorsal and siphon nervous system of the transgenic adult ascidian of Ciona intestinalis Type A (Ciona robusta) in which a Kaede reporter gene is expressed in a pan-neuronal fashion. The fluorescent signal of Kaede revealed the innervation patterns and distribution of neurons in the nervous system of Ciona. Precise microscopic observation demonstrated the clear innervation of the anterior and posterior main nerves to eight and six lobes of the oral and atrial siphons, respectively. Moreover, visceral nerves, previously identified as unpaired nerves, were found to be paired; one nerve was derived from the posterior end of the cerebral ganglion and the other from the right posterior nerve. This study further revealed the full trajectory of the dorsal strand plexus and paired visceral nerves on either side from the cerebral ganglion to the ovary, and precise innervation between the cerebral ganglion and the peripheral organs including the gonoduct, cupular organ, rectum and ovary. The differential innervation patterns of visceral nerves and the dorsal strand plexus indicate that the peripheral organs including the ovary undergo various neural regulations. Collectively, the present anatomical analysis revealed the major innervation of the dorsal and siphon nervous systems of adult Ciona. PMID:28651020

  6. The nervous system of the adult ascidian Ciona intestinalis Type A (Ciona robusta): Insights from transgenic animal models.

    PubMed

    Osugi, Tomohiro; Sasakura, Yasunori; Satake, Honoo

    2017-01-01

    The nervous system of ascidians is an excellent model system to provide insights into the evolutionary process of the chordate nervous system due to their phylogenetic positions as the sister group of vertebrates. However, the entire nervous system of adult ascidians has yet to be functionally and anatomically investigated. In this study, we have revealed the whole dorsal and siphon nervous system of the transgenic adult ascidian of Ciona intestinalis Type A (Ciona robusta) in which a Kaede reporter gene is expressed in a pan-neuronal fashion. The fluorescent signal of Kaede revealed the innervation patterns and distribution of neurons in the nervous system of Ciona. Precise microscopic observation demonstrated the clear innervation of the anterior and posterior main nerves to eight and six lobes of the oral and atrial siphons, respectively. Moreover, visceral nerves, previously identified as unpaired nerves, were found to be paired; one nerve was derived from the posterior end of the cerebral ganglion and the other from the right posterior nerve. This study further revealed the full trajectory of the dorsal strand plexus and paired visceral nerves on either side from the cerebral ganglion to the ovary, and precise innervation between the cerebral ganglion and the peripheral organs including the gonoduct, cupular organ, rectum and ovary. The differential innervation patterns of visceral nerves and the dorsal strand plexus indicate that the peripheral organs including the ovary undergo various neural regulations. Collectively, the present anatomical analysis revealed the major innervation of the dorsal and siphon nervous systems of adult Ciona.

  7. Germ cell mutations of the ascidian Ciona intestinalis with TALE nucleases.

    PubMed

    Yoshida, Keita; Treen, Nicholas; Hozumi, Akiko; Sakuma, Tetsushi; Yamamoto, Takashi; Sasakura, Yasunori

    2014-05-01

    Targeted mutagenesis of genes-of-interest, or gene-knockout, is a powerful method to address the functions of genes. Engineered nucleases have enabled this approach in various organisms because of their ease of use. The ascidian Ciona intestinalis is an excellent organism to analyze gene functions by means of genetic technologies. In our previous study, we reported mutagenesis of Ciona somatic cells with TALE nucleases (TALENs) by electroporating expression constructs. In this study, we report germ cell mutagenesis of Ciona by microinjecting mRNAs encoding TALENs. TALEN mRNAs introduced mutations to target genes in both somatic and germ cells. TALEN-mediated mutations in the germ cell genome were inherited by the next generation. We conclude that knockout lines of Ciona that have disrupted target genes can be established through TALEN-mediated germ cell mutagenesis.

  8. CRISPR/Cas9-mediated gene knockout in the ascidian Ciona intestinalis.

    PubMed

    Sasaki, Haruka; Yoshida, Keita; Hozumi, Akiko; Sasakura, Yasunori

    2014-09-01

    Knockout of genes with CRISPR/Cas9 is a newly emerged approach to investigate functions of genes in various organisms. We demonstrate that CRISPR/Cas9 can mutate endogenous genes of the ascidian Ciona intestinalis, a splendid model for elucidating molecular mechanisms for constructing the chordate body plan. Short guide RNA (sgRNA) and Cas9 mRNA, when they are expressed in Ciona embryos by means of microinjection or electroporation of their expression vectors, introduced mutations in the target genes. The specificity of target choice by sgRNA is relatively high compared to the reports from some other organisms, and a single nucleotide mutation at the sgRNA dramatically reduced mutation efficiency at the on-target site. CRISPR/Cas9-mediated mutagenesis will be a powerful method to study gene functions in Ciona along with another genome editing approach using TALE nucleases.

  9. Huntingtin gene evolution in Chordata and its peculiar features in the ascidian Ciona genus

    PubMed Central

    Gissi, Carmela; Pesole, Graziano; Cattaneo, Elena; Tartari, Marzia

    2006-01-01

    Background To gain insight into the evolutionary features of the huntingtin (htt) gene in Chordata, we have sequenced and characterized the full-length htt mRNA in the ascidian Ciona intestinalis, a basal chordate emerging as new invertebrate model organism. Moreover, taking advantage of the availability of genomic and EST sequences, the htt gene structure of a number of chordate species, including the cogeneric ascidian Ciona savignyi, and the vertebrates Xenopus and Gallus was reconstructed. Results The C. intestinalis htt transcript exhibits some peculiar features, such as spliced leader trans-splicing in the 98 nt-long 5' untranslated region (UTR), an alternative splicing in the coding region, eight alternative polyadenylation sites, and no similarities of both 5' and 3'UTRs compared to homologs of the cogeneric C. savignyi. The predicted protein is 2946 amino acids long, shorter than its vertebrate homologs, and lacks the polyQ and the polyP stretches found in the the N-terminal regions of mammalian homologs. The exon-intron organization of the htt gene is almost identical among vertebrates, and significantly conserved between Ciona and vertebrates, allowing us to hypothesize an ancestral chordate gene consisting of at least 40 coding exons. Conclusion During chordate diversification, events of gain/loss, sliding, phase changes, and expansion of introns occurred in both vertebrate and ascidian lineages predominantly in the 5'-half of the htt gene, where there is also evidence of lineage-specific evolutionary dynamics in vertebrates. On the contrary, the 3'-half of the gene is highly conserved in all chordates at the level of both gene structure and protein sequence. Between the two Ciona species, a fast evolutionary rate and/or an early divergence time is suggested by the absence of significant similarity between UTRs, protein divergence comparable to that observed between mammals and fishes, and different distribution of repetitive elements. PMID:17092333

  10. Transposon-mediated targeted and specific knockdown of maternally expressed transcripts in the ascidian Ciona intestinalis.

    PubMed

    Iitsuka, Takako; Mita, Kaoru; Hozumi, Akiko; Hamada, Mayuko; Satoh, Nori; Sasakura, Yasunori

    2014-05-23

    Maternal mRNAs play crucial roles during early embryogenesis of ascidians, but their functions are largely unknown. In this study, we developed a new method to specifically knockdown maternal mRNAs in Ciona intestinalis using transposon-mediated transgenesis. We found that GFP expression is epigenetically silenced in Ciona intestinalis oocytes and eggs, and this epigenetic silencing of GFP was used to develop the knockdown method. When the 5' upstream promoter and 5' untranslated region (UTR) of a maternal gene are used to drive GFP in eggs, the maternal gene is specifically knocked down together with GFP. The 5' UTR of the maternal gene is the major element that determines the target gene silencing. Zygotic transcription of the target gene is unaffected, suggesting that the observed phenotypes specifically reflect the maternal function of the gene. This new method can provide breakthroughs in studying the functions of maternal mRNAs.

  11. Expression of 17beta-hydroxysteroid dehydrogenease in testis of the ascidian Ciona intestinalis [corrected].

    PubMed

    Kho, Kang Hee; Inaba, Kazuo

    2004-10-31

    Ascidians have been employed as model organisms in investigating spermatogenesis. 17beta-hydroxysteroid dehydrogenase (HSD) is a steroidogenic enzyme essential for invertebrate spermatogenesis. A homologue of HSD was found in the EST database of Ciona intestinalis and cloned. Sequence analysis showed significant homology to zebra fish, sea urchin and human 17beta-HSD. The gene has an open reading frame (ORF) of 918 nucleotides coding for a polypeptide of 306 amino acids and a calculated mass of 35-kDa. Immunoblotting with an antibody raised against HSD recognized a 35-kDa protein purified from the C. intestinalis testis. The HSD protein was localized in steroidogenic cells in the Ciona testis. These results suggest that C. intestinalis 17beta-HSD is equivalent to the enzyme of vertebrate Leydig cells and that 17beta-HSD could be a phylogenetic marker for organisms producing steroids.

  12. Time course for tail regression during metamorphosis of the ascidian Ciona intestinalis.

    PubMed

    Matsunobu, Shohei; Sasakura, Yasunori

    2015-09-01

    In most ascidians, the tadpole-like swimming larvae dramatically change their body-plans during metamorphosis and develop into sessile adults. The mechanisms of ascidian metamorphosis have been researched and debated for many years. Until now information on the detailed time course of the initiation and completion of each metamorphic event has not been described. One dramatic and important event in ascidian metamorphosis is tail regression, in which ascidian larvae lose their tails to adjust themselves to sessile life. In the present study, we measured the time associated with tail regression in the ascidian Ciona intestinalis. Larvae are thought to acquire competency for each metamorphic event in certain developmental periods. We show that the timing with which the competence for tail regression is acquired is determined by the time since hatching, and this timing is not affected by the timing of post-hatching events such as adhesion. Because larvae need to adhere to substrates with their papillae to induce tail regression, we measured the duration for which larvae need to remain adhered in order to initiate tail regression and the time needed for the tail to regress. Larvae acquire the ability to adhere to substrates before they acquire tail regression competence. We found that when larvae adhered before they acquired tail regression competence, they were able to remember the experience of adhesion until they acquired the ability to undergo tail regression. The time course of the events associated with tail regression provides a valuable reference, upon which the cellular and molecular mechanisms of ascidian metamorphosis can be elucidated. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. Transcriptome dynamics in early embryos of the ascidian, Ciona intestinalis.

    PubMed

    Matsuoka, Terumi; Ikeda, Tatsuro; Fujimaki, Kotaro; Satou, Yutaka

    2013-12-15

    Maternally provided mRNAs and proteins direct early development and activate the zygotic genome. Using microarrays, we examined the dynamics of transcriptomes during the early development of a basal chordate, Ciona intestinalis. Microarray analysis of unfertilized eggs, as well as 8-, and 16- and 32-cell embryos revealed that nearly half of the genes encoded in the genome were expressed maternally, and that approximately only one-fourth of these genes were expressed at similar levels among eggs obtained from different individuals. Genes encoding proteins involved in protein phosphorylation were enriched in this latter group. More than 90% of maternal RNAs were not reduced before the 16-cell stage when the zygotic developmental program begins. Additionally we obtained gene expression profiles of individual blastomeres from the 8- and 16-cell embryos. On the basis of these profiles, we concluded that the posterior-most localization, which has been reported for over 20 different transcripts, is the only major localization pattern of maternal transcripts. Our data also showed that maternal factors establish only nine distinct patterns of zygotic gene expression at the 16-cell stage. Therefore, one of the main developmental functions of maternally supplied information is to establish these nine distinct expression patterns in the 16-cell embryo. The dynamics of transcriptomes in early-stage embryos provides a foundation for studying how maternal information starts the zygotic program.

  14. Siphon Regeneration Capacity is Compromised During Aging in the Ascidian Ciona intestinalis

    PubMed Central

    Jeffery, William R.

    2012-01-01

    The ascidian Ciona intestinalis has a short life span and powerful regeneration capacities. The regeneration of the oral siphon (OS) involves wound healing, blastema formation, cell proliferation, and replacement of eight oral pigment organs (OPO), the latter via differentiation and migration of stem/precursor cells from localized siphon niches in the siphon. The restoration of OPO pattern during OS regeneration occurs with a high degree of accuracy through three successive cycles of amputation. It is shown here that oral siphons of the largest and oldest members of a wild Ciona population do not completely regenerate their siphons after amputation. The loss of regeneration capacity was accompanied by reduced cell proliferation. In contrast to arrested OS outgrowth, the stem/precursor cells responsible for OPO replacement “over-differentiate” after OS amputation in the oldest animals, the typical number of OPO is increased from eight to twelve-sixteen, and malformed OPO are produced. Also in contrast to younger animals, the oldest animals of the population show arrested OPO development after two consecutive cycles of amputation and regeneration. We conclude that there is a size and age threshold in Ciona after which the regenerative capacity of the OS is compromised due to effects of aging on cell proliferation. PMID:22935550

  15. Siphon regeneration capacity is compromised during aging in the ascidian Ciona intestinalis.

    PubMed

    Jeffery, William R

    2012-01-01

    The ascidian Ciona intestinalis has a short life span and powerful regeneration capacities. The regeneration of the oral siphon (OS) involves wound healing, blastema formation, cell proliferation, and replacement of 8 oral pigment organs (OPO), the latter via differentiation and migration of stem/precursor cells from localized niches in the siphon. The restoration of OPO pattern during OS regeneration occurs with a high degree of accuracy through three successive cycles of amputation. It is shown here that oral siphons of the largest and oldest members of a wild Ciona population do not completely regenerate their siphons after amputation. The loss of regeneration capacity was accompanied by reduced cell proliferation. In contrast to arrested OS outgrowth, the stem/precursor cells responsible for OPO replacement "over-differentiate" after OS amputation in the oldest animals, the typical number of OPO is increased from 8 to 12-16, and malformed OPO are produced. Also in contrast to younger animals, the oldest animals of the population show arrested OPO development after two consecutive cycles of amputation and regeneration. We conclude that there is a size and age threshold in Ciona after which the regenerative capacity of the OS is compromised due to effects of aging on cell proliferation.

  16. Establishment of enhancer detection lines expressing GFP in the gut of the ascidian Ciona intestinalis.

    PubMed

    Yoshida, Reiko; Sasakura, Yasunori

    2012-01-01

    The gut is a tubular, endodermal organ for digesting food and absorbing nutrients. In this study, we characterized eight enhancer detection lines that express green fluorescent protein (GFP) in the whole or part of the digestive tube of the ascidian Ciona intestinalis. Three enhancer detection lines for the pyloric gland, a structure associated with the digestive tube, were also analyzed. These lines are valuable markers for analyzing the mechanisms of development of the gut. Based on the GFP expression of the enhancer detection lines together with morphological characteristics, the digestive tube of Ciona can be subdivided into at least 10 compartments in which different genetic cascades operate. Causal insertion sites of the enhancer detection lines were identified, and the expression pattern of the genes near the insertion sites were characterized by means of whole-mount in situ hybridization. We have characterized four and two genes that were specifically or strongly expressed in the digestive tube and pyloric gland, respectively. The present data provide the basic information and useful resources for studying gut formation in Ciona.

  17. Spermiotoxicity of nickel nanoparticles in the marine invertebrate Ciona intestinalis (ascidians)

    PubMed Central

    Gallo, Alessandra; Boni, Raffaele; Buttino, Isabella; Tosti, Elisabetta

    2016-01-01

    Abstract Nickel nanoparticles (Ni NPs) are increasingly used in modern industries as catalysts, sensors, and in electronic applications. Due to this large use, their inputs into marine environment have significantly increased; however, the potential ecotoxicological effects in marine environment have so far received little attention. In particular, little is known on the impact of NPs on gamete quality of marine organisms and on the consequences on fertility potential. The present study examines, for the first time, the impact of Ni NPs exposure on sperm quality of the marine invertebrate Ciona intestinalis (ascidian). Several parameters related with sperm status such as plasma membrane lipid peroxidation, mitochondrial membrane potential (MMP), intracellular pH, DNA integrity, and fertilizing ability were assessed as toxicity end points after exposure to different Ni NPs concentrations. Ni NPs generate oxidative stress that in turn induces lipid peroxidation and DNA fragmentation, and alters MMP and sperm morphology. Furthermore, sperm exposure to Ni NPs affects their fertilizing ability and causes developmental anomalies in the offspring. All together, these results reveal a spermiotoxicity of Ni NPs in ascidians suggesting that the application of these NPs should be carefully assessed as to their potential toxic effects on the health of marine organisms that, in turn, may influence the ecological system. This study shows that ascidian sperm represent a suitable and sensitive tool for the investigation of the toxicity of NPs entered into marine environment, for defining the mechanisms of toxic action and for the environmental monitoring purpose. PMID:27080039

  18. Inducible galectins are expressed in the inflamed pharynx of the ascidian Ciona intestinalis.

    PubMed

    Vizzini, Aiti; Parrinello, Daniela; Sanfratello, Maria Antonietta; Salerno, Giuseppina; Cammarata, Matteo; Parrinello, Nicolò

    2012-01-01

    Although ascidians belong to a key group in chordate phylogenesis, amino acid sequences of Ciona intestinalis galectin-CRDs (CiLgals-a and -b) have been retained too divergent from vertebrate galectins. In the present paper, to contribute in disclosing Bi-CRD galectin evolution a novel attempt was carried out on CiLgals-a and -b CRDs phylogenetic analysis, and their involvement in ascidian inflammatory responses was shown. CiLgals resulted aligned with Bi-CRD galectins from vertebrates (Xenopus tropicalis, Gallus gallus, Mus musculus, Homo sapiens), cephalochordates (Branchiostoma floridae), echinoderms (Strongylocentrotus purpuratus) and a mono-CRD galectin from the ascidian Clavelina picta. The CiLgals-a N-terminal and C-terminal CRDs contain the signature sequence involved in carbohydrate binding, whereas the CiLgals-b C-CRD presents only three out of seven key aminoacids and it could not be suitable as sugar binding motif. Sequence similarity between clusters suggests an evolutionary model based on CRD domain gene duplication and sequence diversification. In particular CiLgals-b N-CRD and C-CRD were similar to each other and both grouped with the ascidian C. picta mono-CRD. Homology modeling process shows a CiLgals molecular structure superimposed to chicken and mouse galectins. The CiLgals-a and CiLgals-b genes were upregulated by LPS inoculation suggesting that they are inducible and expressed in the inflamed pharynx as revealed by real-time PCR analysis. Finally, in situ hybridization and immunohistochemical assays showed their localization in the inflamed tissues, while immunoblotting analysis indicated that CiLgals can form oligomers.

  19. Regeneration of oral siphon pigment organs in the ascidian Ciona intestinalis

    PubMed Central

    Auger, Hélène; Sasakura, Yasunori; Joly, Jean-Stéphane; Jeffery, William R.

    2013-01-01

    Ascidians have powerful capacities for regeneration but the underlying mechanisms are poorly understood. Here we examine oral siphon regeneration in the solitary ascidian Ciona intestinalis. Following amputation, the oral siphon rapidly reforms oral pigment organs (OPO) at its distal margin prior to slower regeneration of proximal siphon parts. The early stages of oral siphon reformation include cell proliferation and re-growth of the siphon nerves, although the neural complex (adult brain and associated organs) is not required for regeneration. Young animals reform OPO more rapidly after amputation than old animals indicating that regeneration is age dependent. UV irradiation, microcautery, and cultured siphon explant experiments indicate that OPOs are replaced as independent units based on local differentiation of progenitor cells within the siphon, rather than by cell migration from a distant source in the body. The typical pattern of eight OPOs and siphon lobes is restored with fidelity after distal amputation of the oral siphon, but as many as 16 OPOs and lobes can be reformed following proximal amputation near the siphon base. Thus, the pattern of OPO regeneration is determined by cues positioned along the proximal distal axis of the oral siphon. A model is presented in which columns of siphon tissue along the proximal–distal axis below pre-existing OPO are responsible for reproducing the normal OPO pattern during regeneration. This study reveals previously unknown principles of oral siphon and OPO regeneration that will be important for developing Ciona as a regeneration model in urochordates, which may be the closest living relatives of vertebrates. PMID:20059994

  20. Proteomics of ionomycin-induced ascidian sperm reaction: Released and exposed sperm proteins in the ascidian Ciona intestinalis.

    PubMed

    Nakazawa, Shiori; Shirae-Kurabayashi, Maki; Otsuka, Kei; Sawada, Hitoshi

    2015-12-01

    Sperm proteins mediating sperm-egg interaction should be exhibited on the sperm surface, or exposed or released when sperm approach an egg. In ascidians (protochordates), sperm undergo a sperm reaction, characterized by enhanced sperm motility and mitochondrial swelling and shedding on contact with the vitelline coat (VC) or by treatment with Ca(2+) ionophore. Here, proteomic analysis was conducted on sperm exudates and sperm surface proteins using ionomycin-induced sperm reaction and cell-impermeable labeling in Ciona intestinalis type A (C. robusta). In the exudate from sperm treated with ionomycin, membrane proteins including a possible VC receptor CiUrabin were abundant, indicating the release of membranous compartments during sperm reaction. Among the surface proteins XP_009859314.1 (uncharacterized protein exhibiting homology to HrTTSP-1) was most abundant before the sperm reaction, but XP_004227079.1 (unknown Ig superfamily protein) appears to be most abundantly exposed by the sperm reaction. Moreover, proteins containing a notable set of domains, astacin-like metalloprotease domain and thrombospondin type 1 repeat(s), were found in this fraction. Possible roles in fertilization as well as localizations and behaviors of these proteins are discussed.

  1. Adverse Effect of Antifouling Compounds on the Reproductive Mechanisms of the Ascidian Ciona intestinalis

    PubMed Central

    Gallo, Alessandra; Tosti, Elisabetta

    2013-01-01

    Fertilization and embryo development that occur in sea water are sensitive to xenobiotics from anthropogenic sources. In this work, we evaluated the influence of two antifouling biocides, tributyltin (TBT) and diuron, on the reproductive mechanisms of the marine invertebrate Ciona intestinalis. By using electrophysiological techniques, we examined the impact of these compounds on the electrical properties of the mature oocytes and of events occurring at fertilization. With different toxicity assays, we studied the effect of the two biocides on the gametes by evaluating fertilization rate and embryo development. Results show that sodium (Na+) currents were significantly reduced by either of the two biocides, whereas conductance was significantly increased. The fertilization current frequency and amplitude, fertilization rate and larval development were affected only by TBT. This study suggests that: (i) the two biocides affect either the electrical properties of the oocyte plasma membrane and the reproductive success representing a risk factor for the survival of the species exposed to environmental pollution; (ii) the ascidian Ciona intestinalis may represent a good model organism to test toxicity of marine pollutants. Possible mechanisms of action of the two biocides are discussed. PMID:24065165

  2. Adverse effect of antifouling compounds on the reproductive mechanisms of the ascidian Ciona intestinalis.

    PubMed

    Gallo, Alessandra; Tosti, Elisabetta

    2013-09-20

    Fertilization and embryo development that occur in sea water are sensitive to xenobiotics from anthropogenic sources. In this work, we evaluated the influence of two antifouling biocides, tributyltin (TBT) and diuron, on the reproductive mechanisms of the marine invertebrate Ciona intestinalis. By using electrophysiological techniques, we examined the impact of these compounds on the electrical properties of the mature oocytes and of events occurring at fertilization. With different toxicity assays, we studied the effect of the two biocides on the gametes by evaluating fertilization rate and embryo development. Results show that sodium (Na⁺) currents were significantly reduced by either of the two biocides, whereas conductance was significantly increased. The fertilization current frequency and amplitude, fertilization rate and larval development were affected only by TBT. This study suggests that: (i) the two biocides affect either the electrical properties of the oocyte plasma membrane and the reproductive success representing a risk factor for the survival of the species exposed to environmental pollution; (ii) the ascidian Ciona intestinalis may represent a good model organism to test toxicity of marine pollutants. Possible mechanisms of action of the two biocides are discussed.

  3. Closing the wounds: one hundred and twenty five years of regenerative biology in the ascidian Ciona intestinalis.

    PubMed

    Jeffery, William R

    2015-01-01

    This year marks the 125th anniversary of the beginning of regeneration research in the ascidian Ciona intestinalis. A brief note was published in 1891, reporting the regeneration of the Ciona neural complex and siphons. This launched an active period of Ciona regeneration research culminating in the demonstration of partial body regeneration: the ability of proximal body parts to regenerate distal ones, but not vice versa. In a process resembling regeneration, wounds in the siphon tube were discovered to result in the formation of an ectopic siphon. Ciona regeneration research then lapsed into a period of relative inactivity after the purported demonstration of the inheritance of acquired characters using siphon regeneration as a model. Around the turn of the present century, Ciona regeneration research experienced a new blossoming. The current studies established the morphological and physiological integrity of the regeneration process and its resemblance to ontogeny. They also determined some of the cell types responsible for tissue and organ replacement and their sources in the body. Finally, they showed that regenerative capacity is reduced with age. Many other aspects of regeneration now can be studied at the mechanistic level because of the extensive molecular tools available in Ciona.

  4. Filter-feeding ascidians ( Ciona intestinalis) in a shallow cove: Implications of hydrodynamics for grazing impact

    NASA Astrophysics Data System (ADS)

    Riisgård, Hans Ulrik; Jürgensen, Carsten; Clausen, Torben

    1996-06-01

    The grazing impact by a dense population of filter-feeding ascidians Ciona intestinalis on horizontally flowing water (driven by density circulation) in a shallow cove (Kertinge Nor, Denmark) has been described and quantified by means of a simple one-dimensional numerical model. The agreement between observations and modelled predictions was satisfactory. The applied numerical model has the following analytical solution in the idealized case: Cx = C0e -( fx/ Y2) , where Cx = algal concentration at a downstream distance x, C0 = initial concentration, f = F/ vc; F = area specific population filtration rate; vc = current velocity; Y 2 = depth of mixed layer below halocline. The numerical model quantifies the actual grazing impact while the analytical model illustrates the governing physics in well-known terms. To describe situations with no current ( i.e. stagnant water), we performed simulation studies in the laboratory and measured vertical profiles of algal cells over filter-feeding C. intestinalis. The results showed that phytoplankton became reduced in a near-bottom water layer of 20-30 cm thickness. Such water layers may develop in stagnant water (calm days and no advective currents), thus uncoupling the pelagic food and the filter feeders which within a short time will experience extremely meagre food conditions.

  5. A one-dimensional model of PCP signaling: polarized cell behavior in the notochord of the ascidian Ciona.

    PubMed

    Kourakis, Matthew J; Reeves, Wendy; Newman-Smith, Erin; Maury, Benoit; Abdul-Wajid, Sarah; Smith, William C

    2014-11-01

    Despite its importance in development and physiology the planar cell polarity (PCP) pathway remains one of the most enigmatic signaling mechanisms. The notochord of the ascidian Ciona provides a unique model for investigating the PCP pathway. Interestingly, the notochord appears to be the only embryonic structure in Ciona activating the PCP pathway. Moreover, the Ciona notochord as a single-file array of forty polarized cells is a uniquely tractable system for the study of polarization dynamics and the transmission of the PCP pathway. Here, we test models for propagation of a polarizing signal, interrogating temporal, spatial and signaling requirements. A simple cell-cell relay cascading through the entire length of the notochord is not supported; instead a more complex mechanism is revealed, with interactions influencing polarity between neighboring cells, but not distant ones. Mechanisms coordinating notochord-wide polarity remain elusive, but appear to entrain general (i.e., global) polarity even while local interactions remain important. However, this global polarizer does not appear to act as a localized, spatially-restricted determinant. Coordination of polarity along the long axis of the notochord requires the PCP pathway, a role we demonstrate is temporally distinct from this pathway's earlier role in convergent extension and intercalation. We also reveal polarity in the notochord to be dynamic: a cell's polarity state can be changed and then restored, underscoring the Ciona notochord's amenability for in vivo studies of PCP. Copyright © 2014 Elsevier Inc. All rights reserved.

  6. A one-dimensional model of PCP signaling: polarized cell behavior in the notochord of the ascidian Ciona

    PubMed Central

    Kourakis, Matthew J.; Reeves, Wendy; Newman-Smith, Erin; Maury, Benoit; Abdul-Wajid, Sarah; Smith, William C.

    2014-01-01

    Despite its importance in development and physiology the planar cell polarity (PCP) pathway remains one of the most enigmatic signaling mechanisms. The notochord of the ascidian Ciona provides a unique model for investigating the PCP pathway. Interestingly, the notochord appears to be the only embryonic structure in Ciona activating the PCP pathway. Moreover, the Ciona notochord as a single-file array of forty polarized cells is a uniquely tractable system for the study of polarization dynamics and the transmission of the PCP pathway. Here, we test models for propagation of a polarizing signal, interrogating temporal, spatial and signaling requirements. A simple cell-cell relay cascading through the entire length of the notochord is not supported; instead a more complex mechanism is revealed, with interactions influencing polarity between neighboring cells, but not distant ones. Mechanisms coordinating notochord-wide polarity remain elusive, but appear to entrain general (i.e., global) polarity even while local interactions remain important. However, this global polarizer does not appear to act as a localized, spatially-restricted determinant. Coordination of polarity along the long axis of the notochord requires the PCP pathway, a role we demonstrate is temporally distinct from this pathway’s earlier role in convergent extension and intercalation. We also reveal polarity in the notochord to be dynamic: a cell’s polarity state can be changed and then restored, underscoring the Ciona notochord’s amenability for in vivo studies of PCP. PMID:25173874

  7. Large-scale infection of the ascidian Ciona intestinalis by the gregarine Lankesteria ascidiae in an inland culture system.

    PubMed

    Mita, Kaoru; Kawai, Narudo; Rueckert, Sonja; Sasakura, Yasunori

    2012-11-19

    An important way to keep transgenic and mutant lines of the ascidian Ciona intestinalis, a model system for e.g. genetic functions, in laboratories is via culturing systems. Here we report a disease of C. intestinalis observed in an inland culturing system. The disease, called 'long feces syndrome,' is expressed in affected animals by the following characteristic symptoms of the digestive system: (1) excretion of long and thin feces, (2) pale color of the stomach, and (3) congestion of the digestive tube by digested material. Severely diseased animals usually die within a week after the first symptoms occur, implying a high risk of this disease for ascidian culturing systems. The digestive tubes of the diseased animals are occupied by the gregarine apicomplexan parasite Lankesteria ascidiae, suggesting that large-scale infection by this parasite is the cause of long feces syndrome.

  8. Metamorphosis of the invasive ascidian Ciona savignyi: environmental variables and chemical exposure.

    PubMed

    Cahill, Patrick L; Atalah, Javier; Selwood, Andrew I; Kuhajek, Jeanne M

    2016-01-01

    In this study, the effects of environmental variables on larval metamorphosis of the solitary ascidian Ciona savignyi were investigated in a laboratory setting. The progression of metamorphic changes were tracked under various temperature, photoperiod, substrate, larval density, and vessel size regimes. Metamorphosis was maximised at 18 °C, 12:12 h subdued light:dark, smooth polystyrene substrate, and 10 larvae mL(-1) in a twelve-well tissue culture plate. Eliminating the air-water interface by filling culture vessels to capacity further increased the proportion of metamorphosed larvae; 87 ± 5% of larvae completed metamorphosis within 5 days compared to 45 ± 5% in control wells. The effects of the reference antifouling compounds polygodial, portimine, oroidin, chlorothalonil, and tolylfluanid on C. savignyi were subsequently determined, highlighting (1) the sensitivity of C. savignyi metamorphosis to chemical exposure and (2) the potential to use C. savignyi larvae to screen for bioactivity in an optimised laboratory setting. The compounds were bioactive in the low ng mL(-1) to high µg mL(-1) range. Polygodial was chosen for additional investigations, where it was shown that mean reductions in the proportions of larvae reaching stage E were highly repeatable both within (repeatability = 14 ± 9%) and between (intermediate precision = 17 ± 3%) independent experiments. An environmental extract had no effect on the larvae but exposing larvae to both the extract and polygodial reduced potency relative to polygodial alone. This change in potency stresses the need for caution when working with complex samples, as is routinely implemented when isolating natural compounds from their biological source. Overall, the outcomes of this study highlight the sensitivity of C. savignyi metamorphosis to environmental variations and chemical exposure.

  9. Metamorphosis of the invasive ascidian Ciona savignyi: environmental variables and chemical exposure

    PubMed Central

    Atalah, Javier; Selwood, Andrew I.; Kuhajek, Jeanne M.

    2016-01-01

    In this study, the effects of environmental variables on larval metamorphosis of the solitary ascidian Ciona savignyi were investigated in a laboratory setting. The progression of metamorphic changes were tracked under various temperature, photoperiod, substrate, larval density, and vessel size regimes. Metamorphosis was maximised at 18 °C, 12:12 h subdued light:dark, smooth polystyrene substrate, and 10 larvae mL−1 in a twelve-well tissue culture plate. Eliminating the air-water interface by filling culture vessels to capacity further increased the proportion of metamorphosed larvae; 87 ± 5% of larvae completed metamorphosis within 5 days compared to 45 ± 5% in control wells. The effects of the reference antifouling compounds polygodial, portimine, oroidin, chlorothalonil, and tolylfluanid on C. savignyi were subsequently determined, highlighting (1) the sensitivity of C. savignyi metamorphosis to chemical exposure and (2) the potential to use C. savignyi larvae to screen for bioactivity in an optimised laboratory setting. The compounds were bioactive in the low ng mL−1 to high µg mL−1 range. Polygodial was chosen for additional investigations, where it was shown that mean reductions in the proportions of larvae reaching stage E were highly repeatable both within (repeatability = 14 ± 9%) and between (intermediate precision = 17 ± 3%) independent experiments. An environmental extract had no effect on the larvae but exposing larvae to both the extract and polygodial reduced potency relative to polygodial alone. This change in potency stresses the need for caution when working with complex samples, as is routinely implemented when isolating natural compounds from their biological source. Overall, the outcomes of this study highlight the sensitivity of C. savignyi metamorphosis to environmental variations and chemical exposure. PMID:26966668

  10. Insulin-like genes in ascidians: findings in Ciona and hypotheses on the evolutionary origins of the pancreas

    PubMed Central

    Thompson, Jordan M.; Di Gregorio, Anna

    2014-01-01

    Insulin plays an extensively characterized role in the control of sugar metabolism, growth and homeostasis in a wide range of organisms. In vertebrate chordates, insulin is mainly produced by the beta cells of the endocrine pancreas, while in non-chordate animals insulin-producing cells are mainly found in the nervous system and/or scattered along the digestive tract. However, recent studies have indicated the notochord, the defining feature of the chordate phylum, as an additional site of expression of insulin-like peptides. Here we show that two of the three insulin-like genes identified in Ciona intestinalis, an invertebrate chordate with a dual life cycle, are first expressed in the developing notochord during embryogenesis and transition to distinct areas of the adult digestive tract after metamorphosis. In addition, we present data suggesting that the transcription factor Ciona Brachyury is involved in the control of notochord expression of at least one of these genes, Ciona insulin-like 2. Lastly, we review the information currently available on insulin-producing cells in ascidians and on pancreas-related transcription factors that might control their expression. PMID:25378051

  11. Ciona Genetics

    PubMed Central

    Veeman, Michael T.; Chiba, Shota; Smith, William C.

    2010-01-01

    Ascidians, such as Ciona, are invertebrate chordates with simple embryonic body plans and small, relatively non-redundant genomes. Ciona genetics is in its infancy compared to many other model systems, but it provides a powerful method for studying this important vertebrate outgroup. Here we give basic methods for genetic analysis of Ciona, including protocols for controlled crosses both by natural spawning and by the surgical isolation of gametes; the identification and propagation of mutant lines; and strategies for positional cloning. PMID:21805273

  12. Shared hemocyte- and intestine-dominant expression profiles of intelectin genes in ascidian Ciona intestinalis: insight into the evolution of the innate immune system in chordates.

    PubMed

    Hayashibe, Minoru; Nakayama, Satoshi; Ogasawara, Michio

    2017-06-27

    Intelectin is a soluble lectin known as a pattern-recognition receptor for the innate immune system or as an intestinal lactoferrin receptor. Intelectin genes have been identified in a wide range of chordates and the shared expression pattern in their absorptive intestinal regions has been widely recognized. The chordate intelectins have a shared domain structure with a fibrinogen-related domain and an intelectin domain and an additional sequence has been reported only in ascidian Ciona intestinalis intelectins. However, little is known about the molecular features of the ascidian intelectins, including the distribution of the additional sequence in ascidians. Therefore, we focus on the ascidian species that are available for genome DNA sequence searches and survey intelectin genes with special reference to the additional sequence. We also assess the distribution of Ciona intelectin gene transcripts in transparent juveniles and adult specimens by means of in situ hybridization and reveal hemocyte-dominant expressions as well as stomach-exclusive expression. Comparative gene expression analysis with secretory digestive enzymes and absorption-related proteins in Ciona revealed that intelectin and secretory digestive enzymes were expressed in the same region of the stomach epithelium. Since the domain structure of intelectins and the hemocyte-dominant gene expression of intelectins seem relevant to ficolin, intelectin genes may have evolved from a ficolin-like ancestral gene with hemocytic expression in early chordate evolution.

  13. Evaluation of drug toxicity profiles based on the phenotypes of ascidian Ciona intestinalis.

    PubMed

    Mizotani, Yuji; Itoh, Shun; Hotta, Kohji; Tashiro, Etsu; Oka, Kotaro; Imoto, Masaya

    2015-08-07

    In vivo toxicity evaluation using model organisms is an important step for the development of new drugs. Here, we report that Ciona intestinalis, a chordate invertebrate, is beneficial to drug toxicity evaluation for the following reasons: rapid embryonic and larval development, resemblance to vertebrates, ease of management, low cost, transparent body, and low risk of ethical issues. The dynamic phenotypic change of Ciona larvae during metamorphosis prompted us to examine the effect of cytotoxic drugs on its development by quantifying six toxicity endpoints: degenerated tail size, ampulla length, rotation of body axis, stomach size, heart rate, and body size. As a result, mitochondrial respiratory inhibitors, tubulin polymerization/depolymerization inhibitors, or DNA/RNA synthesis inhibitors showed distinct toxicity profiles against these six endpoints, but drugs with the same targets showed a similar toxicity profile in Ciona. Our results suggest Ciona is an effective animal model for profiling drug toxicity and exploring the mechanisms of drugs with unknown targets.

  14. Distal regeneration involves the age dependent activity of branchial sac stem cells in the ascidian Ciona intestinalis

    PubMed Central

    2014-01-01

    Abstract Tunicates have high capacities for regeneration but the underlying mechanisms and their relationship to life cycle progression are not well understood. Here we investigate the regeneration of distal structures in the ascidian tunicate Ciona intestinalis. Analysis of regenerative potential along the proximal−distal body axis indicated that distal organs, such as the siphons, their pigmented sensory organs, and the neural complex, could only be replaced from body fragments containing the branchial sac. Distal regeneration involves the formation of a blastema composed of cells that undergo cell proliferation prior to differentiation and cells that differentiate without cell proliferation. Both cell types originate in the branchial sac and appear in the blastema at different times after distal injury. Whereas the branchial sac stem cells are present in young animals, they are depleted in old animals that have lost their regeneration capacity. Thus Ciona adults contain a population of age‐related stem cells located in the branchial sac that are a source of precursors for distal body regeneration. PMID:25893097

  15. Distal Regeneration Involves the Age Dependent Activity of Branchial Sac Stem Cells in the Ascidian Ciona intestinalis.

    PubMed

    Jeffery, William R

    2015-02-01

    Tunicates have high capacities for regeneration but the underlying mechanisms and their relationship to life cycle progression are not well understood. Here we investigate the regeneration of distal structures in the ascidian tunicate Ciona intestinalis. Analysis of regenerative potential along the proximal-distal body axis indicated that distal organs, such as the siphons, their pigmented sensory organs, and the neural complex, could only be replaced from body fragments containing the branchial sac. Distal regeneration involves the formation of a blastema composed of cells that undergo cell proliferation prior to differentiation and cells that differentiate without cell proliferation. Both cell types originate in the branchial sac and appear in the blastema at different times after distal injury. Whereas the branchial sac stem cells are present in young animals, they are depleted in old animals that have lost their regeneration capacity. Thus Ciona adults contain a population of age-related stem cells located in the branchial sac that are a source of precursors for distal body regeneration.

  16. Lipid rafts function in Ca2+ signaling responsible for activation of sperm motility and chemotaxis in the ascidian Ciona intestinalis.

    PubMed

    Zhu, Lihong; Inaba, Kazuo

    2011-12-01

    Lipid rafts are specialized membrane microdomains that function as signaling platforms across plasma membranes of many animal and plant cells. Although there are several studies implicating the role of lipid rafts in capacitation of mammalian sperm, the function of these structures in sperm motility activation and chemotaxis remains unknown. In the ascidian Ciona intestinalis, egg-derived sperm activating- and attracting-factor (SAAF) induces both activation of sperm motility and sperm chemotaxis to the egg. Here we found that a lipid raft disrupter, methyl-β-cyclodextrin (MCD), inhibited both SAAF-induced sperm motility activation and chemotaxis. MCD inhibited both SAAF-promoted synthesis of intracellular cyclic AMP and sperm motility induced by ionophore-mediated Ca(2+) entry, but not that induced by valinomycin-mediated hyperpolarization. Ca(2+)-imaging revealed that lipid raft disruption inhibited Ca(2+) influx upon activation of sperm motility. The Ca(2+)-activated adenylyl cyclase was clearly inhibited by MCD in isolated lipid rafts. The results suggest that sperm lipid rafts function in signaling upstream of cAMP synthesis, most likely in SAAF-induced Ca(2+) influx, and are required for Ca(2+)-dependent pathways underlying activation and chemotaxis in Ciona sperm. Copyright © 2011 Wiley Periodicals, Inc.

  17. Toxicity assessment of the antifouling compound zinc pyrithione using early developmental stages of the ascidian Ciona intestinalis.

    PubMed

    Bellas, Juan

    2005-01-01

    This study investigated the toxicity of zinc pyrithione (Zpt) on the early stages of development of the ascidian Ciona intestinalis. Larval morphological abnormalities were studied after the exposure of C. intestinalis embryos at different stages of development. The median effective concentrations (EC50) ranged from 226-590 nM. The larval settlement stage was the most sensitive to Zpt. Toxic effects of Zpt on larval settlement were detected at 9 nM (EC10). The inhibition of C. intestinalis embryonic development was also used to study the loss of toxicity in Zpt solutions exposed to direct sunlight and laboratory UV light. The results showed that the toxicity of Zpt solutions decreased but did not disappear after 4 h exposure to direct sunlight (EC50 = 484 nM) or UV light (EC50 = 453 nM), compared to control Zpt solutions prepared in dark conditions. On the basis of the present data, predicted no effect concentrations of Zpt to C. intestinalis larvae are lower than predicted environmental concentrations of Zpt in certain polluted areas and therefore, may pose a risk to C. intestinalis populations.

  18. Complete mtDNA of Ciona intestinalis reveals extensive gene rearrangement and the presence of an atp8 and an extra trnM gene in ascidians.

    PubMed

    Gissi, Carmela; Iannelli, Fabio; Pesole, Graziano

    2004-04-01

    The complete mitochondrial genome (mtDNA) of the model organism Ciona intestinalis (Urochordata, Ascidiacea) has been amplified by long-PCR using specific primers designed on putative mitochondrial transcripts identified from publicly available mitochondrial-like expressed sequence tags. The C. intestinalis mtDNA encodes 39 genes: 2 rRNAs, 13 subunits of the respiratory complexes, including ATPase subunit 8 ( atp8), and 24 tRNAs, including 2 tRNA-Met with anticodons 5'-UAU-3'and 5'-CAU-3', respectively. All genes are transcribed from the same strand. This gene content seems to be a common feature of ascidian mtDNAs, as we have verified the presence of a previously undetected atp8 and of two trnM genes in the two other sequenced ascidian mtDNAs. Extensive gene rearrangement has been found in C. intestinalis with respect not only to the common Vertebrata/Cephalochordata/Hemichordata gene organization but also to other ascidian mtDNAs, including the cogeneric Ciona savignyi. Other features such as the absence of long noncoding regions, the shortness of rRNA genes, the low GC content (21.4%), and the absence of asymmetric base distribution between the two strands suggest that this genome is more similar to those of some protostomes than to deuterostomes.

  19. Cholinesterase in larvae of the ascidian,Ciona intestinalis, developing from fragments cut from centrifuged eggs.

    PubMed

    Bell, William A; Holland, Nicholas D

    1974-06-01

    UnfertilizedCiona eggs were centrifuged, stratifying their mitochondria and some other cytoplasmic components. Each centrifuged egg had a mitochondria-free, centripetal clear layer that was contiguous with centrifugal layers containing mitochondria. By cutting centrifuged eggs in two at various levels along the centripetal-centrifugal axis, it was possible to obtain centripetal fragments including virtually no mitochondria, about one-tenth of the uncut egg's mitochondria or about one-fourth of the uncut egg's mitochondria. Most of these centripetal fragments, when fertilized, developed into larvae. However, only the centripetal fragments that included about one-fourth of the uncut egg's mitochondria developed into larvae giving the cytochemical reaction for cholinesterase, a convenient indicator of muscle cell differentiation inCiona. Therefore, the inclusion of a minimum number of mitochondria (more than one-tenth but less than one-fourth the number in the uncut egg) is correlated with muscle cell differentiation in larvae developing from the centripetal fragments. The possible influences of mitochondria and of other cytoplasmic components on muscle differentiation are discussed.

  20. Germ cell regeneration-mediated, enhanced mutagenesis in the ascidian Ciona intestinalis reveals flexible germ cell formation from different somatic cells.

    PubMed

    Yoshida, Keita; Hozumi, Akiko; Treen, Nicholas; Sakuma, Tetsushi; Yamamoto, Takashi; Shirae-Kurabayashi, Maki; Sasakura, Yasunori

    2017-03-15

    The ascidian Ciona intestinalis has a high regeneration capacity that enables the regeneration of artificially removed primordial germ cells (PGCs) from somatic cells. We utilized PGC regeneration to establish efficient methods of germ line mutagenesis with transcription activator-like effector nucleases (TALENs). When PGCs were artificially removed from animals in which a TALEN pair was expressed, somatic cells harboring mutations in the target gene were converted into germ cells, this germ cell population exhibited higher mutation rates than animals not subjected to PGC removal. PGC regeneration enables us to use TALEN expression vectors of specific somatic tissues for germ cell mutagenesis. Unexpectedly, cis elements for epidermis, neural tissue and muscle could be used for germ cell mutagenesis, indicating there are multiple sources of regenerated PGCs, suggesting a flexibility of differentiated Ciona somatic cells to regain totipotency. Sperm and eggs of a single hermaphroditic, PGC regenerated animal typically have different mutations, suggesting they arise from different cells. PGCs can be generated from somatic cells even though the maternal PGCs are not removed, suggesting that the PGC regeneration is not solely an artificial event but could have an endogenous function in Ciona. This study provides a technical innovation in the genome-editing methods, including easy establishment of mutant lines. Moreover, this study suggests cellular mechanisms and the potential evolutionary significance of PGC regeneration in Ciona.

  1. Functions of a GnRH receptor heterodimer of the ascidian, Ciona intestinalis.

    PubMed

    Sakai, T; Aoyama, M; Kusakabe, T; Tsuda, M; Satake, H

    2008-01-01

    Gonadotropin-releasing hormone (GnRH) is a ten-amino acid peptide hormone that plays pivotal roles in reproduction in vertebrates and octopus. Recently, six GnRH forms (t-GnRH-3-8) and four GnRH receptor subtypes (Ci-GnRHR-1-4) were identified in the protochordate, Ciona intestinalis. In this study, we show the functional modulation of Ci-GnRHR-1 via heterodimerization with the orphan receptor subtype, Ci-GnRHR-4. The dimerization between Ci-GnRHR-1 and R-4 was detected by co-immunoprecipitation and immunoblot analysis. Binding assays confirmed the binding of t-GnRHs to Ci-GnRHR-1 but not to R-4, and verified no alternation in ligand-binding affinity between Ci-GnRHR-1 homodimer and Ci-GnRHRI&4 heterodimer. The heterodimer was found to stimulate the elevation of intracellular calcium, time-extension of ERK phosphorylation, and up-regulation of cell proliferation, all in a ligand specific manner, compared with the Ci-GnRHR-1 homodimer. In combination, these results indicated that Ci-GnRHR-4 is not an inactive receptor, but a modulatory factor for Ci-GnRHR-1 in C. intestinalis.

  2. Genomewide gene-associated microsatellite markers for the model invasive ascidian, Ciona intestinalis species complex.

    PubMed

    Lin, Yaping; Chen, Yiyong; Xiong, Wei; Zhan, Aibin

    2016-05-01

    The vase tunicate, Ciona intestinalis species complex, has become a good model for ecological and evolutionary studies, especially those focusing on microevolution associated with rapidly changing environments. However, genomewide genetic markers are still lacking. Here, we characterized a large set of genomewide gene-associated microsatellite markers for C. intestinalis spA (=C. robusta). Bioinformatic analysis identified 4654 microsatellites from expressed sequence tags (ESTs), 2126 of which successfully assigned to chromosomes were selected for further analysis. Based on the distribution evenness on chromosomes, function annotation and suitability for primer design, we chose 545 candidate microsatellites for further characterization. After amplification validation and variation assessment, 218 loci were polymorphic in at least one of the two populations collected from the coast of Arenys de Mar, Spain (N = 24-48), and Cape Town, South Africa (N = 24-33). The number of alleles, observed heterozygosity and expected heterozygosity ranged from 2 to 11, 0 to 0.833 and 0.021 to 0.818, and from 2 to 10, 0 to 0.879 and 0.031 to 0.845 for the Spanish and African populations, respectively. When all microsatellites were tested for cross-species utility, only 60 loci (25.8%) could be successfully amplified and all loci were polymorphic in C. intestinalis spB. A high level of genomewide polymorphism is likely responsible for the low transferability. The large set of microsatellite markers characterized here is expected to provide a useful genomewide resource for ecological and evolutionary studies using C. intestinalis as a model.

  3. Proteomic responses to elevated ocean temperature in ovaries of the ascidian Ciona intestinalis.

    PubMed

    Lopez, Chelsea E; Sheehan, Hannah C; Vierra, David A; Azzinaro, Paul A; Meedel, Thomas H; Howlett, Niall G; Irvine, Steven Q

    2017-07-15

    Ciona intestinalis, a common sea squirt, exhibits lower reproductive success at the upper extreme of the water temperatures it experiences in coastal New England. In order to understand the changes in protein expression associated with elevated temperatures, and possible response to global temperature change, we reared C. intestinalis from embryos to adults at 18°C (a temperature at which they reproduce normally at our collection site in Rhode Island) and 22°C (the upper end of the local temperature range). We then dissected ovaries from animals at each temperature, extracted protein, and measured proteomic levels using shotgun mass spectrometry (LC-MS/MS). 1532 proteins were detected at a 1% false discovery rate present in both temperature groups by our LC-MS/MS method. 62 of those proteins are considered up- or down-regulated according to our statistical criteria. Principal component analysis shows a clear distinction in protein expression pattern between the control (18°C) group and high temperature (22°C) group. Similar to previous studies, cytoskeletal and chaperone proteins are upregulated in the high temperature group. Unexpectedly, we find evidence that proteolysis is downregulated at the higher temperature. We propose a working model for the high temperature response in C. intestinalis ovaries whereby increased temperature induces upregulation of signal transduction pathways involving PTPN11 and CrkL, and activating coordinated changes in the proteome especially in large lipid transport proteins, cellular stress responses, cytoskeleton, and downregulation of energy metabolism. © 2017. Published by The Company of Biologists Ltd.

  4. Trunk lateral cells are neural crest-like cells in the ascidian Ciona intestinalis: insights into the ancestry and evolution of the neural crest.

    PubMed

    Jeffery, William R; Chiba, Takuto; Krajka, Florian Razy; Deyts, Carole; Satoh, Nori; Joly, Jean-Stéphane

    2008-12-01

    Neural crest-like cells (NCLC) that express the HNK-1 antigen and form body pigment cells were previously identified in diverse ascidian species. Here we investigate the embryonic origin, migratory activity, and neural crest related gene expression patterns of NCLC in the ascidian Ciona intestinalis. HNK-1 expression first appeared at about the time of larval hatching in dorsal cells of the posterior trunk. In swimming tadpoles, HNK-1 positive cells began to migrate, and after metamorphosis they were localized in the oral and atrial siphons, branchial gill slits, endostyle, and gut. Cleavage arrest experiments showed that NCLC are derived from the A7.6 cells, the precursors of trunk lateral cells (TLC), one of the three types of migratory mesenchymal cells in ascidian embryos. In cleavage arrested embryos, HNK-1 positive TLC were present on the lateral margins of the neural plate and later became localized adjacent to the posterior sensory vesicle, a staging zone for their migration after larval hatching. The Ciona orthologues of seven of sixteen genes that function in the vertebrate neural crest gene regulatory network are expressed in the A7.6/TLC lineage. The vertebrate counterparts of these genes function downstream of neural plate border specification in the regulatory network leading to neural crest development. The results suggest that NCLC and neural crest cells may be homologous cell types originating in the common ancestor of tunicates and vertebrates and support the possibility that a putative regulatory network governing NCLC development was co-opted to produce neural crest cells during vertebrate evolution.

  5. Monitoring the magnitude of marine vessel infestation by non-indigenous ascidians in the Mediterranean.

    PubMed

    Gewing, Mey-Tal; Shenkar, Noa

    2017-08-15

    Invasive ascidians (Chordata, Tunicata) are dominant nuisance organisms. The current study investigated the role of marine vessels in their dispersal and introduction. An examination of 45 dry-docked marine vessels, comprising recreational, commercial, and military craft, in five Israeli shipyards along the Mediterranean coast, revealed non-indigenous ascidians (NIA) on every second vessel investigated. Military vessels featured the highest ascidian abundance and richness, potentially related to their maintenance routine. Niche areas on the vessels such as sea chests and the propeller exhibited the highest occurrence of ascidians. Overall, these findings provide strong evidence that marine vessels play an acute role in NIA introduction and dispersal, with military vessels and niche areas on all the vessels being more susceptible to serving as vectors. A discovery of a new introduced species during the surveys suggests that the monitoring of marine vessels can serve as an effective tool for the early detection of NIA. Copyright © 2017 Elsevier Ltd. All rights reserved.

  6. The diatom-derived aldehyde decadienal affects life cycle transition in the ascidian Ciona intestinalis through nitric oxide/ERK signalling

    PubMed Central

    Castellano, Immacolata; Ercolesi, Elena; Romano, Giovanna; Ianora, Adrianna; Palumbo, Anna

    2015-01-01

    Polyunsaturated aldehydes (PUAs) are fatty-acid-derived metabolites produced by some microalgae, including different diatom species. PUAs are mainly produced as a wound-activated defence mechanism against microalgal predators or released from senescent cells at the end of a bloom. PUAs, including 2,4-trans-decadienal (DD), induce deleterious effects on embryonic and larval development of several planktonic and benthic organisms. Here, we report on the effects of DD on larval development and metamorphosis of the ascidian Ciona intestinalis. Ciona larval development is regulated by the cross-talking of different molecular events, including nitric oxide (NO) production, ERK activation and caspase 3-dependent apoptosis. We report that treatment with DD at the competence larval stage results in a delay in metamorphosis. DD affects redox balance by reducing total glutathione and NO levels. By biochemical and quantitative gene expression analysis, we identify the NO-signalling network affected by DD, including the upregulation of ERK phosphatase mkp1 and consequent reduction of ERK phosphorylation, with final changes in the expression of downstream ERK target genes. Overall, these results give new insights into the molecular pathways induced in marine organisms after exposure to PUAs during larval development, demonstrating that this aldehyde affects key checkpoints of larval transition from the vegetative to the reproductive life stage. PMID:25788553

  7. The central nervous system of the ascidian larva: mitotic history of cells forming the neural tube in late embryonic Ciona intestinalis.

    PubMed

    Cole, Alison G; Meinertzhagen, Ian A

    2004-07-15

    Ascidian larvae develop after an invariant pattern of embryonic cleavage. Fewer than 400 cells constitute the larval central nervous system (CNS), which forms without either extensive migration or cell death. We catalogue the mitotic history of these cells in Ciona intestinalis, using confocal microscopy of whole-mount embryos at stages from neurulation until hatching. The positions of cells contributing to the CNS were reconstructed from confocal image stacks of embryonic nuclei, and maps of successive stages were used to chart the mitotic descent, thereby creating a cell lineage for each cell. The entire CNS is formed from 10th- to 14th-generation cells. Although minor differences exist in cell position, lineage is invariant in cells derived from A-line blastomeres, which form the caudal nerve cord and visceral ganglion. We document the lineage of five pairs of presumed motor neurons within the visceral ganglion: one pair arises from A/A 10.57, and four from progeny of A/A 9.30. The remaining cells of the visceral ganglion are in their 13th and 14th generations at hatching, with most mitotic activity ceasing around 85% of embryonic development. Of the approximately 330 larval cells previously reported in the CNS of Ciona, we document the lineage of 226 that derive predominantly from A-line blastomeres.

  8. Structural and functional properties of CiNTH, an endonuclease III homologue of the ascidian Ciona intestinalis: critical role of N-terminal region.

    PubMed

    Kato, Seiji; Hashiguchi, Kazunari; Igarashi, Kento; Moriwaki, Takahito; Yonekura, Shin-Ichiro; Zhang-Akiyama, Qiu-Mei

    2012-01-01

    Oxidatively damaged bases in DNA can cause cell death, mutation and/or cancer induction. To overcome such deleterious effects of DNA base oxidation, cells are equipped with base excision repair (BER) initiated by DNA glycosylases. Endonuclease III (Nth), a major DNA glycosylase, mainly excises oxidatively damaged pyrimidines from DNA. The aims of this study were to obtain an overview of the repair mechanism of oxidatively damaged bases and to elucidate the function of BER in maintaining genome stability during embryogenesis and development. In this study, we used the ascidian Ciona intestinalis because at every developmental stage it is possible to observe the phenotype of individuals with DNA damage or mutations. Sequence alignment analysis revealed that the amino acid sequence of Ciona intestinalis Nth homologue (CiNTH) had high homology with those of Escherichia coli, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Caenorhabditis elegans and human Nth homologues. It was evident that two domains, the Helix-hairpin-Helix and 4Fe-4S cluster domains that are critical regions for the Nth activity, are well conserved in CiNTH. CiNTH efficiently complemented the sensitivity of E. coli nth nei mutant to H(2)O(2). CiNTH was bifunctional, with DNA glycosylase and AP lyase activities. It removed thymine glycol, 5-formyluracil and 8-oxoguanine paired with G from DNA via a β-elimination reaction. Interestingly, the N-terminal 44 amino acids were essential for the DNA glycosylase activity of CiNTH.

  9. Nitric oxide affects ERK signaling through down-regulation of MAP kinase phosphatase levels during larval development of the ascidian Ciona intestinalis.

    PubMed

    Castellano, Immacolata; Ercolesi, Elena; Palumbo, Anna

    2014-01-01

    In the ascidian Ciona intestinalis larval development and metamorphosis require a complex interplay of events, including nitric oxide (NO) production, MAP kinases (ERK, JNK) and caspase-3 activation. We have previously shown that NO levels affect the rate of metamorphosis, regulate caspase activity and promote an oxidative stress pathway, resulting in protein nitration. Here, we report that NO down-regulates MAP kinase phosphatases (mkps) expression affecting positively ERK signaling. By pharmacological approach, we observed that the reduction of endogenous NO levels caused a decrease of ERK phosphorylation, whereas increasing levels of NO induced ERK activation. We have also identified the ERK gene network affected by NO, including mpk1, mpk3 and some key developmental genes by quantitative gene expression analysis. We demonstrate that NO induces an ERK-independent down-regulation of mkp1 and mkp3, responsible for maintaining the ERK phosphorylation levels necessary for transcription of key metamorphic genes, such as the hormone receptor rev-erb and the van willebrand protein vwa1c. These results add new insights into the role played by NO during larval development and metamorphosis in Ciona, highlighting the cross-talk between different signaling pathways.

  10. The diatom-derived aldehyde decadienal affects life cycle transition in the ascidian Ciona intestinalis through nitric oxide/ERK signalling.

    PubMed

    Castellano, Immacolata; Ercolesi, Elena; Romano, Giovanna; Ianora, Adrianna; Palumbo, Anna

    2015-03-01

    Polyunsaturated aldehydes (PUAs) are fatty-acid-derived metabolites produced by some microalgae, including different diatom species. PUAs are mainly produced as a wound-activated defence mechanism against microalgal predators or released from senescent cells at the end of a bloom. PUAs, including 2,4-trans-decadienal (DD), induce deleterious effects on embryonic and larval development of several planktonic and benthic organisms. Here, we report on the effects of DD on larval development and metamorphosis of the ascidian Ciona intestinalis. Ciona larval development is regulated by the cross-talking of different molecular events, including nitric oxide (NO) production, ERK activation and caspase 3-dependent apoptosis. We report that treatment with DD at the competence larval stage results in a delay in metamorphosis. DD affects redox balance by reducing total glutathione and NO levels. By biochemical and quantitative gene expression analysis, we identify the NO-signalling network affected by DD, including the upregulation of ERK phosphatase mkp1 and consequent reduction of ERK phosphorylation, with final changes in the expression of downstream ERK target genes. Overall, these results give new insights into the molecular pathways induced in marine organisms after exposure to PUAs during larval development, demonstrating that this aldehyde affects key checkpoints of larval transition from the vegetative to the reproductive life stage.

  11. A pipeline for the systematic identification of non-redundant full-ORF cDNAs for polymorphic and evolutionary divergent genomes: Application to the ascidian Ciona intestinalis

    SciTech Connect

    Gilchrist, Michael J.; Sobral, Daniel; Khoueiry, Pierre; Daian, Fabrice; Laporte, Batiste; Patrushev, Ilya; Matsumoto, Jun; Dewar, Ken; Hastings, Kenneth E. M.; Satou, Yutaka; Lemaire, Patrick; Rothbächer, Ute

    2015-05-27

    Genome-wide resources, such as collections of cDNA clones encoding for complete proteins (full-ORF clones), are crucial tools for studying the evolution of gene function and genetic interactions. Non-model organisms, in particular marine organisms, provide a rich source of functional diversity. Marine organism genomes are, however, frequently highly polymorphic and encode proteins that diverge significantly from those of well-annotated model genomes. The construction of full-ORF clone collections from non-model organisms is hindered by the difficulty of predicting accurately the N-terminal ends of proteins, and distinguishing recent paralogs from highly polymorphic alleles. We also report a computational strategy that overcomes these difficulties, and allows for accurate gene level clustering of transcript data followed by the automated identification of full-ORFs with correct 5'- and 3'-ends. It is robust to polymorphism, includes paralog calling and does not require evolutionary proximity to well annotated model organisms. Here, we developed this pipeline for the ascidian Ciona intestinalis, a highly polymorphic member of the divergent sister group of the vertebrates, emerging as a powerful model organism to study chordate gene function, Gene Regulatory Networks and molecular mechanisms underlying human pathologies. Furthermore, using this pipeline we have generated the first full-ORF collection for a highly polymorphic marine invertebrate. It contains 19,163 full-ORF cDNA clones covering 60% of Ciona coding genes, and full-ORF orthologs for approximately half of curated human disease-associated genes.

  12. A pipeline for the systematic identification of non-redundant full-ORF cDNAs for polymorphic and evolutionary divergent genomes: Application to the ascidian Ciona intestinalis.

    PubMed

    Gilchrist, Michael J; Sobral, Daniel; Khoueiry, Pierre; Daian, Fabrice; Laporte, Batiste; Patrushev, Ilya; Matsumoto, Jun; Dewar, Ken; Hastings, Kenneth E M; Satou, Yutaka; Lemaire, Patrick; Rothbächer, Ute

    2015-08-15

    Genome-wide resources, such as collections of cDNA clones encoding for complete proteins (full-ORF clones), are crucial tools for studying the evolution of gene function and genetic interactions. Non-model organisms, in particular marine organisms, provide a rich source of functional diversity. Marine organism genomes are, however, frequently highly polymorphic and encode proteins that diverge significantly from those of well-annotated model genomes. The construction of full-ORF clone collections from non-model organisms is hindered by the difficulty of predicting accurately the N-terminal ends of proteins, and distinguishing recent paralogs from highly polymorphic alleles. We report a computational strategy that overcomes these difficulties, and allows for accurate gene level clustering of transcript data followed by the automated identification of full-ORFs with correct 5'- and 3'-ends. It is robust to polymorphism, includes paralog calling and does not require evolutionary proximity to well annotated model organisms. We developed this pipeline for the ascidian Ciona intestinalis, a highly polymorphic member of the divergent sister group of the vertebrates, emerging as a powerful model organism to study chordate gene function, Gene Regulatory Networks and molecular mechanisms underlying human pathologies. Using this pipeline we have generated the first full-ORF collection for a highly polymorphic marine invertebrate. It contains 19,163 full-ORF cDNA clones covering 60% of Ciona coding genes, and full-ORF orthologs for approximately half of curated human disease-associated genes.

  13. Evolution of the Chordate Regeneration Blastema: Differential Gene Expression and Conserved Role of Notch Signaling During Siphon Regeneration in the Ascidian Ciona

    PubMed Central

    Hamada, Mayuko; Goricki, Spela; Byerly, Mardi S.; Satoh, Noriyuki; Jeffery, William R.

    2015-01-01

    The regeneration of the oral siphon (OS) and other distal structures in the ascidian Ciona intestinalis occurs by epimorphosis involving the formation of a blastema of proliferating cells. Despite the longstanding use of Ciona as a model in molecular developmental biology, regeneration in this system has not been previously explored by molecular analysis. Here we have employed microarray analysis and quantitative real time RT-PCR to identify genes with differential expression profiles during OS regeneration. The majority of differentially expressed genes were downregulated during OS regeneration, suggesting roles in normal growth and homeostasis. However, a subset of differentially expressed genes was upregulated in the regenerating OS, suggesting functional roles during regeneration. Among the upregulated genes were key members of the Notch signaling pathway, including those encoding the delta and jagged ligands, two fringe modulators, and to a lesser extent the notch receptor. In situ hybridization showed a complementary pattern of delta1 and notch gene expression in the blastema of the regenerating OS. Chemical inhibition of the Notch signaling pathway reduced the levels of cell proliferation in the branchial sac, a stem cell niche that contributes progenitor cells to the regenerating OS, and in the OS regeneration blastema, where siphon muscle fibers eventually re-differentiate. Chemical inhibition also prevented the replacement of oral siphon pigment organs, sensory receptors rimming the entrance of the OS, and siphon muscle fibers, but had no effects on the formation of the wound epidermis. Since Notch signaling is involved in the maintenance of proliferative activity in both the Ciona and vertebrate regeneration blastema, the results suggest a conserved evolutionary role of this signaling pathway in chordate regeneration. The genes identified in this investigation provide the foundation for future molecular analysis of OS regeneration. PMID:26206613

  14. Transposon-mediated enhancer detection reveals the location, morphology and development of the cupular organs, which are putative hydrodynamic sensors, in the ascidian Ciona intestinalis.

    PubMed

    Ohta, Naoyuki; Horie, Takeo; Satoh, Nori; Sasakura, Yasunori

    2010-11-01

    The adult of the ascidian Ciona intestinalis has cupular organs, i.e., putative hydrodynamic sensors, at the atrial epithelium. The cupular organ consists of support cells and sensory neurons, and it extends a gelatinous matrix, known as a cupula, toward the atrial cavity. These characteristics are shared with sensory hair cells in the vertebrate inner ear and lateral line neuromasts in fish and amphibians, which suggests an evolutionary link between the cupular organ and these vertebrate hydrodynamic sensors. In the present study, we have isolated and investigated two transposon-mediated enhancer detection lines that showed GFP expression in support cells of the cupular organs. Using the enhancer detection lines and neuron marker transgenic lines, we describe the position, morphology, and development of the cupular organs. Cupular organs were found at the atrial epithelium, but not in the branchial epithelium. We found that cupular organs are also present along the dorsal fold and the gonoducts. The cells lining the pre-atrial opening in juveniles are presumably precursor cells of the cupular organ. To our knowledge, the present study is the first precise description of the ascidian cupular organ, providing evidence that may help to resolve discrepancies among previous studies on the organ.

  15. Effects of the azole fungicide Imazalil on the development of the ascidian Ciona intestinalis (Chordata, Tunicata): morphological and molecular characterization of the induced phenotype.

    PubMed

    Zega, Giuliana; De Bernardi, Fiorenza; Groppelli, Silvia; Pennati, Roberta

    2009-02-19

    Imazalil (IMA) is a fungicide that is used extensively in fruit plantations and post-harvest treatments, but has teratogenic effects on vertebrate development, possibly due to the perturbation of retinoic acid (RA) levels in the embryo. Ascidians are sessile marine invertebrate chordates that develop through a tadpole larva, with a body plan that shares basic homologies with vertebrates. In this work, we tested the effects of IMA on the development of the solitary ascidian Ciona intestinalis by treating two-cell stage embryos with a range of concentrations (0.1, 0.5, 1, 2.5, 5, 10, 20 and 50microThe fungicide significantly altered ascidian development even at low concentrations and its effects were dose-dependent. Probit analysis revealed that the median lethal concentration, LC(50), was 4.87microM and the median teratogenic concentration, TC(50), was 0.73microM. Larvae developing from embryos exposed to IMA showed malformations of the anterior structures, which became more severe as IMA concentration increased. In particular, the anterior nervous system and the sensory vesicle were reduced, and the pigmented organs (the ocellus and the otolith) progressively lost their pigmentation. The larval phenotype induced by 5microM IMA exposure was further characterized by means of molecular analysis, through whole mount in situ hybridization with probes for genes related to the nervous system: Ci-Otp, Ci-GAD, Ci-POU IV, which are markers of the anterior neuro-ectoderm, the central nervous system and the peripheral nervous system respectively, and Ci-Hox-1, a gene specifically activated by RA, and Ci-Aldh2, a gene for aldehyde dehydrogenase, which is involved in RA synthesis. The altered expression of Ci-Otp, Ci-GAD, Ci-POU IV in 5microM IMA-exposed larvae compared to control larvae showed that this fungicide could affect the differentiation of the anterior nervous system, particularly of the sensory vesicle neurons. Recent studies suggest a similarity between IMA- and RA

  16. cDNA cloning, localization, and candidate binding partners of acid-extractable vitelline-coat protein Ci-v-Themis-like in the ascidian Ciona intestinalis.

    PubMed

    Otsuka, Kei; Yamada, Lixy; Sawada, Hitoshi

    2013-10-01

    Ascidians are hermaphrodites, although several ascidian species show self-sterility because of the occurrence of a self/nonself-recognition system called the self-incompatibility system. We previously reported that two pairs of sperm polycystin 1-like receptors, s-Themis-A and s-Themis-B, and egg fibrinogen-like ligands, v-Themis-A and v-Themis-B, are responsible for self-incompatibility in the ascidian Ciona intestinalis. Our previous results showed that v-Themis-A and v-Themis-B were hardly extracted from the vitelline coat (VC) by acid treatment, which is not in accordance with a report that an acid-extractable VC factor has the ability to distinguish self- from nonself-sperm. These results led us to explore a novel factor from acid-extractable VC proteins that could be involved in self-incompatibility. Here, we report cDNA cloning, expression, and localization of Ci-v-Themis-like, a major acid-extractable VC protein. This protein has a fibrinogen-like domain, as do v-Themis-A and v-Themis-B, but it showed no polymorphisms. Phylogenic analysis suggested that Ci-v-Themis-like is an ancestral protein of v-Themis-A and v-Themis-B. Whole mount in situ hybridization revealed that Ci-v-Themis-like mRNA is expressed in the ovary and testis. Western blotting and immunocytochemistry showed the occurrence of Ci-v-Themis-like in developing oocytes and on the VC of mature eggs. Yeast two-hybrid screenings using testis and ovary libraries revealed candidate interacting proteins; among these candidates, we succeeded in identifying several testis-specific proteins, including sperm proteases and coiled-coil-domain-containing proteins. The results suggest that Ci-v-Themis-like and its binding partners are involved in sperm binding to the VC prior to the allorecognition process during C. intestinalis fertilization.

  17. Differential gene regulation by V(IV) and V (V) ions in the branchial sac, intestine, and blood cells of a vanadium-rich ascidian, Ciona intestinalis.

    PubMed

    Kume, Satoshi; Ueki, Tatsuya; Matsuoka, Hiroki; Hamada, Mayuko; Satoh, Nori; Michibata, Hitoshi

    2012-10-01

    Ascidians are hyperaccumulators that have been studied in detail. Proteins and genes involved in the accumulation process have been identified, but regulation of gene expression related to vanadium accumulation remains unknown. To gain insights into the regulation of gene expression by vanadium in a genome-wide manner, we performed a comprehensive study on the effect of excess vanadium ions on a vanadium-rich ascidian, Ciona intestinalis, using a microarray. RT-PCR and enzyme activity assay were performed from the perspective of redox and accumulation of metal ions in each tissue. Glutathione metabolism-related proteins were significantly up-regulated by V(IV) treatment. Several genes involved in the transport of vanadium and protons, such as Nramp and V-ATPase, were significantly up-regulated by V(IV) treatment. We observed significant up-regulation of glutathione synthesis and degradation pathways in the intestine and branchial sac. In blood cells, expression of Ci-Vanabin4, glutathione reductase activity, glutathione levels, and vanadium concentration increased after V(IV) treatment. V(IV) treatment induced significant changes related to vanadium exclusion, seclusion, and redox pathways in the intestine and branchial sac. It also induced an enhancement of the vanadium reduction and accumulation cascade in blood cells. These differential responses in each tissue in the presence of excess vanadium ions suggest that vanadium accumulation and reduction may have regulatory functions. This is the first report on the gene regulation by the treatment of vanadium-rich ascidians with excess vanadium ions. It provided much information for the mechanism of regulation of gene expression related to vanadium accumulation.

  18. A pipeline for the systematic identification of non-redundant full-ORF cDNAs for polymorphic and evolutionary divergent genomes: Application to the ascidian Ciona intestinalis

    DOE PAGES

    Gilchrist, Michael J.; Sobral, Daniel; Khoueiry, Pierre; ...

    2015-05-27

    Genome-wide resources, such as collections of cDNA clones encoding for complete proteins (full-ORF clones), are crucial tools for studying the evolution of gene function and genetic interactions. Non-model organisms, in particular marine organisms, provide a rich source of functional diversity. Marine organism genomes are, however, frequently highly polymorphic and encode proteins that diverge significantly from those of well-annotated model genomes. The construction of full-ORF clone collections from non-model organisms is hindered by the difficulty of predicting accurately the N-terminal ends of proteins, and distinguishing recent paralogs from highly polymorphic alleles. We also report a computational strategy that overcomes these difficulties,more » and allows for accurate gene level clustering of transcript data followed by the automated identification of full-ORFs with correct 5'- and 3'-ends. It is robust to polymorphism, includes paralog calling and does not require evolutionary proximity to well annotated model organisms. Here, we developed this pipeline for the ascidian Ciona intestinalis, a highly polymorphic member of the divergent sister group of the vertebrates, emerging as a powerful model organism to study chordate gene function, Gene Regulatory Networks and molecular mechanisms underlying human pathologies. Furthermore, using this pipeline we have generated the first full-ORF collection for a highly polymorphic marine invertebrate. It contains 19,163 full-ORF cDNA clones covering 60% of Ciona coding genes, and full-ORF orthologs for approximately half of curated human disease-associated genes.« less

  19. Molecular and functional characterization of cionin receptors in the ascidian, Ciona intestinalis: the evolutionary origin of the vertebrate cholecystokinin/gastrin family.

    PubMed

    Sekiguchi, Toshio; Ogasawara, Michio; Satake, Honoo

    2012-04-01

    Cholecystokinin (CCK) and gastrin are vertebrate brain-gut peptides featured by a sulfated tyrosine residue and a C-terminally amidated tetrapeptide consensus sequence. Cionin, identified in the ascidian, Ciona intestinalis, the closest species to vertebrates, harbors two sulfated tyrosines and the CCK/gastrin consensus tetrapeptide sequence. While a putative cionin receptor, cior, was cloned, the ligand-receptor relationship between cionin and CioR remains unidentified. Here, we identify two cionin receptors, CioR1 and CioR2, which are the aforementioned putative cionin receptor and its novel paralog respectively. Phylogenetic analysis revealed that CioRs are homologous to vertebrate CCK receptors (CCKRs) and diverged from a common ancestor in the Ciona-specific lineage. Cionin activates intracellular calcium mobilization in cultured cells expressing CioR1 or CioR2. Monosulfated and nonsulfated cionin exhibited less potent or no activity, indicating that CioRs possess pharmacological features similar to the vertebrate CCK-specific receptor CCK1R, rather than its subtype CCK2R, given that a sulfated tyrosine in CCK is required for binding to CCK1R, but not to CCK2R. Collectively, the present data reveal that CioRs share a common ancestor with vertebrate CCKRs and indicate that CCK and CCK1R form the ancestral ligand-receptor pair in the vertebrate CCK/gastrin system. Cionin is expressed in the neural complex, digestive organs, oral siphon and atrial siphons, whereas the expression of ciors was detected mainly in these tissues and the ovary. Furthermore, cioninergic neurons innervate both of the siphons. These results suggest that cionin is involved in the regulation of siphonal functions.

  20. Characterization and metal-induced gene transcription of two new copper zinc superoxide dismutases in the solitary ascidian Ciona intestinalis.

    PubMed

    Ferro, Diana; Franchi, Nicola; Mangano, Valentina; Bakiu, Rigers; Cammarata, Matteo; Parrinello, Nicolò; Santovito, Gianfranco; Ballarin, Loriano

    2013-09-15

    Antioxidant enzymes are known to protect living organisms against the oxidative stress risk, also induced by metals. In the present study, we describe the purification and molecular characterization of two Cu,Zn superoxide dismutases (SODs), referred to as Ci-SODa and Ci-SODb, from Ciona intestinalis, a basal chordate widely distributed in temperate shallow seawater. The putative amino acid sequences were compared with Cu,Zn SODs from other metazoans and phylogenetic analyses indicate that the two putative Ci-SODs are more related to invertebrate SODs than vertebrate ones. Both phylogenetic and preliminary homology modeling analyses suggest that Ci-SODa and Ci-SODb are extracellular and intracellular isoform, respectively. The mRNA of the two Cu,Zn SODs was localized in hemocytes and in ovarian follicular cells, as revealed by in situ hybridization. The time course of SOD mRNA levels in the presence of three different metals showed upregulation of ci-soda and inhibition of ci-sodb. Spectrophotometric analysis confirms the presence of SOD activity in Ciona tissues. Our in silico analyses of the ci-soda promoter region revealed putative consensus sequences similar to mammalian metal-responsive elements (MRE), suggesting that the transcription of these genes directly depends on metals. These data emphasize the importance of complex metal regulation of ci-soda and ci-sodb transcription, as components of an efficient detoxification pathway allowing the survival of C. intestinalis in continued, elevated presence of metals in the environment.

  1. Reference gene selection for quantitative gene expression studies during biological invasions: A test on multiple genes and tissues in a model ascidian Ciona savignyi.

    PubMed

    Huang, Xuena; Gao, Yangchun; Jiang, Bei; Zhou, Zunchun; Zhan, Aibin

    2016-01-15

    As invasive species have successfully colonized a wide range of dramatically different local environments, they offer a good opportunity to study interactions between species and rapidly changing environments. Gene expression represents one of the primary and crucial mechanisms for rapid adaptation to local environments. Here, we aim to select reference genes for quantitative gene expression analysis based on quantitative Real-Time PCR (qRT-PCR) for a model invasive ascidian, Ciona savignyi. We analyzed the stability of ten candidate reference genes in three tissues (siphon, pharynx and intestine) under two key environmental stresses (temperature and salinity) in the marine realm based on three programs (geNorm, NormFinder and delta Ct method). Our results demonstrated only minor difference for stability rankings among the three methods. The use of different single reference gene might influence the data interpretation, while multiple reference genes could minimize possible errors. Therefore, reference gene combinations were recommended for different tissues - the optimal reference gene combination for siphon was RPS15 and RPL17 under temperature stress, and RPL17, UBQ and TubA under salinity treatment; for pharynx, TubB, TubA and RPL17 were the most stable genes under temperature stress, while TubB, TubA and UBQ were the best under salinity stress; for intestine, UBQ, RPS15 and RPL17 were the most reliable reference genes under both treatments. Our results suggest that the necessity of selection and test of reference genes for different tissues under varying environmental stresses. The results obtained here are expected to reveal mechanisms of gene expression-mediated invasion success using C. savignyi as a model species.

  2. Identification and localization of the sperm CRISP family protein CiUrabin involved in gamete interaction in the ascidian Ciona intestinalis.

    PubMed

    Yamaguchi, Akira; Saito, Takako; Yamada, Lixy; Taniguchi, Hisaaki; Harada, Yoshito; Sawada, Hitoshi

    2011-07-01

    Ascidians are hermaphrodites, and most release sperm and eggs nearly simultaneously. Many species, including Halocynthia roretzi and Ciona intestinalis, are self-sterile. We previously reported that the interaction between a 12 EGF-like repeat-containing vitelline-coat (VC) protein, HrVC70, and a sperm GPI-anchored CRISP, HrUrabin, in lipid rafts plays a key role in self-/nonself-recognizable gamete interaction in H. roretzi. On the other hand, we recently identified two pairs of polymorphic genes responsible for self-incompatibility in C. intestinalis by positional cloning: The sperm polycystin 1-like receptors s-Themis-A/B and its fibrinogen-like ligand v-Themis-A/B on the VC. However, it is not known if the orthologs of HrVC70 and HrUrabin also participate in gamete interaction in C. intestinalis since they are from different orders. Here, we tested for a C. intestinalis ortholog (CiUrabin) of HrUrabin by searching the genome database and proteomes of sperm lipid rafts. The identified CiUrabin belongs to the CRISP family, with a PR domain and a GPI-anchor-attachment site. CiUrabin appears to be specifically expressed in the testis and localized at the surface of the sperm head, as revealed by Northern blotting and immunocytochemistry, respectively. The specific interaction between CiVC57, a C. intestinalis ortholog of HrVC70, and CiUrabin was confirmed by Far Western analysis, similarly to the interaction between HrVC70 and HrUrabin. The molecular interaction between CiVC57 and CiUrabin may be involved in the primary binding of sperm to the VC prior to the allorecognition process, mediated by v-Themis-A/B and s-Themis-A/B, during fertilization of C. intestinalis.

  3. Decoding cis-regulatory systems in ascidians.

    PubMed

    Kusakabe, Takehiro

    2005-02-01

    Ascidians, or sea squirts, are lower chordates, and share basic gene repertoires and many characteristics, both developmental and physiological, with vertebrates. Therefore, decoding cis-regulatory systems in ascidians will contribute toward elucidating the genetic regulatory systems underlying the developmental and physiological processes of vertebrates. cis-Regulatory DNAs can also be used for tissue-specific genetic manipulation, a powerful tool for studying ascidian development and physiology. Because the ascidian genome is compact compared with vertebrate genomes, both intergenic regions and introns are relatively small in ascidians. Short upstream intergenic regions contain a complete set of cis-regulatory elements for spatially regulated expression of a majority of ascidian genes. These features of the ascidian genome are a great advantage in identifying cis-regulatory sequences and in analyzing their functions. Function of cis-regulatory DNAs has been analyzed for a number of tissue-specific and developmentally regulated genes of ascidians by introducing promoter-reporter fusion constructs into ascidian embryos. The availability of the whole genome sequences of the two Ciona species, Ciona intestinalis and Ciona savignyi, facilitates comparative genomics approaches to identify cis-regulatory DNAs. Recent studies demonstrate that computational methods can help identify cis-regulatory elements in the ascidian genome. This review presents a comprehensive list of ascidian genes whose cis-regulatory regions have been subjected to functional analysis, and highlights the recent advances in bioinformatics and comparative genomics approaches to cis-regulatory systems in ascidians.

  4. Phylogenetics, biogeography and population genetics of the ascidian Botryllus schlosseri in the Mediterranean Sea and beyond.

    PubMed

    Reem, Eitan; Douek, Jacob; Paz, Guy; Katzir, Gadi; Rinkevich, Baruch

    2017-02-01

    The wide distribution of the ascidian Botryllus schlosseri along the Mediterranean coasts has been documented since the eighteenth century. However, despite copious documentation, analyses of dispersal modes and genetic profiles were limited to local populations or restricted regions. In order to get a pan-Mediterranean overview, 288 specimens from 11 populations of B. schlosseri from the western and eastern Mediterranean basins were sampled and analyzed using five microsatellite loci and COI sequences. Both molecular markers revealed high polymorphisms, with 182 microsatellites alleles and 54 COI haplotypes. Overall, Fst, Dest, and COI Фpt values were 0.146, 0.635 and 0.322, respectively, reflecting a high genetic diversity and a significant genetic structure as compared to other B. schlosseri populations worldwide, reflected by substantially higher values for effective number of alleles (Ne) in the Mediterranean. A phylogenetic analysis of the COI sequences resulted in four distinct clades and two molecular operational taxonomic units (OTUs). We recorded a stronger genetic structure among the populations of the eastern basin compared to the western basin (microsatellites Fst=0.217 versus 0.082; COI Фpt=0.416 versus 0.171), suggesting either a restricted connectivity between the basins or a stronger genetic drift in each basin. The occurrence of two OTUs and different ecological conditions may also contribute to this finding. Mean Nei's genetic distance in the eastern Mediterranean populations was more than three times higher compared to the western basin. No correlation was observed between geographic and genetic distances (Mantel test), suggesting that maritime transport is the main dispersal vector of B. schlosseri colonies. The possibility that the Mediterranean is a center of diversity for B. schlosseri, and probably its site of origin, is further discussed.

  5. How Fast Is the Sessile Ciona?

    PubMed Central

    Berná, Luisa; Alvarez-Valin, Fernando; D'Onofrio, Giuseppe

    2009-01-01

    Genomewide analyses of distances between orthologous gene pairs from the ascidian species Ciona intestinalis and Ciona savignyi were compared with those of vertebrates. Combining this data with a detailed and careful use of vertebrate fossil records, we estimated the time of divergence between the two ascidians nearly 180 My. This estimation was obtained after correcting for the different substitution rates found comparing several groups of chordates; indeed we determine here that on average Ciona species evolve 50% faster than vertebrates. PMID:20052388

  6. Ciona intestinalis: chordate development made simple.

    PubMed

    Passamaneck, Yale J; Di Gregorio, Anna

    2005-05-01

    Thanks to their transparent and rapidly developing mosaic embryos, ascidians (or sea squirts) have been a model system for embryological studies for over a century. Recently, ascidians have entered the postgenomic era, with the sequencing of the Ciona intestinalis genome and the accumulation of molecular resources that rival those available for fruit flies and mice. One strength of ascidians as a model system is their close similarity to vertebrates. Literature reporting molecular homologies between vertebrate and ascidian tissues has flourished over the past 15 years, since the first ascidian genes were cloned. However, it should not be forgotten that ascidians diverged from the lineage leading to vertebrates over 500 million years ago. Here, we review the main similarities and differences so far identified, at the molecular level, between ascidian and vertebrate tissues and discuss the evolution of the compact ascidian genome. Copyright 2005 Wiley-Liss, Inc.

  7. Stress response in the ascidian Ciona intestinalis: transcriptional profiling of genes for the heat shock protein 70 chaperone system under heat stress and endoplasmic reticulum stress.

    PubMed

    Fujikawa, Tetsuya; Munakata, Takeo; Kondo, Shin-ichi; Satoh, Nori; Wada, Shuichi

    2010-03-01

    The genome of Ciona intestinalis contains eight genes for HSP70 superfamily proteins, 36 genes for J-proteins, a gene for a J-like protein, and three genes for BAG family proteins. To understand the stress responses of genes in the HSP70 chaperone system comprehensively, the transcriptional profiles of these 48 genes under heat stress and endoplasmic reticulum (ER) stress were studied using real-time reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. Heat stress treatment increased the messenger RNA (mRNA) levels of six HSP70 superfamily genes, eight J-protein family genes, and two BAG family genes. In the cytoplasmic group of the DnaK subfamily of the HSP70 family, Ci-HSPA1/6/7-like was the only heat-inducible gene and Ci-HSPA2/8 was the only constitutively active gene which showed striking simplicity in comparison with other animals that have been examined genome-wide so far. Analyses of the time course and temperature dependency of the heat stress responses showed that the induction of Ci-HSPA1/6/7-like expression rises to a peak after heat stress treatment at 28 degrees C (10 degrees C upshift from control temperature) for 1 h. ER stress treatment with Brefeldin A, a drug that is known to act as ER stress inducer, increased the mRNA levels of four HSP70 superfamily genes and four J-protein family genes. Most stress-inducible genes are conserved between Ciona and vertebrates, as expected from a close evolutionary relationship between them. The present study characterized the stress responses of HSP70 chaperone system genes in Ciona for the first time and provides essential data for comprehensive understanding of the functions of the HSP70 chaperone system.

  8. Cell lineage and cis-regulation for a unique GABAergic/glycinergic neuron type in the larval nerve cord of the ascidian Ciona intestinalis.

    PubMed

    Nishitsuji, Koki; Horie, Takeo; Ichinose, Aoi; Sasakura, Yasunori; Yasuo, Hitoyoshi; Kusakabe, Takehiro G

    2012-02-01

    The tunicate Ciona intestinalis larva has a simple central nervous system (CNS), consisting of fewer than 400 cells, which is homologous to the vertebrate CNS. Recent studies have revealed neuronal types and networks in the larval CNS of C. intestinalis, yet their cell lineage and the molecular mechanism by which particular types of neurons are specified and differentiate remain poorly understood. Here, we report cell lineage origin and a cis-regulatory module for the anterior caudal inhibitory neurons (ACINs), a putative component of the central pattern generator regulating swimming locomotion. The vesicular GABA⁄ glycine transporter gene Ci-VGAT, a specific marker for GABAergic ⁄ glycinergic neurons, is expressed in distinct sets of neurons, including ACINs of the tail nerve cord and others in the brain vesicle and motor ganglion. Comparative genomics analysis between C. intestinalis and Ciona savignyi and functional analysis in vivo identified the cis-regulatory module responsible for Ci-VGAT expression in ACINs. Our cell lineage analyses inferred that ACINs derive from A11.116 cells, which have been thought to solely give rise to glial ependymal cells of the lateral wall of the nerve cord. The present findings will provide a solid basis for future studies addressing the molecular mechanism underlying specification of ACINs, which play a critical role in controlling larval locomotion

  9. Stress response in the ascidian Ciona intestinalis: transcriptional profiling of genes for the heat shock protein 70 chaperone system under heat stress and endoplasmic reticulum stress

    PubMed Central

    Fujikawa, Tetsuya; Munakata, Takeo; Kondo, Shin-ichi; Satoh, Nori

    2009-01-01

    The genome of Ciona intestinalis contains eight genes for HSP70 superfamily proteins, 36 genes for J-proteins, a gene for a J-like protein, and three genes for BAG family proteins. To understand the stress responses of genes in the HSP70 chaperone system comprehensively, the transcriptional profiles of these 48 genes under heat stress and endoplasmic reticulum (ER) stress were studied using real-time reverse transcriptase–polymerase chain reaction (RT-PCR) analysis. Heat stress treatment increased the messenger RNA (mRNA) levels of six HSP70 superfamily genes, eight J-protein family genes, and two BAG family genes. In the cytoplasmic group of the DnaK subfamily of the HSP70 family, Ci-HSPA1/6/7-like was the only heat-inducible gene and Ci-HSPA2/8 was the only constitutively active gene which showed striking simplicity in comparison with other animals that have been examined genome-wide so far. Analyses of the time course and temperature dependency of the heat stress responses showed that the induction of Ci-HSPA1/6/7-like expression rises to a peak after heat stress treatment at 28°C (10°C upshift from control temperature) for 1 h. ER stress treatment with Brefeldin A, a drug that is known to act as ER stress inducer, increased the mRNA levels of four HSP70 superfamily genes and four J-protein family genes. Most stress-inducible genes are conserved between Ciona and vertebrates, as expected from a close evolutionary relationship between them. The present study characterized the stress responses of HSP70 chaperone system genes in Ciona for the first time and provides essential data for comprehensive understanding of the functions of the HSP70 chaperone system. Electronic supplementary material The online version of this article (doi:10.1007/s12192-009-0133-x) contains supplementary material, which is available to authorized users. PMID:19629754

  10. Application of the cis-regulatory region of a heat-shock protein 70 gene to heat-inducible gene expression in the ascidian Ciona intestinalis.

    PubMed

    Kawaguchi, Akane; Utsumi, Nanami; Morita, Maki; Ohya, Aya; Wada, Shuichi

    2015-01-01

    Temporally controlled induction of gene expression is a useful technique for analyzing gene function. To make such a technique possible in Ciona intestinalis embryos, we employed the cis-regulatory region of the heat-shock protein 70 (HSP70) gene Ci-HSPA1/6/7-like for heat-inducible gene expression in C. intestinalis embryos. We showed that Ci-HSPA1/6/7-like becomes heat shock-inducible by the 32-cell stage during embryogenesis. The 5'-upstream region of Ci-HSPA1/6/7-like, which contains heat-shock elements indispensable for heat-inducible gene expression, induces the heat shock-dependent expression of a reporter gene in the whole embryo from the 32-cell to the middle gastrula stages and in progressively restricted areas of embryos in subsequent stages. We assessed the effects of heat-shock treatments in different conditions on the normality of embryos and induction of transgene expression. We evaluated the usefulness of this technique through overexpression experiments on the well-characterized, developmentally relevant gene, Ci-Bra, and showed that this technique is applicable for inferring the gene function in C. intestinalis.

  11. Two Decades of Ascidian Developmental Biology: A Personal Research Story.

    PubMed

    Satoh, Noriyuki

    2016-01-01

    Ascidians are the closest relatives of vertebrates. Their utility in experimental embryology has been well recognized because of their simple mode of embryogenesis to form tadpole larvae with a basal chordate body plan. Approximately two decades of research, including decoding of the Ciona genome, have promoted ascidians as one of the best systems for exploring genome-wide mechanisms of developmental transcriptional control and chordate evolution.

  12. Mutation studies in ascidians: a review.

    PubMed

    Crocetta, Fabio; Marino, Rita; Cirino, Paola; Macina, Alberto; Staiano, Leopoldo; Esposito, Rosaria; Pezzotti, Maria Rosa; Racioppi, Claudia; Toscano, Francesco; De Felice, Elena; Locascio, Annamaria; Ristoratore, Filomena; Spagnuolo, Antonietta; Zanetti, Laura; Branno, Margherita; Sordino, Paolo

    2015-01-01

    Historically, mutations have had a significant impact on the study of developmental processes and phenotypic evolution. Lesions in DNA are created by artificial methods or detected by natural genetic variation. Random mutations are then ascribed to genetic change by direct sequencing or positional cloning. Tunicate species of the ascidian genus Ciona represent nearly fully realized model systems in which gene function can be investigated in depth. Additionally, tunicates are valuable organisms for the study of naturally occurring mutations due to the capability to exploit genetic variation down to the molecular level. Here, we summarize the available information about how mutations are studied in ascidians with examples of insights that have resulted from these applications. We also describe notions and methodologies that might be useful for the implementation of easy and tight procedures for mutations studies in Ciona.

  13. Genomics and developmental approaches to an ascidian adenohypophysis primordium.

    PubMed

    Kano, Shungo

    2010-07-01

    Ascidians, which are the closest phylogenetic relatives to vertebrates, lack a distinct pituitary gland, which is the major endocrine gland in vertebrates. Nevertheless, for the past 130 years, it has been debated that the ascidian neural complex (NC) is homologous to the pituitary. Of the three major components of the NC, the neural gland (NG) has mainly been thought to be the ascidian counterpart of the pituitary. Recently, however, the ciliated funnel, and not the NG, was postulated to be the adenohypophysis (AH) primordium because it is likely derived from oral ectoderm, and because the expression of several placodal genes is comparable to their expression in vertebrates. An extensive in silico survey of the Ciona intestinalis genome sequence revealed that genes encoding pituitary hormones are absent in ascidians. Under the circumstances, this thesis attempts to find a path that shows that the AH primordium is recognizable in the ascidian by revisiting molecular and developmental data from recent public resources on C. intestinalis, and through the use of advanced bio-imaging techniques. A putative Ciona genetic pathway, which was constructed by referring to data from mammals, shows that only a patchwork of the genetic network exists to achieve terminal differentiation of the AH endocrine cells in the Ciona genome. Re-annotation on glycoprotein hormone related proteins, a GPA2/ARP and two GPB5/BRP ones previously reported, reveals that the GPA2 locus contains two splicing variants, and one variant likely formed a three-dimensional conformation similar to that of human GPA2. No clone of the GPB5/BRP1 locus has been isolated, and another candidate, BRP2, is unlikely to be a GPB5. Next, I argued a possibility that endocrine activities of Ciona species could be specialized in association with its short generation time, and I suggest that not only Ciona species but also other ascidians should be studied in order to understand ascidian endocrinology. Confocal images

  14. The Ciona intestinalis cleavage clock is independent of DNA methylation.

    PubMed

    Suzuki, Miho M; Mori, Tomoko; Satoh, Noriyuki

    2016-10-01

    The initiation of embryonic gene expression in ascidian embryos appears to be tightly regulated by the number of DNA replication cycles. DNA methylation is thought to contribute to the clock mechanism that counts the rounds of DNA replication. We used mass spectrometry and whole genome bisulfite sequencing to characterize DNA methylation changes that occur in early developmental stages of the ascidian, Ciona intestinalis. We found that global DNA methylation in early Ciona development was static, and a base-wise comparison between the genomes of consecutive developmental stages found no DNA demethylation that was related to zygotic gene activation. Additionally, 5hmC was hardly detected by mass spectrometry in the developmental samples, suggesting a lack of demethylation mediated by ten eleven translocation (TET) methylcytosine dioxygenase in C. intestinalis. We conclude that DNA methylation is not involved in regulating DNA replication-dependent transcriptional activation.

  15. Computational analysis of Ciona intestinalis operons.

    PubMed

    Zeller, Robert W

    2010-07-01

    Operons are clusters of genes that are co-regulated from a common promoter. Operons are typically associated with prokaryotes, although a small number of eukaryotes have been shown to possess them. Among metazoans, operons have been extensively characterized in the nematode Caenorhabditis elegans in which ∼15% of the total genes are organized into operons. The most recent genome assembly for the ascidian Ciona intestinalis placed ∼20% of the genes (2909 total) into 1310 operons. The majority of these operons are composed of two genes, while the largest are composed of six. Here is reported a computational analysis of the genes that comprise the Ciona operons. Gene ontology (GO) terms were identified for about two-thirds of the operon-encoded genes. Using the extensive collection of public EST libraries, estimates of temporal patterns of gene expression were generated for the operon-encoded genes. Lastly, conservation of operons was analyzed by determining how many operon-encoded genes were present in the ascidian Ciona savignyi and whether these genes were organized in orthologous operons. Over 68% of the operon-encoded genes could be assigned one or more GO terms and 697 of the 1310 operons contained genes in which all genes had at least one GO term. Of these 697 operons, GO terms were shared by all of the genes within 146 individual operons, suggesting that most operons encode genes with unrelated functions. An analysis of operon gene expression from nine different EST libraries indicated that for 587 operons, all of the genes that comprise an individual operon were expressed together in at least one EST library, suggesting that these genes may be co-regulated. About 50% (74/146) of the operons with shared GO terms also showed evidence of gene co-regulation. Comparisons with the C. savignyi genome identified orthologs for 1907 of 2909 operon genes. About 38% (504/1310) of the operons are conserved between the two Ciona species. These results suggest that like

  16. Identification of autophagy genes in Ciona intestinalis: a new experimental model to study autophagy mechanism.

    PubMed

    Godefroy, Nelly; Hoa, Céline; Tsokanos, Foivos; Le Goff, Emilie; Douzery, Emmanuel J P; Baghdiguian, Stephen; Martinand-Mari, Camille

    2009-08-01

    Programmed cell death (PCD) is a mechanism implicated in many physiological and pathological processes. Until recently, apoptosis (self-killing) was the most largely studied mechanism of PCD but a growing number of laboratories are now interested in autophagy (self-eating). In the past few years data showing a tight link between both pathways has accumulated. Until now our laboratory used Ciona intestinalis, a chordate model in which in vivo experiments are possible, to study apoptosis. Recently, we showed that autophagy also occurs in the development of Ciona intestinalis and that the specific markers of both types of death are found in the same tissues and/or in the same cells. These results drove us to postulate that Ciona intestinalis can be a good model to study the link between apoptosis and autophagy. In this article, we conducted an in silico study of autophagy genes. We explored the genomes of Ciona intestinalis, of the second ascidian Ciona savignyi, and those of the classical biological models (Saccharomyces cerevisiae, Drosophila melanogaster, Caenorhabditis elegans and Homo sapiens) to extract and compare autophagy gene sequences. This genomic study was completed by an analysis of: (i) mRNA profile expression during development and (ii) the localization of Beclin protein by immunofluorescent staining in the Ciona intestinalis larvae. Taken together, the results allowed us to conclude that a complex autophagic machinery is present in Ciona intestinalis. Actually, the number of autophagy genes in Ciona intestinalis is comparable to the number of autophagy genes in human.

  17. Ascidian fauna (Tunicata, Ascidiacea) of subantarctic and temperate regions of Chile.

    PubMed

    Turon, Xavier; Cañete, Juan I; Sellanes, Javier; Rocha, Rosana M; López-Legentil, Susanna

    2016-03-21

    We studied the ascidian fauna from two zones located in subantarctic (Punta Arenas, latitude 53º) and temperate Chile (Coquimbo, latitude 29º). The different oceanographic features of the two zones, with influence of the Humboldt Current in the north and the Cape Horn Current System and freshwater inputs in the south, led to markedly different ascidian faunas. A total of 22 species were recorded, with no shared species across the two areas (11 species each). The new species Polyzoa iosune is described, Lissoclinum perforatum is found for the first time in the Pacific Ocean, and Synoicum georgianum and Polyzoa minor are new to the Chilean fauna. The populations of Ciona in the Coquimbo area (formerly attributed to Ciona intestinalis) correspond to the species Ciona robusta. A total of 35 Cytochrome oxidase (COI) sequences of the standard barcode region have been obtained for 17 of the 22 species reported.

  18. Peptidomic analysis of the central nervous system of the protochordate, Ciona intestinalis: homologs and prototypes of vertebrate peptides and novel peptides.

    PubMed

    Kawada, Tsuyoshi; Ogasawara, Michio; Sekiguchi, Toshio; Aoyama, Masato; Hotta, Kohji; Oka, Kotaro; Satake, Honoo

    2011-06-01

    The phylogenetic position of ascidians as the chordate invertebrates closest to vertebrates suggests that they might possess homologs and/or prototypes of vertebrate peptide hormones and neuropeptides as well as ascidian-specific peptides. However, only a small number of peptides have so far been identified in ascidians. In the present study, we have identified various peptides in the ascidian, Ciona intestinalis. Mass spectrometry-based peptidomic analysis detected 33 peptides, including 26 novel peptides, from C. intestinalis. The ascidian peptides are largely classified into three categories: 1) prototypes and homologs of vertebrate peptides, such as galanin/galanin-like peptide, which have never been identified in any invertebrates; 2) peptides partially homologous with vertebrate peptides, including novel neurotesin-like peptides; 3) novel peptides. These results not only provide evidence that C. intestinalis possesses various homologs and prototypes of vertebrate neuropeptides and peptide hormones but also suggest that several of these peptides might have diverged in the ascidian-specific evolutionary lineage. All Ciona peptide genes were expressed in the neural complex, whereas several peptide gene transcripts were also distributed in peripheral tissues, including the ovary. Furthermore, a Ciona neurotensin-like peptide, C. intestinalis neurotensin-like peptide 6, was shown to down-regulate growth of Ciona vitellogenic oocytes. These results suggest that the Ciona peptides act not only as neuropeptides in the neural tissue but also as hormones in nonneuronal tissues and that ascidians, unlike other invertebrates, such as nematodes, insects, and sea urchins, established an evolutionary origin of the peptidergic neuroendocrine, endocrine, and nervous systems of vertebrates with certain specific molecular diversity.

  19. Seasonal heterogeneity provides a niche opportunity for ascidian invasion in subtropical marine communities.

    PubMed

    Astudillo, Juan C; Leung, Kenneth M Y; Bonebrake, Timothy C

    2016-12-01

    Implications of changes in environmental conditions caused by seasonality and human alterations on the recruitment of non-native species and their biotic resistance to predation are poorly understood. Here, through the use of experimental recruitment panels and predation exclusion cages, we examined 1) whether a subtropical seasonality (i.e., tropical and temperate conditions) affects the recruitment and abundance of the non-native ascidian Ciona intestinalis, the cryptogenic Styela plicata and Ascidia sydneiensis, and native Hermandia momus in fouling communities in Hong Kong, 2) whether human environmental alterations (i.e., typhoon shelters and sheltered bays with different habitat alteration and seawater quality) affect the abundance of the ascidians, and 3) whether predation reduces the abundance of ascidians under different environmental conditions caused by seasonality and human alteration. Our experimental results indicate that seasonality provides a temporal niche for the recruitment of the ascidians; C. intestinalis and S. plicata recruited mostly in winter, whereas A. sydneiensis and H. momus recruited in summer. Ciona intestinalis was the only ascidian that prospered in anthropogenically altered environments where it monopolized communities. The marked seasonal recruitment of the ascidians obscured the effect of predation between seasons, whereas human alteration did not affect predation. The recruitment of the ascidians in subtropical communities appeared to correspond to their original temperate or tropical distributions, hence Ciona intestinalis, with a temperate native distribution, benefits from a temporal niche opportunity during winter conditions. We argue that seasonality, as an important ecological factor for recruitment and community ecology dynamics, must also be considered in the context of biological invasion. Copyright © 2016 Elsevier Ltd. All rights reserved.

  20. An Otx/Nodal Regulatory Signature for Posterior Neural Development in Ascidians

    PubMed Central

    Roure, Agnès; Lemaire, Patrick; Darras, Sébastien

    2014-01-01

    In chordates, neural induction is the first step of a complex developmental process through which ectodermal cells acquire a neural identity. In ascidians, FGF-mediated neural induction occurs at the 32-cell stage in two blastomere pairs, precursors respectively of anterior and posterior neural tissue. We combined molecular embryology and cis-regulatory analysis to unveil in the ascidian Ciona intestinalis the remarkably simple proximal genetic network that controls posterior neural fate acquisition downstream of FGF. We report that the combined action of two direct FGF targets, the TGFβ factor Nodal, acting via Smad- and Fox-binding sites, and the transcription factor Otx suffices to trigger ascidian posterior neural tissue formation. Moreover, we found that this strategy is conserved in the distantly related ascidian Phallusia mammillata, in spite of extreme sequence divergence in the cis-regulatory sequences involved. Our results thus highlight that the modes of gene regulatory network evolution differ with the evolutionary scale considered. Within ascidians, developmental regulatory networks are remarkably robust to genome sequence divergence. Between ascidians and vertebrates, major fate determinants, such as Otx and Nodal, can be co-opted into different networks. Comparative developmental studies in ascidians with divergent genomes will thus uncover shared ascidian strategies, and contribute to a better understanding of the diversity of developmental strategies within chordates. PMID:25121599

  1. Gene expression profiles of FABP genes in protochordates, Ciona intestinalis and Branchiostoma belcheri.

    PubMed

    Orito, Wataru; Ohhira, Fuyuko; Ogasawara, Michio

    2015-11-01

    Fatty-acid-binding proteins (FABPs) are small intracellular proteins associated with the transportation of fatty acids. Members of the FABPs share similar amino acid sequences and tertiary structures and form, together with a member of the cellular retinol-binding proteins (CRBPs), the intracellular-lipid-binding protein (iLBP) family. In vertebrates, several types of FABP have been isolated and classified into three subfamilies: 2-4. In invertebrates, several FABP-related proteins have been reported in protostomes and amphioxus; however, little is known about the relationship between their phylogenetic positions and expression patterns. We have performed a genome-wide survey of FABP-related genes in protochordates: amphioxus Branchiostoma belcheri and the ascidian Ciona intestinalis. Comprehensive BLAST searches in NCBI and the Ciona Ghost Database by using amino acid sequences of all FABPs have revealed that the ascidian C. intestinalis and amphioxus B. belcheri contain six and seven FABP-related genes in their haploid genomes, respectively. Expression pattern analyses by whole-mount in situ hybridization in Ciona transparent juveniles and serial-section in situ hybridizations in adult amphioxus have revealed that all genes are mainly expressed in the postpharyngeal digestive tract. In particular, the expression of FABP-related genes of subfamily-2 (liver/ileum type) and subfamily-3 (intestinal type) in the ascidian pyloric gland and amphioxus hepatic cecum provides insight into the evolution of hepatic-related structures of chordates and FABP-related genes.

  2. Ciona as a Simple Chordate Model for Heart Development and Regeneration

    PubMed Central

    Anderson, Heather Evans; Christiaen, Lionel

    2016-01-01

    Cardiac cell specification and the genetic determinants that govern this process are highly conserved among Chordates. Recent studies have established the importance of evolutionarily-conserved mechanisms in the study of congenital heart defects and disease, as well as cardiac regeneration. As a basal Chordate, the Ciona model system presents a simple scaffold that recapitulates the basic blueprint of cardiac development in Chordates. Here we will focus on the development and cellular structure of the heart of the ascidian Ciona as compared to other Chordates, principally vertebrates. Comparison of the Ciona model system to heart development in other Chordates presents great potential for dissecting the genetic mechanisms that underlie congenital heart defects and disease at the cellular level and might provide additional insight into potential pathways for therapeutic cardiac regeneration. PMID:27642586

  3. Cloning of ascidian homeobox genes provides evidence for a primordial chordate cluster.

    PubMed

    Di Gregorio, A; Spagnuolo, A; Ristoratore, F; Pischetola, M; Aniello, F; Branno, M; Cariello, L; Di Lauro, R

    1995-04-24

    In order to isolate genes important in controlling embryonic development in Tunicates, a genomic library from the ascidian Ciona intestinalis was screened with a degenerate oligodeoxyribonucleotide encoding the third helix of Antennapedia-type homeoboxes. Fourteen C. intestinalis homeobox genes, corresponding to several classes of homeodomains, have been identified. Five of the isolated homeoboxes show their highest homology to members of the Vertebrate HOX clusters. mRNAs for two of the isolated homeoboxes are present in unfertilized C. intestinalis eggs.

  4. Divergent mechanisms regulate conserved cardiopharyngeal development and gene expression in distantly related ascidians

    PubMed Central

    Stolfi, Alberto; Lowe, Elijah K; Racioppi, Claudia; Ristoratore, Filomena; Brown, C Titus; Swalla, Billie J; Christiaen, Lionel

    2014-01-01

    Ascidians present a striking dichotomy between conserved phenotypes and divergent genomes: embryonic cell lineages and gene expression patterns are conserved between distantly related species. Much research has focused on Ciona or Halocynthia spp. but development in other ascidians remains poorly characterized. In this study, we surveyed the multipotent myogenic B7.5 lineage in Molgula spp. Comparisons to the homologous lineage in Ciona revealed identical cell division and fate specification events that result in segregation of larval, cardiac, and pharyngeal muscle progenitors. Moreover, the expression patterns of key regulators are conserved, but cross-species transgenic assays uncovered incompatibility, or ‘unintelligibility’, of orthologous cis-regulatory sequences between Molgula and Ciona. These sequences drive identical expression patterns that are not recapitulated in cross-species assays. We show that this unintelligibility is likely due to changes in both cis- and trans-acting elements, hinting at widespread and frequent turnover of regulatory mechanisms underlying otherwise conserved aspects of ascidian embryogenesis. DOI: http://dx.doi.org/10.7554/eLife.03728.001 PMID:25209999

  5. A chemoattractant for ascidian spermatozoa is a sulfated steroid

    PubMed Central

    Yoshida, Manabu; Murata, Michio; Inaba, Kazuo; Morisawa, Masaaki

    2002-01-01

    Sperm chemotaxis toward eggs before fertilization has been demonstrated in many animals and plants, and several peptides and small organic compounds acting as chemoattractants have been identified. We previously showed that sperm of the ascidians Ciona intestinalis and Ciona savignyi are activated and then attracted toward the egg by a common factor released from the egg. In this study, we purified sperm-activating and -attracting factor (SAAF) from the egg-conditioning medium of C. intestinalis by using several steps of column chromatography. Determination of the molecular structure by NMR and MS/MS analysis revealed that SAAF is a previously uncharacterized sulfated steroid: 3,4,7,26-tetrahydroxycholestane-3,26-disulfate. Furthermore, it was shown that the SAAF of C. savignyi was indistinguishable from that of C. intestinalis in terms of the chromatographic behavior and molecular weight, indicating that the same compound might be responsible for sperm activation and chemotaxis in both the species. Furthermore, we established a method for quantitative analysis of sperm chemotaxis and showed that the chemotactic behavior of Ciona sperm is controlled by the “chemotactic turn” associated with decrease in the concentration of SAAF. PMID:12411583

  6. Metamorphosis in solitary ascidians.

    PubMed

    Karaiskou, Anthi; Swalla, Billie J; Sasakura, Yasunori; Chambon, Jean-Philippe

    2015-01-01

    Embryonic and postembryonic development in ascidians have been studied for over a century, but it is only in the last 10 years that the complex molecular network involved in coordinating postlarval development and metamorphosis has started to emerge. In most ascidians, the transition from the larval to the sessile juvenile/adult stage, or metamorphosis, requires a combination of environmental and endogenous signals and is characterized by coordinated global morphogenetic changes that are initiated by the adhesion of the larvae. Cloney was the first to describe cellular events of ascidians' metamorphosis in 1978 and only recently elements of the molecular regulation of this crucial developmental step have been revealed. This review aims to present a thorough view of this crucial developmental step by combining recent molecular data to the already established cellular events. © 2014 Wiley Periodicals, Inc.

  7. Diverse ETS transcription factors mediate FGF signaling in the Ciona anterior neural plate.

    PubMed

    Gainous, T Blair; Wagner, Eileen; Levine, Michael

    2015-03-15

    The ascidian Ciona intestinalis is a marine invertebrate belonging to the sister group of the vertebrates, the tunicates. Its compact genome and simple, experimentally tractable embryos make Ciona well-suited for the study of cell-fate specification in chordates. Tunicate larvae possess a characteristic chordate body plan, and many developmental pathways are conserved between tunicates and vertebrates. Previous studies have shown that FGF signals are essential for neural induction and patterning at sequential steps of Ciona embryogenesis. Here we show that two different ETS family transcription factors, Ets1/2 and Elk1/3/4, have partially redundant activities in the anterior neural plate of gastrulating embryos. Whereas Ets1/2 promotes pigment cell formation in lateral lineages, both Ets1/2 and Elk1/3/4 are involved in the activation of Myt1L in medial lineages and the restriction of Six3/6 expression to the anterior-most regions of the neural tube. We also provide evidence that photoreceptor cells arise from posterior regions of the presumptive sensory vesicle, and do not depend on FGF signaling. Cells previously identified as photoreceptor progenitors instead form ependymal cells and neurons of the larval brain. Our results extend recent findings on FGF-dependent patterning of anterior-posterior compartments in the Ciona central nervous system. Copyright © 2015. Published by Elsevier Inc.

  8. Functional studies of the Ciona intestinalis myogenic regulatory factor reveal conserved features of chordate myogenesis.

    PubMed

    Izzi, Stephanie A; Colantuono, Bonnie J; Sullivan, Kelly; Khare, Parul; Meedel, Thomas H

    2013-04-15

    Ci-MRF is the sole myogenic regulatory factor (MRF) of the ascidian Ciona intestinalis, an invertebrate chordate. In order to investigate its properties we developed a simple in vivo assay based on misexpressing Ci-MRF in the notochord of Ciona embryos. We used this assay to examine the roles of three structural motifs that are conserved among MRFs: an alanine-threonine (Ala-Thr) dipeptide of the basic domain that is known in vertebrates as the myogenic code, a cysteine/histidine-rich (C/H) domain found just N-terminal to the basic domain, and a carboxy-terminal amphipathic α-helix referred to as Helix III. We show that the Ala-Thr dipeptide is necessary for normal Ci-MRF function, and that while eliminating the C/H domain or Helix III individually has no demonstrable effect on Ci-MRF, simultaneous loss of both motifs significantly reduces its activity. Our studies also indicate that direct interaction between CiMRF and an essential E-box of Ciona Troponin I is required for the expression of this muscle-specific gene and that multiple classes of MRF-regulated genes exist in Ciona. These findings are consistent with substantial conservation of MRF-directed myogenesis in chordates and demonstrate for the first time that the Ala/Thr dipeptide of the basic domain of an invertebrate MRF behaves as a myogenic code.

  9. Nonreproductive role of gonadotropin-releasing hormone in the control of ascidian metamorphosis.

    PubMed

    Kamiya, Chisato; Ohta, Naoyuki; Ogura, Yosuke; Yoshida, Keita; Horie, Takeo; Kusakabe, Takehiro G; Satake, Honoo; Sasakura, Yasunori

    2014-12-01

    Gonadotropin-releasing hormones (GnRHs) are neuropeptides that play central roles in the reproduction of vertebrates. In the ascidian Ciona intestinalis, GnRHs and their receptors are expressed in the nervous systems at the larval stage, when animals are not yet capable of reproduction, suggesting that the hormones have non-reproductive roles. We showed that GnRHs in Ciona are involved in the animal's metamorphosis by regulating tail absorption and adult organ growth. Absorption of the larval tail and growth of the adult organs are two major events in the metamorphosis of ascidians. When larvae were treated with GnRHs, they completed tail absorption more frequently than control larvae. cAMP was suggested to be a second messenger for the induction of tail absorption by GnRHs. tGnRH-3 and tGnRH-5 (the "t" indicates "tunicate") inhibited the growth of adult organs by arresting cell cycle progression in parallel with the promotion of tail absorption. This study provides new insights into the molecular mechanisms of ascidian metamorphosis conducted by non-reproductive GnRHs. © 2014 Wiley Periodicals, Inc.

  10. The significance of Ciona intestinalis as a stem organism in integrative studies of functional evolution of the chordate endocrine, neuroendocrine, and nervous systems.

    PubMed

    Matsubara, Shin; Kawada, Tsuyoshi; Sakai, Tsubasa; Aoyama, Masato; Osugi, Tomohiro; Shiraishi, Akira; Satake, Honoo

    2016-02-01

    Ascidians are the closest phylogenetic neighbors to vertebrates and are believed to conserve the evolutionary origin in chordates of the endocrine, neuroendocrine, and nervous systems involving neuropeptides and peptide hormones. Ciona intestinalis harbors various homologs or prototypes of vertebrate neuropeptides and peptide hormones including gonadotropin-releasing hormones (GnRHs), tachykinins (TKs), and calcitonin, as well as Ciona-specific neuropeptides such as Ciona vasopressin, LF, and YFV/L peptides. Moreover, molecular and functional studies on Ciona tachykinin (Ci-TK) have revealed the novel molecular mechanism of inducing oocyte growth via up-regulation of vitellogenesis-associated protease activity, which is expected to be conserved in vertebrates. Furthermore, a series of studies on Ciona GnRH receptor paralogs have verified the species-specific regulation of GnRHergic signaling including unique signaling control via heterodimerization among multiple GnRH receptors. These findings confirm the remarkable significance of ascidians in investigations of the evolutionary processes of the peptidergic systems in chordates, leading to the promising advance in the research on Ciona peptides in the next stage based on the recent development of emerging technologies including genome-editing techniques, peptidomics-based multi-color staining, machine-learning prediction, and next-generation sequencing. These technologies and bioinformatic integration of the resultant "multi-omics" data will provide unprecedented insights into the comprehensive understanding of molecular and functional regulatory mechanisms of the Ciona peptides, and will eventually enable the exploration of both conserved and diversified endocrine, neuroendocrine, and nervous systems in the evolutionary lineage of chordates. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. Monoaminergic modulation of photoreception in ascidian: evidence for a proto-hypothalamo-retinal territory

    PubMed Central

    2012-01-01

    Background The retina of craniates/vertebrates has been proposed to derive from a photoreceptor prosencephalic territory in ancestral chordates, but the evolutionary origin of the different cell types making the retina is disputed. Except for photoreceptors, the existence of homologs of retinal cells remains uncertain outside vertebrates. Methods The expression of genes expressed in the sensory vesicle of the ascidian Ciona intestinalis including those encoding components of the monoaminergic neurotransmission systems, was analyzed by in situ hybridization or in vivo transfection of the corresponding regulatory elements driving fluorescent reporters. Modulation of photic responses by monoamines was studied by electrophysiology combined with pharmacological treatments. Results We show that many molecular characteristics of dopamine-synthesizing cells located in the vicinity of photoreceptors in the sensory vesicle of the ascidian Ciona intestinalis are similar to those of amacrine dopamine cells of the vertebrate retina. The ascidian dopamine cells share with vertebrate amacrine cells the expression of the key-transcription factor Ptf1a, as well as that of dopamine-synthesizing enzymes. Surprisingly, the ascidian dopamine cells accumulate serotonin via a functional serotonin transporter, as some amacrine cells also do. Moreover, dopamine cells located in the vicinity of the photoreceptors modulate the light-off induced swimming behavior of ascidian larvae by acting on alpha2-like receptors, instead of dopamine receptors, supporting a role in the modulation of the photic response. These cells are located in a territory of the ascidian sensory vesicle expressing genes found both in the retina and the hypothalamus of vertebrates (six3/6, Rx, meis, pax6, visual cycle proteins). Conclusion We propose that the dopamine cells of the ascidian larva derive from an ancestral multifunctional cell population located in the periventricular, photoreceptive field of the anterior

  12. Expression of a single prominin homolog in the embryo of the model chordate Ciona intestinalis.

    PubMed

    Russo, Monia T; Racioppi, Claudia; Zanetti, Laura; Ristoratore, Filomena

    2014-05-01

    Prominins are a family of pentaspan transmembrane glycoproteins, expressed in various types of cells, including stem and cancer stem cells in mammals. Prominin-1 is critical in generating and maintaining the structure of the photoreceptors in the eye since mutations in the PROM1 gene are associated with retinal and macular degeneration in human. In this study, we identified a single prominin homolog, Ci-prom1/2, in the model chordate the ascidian Ciona intestinalis and characterized Ci-prom1/2 expression profile in relation to photoreceptor differentiation during Ciona embryonic development. In situ hybridization experiments show Ci-prom1/2 transcripts localized in the developing central nervous system, predominantly in photoreceptor cell precursors as early as neurula stage and expression is maintained through larva stage in photoreceptor cells around the simple eye. We also isolated the regulatory region responsible for the specific spatio-temporal expression of the Ci-prom1/2 in photoreceptor cell lineage. Collectively, we report that Ci-prom1/2 is a novel molecular marker for ascidian photoreceptor cells and might represent a potential source to enlarge the knowledge about the function of prominin family in photoreceptor cell evolution and development.

  13. ANISEED 2015: a digital framework for the comparative developmental biology of ascidians

    PubMed Central

    Brozovic, Matija; Martin, Cyril; Dantec, Christelle; Dauga, Delphine; Mendez, Mickaël; Simion, Paul; Percher, Madeline; Laporte, Baptiste; Scornavacca, Céline; Di Gregorio, Anna; Fujiwara, Shigeki; Gineste, Mathieu; Lowe, Elijah K.; Piette, Jacques; Racioppi, Claudia; Ristoratore, Filomena; Sasakura, Yasunori; Takatori, Naohito; Brown, Titus C.; Delsuc, Frédéric; Douzery, Emmanuel; Gissi, Carmela; McDougall, Alex; Nishida, Hiroki; Sawada, Hitoshi; Swalla, Billie J.; Yasuo, Hitoyoshi; Lemaire, Patrick

    2016-01-01

    Ascidians belong to the tunicates, the sister group of vertebrates and are recognized model organisms in the field of embryonic development, regeneration and stem cells. ANISEED is the main information system in the field of ascidian developmental biology. This article reports the development of the system since its initial publication in 2010. Over the past five years, we refactored the system from an initial custom schema to an extended version of the Chado schema and redesigned all user and back end interfaces. This new architecture was used to improve and enrich the description of Ciona intestinalis embryonic development, based on an improved genome assembly and gene model set, refined functional gene annotation, and anatomical ontologies, and a new collection of full ORF cDNAs. The genomes of nine ascidian species have been sequenced since the release of the C. intestinalis genome. In ANISEED 2015, all nine new ascidian species can be explored via dedicated genome browsers, and searched by Blast. In addition, ANISEED provides full functional gene annotation, anatomical ontologies and some gene expression data for the six species with highest quality genomes. ANISEED is publicly available at: http://www.aniseed.cnrs.fr. PMID:26420834

  14. ANISEED 2015: a digital framework for the comparative developmental biology of ascidians.

    PubMed

    Brozovic, Matija; Martin, Cyril; Dantec, Christelle; Dauga, Delphine; Mendez, Mickaël; Simion, Paul; Percher, Madeline; Laporte, Baptiste; Scornavacca, Céline; Di Gregorio, Anna; Fujiwara, Shigeki; Gineste, Mathieu; Lowe, Elijah K; Piette, Jacques; Racioppi, Claudia; Ristoratore, Filomena; Sasakura, Yasunori; Takatori, Naohito; Brown, Titus C; Delsuc, Frédéric; Douzery, Emmanuel; Gissi, Carmela; McDougall, Alex; Nishida, Hiroki; Sawada, Hitoshi; Swalla, Billie J; Yasuo, Hitoyoshi; Lemaire, Patrick

    2016-01-04

    Ascidians belong to the tunicates, the sister group of vertebrates and are recognized model organisms in the field of embryonic development, regeneration and stem cells. ANISEED is the main information system in the field of ascidian developmental biology. This article reports the development of the system since its initial publication in 2010. Over the past five years, we refactored the system from an initial custom schema to an extended version of the Chado schema and redesigned all user and back end interfaces. This new architecture was used to improve and enrich the description of Ciona intestinalis embryonic development, based on an improved genome assembly and gene model set, refined functional gene annotation, and anatomical ontologies, and a new collection of full ORF cDNAs. The genomes of nine ascidian species have been sequenced since the release of the C. intestinalis genome. In ANISEED 2015, all nine new ascidian species can be explored via dedicated genome browsers, and searched by Blast. In addition, ANISEED provides full functional gene annotation, anatomical ontologies and some gene expression data for the six species with highest quality genomes. ANISEED is publicly available at: http://www.aniseed.cnrs.fr. © The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

  15. Evolutionary origins of the vertebrate heart: Specification of the cardiac lineage in Ciona intestinalis

    PubMed Central

    Davidson, Brad; Levine, Michael

    2003-01-01

    Here we exploit the extensive cell lineage information and streamlined genome of the ascidian, Ciona intestinalis, to investigate heart development in a basal chordate. Several cardiac genes were analyzed, including the sole Ciona ortholog of the Drosophila tinman gene, and tissue-specific enhancers were isolated for some of the genes. Conserved sequence motifs within these enhancers facilitated the isolation of a heart enhancer for the Ciona Hand-like gene. Altogether, these studies provide a regulatory framework for the differentiation of the cardiac mesoderm, beginning at the 110-cell stage, and extending through the fusion of cardiac progenitors during tail elongation. The cardiac lineage shares a common origin with the germ line, and zygotic transcription is first detected in the heart progenitors only after its separation from the germ line at the 64-cell stage. We propose that germ-line determinants influence the specification of the cardiac mesoderm, both by inhibiting inductive signals required for the development of noncardiac mesoderm lineages, and by providing a localized source of Wnt-5 and other signals required for heart development. We discuss the possibility that the germ line also influences the specification of the vertebrate heart. PMID:14500781

  16. Transcriptional regulation of the peripheral nervous system in Ciona intestinalis.

    PubMed

    Joyce Tang, W; Chen, Jerry S; Zeller, Robert W

    2013-06-15

    The formation of the sensory organs and cells that make up the peripheral nervous system (PNS) relies on the activity of transcription factors encoded by proneural genes (PNGs). Although PNGs have been identified in the nervous systems of both vertebrates and invertebrates, the complexity of their interactions has complicated efforts to understand their function in the context of their underlying regulatory networks. To gain insight into the regulatory network of PNG activity in chordates, we investigated the roles played by PNG homologs in regulating PNS development of the invertebrate chordate Ciona intestinalis. We discovered that in Ciona, MyT1, Pou4, Atonal, and NeuroD-like are expressed in a sequential regulatory cascade in the developing epidermal sensory neurons (ESNs) of the PNS and act downstream of Notch signaling, which negatively regulates these genes and the number of ESNs along the tail midlines. Transgenic embryos mis-expressing any of these proneural genes in the epidermis produced ectopic midline ESNs. In transgenic embryos mis-expressing Pou4, and MyT1 to a lesser extent, numerous ESNs were produced outside of the embryonic midlines. In addition we found that the microRNA miR-124, which inhibits Notch signaling in ESNs, is activated downstream of all the proneural factors we tested, suggesting that these genes operate collectively in a regulatory network. Interestingly, these factors are encoded by the same genes that have recently been demonstrated to convert fibroblasts into neurons. Our findings suggest the ascidian PNS can serve as an in vivo model to study the underlying regulatory mechanisms that enable the conversion of cells into sensory neurons.

  17. Ordered expression pattern of Hox and ParaHox genes along the alimentary canal in the ascidian juvenile.

    PubMed

    Nakayama, Satoshi; Satou, Kunihiro; Orito, Wataru; Ogasawara, Michio

    2016-07-01

    The Hox and ParaHox genes of bilateria share a similar expression pattern along the body axis and are known to be associated with anterior-posterior patterning. In vertebrates, the Hox genes are also expressed in presomitic mesoderm and gut endoderm and the ParaHox genes show a restricted expression pattern in the gut-related derivatives. Regional expression patterns in the embryonic central nervous system of the basal chordates amphioxus and ascidian have been reported; however, little is known about their endodermal expression in the alimentary canal. We focus on the Hox and ParaHox genes in the ascidian Ciona intestinalis and investigate the gene expression patterns in the juvenile, which shows morphological regionality in the alimentary canal. Gene expression analyses by using whole-mount in situ hybridization reveal that all Hox genes have a regional expression pattern along the alimentary canal. Expression of Hox1 to Hox4 is restricted to the posterior region of pharyngeal derivatives. Hox5 to Hox13 show an ordered expression pattern correlated with each Hox gene number along the postpharyngeal digestive tract. This expression pattern along the anterior-posterior axis has also been observed in Ciona ParaHox genes. Our observations suggest that ascidian Hox and ParaHox clusters are dispersed; however, the ordered expression patterns along the alimentary canal appear to be conserved among chordates.

  18. The ascidian mouth opening is derived from the anterior neuropore: reassessing the mouth/neural tube relationship in chordate evolution.

    PubMed

    Veeman, Michael T; Newman-Smith, Erin; El-Nachef, Danny; Smith, William C

    2010-08-01

    The relative positions of the brain and mouth are of central importance for models of chordate evolution. The dorsal hollow neural tube and the mouth have often been thought of as developmentally distinct structures that may have followed independent evolutionary paths. In most chordates however, including vertebrates and ascidians, the mouth primordia have been shown to fate to the anterior neural boundary. In ascidians such as Ciona there is a particularly intimate relationship between brain and mouth development, with a thin canal connecting the neural tube lumen to the mouth primordium at larval stages. This so-called neurohypophyseal canal was previously thought to be a secondary connection that formed relatively late, after the independent formation of the mouth primordium and the neural tube. Here we show that the Ciona neurohypophyseal canal is present from the end of neurulation and represents the anteriormost neural tube, and that the future mouth opening is actually derived from the anterior neuropore. The mouth thus forms at the anterior midline transition between neural tube and surface ectoderm. In the vertebrate Xenopus, we find that although the mouth primordium is not topologically continuous with the neural tube lumen, it nonetheless forms at this same transition point. This close association between the mouth primordium and the anterior neural tube in both ascidians and amphibians suggests that the evolution of these two structures may be more closely linked than previously appreciated. Copyright (c) 2010 Elsevier Inc. All rights reserved.

  19. Inverse correlation of population similarity and introduction date for invasive ascidians.

    PubMed

    Silva, Nathan; Smith, William C

    2008-06-25

    The genomes of many marine invertebrates, including the purple sea urchin and the solitary ascidians Ciona intestinalis and Ciona savignyi, show exceptionally high levels of heterozygosity, implying that these populations are highly polymorphic. Analysis of the C. savignyi genome found little evidence to support an elevated mutation rate, but rather points to a large population size contributing to the polymorphism level. In the present study, the relative genetic polymorphism levels in sampled populations of ten different ascidian species were determined using a similarity index generated by AFLP analysis. The goal was to determine the range of polymorphism within the populations of different species, and to uncover factors that may contribute to the high level of polymorphism. We observe that, surprisingly, the levels of polymorphism within these species show a negative correlation with the reported age of invasive populations, and that closely related species show substantially different levels of genetic polymorphism. These findings show exceptions to the assumptions that invasive species start with a low level of genetic polymorphism that increases over time and that closely related species have similar levels of genetic polymorphism.

  20. Screening of ovarian steroidogenic pathway in Ciona intestinalis and its modulation after tributyltin exposure

    SciTech Connect

    Cangialosi, Maria Vittoria; Puccia, Egidio; Mazzola, Antonio; Mansueto, Valentina; Arukwe, Augustine

    2010-05-15

    In this study, we have identified several ovarian steroids in Ciona with high similarity to vertebrate steroids and showed that cholesterol, corticosterone, dehydroepiandrosterone, estrone, estradiol-17beta, testosterone, pregnenolone, progesterone, have identical molecular spectra with vertebrate steroids. In addition, we have studied the effects of an endocrine disruptor (tributyltin: TBT) on these sex hormones and their precursors, ovarian morphology, and gene expression of some key enzymes in steroidogenic pathway in the ovary of Ciona. Ovarian specimens were cultured in vitro using different concentrations of TBT (10{sup -5}, 10{sup -4} and 10{sup -3} M). Ethanol was used as solvent control. Gene expression analysis was performed for adrenodoxin (ADREN) and adrenodoxin reductase (ADOX) (mediators of acute steroidogenesis) and 17beta-hydroxysteroid dehydrogenase (17beta-HSD). These transcripts were detected and measured by quantitative (real-time) polymerase chain reaction (qPCR). Sex steroids and their precursors were identified and quantified by a gas chromatography-mass spectroscopy (GC-MS) method. Exposure of Ciona ovaries to TBT produced modulations (either increased or decreased) of sterols and sex steroid levels, whereas no significant differences in ADREN, ADOX or 17beta-HSD mRNA expression patterns were observed. Histological analysis shows that TBT produced several modifications on Ciona ovarian morphology that includes irregular outline of nuclear membrane, less compacted cytoplasm, in addition to test and granulosa cells that were detached from the oocyte membrane. Given that the ascidians represent very simple experimental models for the study of endocrine disruption by environmental contaminants, our findings provide excellent models for multiple identification and quantification of sex steroid and their precursors in biological samples exposed to endocrine-disrupting chemicals and for direct extrapolation of such effects across taxonomic groups

  1. Screening of ovarian steroidogenic pathway in Ciona intestinalis and its modulation after tributyltin exposure.

    PubMed

    Cangialosi, Maria Vittoria; Puccia, Egidio; Mazzola, Antonio; Mansueto, Valentina; Arukwe, Augustine

    2010-05-15

    In this study, we have identified several ovarian steroids in Ciona with high similarity to vertebrate steroids and showed that cholesterol, corticosterone, dehydroepiandrosterone, estrone, estradiol-17beta, testosterone, pregnenolone, progesterone, have identical molecular spectra with vertebrate steroids. In addition, we have studied the effects of an endocrine disruptor (tributyltin: TBT) on these sex hormones and their precursors, ovarian morphology, and gene expression of some key enzymes in steroidogenic pathway in the ovary of Ciona. Ovarian specimens were cultured in vitro using different concentrations of TBT (10(-5), 10(-4) and 10(-3)M). Ethanol was used as solvent control. Gene expression analysis was performed for adrenodoxin (ADREN) and adrenodoxin reductase (ADOX) (mediators of acute steroidogenesis) and 17beta-hydroxysteroid dehydrogenase (17beta-HSD). These transcripts were detected and measured by quantitative (real-time) polymerase chain reaction (qPCR). Sex steroids and their precursors were identified and quantified by a gas chromatography-mass spectroscopy (GC-MS) method. Exposure of Ciona ovaries to TBT produced modulations (either increased or decreased) of sterols and sex steroid levels, whereas no significant differences in ADREN, ADOX or 17beta-HSD mRNA expression patterns were observed. Histological analysis shows that TBT produced several modifications on Ciona ovarian morphology that includes irregular outline of nuclear membrane, less compacted cytoplasm, in addition to test and granulosa cells that were detached from the oocyte membrane. Given that the ascidians represent very simple experimental models for the study of endocrine disruption by environmental contaminants, our findings provide excellent models for multiple identification and quantification of sex steroid and their precursors in biological samples exposed to endocrine-disrupting chemicals and for direct extrapolation of such effects across taxonomic groups and phyla. In

  2. Characterization of a Novel Rhamnose-containing Acidic Glycosphingolipid from the Ascidian Halocynthia aurantium.

    PubMed

    Kojima, Hisao; Shinohara, Ryota; Itonori, Saki; Ito, Masahiro

    2017-03-01

    Halocynthia aurantium, an edible ascidian species belonging to Urochordata, was subjected to structural characterization of acidic glycosphingolipids to investigate these molecules in ascidians: sulfatide from Ciona intestinalis and the glucuronic acid-containing acidic glycosphingolipid from H. roretzi. Acidic glycosphingolipids containing three or five sugars were isolated from soft parts of the ascidian H. aurantium by chloroform-methanol extraction, mild-alkaline hydrolysis, precipitation with cold acetone, and subsequent column chromatography using a DEAE-Sephadex A-25 column, a Florisil column, and an Iatrobead column. The structures of these glycosphingolipids were determined by methylation studies, sugar analysis, fatty acid analysis, sphingoid analysis, mass spectrometry, and proton nuclear magnetic resonance spectroscopy. A novel glucuronic acid-containing glycosphingolipid having a rhamnose residue was identified as Rhaα1-3GlcNAcβ1-3Galβ1-4(Fucα1-3)GlcAβ1-Cer (UGL-2). This novel structure is particularly unusual given that it contains both a rhamnose residue and a reducing terminal glucuronic acid residue within a single molecule. Rhamnose is a characteristic sugar, which is a component of cell wall pectin in plants and exopolysaccharides in bacteria. Ascidians acquired the cellulose synthase gene via lateral gene transfer, and therefore, it can be speculated that they also acquired the rhamnosyltransferase gene in the same manner. We also detected Galβ1-4(Fucα1-3)GlcAβ1-Cer (UGL-1), which was already identified in another ascidian, H. roretzi.

  3. The Ciona intestinalis genome: when the constraints are off

    NASA Technical Reports Server (NTRS)

    Holland, Linda Z.; Gibson-Brown, Jeremy J.

    2003-01-01

    The recent genome sequencing of a non-vertebrate deuterostome, the ascidian tunicate Ciona intestinalis, makes a substantial contribution to the fields of evolutionary and developmental biology.1 Tunicates have some of the smallest bilaterian genomes, embryos with relatively few cells, fixed lineages and early determination of cell fates. Initial analyses of the C. intestinalis genome indicate that it has been evolving rapidly. Comparisons with other bilaterians show that C. intestinalis has lost a number of genes, and that many genes linked together in most other bilaterians have become uncoupled. In addition, a number of independent, lineage-specific gene duplications have been detected. These new results, although interesting in themselves, will take on a deeper significance once the genomes of additional invertebrate deuterostomes (e.g. echinoderms, hemichordates and amphioxus) have been sequenced. With such a broadened database, comparative genomics can begin to ask pointed questions about the relationship between the evolution of genomes and the evolution of body plans. Copyright 2003 Wiley Periodicals, Inc.

  4. Fgf genes in the basal chordate Ciona intestinalis.

    PubMed

    Satou, Yutaka; Imai, Kaoru S; Satoh, Nori

    2002-10-01

    In vertebrates, a number of fibroblast growth factors (FGFs) have been shown to play important roles in developing embryos and adult organisms. However, the molecular relationships of the vertebrate FGFs are not yet completely understood, partly due to the divergence of their amino acid sequences. To solve this problem, we have identified six FGF genes in a basal chordate, the ascidian Ciona intestinalis. A phylogenetic analysis confidently assigned two of them to vertebrate FGF8/17/18 and FGF11/12/13/14, respectively. Based on the presence of the conserved domains within or outside of the FGF domains, we speculate that three of the other genes are orthologous to vertebrate FGF3/7/10/22, FGF4/5/6 and FGF9/16/20, respectively, although we cannot assign the sixth member to any of the vertebrate FGFs. A survey of the raw whole genome shotgun sequences of C. intestinalis demonstrated the presence of no FGF genes other than the six genes in the genome. The identification of these six FGF genes in the basal chordate gave us an insight into the diversification of specific subfamilies of vertebrate FGFs.

  5. The Ciona intestinalis genome: when the constraints are off

    NASA Technical Reports Server (NTRS)

    Holland, Linda Z.; Gibson-Brown, Jeremy J.

    2003-01-01

    The recent genome sequencing of a non-vertebrate deuterostome, the ascidian tunicate Ciona intestinalis, makes a substantial contribution to the fields of evolutionary and developmental biology.1 Tunicates have some of the smallest bilaterian genomes, embryos with relatively few cells, fixed lineages and early determination of cell fates. Initial analyses of the C. intestinalis genome indicate that it has been evolving rapidly. Comparisons with other bilaterians show that C. intestinalis has lost a number of genes, and that many genes linked together in most other bilaterians have become uncoupled. In addition, a number of independent, lineage-specific gene duplications have been detected. These new results, although interesting in themselves, will take on a deeper significance once the genomes of additional invertebrate deuterostomes (e.g. echinoderms, hemichordates and amphioxus) have been sequenced. With such a broadened database, comparative genomics can begin to ask pointed questions about the relationship between the evolution of genomes and the evolution of body plans. Copyright 2003 Wiley Periodicals, Inc.

  6. Harbor networks as introduction gateways: contrasting distribution patterns of native and introduced ascidians.

    PubMed

    López-Legentil, Susanna; Legentil, Miquel L; Erwin, Patrick M; Turon, Xavier

    Harbors and marinas are well known gateways for species introductions in marine environments but little work has been done to ascertain relationships between species diversity, harbor type, and geographic distance to uncover patterns of secondary spread. Here, we sampled ascidians from 32 harbors along ca. 300 km of the NW Mediterranean coast and investigated patterns of distribution and spread related to harbor type (marina, fishing, commercial) and geographic location using multivariate techniques. In total, 28 ascidians were identified at the species level and another 9 at the genus level based on morphology and genetic barcoding. Eight species were assigned to introduced forms, 15 were given native status and 5 were classified as cryptogenic. Aplidium accarense was reported for the first time in the Mediterranean Sea and was especially abundant in 23 of the harbors. Introduced and cryptogenic species were abundant in most of the surveyed harbors, while native forms were rare and restricted to a few harbors. Significant differences in the distribution of ascidians according to harbor type and latitudinal position were observed. These differences were due to the distribution of introduced species. We obtained a significant correlation between geographic distance and ascidian composition, indicating that closely located harbors shared more ascidian species among them. This study showed that harbors act as dispersal strongholds for introduced species, with native species only appearing sporadically, and that harbor type and geographic location should also be considered when developing management plans to constrain the spread of non-indigenous species in highly urbanized coastlines.

  7. Temporal stability of bacterial symbionts in a temperate ascidian

    PubMed Central

    López-Legentil, Susanna; Turon, Xavier; Espluga, Roger; Erwin, Patrick M.

    2015-01-01

    In temperate seas, both bacterioplankton communities and invertebrate lifecycles follow a seasonal pattern. To investigate whether the bacterial community associated with the Mediterranean ascidian Didemnum fulgens exhibited similar variations, we monitored its bacterial community structure monthly for over a year using terminal restriction fragment length polymorphism and clone library analyses based on a nearly full length fragment of the 16S rRNA gene. D. fulgens harbored a bacterial consortium typical of ascidians, including numerous members of the phylum Proteobacteria, and a few members of the phyla Cyanobacteria and Acidobacteria. The overall bacterial community in D. fulgens had a distinct signature from the surrounding seawater and was stable over time and across seasonal fluctuations in temperature. Bacterial symbionts were also observed around animal cells in the tunic of adult individuals and in the inner tunic of D. fulgens larvae by transmission electron microscopy. Our results suggest that, as seen for sponges and corals, some species of ascidians host stable and unique bacterial communities that are at least partially inherited by their progeny by vertical transmission. PMID:26441944

  8. Identification of a retinoic acid-responsive neural enhancer in the Ciona intestinalis Hox1 gene.

    PubMed

    Kanda, Miyuki; Ikeda, Taku; Fujiwara, Shigeki

    2013-02-01

    The Hox1 gene in the urochordate ascidian Ciona intestinalis (Ci-Hox1) is expressed in the nerve cord and epidermis. We identified a nerve cord enhancer in the second intron of Ci-Hox1, and demonstrated that retinoic acid (RA) plays a major role in activating this enhancer. The enhancer contained a putative retinoic acid-response element (RARE). Mutation of the RARE in the Ci-Hox1 nerve cord enhancer only partially abolished the enhancer activity. Genes encoding RA synthase and the RA receptor were knocked down using specific antisense morpholino oligos (MOs), and injection of embryos with these MOs resulted in the complete disappearance of epidermal expression of Ci-Hox1 and reduction of neural expression. However, nerve cord expression was not completely repressed. These results suggest that the nerve cord enhancer is activated by two partially redundant pathways; one RA-dependent and one RA-independent.

  9. Ciona intestinalis interleukin 17-like genes expression is upregulated by LPS challenge.

    PubMed

    Vizzini, Aiti; Di Falco, Felicia; Parrinello, Daniela; Sanfratello, Maria Antonietta; Mazzarella, Claudia; Parrinello, Nicolò; Cammarata, Matteo

    2015-01-01

    In humans, IL-17 is a proinflammatory cytokine that plays a key role in the clearance of extracellular bacteria promoting cell infiltration and production of several cytokines and chemokines. Here, we report on three Ciona intestinalis IL-17 homologues (CiIL17-1, CiIL17-2, CiIL17-3). The gene organization, phylogenetic tree and modeling supported the close relationship with the mammalian IL-17A and IL-17F suggesting that the C. intestinalis IL-17 genes share a common ancestor in the chordate lineages. Real time PCR analysis showed a prompt expression induced by LPS inoculation suggesting that they are involved in the first phase of inflammatory response. In situ hybridization assays disclosed that the genes transcription was upregulated in the pharynx, the main organ of the ascidian immune system, and expressed by hemocytes (granulocytes and univacuolar refractile granulocyte) inside the pharynx vessels.

  10. Further EST analysis of endocrine genes that are preferentially expressed in the neural complex of Ciona intestinalis: receptor and enzyme genes associated with endocrine system in the neural complex.

    PubMed

    Sekiguchi, Toshio; Kawashima, Takeshi; Satou, Yutaka; Satoh, Nori

    2007-01-15

    Identification of orthologs of vertebrate neuropeptides and hypothalamic hormones in the neural complex of ascidians suggests integral roles of the ascidian neural complex in the endocrine system. In the present study, we investigated endocrine-related genes expressed in the neural complex of Ciona intestinalis. Comprehensive analyses of 3'-end sequences of the neural complex cDNAs placed 10,029 clones into 4051 independent clusters or genes, 1524 of them being expressed preferentially in this organ. Comparison of the 1524 genes with the human proteome databank demonstrated that 476 matched previously identified human proteins with distinct functions. Further analyses of sequence similarity of the 476 genes demonstrated that 21 genes are candidates for those involved in the endocrine system. Although we cannot detect hormone or peptide candidates, we found 21 genes such as receptors for peptide ligands, receptor-modulating proteins, and processing enzymes. We then characterized the Ciona prohormone convertase 2 (Ci-PC2) and carboxypeptidase E (Ci-CPE), which are associated with endoproteolytic processing of peptide hormone precursors. Furthermore, genes encoding these transcripts are expressed specifically in the neural complex of young adult ascidians. These data provide the molecular basis for further functional studies of the endocrine role of the neural complex of ascidians.

  11. 3D-Printed Microwell Arrays for Ciona Microinjection and Timelapse Imaging

    PubMed Central

    Gregory, Clint; Veeman, Michael

    2013-01-01

    Ascidians such as Ciona are close chordate relatives of the vertebrates with small, simple embryonic body plans and small, simple genomes. The tractable size of the embryo offers considerable advantages for in toto imaging and quantitative analysis of morphogenesis. For functional studies, Ciona eggs are considerably more challenging to microinject than the much larger eggs of other model organisms such as zebrafish and Xenopus. One of the key difficulties is in restraining the eggs so that the microinjection needle can be easily introduced and withdrawn. Here we develop and test a device to cast wells in agarose that are each sized to hold a single egg. This injection mold is fabricated by micro-resolution stereolithography with a grid of egg-sized posts that cast corresponding wells in agarose. This 3D printing technology allows the rapid and inexpensive testing of iteratively refined prototypes. In addition to their utility in microinjection, these grids of embryo-sized wells are also valuable for timelapse imaging of multiple embryos. PMID:24324769

  12. Three-dimensional anatomy of the Ciona intestinalis tailbud embryo at single-cell resolution.

    PubMed

    Nakamura, Mitsuru J; Terai, Jun; Okubo, Reiko; Hotta, Kohji; Oka, Kotaro

    2012-12-15

    During embryogenesis, chordates pass through a tailbud stage in which the larval tail is formed. Since acquisition of a tadpole-like tail during tailbud stage is one of the key events in the evolution of chordates, understanding the anatomy of the tailbud stage chordate embryo is of special interest. In this study, to understand comprehensively the anatomy of the tailbud embryo at single-cell-level, real microscopic image stacks of the tailbud embryo in Ciona intestinalis were reconstructed into a 3D computer model. This comprehensive 3D model of the ascidian tailbud embryo was based on real images of confocal laser scanning microscope (CLSM) and therefore, cell shape, location and cell arrangement reflect real geometries of the tailbud embryo. We found that the tailbud embryo consists of 1579 cells, including 836 epidermal cells, 228 cells in the central nervous system, 218 mesenchymal cells, four trunk ventral cells, two B/B(⁎)8.11 cells, 36 muscle cells, 40 notochord cells, four primordial germ cells, and 199 endodermal cells. Moreover, we identified for the first time two populations of previously undefined cells (a total of 12 cells) in Ciona: one located in the lateral trunk and the other located under the tail dorsal epidermis. This information provides a first step for understanding how the body plan of the chordate tailbud embryo formed and evolved.

  13. A transiently expressed connexin is essential for anterior neural plate development in Ciona intestinalis.

    PubMed

    Hackley, Christopher; Mulholland, Erin; Kim, Gil Jung; Newman-Smith, Erin; Smith, William C

    2013-01-01

    A forward genetic screen in the ascidian Ciona intestinalis identified a mutant line (frimousse) with a profound disruption in neural plate development. In embryos with the frimousse mutation, the anteriormost neural plate cells, which are products of an FGF induction at the blastula and gastrula stages, initially express neural plate-specific genes but fail to maintain the induced state and ultimately default to epidermis. The genetic lesion in the frimousse mutant lies within a connexin gene (cx-11) that is transiently expressed in the developing neural plate in a temporal window corresponding to the period of a-lineage neural induction. Using a genetically encoded calcium indicator we observed multiple calcium transients throughout the developing neural plate in wild-type embryos, but not in mutant embryos. A series of treatments at the gastrula and neurula stages that block the calcium transients, including gap junction inhibition and calcium depletion, were also found to disrupt the development of the anterior neural plate in a similar way to the frimousse mutation. The requirement for cx-11 for anterior neural fate points to a crucial role for intercellular communication via gap junctions, probably through mediation of Ca(2+) transients, in Ciona intestinalis neural induction.

  14. Feeding cessation alters host morphology and bacterial communities in the ascidian Pseudodistoma crucigaster.

    PubMed

    López-Legentil, Susanna; Turon, Xavier; Erwin, Patrick M

    2016-01-01

    Ascidians can associate with abundant and diverse consortia of microbial symbionts, yet these communities remain unexamined for the majority of host ascidians and little is known about host-symbiont interactions. We coupled electron microscopy and 16S rRNA gene tag pyrosequencing to investigate the bacterial communities associated with the colonial ascidian Pseudodistoma crucigaster, a species endemic to the Mediterranean Sea that has a life cycle with two phases: actively-filtering (active) and non-filtering (resting) forms. Resting colonies exhibited a reduced branchial sac (feeding apparatus) and a thickened cuticle. Electron microscope images also suggested higher abundance of colonizing microorganisms on surfaces of resting colonies. Accordingly, bacterial sequences associated with environmental sources (sediment and biofilms, >99 % similarity) were detected exclusively in resting colonies. Bacterial communities of P. crucigaster colonies (active and resting) were dominated by 3 core taxa affiliated (>94 % similarity) with previously described symbiotic Alphaproteobacteria in marine invertebrates. Shifts in rare bacteria were detected when ascidians entered the resting phase, including the appearance of strictly anaerobic lineages and nitrifying bacterial guilds. These findings suggest that physical (thickened cuticle) and metabolic (feeding cessation) changes in host ascidians have cascading effects on associated bacteria, where modified oxygen concentrations and chemical substrates for microbial metabolism may create anaerobic microhabitats and promote colonization by environmental microorganisms.

  15. Genetic signatures of natural selection in a model invasive ascidian

    PubMed Central

    Lin, Yaping; Chen, Yiyong; Yi, Changho; Fong, Jonathan J.; Kim, Won; Rius, Marc; Zhan, Aibin

    2017-01-01

    Invasive species represent promising models to study species’ responses to rapidly changing environments. Although local adaptation frequently occurs during contemporary range expansion, the associated genetic signatures at both population and genomic levels remain largely unknown. Here, we use genome-wide gene-associated microsatellites to investigate genetic signatures of natural selection in a model invasive ascidian, Ciona robusta. Population genetic analyses of 150 individuals sampled in Korea, New Zealand, South Africa and Spain showed significant genetic differentiation among populations. Based on outlier tests, we found high incidence of signatures of directional selection at 19 loci. Hitchhiking mapping analyses identified 12 directional selective sweep regions, and all selective sweep windows on chromosomes were narrow (~8.9 kb). Further analyses indentified 132 candidate genes under selection. When we compared our genetic data and six crucial environmental variables, 16 putatively selected loci showed significant correlation with these environmental variables. This suggests that the local environmental conditions have left significant signatures of selection at both population and genomic levels. Finally, we identified “plastic” genomic regions and genes that are promising regions to investigate evolutionary responses to rapid environmental change in C. robusta. PMID:28266616

  16. Genetic signatures of natural selection in a model invasive ascidian

    NASA Astrophysics Data System (ADS)

    Lin, Yaping; Chen, Yiyong; Yi, Changho; Fong, Jonathan J.; Kim, Won; Rius, Marc; Zhan, Aibin

    2017-03-01

    Invasive species represent promising models to study species’ responses to rapidly changing environments. Although local adaptation frequently occurs during contemporary range expansion, the associated genetic signatures at both population and genomic levels remain largely unknown. Here, we use genome-wide gene-associated microsatellites to investigate genetic signatures of natural selection in a model invasive ascidian, Ciona robusta. Population genetic analyses of 150 individuals sampled in Korea, New Zealand, South Africa and Spain showed significant genetic differentiation among populations. Based on outlier tests, we found high incidence of signatures of directional selection at 19 loci. Hitchhiking mapping analyses identified 12 directional selective sweep regions, and all selective sweep windows on chromosomes were narrow (~8.9 kb). Further analyses indentified 132 candidate genes under selection. When we compared our genetic data and six crucial environmental variables, 16 putatively selected loci showed significant correlation with these environmental variables. This suggests that the local environmental conditions have left significant signatures of selection at both population and genomic levels. Finally, we identified “plastic” genomic regions and genes that are promising regions to investigate evolutionary responses to rapid environmental change in C. robusta.

  17. Porcine relaxin, a 500 million-year-old hormone? the tunicate Ciona intestinalis has porcine relaxin.

    PubMed

    Georges, D; Schwabe, C

    1999-07-01

    The fossil record of tunicates reaches back to the upper Cambrian period. Ascidians have mobile, tadpole-like juvenile forms with a notochord, which inspired the classification of tunicates as Urochordata, i.e., predecessors of vertebrates. The genome of the tunicate Ciona intestinalis contains a relaxin coding region that is organized like a mammalian gene, i.e., signal peptide, B-chain domain, connecting peptide domain, followed by the A-chain domain with a stop codon after cysteine A-22. RNA-derived cDNA encodes a relaxin that is identical to the circulating form of the porcine hormone. In contrast to the porcine gene, the ascidian gene has no intron in the C-peptide domain, and in that respect is similar to the bombyxin gene of the silkworm. During the spawning period, only enough relaxin could be extracted and isolated from gonads of C. intestinalis for a partial sequence analysis. Remarkable as it may be, these findings suggest that relaxin is identical in pigs, whales, and the tunicate C. intestinalis.

  18. A standardisation of Ciona intestinalis (Chordata, Ascidiacea) embryo-larval bioassay for ecotoxicological studies.

    PubMed

    Bellas, Juan; Beiras, Ricardo; Vázquez, Elsa

    2003-11-01

    A standardisation of the ascidian Ciona intestinalis embryo-larval bioassay for marine pollution assessment has been developed. The minimum percentage of embryogenesis success was established to assess the quality of the biological material used; minimum sample size and number of replicates per treatment were also estimated. The suitability of artificial and natural seawater for the incubation of ascidian embryos and larvae was compared, and the optimum conditions of temperature, salinity, pH, density of embryos in the vials and the sperm/egg ratio were investigated. On the basis of the 10th percentile of the distribution of larval abnormalities, we proposed a threshold of 50% normal larvae in the control in order to consider the test of acceptable biological quality. According to our results n=5 is a sufficiently high replication to detect 5% differences among treatment means with a power of P=90% and alpha=0.05, and a sampling size >/=222 allows a 95% confidence in the estimate with an error of 0.05. Egg density did not affect larval development within the range 1-20 eggs/ml, and the optimum sperm/egg ratio which fertilise 100% of the eggs was 3000-30,000 sperm/egg (i.e. 10(8)-10(7) sperm/ml). There were not significant differences between the two water types tested, and the optimum tolerance ranges were 18-23 degrees C temperature, 34-42 ppt salinity (42 ppt was the highest salinity tested), and 7.4-8.8 pH. The median effective concentration (EC(50)) of copper (Cu) causing a 50% reduction of normal hatched larvae was 54.2 microg/l (0.85 microM), which shows a sensitivity of this species similar to the commonly used bivalve and sea-urchin tests. The ascidian embryo-larval bioassay is an accurate, reliable, simple and rapid method that can be used in ecotoxicological studies.

  19. Ciona intestinalis as an emerging model organism: its regeneration under controlled conditions and methodology for egg dechorionation.

    PubMed

    Liu, Li-ping; Xiang, Jian-hai; Dong, Bo; Natarajan, Pavanasam; Yu, Kui-jie; Cai, Nan-er

    2006-06-01

    The ascidian Ciona intestinalis is a model organism of developmental and evolutionary biology and may provide crucial clues concerning two fundamental matters, namely, how chordates originated from the putative deuterostome ancestor and how advanced chordates originated from the simplest chordates. In this paper, a whole-life-span culture of C. intestinalis was conducted. Fed with the diet combination of dry Spirulina, egg yolk, Dicrateria sp., edible yeast and weaning diet for shrimp, C. intestinalis grew up to average 59 mm and matured after 60 d cultivation. This culture process could be repeated using the artificially cultured mature ascidians as material. When the fertilized eggs were maintained under 10, 15, 20, 25 degrees C, they hatched within 30 h, 22 h, 16 h and 12 h 50 min respectively experiencing cleavage, blastulation, gastrulation, neurulation, tailbud stage and tadpole stage. The tadpole larvae were characterized as typical but simplified chordates because of their dorsal nerve cord, notochord and primordial brain. After 8 - 24 h freely swimming, the tadpole larvae settled on the substrates and metamorphosized within 1- 2 d into filter feeding sessile juvenile ascidians. In addition, unfertilized eggs were successfully dechorionated in filtered seawater containing 1% Tripsin, 0.25% EDTA at pH of 10.5 within 40 min. After fertilization, the dechorionated eggs developed well and hatched at normal hatching rate. In conclusion, this paper presented feasible methodology for rearing the tadpole larvae of C. intestinalis into sexual maturity under controlled conditions and detailed observations on the embryogenesis of the laboratory cultured ascidians, which will facilitate developmental and genetic research using this model system.

  20. Transcript Mapping and Genome Annotation of Ascidian mtDNA Using EST Data

    PubMed Central

    Gissi, Carmela; Pesole, Graziano

    2003-01-01

    Mitochondrial transcripts of two ascidian species were reconstructed through sequence assembly of publicly available ESTs resembling mitochondrial DNA sequences (mt-ESTs). This strategy allowed us to analyze processing and mapping of the mitochondrial transcripts and to investigate the gene organization of a previously uncharacterized mitochondrial genome (mtDNA). This new strategy would greatly facilitate the sequencing and annotation of mtDNAs. In Ciona intestinalis, the assembled mt-ESTs covered 22 mitochondrial genes (∼12,000 bp) and provided the partial sequence of the mtDNA and the prediction of its gene organization. Such sequences were confirmed by amplification and sequencing of the entire Ciona mtDNA. For Halocynthia roretzi, for which the mtDNA sequence was already available, the inferred mt transcripts allowed better definition of gene boundaries (16S rRNA, ND1, ATP6, and tRNA-Ser genes) and the identification of a new gene (an additional Phe-tRNA). In both species, polycistronic and immature transcripts, creation of stop codons by polyadenylation, tRNA signal processing, and rRNA transcript termination signals were identified, thus suggesting that the main features of mitochondrial transcripts are conserved in Chordata. PMID:12915488

  1. Ascidians (Tunicata) of the French Guiana Expedition.

    PubMed

    Monniot, Françoise

    2016-05-24

    Ascidians were collected along the shore of Iles du Salut and deeper on the slope in a program of evaluation of the biodiversity in Guiana. Most of the samples belong to already known species from the Caribbean area and Brazilian coast. The colonial forms dominate. The 6 new species have been dredged deeper than 50m but not found by SCUBA divers. In spite of an intensive sampling, the ascidian diversity in Guiana is low with 36 species recorded. This is the result of the abundance of sediment suspended in the water and uniformly deposited on all substrates which affects filtration rate and limits the settling of the ascidian larvae.

  2. Ascidian Mitogenomics: Comparison of Evolutionary Rates in Closely Related Taxa Provides Evidence of Ongoing Speciation Events

    PubMed Central

    Griggio, Francesca; Voskoboynik, Ayelet; Iannelli, Fabio; Justy, Fabienne; Tilak, Marie-Ka; Xavier, Turon; Pesole, Graziano; Douzery, Emmanuel J.P.; Mastrototaro, Francesco; Gissi, Carmela

    2014-01-01

    Ascidians are a fascinating group of filter-feeding marine chordates characterized by rapid evolution of both sequences and structure of their nuclear and mitochondrial genomes. Moreover, they include several model organisms used to investigate complex biological processes in chordates. To study the evolutionary dynamics of ascidians at short phylogenetic distances, we sequenced 13 new mitogenomes and analyzed them, together with 15 other available mitogenomes, using a novel approach involving detailed whole-mitogenome comparisons of conspecific and congeneric pairs. The evolutionary rate was quite homogeneous at both intraspecific and congeneric level, and the lowest congeneric rates were found in cryptic (morphologically undistinguishable) and in morphologically very similar species pairs. Moreover, congeneric nonsynonymous rates (dN) were up to two orders of magnitude higher than in intraspecies pairs. Overall, a clear-cut gap sets apart conspecific from congeneric pairs. These evolutionary peculiarities allowed easily identifying an extraordinary intraspecific variability in the model ascidian Botryllus schlosseri, where most pairs show a dN value between that observed at intraspecies and congeneric level, yet consistently lower than that of the Ciona intestinalis cryptic species pair. These data suggest ongoing speciation events producing genetically distinct B. schlosseri entities. Remarkably, these ongoing speciation events were undetectable by the cox1 barcode fragment, demonstrating that, at low phylogenetic distances, the whole mitogenome has a higher resolving power than cox1. Our study shows that whole-mitogenome comparative analyses, performed on a suitable sample of congeneric and intraspecies pairs, may allow detecting not only cryptic species but also ongoing speciation events. PMID:24572017

  3. The CNS connectome of a tadpole larva of Ciona intestinalis (L.) highlights sidedness in the brain of a chordate sibling.

    PubMed

    Ryan, Kerrianne; Lu, Zhiyuan; Meinertzhagen, Ian A

    2016-12-06

    Left-right asymmetries in brains are usually minor or cryptic. We report brain asymmetries in the tiny, dorsal tubular nervous system of the ascidian tadpole larva, Ciona intestinalis. Chordate in body plan and development, the larva provides an outstanding example of brain asymmetry. Although early neural development is well studied, detailed cellular organization of the swimming larva's CNS remains unreported. Using serial-section EM we document the synaptic connectome of the larva's 177 CNS neurons. These formed 6618 synapses including 1772 neuromuscular junctions, augmented by 1206 gap junctions. Neurons are unipolar with at most a single dendrite, and few synapses. Some synapses are unpolarised, others form reciprocal or serial motifs; 922 were polyadic. Axo-axonal synapses predominate. Most neurons have ciliary organelles, and many features lack structural specialization. Despite equal cell numbers on both sides, neuron identities and pathways differ left/right. Brain vesicle asymmetries include a right ocellus and left coronet cells.

  4. Upregulated transcription of phenoloxidase genes in the pharynx and endostyle of Ciona intestinalis in response to LPS.

    PubMed

    Vizzini, Aiti; Parrinello, Daniela; Sanfratello, Maria Antonietta; Trapani, Maria Rosa; Mangano, Valentina; Parrinello, Nicolò; Cammarata, Matteo

    2015-03-01

    We investigated the role of phenoloxidases (POs) in ascidians inflammatory reaction, a components of a copper-containing protein family involved in invertebrate immune system. In Ciona intestinalis two phenoloxidases (CinPO-1, CinPO-2) have been sequenced. In the present study, real time PCR analysis showed that both CinPO-1 and CinPO-2 genes were modulated by LPS inoculation suggesting that they are inducible and highly expressed in the inflamed pharynx. In situ hybridization disclosed CinPO-1 and CinPO-2 transcripts in pharynx hemocytes (granulocytes) and, mainly, in unilocular refractile granulocytes (URG) which mainly populated the inflamed tunic matrix. Interestingly, the genes are also upregulated by LPS in the endostyle (zones 7, 8 and 9) that is considered homolog to the vertebrate thyroid.

  5. Functional Brachyury Binding Sites Establish a Temporal Read-out of Gene Expression in the Ciona Notochord

    PubMed Central

    Passamaneck, Yale J.; Gazdoiu, Stefan; José-Edwards, Diana S.; Kugler, Jamie E.; Oda-Ishii, Izumi; Imai, Janice H.; Nibu, Yutaka; Di Gregorio, Anna

    2013-01-01

    The appearance of the notochord represented a milestone in Deuterostome evolution. The notochord is necessary for the development of the chordate body plan and for the formation of the vertebral column and numerous organs. It is known that the transcription factor Brachyury is required for notochord formation in all chordates, and that it controls transcription of a large number of target genes. However, studies of the structure of the cis-regulatory modules (CRMs) through which this control is exerted are complicated in vertebrates by the genomic complexity and the pan-mesodermal expression territory of Brachyury. We used the ascidian Ciona, in which the single-copy Brachyury is notochord-specific and CRMs are easily identifiable, to carry out a systematic characterization of Brachyury-downstream notochord CRMs. We found that Ciona Brachyury (Ci-Bra) controls most of its targets directly, through non-palindromic binding sites that function either synergistically or individually to activate early- and middle-onset genes, respectively, while late-onset target CRMs are controlled indirectly, via transcriptional intermediaries. These results illustrate how a transcriptional regulator can efficiently shape a shallow gene regulatory network into a multi-tiered transcriptional output, and provide insights into the mechanisms that establish temporal read-outs of gene expression in a fast-developing chordate embryo. PMID:24204212

  6. Enhancer of zeste acts as a major developmental regulator of Ciona intestinalis embryogenesis.

    PubMed

    Le Goff, Emilie; Martinand-Mari, Camille; Martin, Marianne; Feuillard, Jérôme; Boublik, Yvan; Godefroy, Nelly; Mangeat, Paul; Baghdiguian, Stephen; Cavalli, Giacomo

    2015-08-14

    The paradigm of developmental regulation by Polycomb group (PcG) proteins posits that they maintain silencing outside the spatial expression domains of their target genes, particularly of Hox genes, starting from mid embryogenesis. The Enhancer of zeste [E(z)] PcG protein is the catalytic subunit of the PRC2 complex, which silences its targets via deposition of the H3K27me3 mark. Here, we studied the ascidian Ciona intestinalis counterpart of E(z). Ci-E(z) is detected by immunohistochemistry as soon as the 2- and 4-cell stages as a cytoplasmic form and becomes exclusively nuclear thereafter, whereas the H3K27me3 mark is detected starting from the gastrula stage and later. Morpholino invalidation of Ci-E(z) leads to the total disappearance of both Ci-E(z) protein and its H3K27me3 mark. Ci-E(z) morphants display a severe phenotype. Strikingly, the earliest defects occur at the 4-cell stage with the dysregulation of cell positioning and mitotic impairment. At later stages, Ci-E(z)-deficient embryos are affected by terminal differentiation defects of neural, epidermal and muscle tissues, by the failure to form a notochord and by the absence of caudal nerve. These major phenotypic defects are specifically rescued by injection of a morpholino-resistant Ci-E(z) mRNA, which restores expression of Ci-E(z) protein and re-deposition of the H3K27me3 mark. As observed by qPCR analyses, Ci-E(z) invalidation leads to the early derepression of tissue-specific developmental genes, whereas late-acting developmental genes are generally down-regulated. Altogether, our results suggest that Ci-E(z) plays a major role during embryonic development in Ciona intestinalis by silencing early-acting developmental genes in a Hox-independent manner.

  7. miR-124 function during Ciona intestinalis neuronal development includes extensive interaction with the Notch signaling pathway.

    PubMed

    Chen, Jerry S; Pedro, Matthew San; Zeller, Robert W

    2011-11-01

    The nervous system-enriched microRNA miR-124 is necessary for proper nervous system development, although the mechanism remains poorly understood. Here, through a comprehensive analysis of miR-124 and its gene targets, we demonstrate that, in the chordate ascidian Ciona intestinalis, miR-124 plays an extensive role in promoting nervous system development. We discovered that feedback interaction between miR-124 and Notch signaling regulates the epidermal-peripheral nervous system (PNS) fate choice in tail midline cells. Notch signaling silences miR-124 in epidermal midline cells, whereas in PNS midline cells miR-124 silences Notch, Neuralized and all three Ciona Hairy/Enhancer-of-Split genes. Furthermore, ectopic expression of miR-124 is sufficient to convert epidermal midline cells into PNS neurons, consistent with a role in modulating Notch signaling. More broadly, genome-wide target extraction with validation using an in vivo tissue-specific sensor assay indicates that miR-124 shapes neuronal progenitor fields by downregulating non-neural genes, notably the muscle specifier Macho-1 and 50 Brachyury-regulated notochord genes, as well as several anti-neural factors including SCP1 and PTBP1. 3'UTR conservation analysis reveals that miR-124 targeting of SCP1 is likely to have arisen as a shared, derived trait in the vertebrate/tunicate ancestor and targeting of PTBP1 is conserved among bilaterians except for ecdysozoans, while extensive Notch pathway targeting appears to be Ciona specific. Altogether, our results provide a comprehensive insight into the specific mechanisms by which miR-124 promotes neuronal development.

  8. Revised lineage of larval photoreceptor cells in Ciona reveals archetypal collaboration between neural tube and neural crest in sensory organ formation.

    PubMed

    Oonuma, Kouhei; Tanaka, Moeko; Nishitsuji, Koki; Kato, Yumiko; Shimai, Kotaro; Kusakabe, Takehiro G

    2016-12-01

    The Ciona intestinalis larva has two distinct photoreceptor organs, a conventional pigmented ocellus and a nonpigmented ocellus, that are asymmetrically situated in the brain. The ciliary photoreceptor cells of these ocelli resemble visual cells of the vertebrate retina. Precise elucidation of the lineage of the photoreceptor cells will be key to understanding the developmental mechanisms of these cells as well as the evolutionary relationships between the photoreceptor organs of ascidians and vertebrates. Photoreceptor cells of the pigmented ocellus have been thought to develop from anterior animal (a-lineage) blastomeres, whereas the developmental origin of the nonpigmented ocellus has not been determined. Here, we show that the photoreceptor cells of both ocelli develop from the right anterior vegetal hemisphere: those of the pigmented ocellus from the right A9.14 cell and those of the nonpigmented ocellus from the right A9.16 cell. The pigmented ocellus is formed by a combination of two lineages of cells with distinct embryonic origins: the photoreceptor cells originate from a medial portion of the A-lineage neural plate, while the pigment cell originates from the lateral edge of the a-lineage neural plate. In light of the recently proposed close evolutionary relationship between the ocellus pigment cell of ascidians and the cephalic neural crest of vertebrates, the ascidian ocellus may represent a prototypic contribution of the neural crest to a cranial sensory organ. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Chondroitin 4-O-sulfotransferases are required for cell adhesion and morphogenesis in the Ciona intestinalis embryo.

    PubMed

    Nakamura, Jun; Tetsukawa, Akira; Fujiwara, Shigeki

    2015-01-01

    Chondroitin sulfate (CS) is a carbohydrate component of proteoglycans. Several types of sulfotransferases determine the pattern of CS sulfation, and thus regulate the biological functions of proteoglycans. The protochordate ascidians are the closest relatives of vertebrates, but the functions of their sulfotransferases have not been investigated. Here, we show that two chondroitin 4-O-sulfotransferases (C4STs) play important roles in the embryonic morphogenesis of the ascidian Ciona intestinalis. Ci-C4ST-like1 is predominantly expressed in the epidermis and muscle. Epidermal and muscle cells became spherical upon the injection of a Ci-C4ST-like1-specific morpholino oligo (MO), thus suggesting weakened cell adhesion. Co-injection of a Ci-C4ST-like1-expressing transgene rescued the phenotype, suggesting that the effects of the MO were specific. Ci-C4ST-like3 was expressed in the central nervous system, muscle, and mesenchyme. A specific MO appeared to affect cell adhesion in the epidermis and muscle. Convergent extension movement of notochordal cells was also impaired. Forced expression of Ci-C4ST-like3 restored normal morphogenesis, suggesting that the effects of the MO were specific. The present study suggests that Ci-C4ST-like1 and Ci-C4ST-like3 are required for cell adhesion mainly in the epidermis and muscle.

  10. Developmental toxicity of benzotriazole in the protochordate Ciona intestinalis (Chordata, Ascidiae).

    PubMed

    Kadar, Eniko; Dashfield, Sarah; Hutchinson, Thomas H

    2010-01-01

    Benzotriazoles (BT) are applied as anticorrosive and de-icing agents and have been detected in a variety of aquatic ecosystems and municipal wastewater effluents. We have assessed the developmental effects of benzotriazole (CAS number 95-14-7) to the marine invertebrate Ciona intestinalis (Chordata, Ascidiae). At 15 +/- 1 degrees C, the 24 h No-Observed Effect Concentration (NOEC) and Lowest Observed Effect Concentration (LOEC) values based on embryo morphological development were 100 and >100 mg L(-1), respectively (nominal concentration under static conditions). After 48 h, the NOEC and LOEC values were 10 and 32 mg L(-1), respectively. Light and electron microscopy studies on benzotriazole-exposed larva indicated that the primary target cells were the extra-embryonic test cells, which are known to play a significant apoptotic role during ascidian metamorphosis. The visible decline of test cells in benzotriazole-exposed larvae raises the possibility that the compound interferes with the regulation of embryo development in protochordates such as C. intestinalis. Further research is warranted to assess the potential longer term sublethal impacts of benzotriazole in aquatic organisms.

  11. Ammonium channel expression is essential for brain development and function in the larva of Ciona intestinalis.

    PubMed

    Marino, Rita; Melillo, Daniela; Di Filippo, Miriam; Yamada, Atsuko; Pinto, Maria Rosaria; De Santis, Rosaria; Brown, Euan R; Matassi, Giorgio

    2007-07-01

    Ammonium uptake into the cell is known to be mediated by ammonium transport (Amt) proteins, which are present in all domains of life. The physiological role of Amt proteins remains elusive; indeed, loss-of-function experiments suggested that Amt proteins do not play an essential role in bacteria, yeast, and plants. Here we show that the reverse holds true in the tunicate Ciona intestinalis. The genome of C. intestinalis contains two AMT genes, Ci-AMT1a and Ci-AMT1b, which we show derive from an ascidian-specific gene duplication. We analyzed Ci-AMT expression during embryo development. Notably, Ci-AMT1a is expressed in the larval brain in a small number of cells defining a previously unseen V-shaped territory; these cells connect the brain cavity to the external environment. We show that the knockdown of Ci-AMT1a impairs the formation of the brain cavity and consequently the function of the otolith, the gravity-sensing organ contained in it. We speculate that the normal mechanical functioning (flotation and free movement) of the otolith may require a close regulation of ammonium salt(s) concentration in the brain cavity, because ammonium is known to affect both fluid density and viscosity; the cells forming the V territory may act as a conduit in achieving such a regulation.

  12. Stochasticity and stereotypy in the Ciona notochord.

    PubMed

    Carlson, Maia; Reeves, Wendy; Veeman, Michael

    2015-01-15

    Fate mapping with single cell resolution has typically been confined to embryos with completely stereotyped development. The lineages giving rise to the 40 cells of the Ciona notochord are invariant, but the intercalation of those cells into a single-file column is not. Here we use genetic labeling methods to fate map the Ciona notochord with both high resolution and large sample sizes. We find that the ordering of notochord cells into a single column is not random, but instead shows a distinctive signature characteristic of mediolaterally-biased intercalation. We find that patterns of cell intercalation in the notochord are somewhat stochastic but far more stereotyped than previously believed. Cell behaviors vary by lineage, with the secondary notochord lineage being much more constrained than the primary lineage. Within the primary lineage, patterns of intercalation reflect the geometry of the intercalating tissue. We identify the latest point at which notochord morphogenesis is largely stereotyped, which is shortly before the onset of mediolateral intercalation and immediately after the final cell divisions in the primary lineage. These divisions are consistently oriented along the AP axis. Our results indicate that the interplay between stereotyped and stochastic cell behaviors in morphogenesis can only be assessed by fate mapping experiments that have both cellular resolution and large sample sizes. Copyright © 2014 Elsevier Inc. All rights reserved.

  13. Gene regulatory networks in the early ascidian embryo.

    PubMed

    Satou, Yutaka; Satoh, Nori; Imai, Kaoru S

    2009-04-01

    Ascidians, or sea squirts, are tunicates that diverged from the vertebrate lineage early in the chordate evolution. The compact and simple organization of the ascidian genome makes this organism an ideal model system for analyzing gene regulatory networks in embryonic development. Embryos contain relatively few cells and gene activities by individual cells have been determined. Here we review and discuss advances in our understanding of the ascidian embryogenesis emerging from genomic expression studies and analyses at the single cell level.

  14. Reprotoxicity of the Antifoulant Chlorothalonil in Ascidians: An Ecological Risk Assessment

    PubMed Central

    Gallo, Alessandra; Tosti, Elisabetta

    2015-01-01

    Chlorothalonil is a widely used biocide in antifouling paint formulation that replaces tin-based compounds after their definitive ban. Although chlorothalonil inputs into the marine environment have significantly increased in recent years, little is known about its effect on marine animals and in particular on their reproductive processes. In this line, the aim of the present study was to investigate the effects of chlorothalonil exposure on the gamete physiology, fertilization rate and transmissible damage to offspring in the marine invertebrate Ciona intestinalis (ascidians). To identify a possible mechanism of action of chlorothalonil, electrophysiological techniques were used to study the impact on oocyte membrane excitability and on the electrical events occurring at fertilization. The pre-exposure of spermatozoa and oocytes to chlorothalonil did not affect the fertilization rate but caused damage to the offspring by inducing larval malformation. The highest toxicity was observed when fertilization was performed in chlorothalonil solutions with the lowest EC50 value. In particular, it was observed that low chlorothalonil concentrations interfered with embryo development and led to abnormal larvae, whereas high concentrations arrested embryo formation. In mature oocytes, a decrease in the amplitudes of the sodium and fertilization currents was observed, suggesting an involvement of plasma membrane ion currents in the teratogenic mechanism of chlorothalonil action. The risk estimation confirmed that the predicted no-effect concentration (PNEC) exceeded the predicted effect concentration (PEC), thus indicating that chlorothalonil may pose a risk to aquatic species. PMID:25875759

  15. Cortical anchorages and cell type segregations of maternal postplasmic/PEM RNAs in ascidians.

    PubMed

    Paix, Alexandre; Yamada, Lixy; Dru, Philippe; Lecordier, Helene; Pruliere, Gerard; Chenevert, Janet; Satoh, Nori; Sardet, Christian

    2009-12-01

    Ascidian postplasmic/PEM RNAs constitute a large class of cortical maternal RNAs which include developmental determinants (macho-1 and pem-1). We have analyzed the localization, cortical anchorage and cell type segregation of postplasmic/PEM RNAs in Ciona intestinalis and Phallusia mammillata using very high-resolution fluorescent in situ hybridization. We also compared RNAs extracted from whole oocytes and from isolated cortices using microarrays and localized RNAs possessing clusters of xCACx motifs in their 3'UTRs. Based on these combined approaches we conclude that: (1) the vast majority of the 39 postplasmic/PEM RNAs (including vasa) are localized in the egg cortex. (2) Many postplasmic/PEM RNAs 3'UTR are enriched in xCACx motifs, allowing us to identify 2 novel postplasmic/PEM RNAs (PSD and MnK). (3) Postplasmic/PEM RNAs anchored to cortical Endoplasmic Reticulum (cER) and those associated with granules have different cell destinations. We propose that there are 2 distinct categories of postplasmic/PEM RNAs on the basis of their cortical anchorages and cell destinations: (1) macho-1-like postplasmic/PEM RNAs anchored to cER which segregate into somatic B8.11 cells. (2) vasa-like postplasmic/PEM RNAs associated with granules which in addition to B8.11 cells segregate into B8.12 germ cells.

  16. Auto and cross regulatory elements control Onecut expression in the ascidian nervous system.

    PubMed

    Pezzotti, Maria Rosa; Locascio, Annamaria; Racioppi, Claudia; Fucci, Laura; Branno, Margherita

    2014-06-15

    The expression pattern of Onecut genes in the central and peripheral nervous systems is highly conserved in invertebrates and vertebrates but the regulatory networks in which they are involved are still largely unknown. The presence of three gene copies in vertebrates has revealed the functional roles of the Onecut genes in liver, pancreas and some populations of motor neurons. Urochordates have only one Onecut gene and are the closest living relatives of vertebrates and thus represent a good model system to understand its regulatory network and involvement in nervous system formation. In order to define the Onecut genetic cascade, we extensively characterized the Onecut upstream cis-regulatory DNA in the ascidian Ciona intestinalis. Electroporation experiments using a 2.5kb genomic fragment and of a series of deletion constructs identified a small region of 262bp able to reproduce most of the Onecut expression profile during embryonic development. Further analyses, both bioinformatic and in vivo using transient transgenes, permitted the identification of transcription factors responsible for Onecut endogenous expression. We provide evidence that Neurogenin is a direct activator of Onecut and that an autoregulatory loop is responsible for the maintenance of its expression. Furthermore, for the first time we propose the existence of a direct connection among Neurogenin, Onecut and Rx transcription factors in photoreceptor cell formation. Copyright © 2014 Elsevier Inc. All rights reserved.

  17. Improved genome assembly and evidence-based global gene model set for the chordate Ciona intestinalis: new insight into intron and operon populations

    PubMed Central

    Satou, Yutaka; Mineta, Katsuhiko; Ogasawara, Michio; Sasakura, Yasunori; Shoguchi, Eiichi; Ueno, Keisuke; Yamada, Lixy; Matsumoto, Jun; Wasserscheid, Jessica; Dewar, Ken; Wiley, Graham B; Macmil, Simone L; Roe, Bruce A; Zeller, Robert W; Hastings, Kenneth EM; Lemaire, Patrick; Lindquist, Erika; Endo, Toshinori; Hotta, Kohji; Inaba, Kazuo

    2008-01-01

    Background The draft genome sequence of the ascidian Ciona intestinalis, along with associated gene models, has been a valuable research resource. However, recently accumulated expressed sequence tag (EST)/cDNA data have revealed numerous inconsistencies with the gene models due in part to intrinsic limitations in gene prediction programs and in part to the fragmented nature of the assembly. Results We have prepared a less-fragmented assembly on the basis of scaffold-joining guided by paired-end EST and bacterial artificial chromosome (BAC) sequences, and BAC chromosomal in situ hybridization data. The new assembly (115.2 Mb) is similar in length to the initial assembly (116.7 Mb) but contains 1,272 (approximately 50%) fewer scaffolds. The largest scaffold in the new assembly incorporates 95 initial-assembly scaffolds. In conjunction with the new assembly, we have prepared a greatly improved global gene model set strictly correlated with the extensive currently available EST data. The total gene number (15,254) is similar to that of the initial set (15,582), but the new set includes 3,330 models at genomic sites where none were present in the initial set, and 1,779 models that represent fusions of multiple previously incomplete models. In approximately half, 5'-ends were precisely mapped using 5'-full-length ESTs, an important refinement even in otherwise unchanged models. Conclusion Using these new resources, we identify a population of non-canonical (non-GT-AG) introns and also find that approximately 20% of Ciona genes reside in operons and that operons contain a high proportion of single-exon genes. Thus, the present dataset provides an opportunity to analyze the Ciona genome much more precisely than ever. PMID:18854010

  18. High-throughput sequence analysis of Ciona intestinalis SL trans-spliced mRNAs: alternative expression modes and gene function correlates.

    PubMed

    Matsumoto, Jun; Dewar, Ken; Wasserscheid, Jessica; Wiley, Graham B; Macmil, Simone L; Roe, Bruce A; Zeller, Robert W; Satou, Yutaka; Hastings, Kenneth E M

    2010-05-01

    Pre-mRNA 5' spliced-leader (SL) trans-splicing occurs in some metazoan groups but not in others. Genome-wide characterization of the trans-spliced mRNA subpopulation has not yet been reported for any metazoan. We carried out a high-throughput analysis of the SL trans-spliced mRNA population of the ascidian tunicate Ciona intestinalis by 454 Life Sciences (Roche) pyrosequencing of SL-PCR-amplified random-primed reverse transcripts of tailbud embryo RNA. We obtained approximately 250,000 high-quality reads corresponding to 8790 genes, approximately 58% of the Ciona total gene number. The great depth of this data revealed new aspects of trans-splicing, including the existence of a significant class of "infrequently trans-spliced" genes, accounting for approximately 28% of represented genes, that generate largely non-trans-spliced mRNAs, but also produce trans-spliced mRNAs, in part through alternative promoter use. Thus, the conventional qualitative dichotomy of trans-spliced versus non-trans-spliced genes should be supplanted by a more accurate quantitative view recognizing frequently and infrequently trans-spliced gene categories. Our data include reads representing approximately 80% of Ciona frequently trans-spliced genes. Our analysis also revealed significant use of closely spaced alternative trans-splice acceptor sites which further underscores the mechanistic similarity of cis- and trans-splicing and indicates that the prevalence of +/-3-nt alternative splicing events at tandem acceptor sites, NAGNAG, is driven by spliceosomal mechanisms, and not nonsense-mediated decay, or selection at the protein level. The breadth of gene representation data enabled us to find new correlations between trans-splicing status and gene function, namely the overrepresentation in the frequently trans-spliced gene class of genes associated with plasma/endomembrane system, Ca(2+) homeostasis, and actin cytoskeleton.

  19. High-throughput sequence analysis of Ciona intestinalis SL trans-spliced mRNAs: Alternative expression modes and gene function correlates

    PubMed Central

    Matsumoto, Jun; Dewar, Ken; Wasserscheid, Jessica; Wiley, Graham B.; Macmil, Simone L.; Roe, Bruce A.; Zeller, Robert W.; Satou, Yutaka; Hastings, Kenneth E.M.

    2010-01-01

    Pre-mRNA 5′ spliced-leader (SL) trans-splicing occurs in some metazoan groups but not in others. Genome-wide characterization of the trans-spliced mRNA subpopulation has not yet been reported for any metazoan. We carried out a high-throughput analysis of the SL trans-spliced mRNA population of the ascidian tunicate Ciona intestinalis by 454 Life Sciences (Roche) pyrosequencing of SL-PCR-amplified random-primed reverse transcripts of tailbud embryo RNA. We obtained ∼250,000 high-quality reads corresponding to 8790 genes, ∼58% of the Ciona total gene number. The great depth of this data revealed new aspects of trans-splicing, including the existence of a significant class of “infrequently trans-spliced” genes, accounting for ∼28% of represented genes, that generate largely non-trans-spliced mRNAs, but also produce trans-spliced mRNAs, in part through alternative promoter use. Thus, the conventional qualitative dichotomy of trans-spliced versus non-trans-spliced genes should be supplanted by a more accurate quantitative view recognizing frequently and infrequently trans-spliced gene categories. Our data include reads representing ∼80% of Ciona frequently trans-spliced genes. Our analysis also revealed significant use of closely spaced alternative trans-splice acceptor sites which further underscores the mechanistic similarity of cis- and trans-splicing and indicates that the prevalence of ±3-nt alternative splicing events at tandem acceptor sites, NAGNAG, is driven by spliceosomal mechanisms, and not nonsense-mediated decay, or selection at the protein level. The breadth of gene representation data enabled us to find new correlations between trans-splicing status and gene function, namely the overrepresentation in the frequently trans-spliced gene class of genes associated with plasma/endomembrane system, Ca2+ homeostasis, and actin cytoskeleton. PMID:20212022

  20. Ascidian introductions through the Suez Canal: The case study of an Indo-Pacific species.

    PubMed

    Rius, Marc; Shenkar, Noa

    2012-10-01

    Although marine biological invasions via the Suez Canal have been extensively documented, little is known about the introduction of non-indigenous ascidians (Chordata, Ascidiacea), a group containing particularly aggressive invasive species. Here, we used a multidisciplinary approach to study the introduction of the ascidian Herdmania momus into the Mediterranean Sea. We reviewed its taxonomy and global distribution, and analyzed how genetic variation is partitioned between sides of the Suez Canal. The taxonomic revision showed that H. momus currently has a wide Indo-Pacific distribution. Genetic data indicated two well-differentiated colonization histories across the eastern Mediterranean. Our findings suggest that the range expansion of H. momus has been greatly facilitated by the combined effect of human-mediated transport and the species' ability to adapt to different environments. The integrative approach presented here is critical to attain a holistic understanding of marine biological invasions, especially when studying groups with a poorly resolved taxonomy. Copyright © 2012 Elsevier Ltd. All rights reserved.

  1. Transmission and Scanning Electron Microscopy of the Accessory Cells and Chorion During Development of Ciona intestinalis Type B Embryos and the Impact of Their Removal on Cell Morphology.

    PubMed

    Thompson, Helen; Shimeld, Sebastian M

    2015-06-01

    Spawned ascidian oocytes are surrounded by a membrane called the chorion (or vitelline coat) and associated with two populations of maternally-supplied cells. Outside the chorion are follicle cells, which may affect the buoyancy of eggs. Inside the chorion are test cells, which during oogenesis provision the egg and which after fertilisation contribute to the larval tunic. The structure of maternal cells may vary between species. The model ascidian Ciona intestinalis has been recently split into two species, currently named type A and type B. The ultrastructure of extraembryonic cells and structures from type A embryos has been reported. Here we describe the ultrastructure of follicle and test cells from C. intestinalis type B embryos. Test cells are about 5 µm in diameter and line the inside of the chorion of developing embryos in a dense sheet. Follicle cells are large (> 100 µm long) and spike-shaped, with many large vesicles. Terminal electron dense granules are found towards the tips of spikes, adjacent to cytoplasm containing numerous small electron dense bodies connected by filaments. These are probably vesicles containing material for the terminal granules. Removal of maternal structures and cells just after fertilisation, as commonly used in many experiments manipulating C. intestinalis development, has been reported to affect embryonic patterning. We examined the impact of this on embryonic ectoderm cells by scanning electron microscopy. Cells of embryos that developed without maternal structures still developed cilia, but had indistinct cell boundaries and a more flattened appearance than those that developed within the chorion.

  2. The Ciona intestinalis immune-related galectin genes (CiLgals-a and CiLgals-b) are expressed by the gastric epithelium.

    PubMed

    Parrinello, Daniela; Sanfratello, Maria Antonietta; Vizzini, Aiti; Testasecca, Lelia; Parrinello, Nicolò; Cammarata, Matteo

    2017-03-01

    The transcription of two Ciona intestinalis galectin genes (CiLgals-a and CiLgals-b) is uparegulated by LPS in the pharynxis (hemocytes, vessel epithelium, endostilar zones) which is retained the main organ of the immunity. In this ascidian, for the first time we show, by immunohistochemistry and in situ hybridization methods, that these two immune-related genes are expressed in the gastric epithelium of naïve ascidians, whereas the galectins appear to be only contained in the intestine columnar epithelium. In addition, according to previous results on the pharynx, the genes are also expressed and galectins produced by hemocytes scattered in the connective tissue surrounding the gut. The genes expression and galectin localization in several tissues, including the previous findings on the transcription upregulation, the constitutive expression of these genes by endostylar zones and by the gastric epithelium suggest a potential multifunctional role of these galectins. In this respect, it is of interest to define where the CiLgals are normally found as related to the tissue functions. Such an approach should be a starting point for further investigations.

  3. Ascidians: An Emerging Marine Model for Drug Discovery and Screening.

    PubMed

    Dumollard, Remi; Gazo, Ievgeniia; Gomes, Isa D L; Besnardeau, Lydia; McDougall, Alex

    2017-01-01

    Ascidians (tunicates; sea squirts) are marine animals which provide a source of diverse, bioactive natural products, and a model for toxicity screenings. Compounds isolated from ascidians comprise an approved anti-tumor drug and many others are potent drug leads. Furthermore, the use of invertebrate embryos for toxicological screening tests or analysis offers the possibility to image a large number of samples for high throughput screens. Ascidians are members of a sister clade to the vertebrates and make a vertebrate-like tadpole larva composed of less than 3000 cells in 18 hours. The neural complex of the ascidian larva is made of only 350 cells (of which 100 are neurons) and functional genomic studies have now uncovered numerous GRNs underpinning neural specification and differentiation. Numerous studies showed that brain formation in ascidians is sensitive to toxic insults especially from endocrine disruptors making them a suitable model to study neurodevelopmental defects. Modern techniques available for ascidians, including transgenic embryos where 3D time lapse imaging of GFPexpressing reporter constructs can be analyzed, now permit numerous end-points to be evaluated in order to test the specific mode of action of many compounds. This review summarizes the key evidence suggesting that ascidian embryos are a favorable embryological model to study neurodevelopmental toxicity of different compounds with molecular and cellular end-points. We predict that ascidians may become a significant source of marine blue biotechnologies in the 21st century. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

  4. Linking chordate gene networks to cellular behavior in ascidians.

    PubMed

    Davidson, Brad; Christiaen, Lionel

    2006-01-27

    Embryos of simple chordates called ascidians (sea squirts) have few cells, develop rapidly, and are transparent, enabling the in vivo fluorescent imaging of labeled cell lineages. Ascidians are also simple genetically, with limited redundancy and compact regulatory regions. This cellular and genetic simplicity is now being exploited to link comprehensive gene networks to the cellular events underlying morphogenesis.

  5. On the sterols of some ascidians.

    PubMed

    Voogt, P A; van Rheenen, J W

    1975-08-01

    The lipid content of sea squirts is low, namely less than a half percent of the fresh weight. Lipids consist of about seventy percent of saponifiable lipids and of about twenty percent of non-saponifiable lipids. Both types of these lipids, including sterols, can be synthesized from acetate by these animals. Small amounts of C30 sterols were observed only in Microcosmus sulcatus and Halocynthia papillosa, the species with a low content of C27 sterols and a high content of C28 sterols. In addition these species contained considerable higher amounts of sterols with a double bond at the C22 position than Ciona intestinalis and Styela plicata did.

  6. Computational identification of Ciona intestinalis microRNAs.

    PubMed

    Keshavan, Raja; Virata, Michael; Keshavan, Anisha; Zeller, Robert W

    2010-02-01

    MicroRNAs (miRNAs) are conserved non-coding small RNAs with potent post-transcriptional gene regulatory functions. Recent computational approaches and sequencing of small RNAs had indicated the existence of about 80 Ciona intestinalis miRNAs, although it was not clear whether other miRNA genes were present in the genome. We undertook an alternative computational approach to look for Ciona miRNAs. Conserved non-coding sequences from the C. intestinalis genome were extracted and computationally folded to identify putative hairpin-like structures. After applying additional criteria, we obtained 458 miRNA candidates whose sequences were used to design a custom microarray. Over 100 of our predicted hairpins were identified in this array when probed with RNA from various Ciona stages. We also compared our predictions to recently deposited sequences of Ciona small RNAs and report that 170 of our predicted hairpins are represented in this data set. Altogether, about 250 of our 458 predicted miRNAs were represented in either our array data or the small-RNA sequence database. These results suggest that Ciona has a large number of genomically encoded miRNAs that play an important role in modulating gene activity in developing embryos and adults.

  7. Species specificity of symbiosis and secondary metabolism in ascidians

    PubMed Central

    Tianero, Ma Diarey B; Kwan, Jason C; Wyche, Thomas P; Presson, Angela P; Koch, Michael; Barrows, Louis R; Bugni, Tim S; Schmidt, Eric W

    2015-01-01

    Ascidians contain abundant, diverse secondary metabolites, which are thought to serve a defensive role and which have been applied to drug discovery. It is known that bacteria in symbiosis with ascidians produce several of these metabolites, but very little is known about factors governing these ‘chemical symbioses'. To examine this phenomenon across a wide geographical and species scale, we performed bacterial and chemical analyses of 32 different ascidians, mostly from the didemnid family from Florida, Southern California and a broad expanse of the tropical Pacific Ocean. Bacterial diversity analysis showed that ascidian microbiomes are highly diverse, and this diversity does not correlate with geographical location or latitude. Within a subset of species, ascidian microbiomes are also stable over time (R=−0.037, P-value=0.499). Ascidian microbiomes and metabolomes contain species-specific and location-specific components. Location-specific bacteria are found in low abundance in the ascidians and mostly represent strains that are widespread. Location-specific metabolites consist largely of lipids, which may reflect differences in water temperature. By contrast, species-specific bacteria are mostly abundant sequenced components of the microbiomes and include secondary metabolite producers as major components. Species-specific chemicals are dominated by secondary metabolites. Together with previous analyses that focused on single ascidian species or symbiont type, these results reveal fundamental properties of secondary metabolic symbiosis. Different ascidian species have established associations with many different bacterial symbionts, including those known to produce toxic chemicals. This implies a strong selection for this property and the independent origin of secondary metabolite-based associations in different ascidian species. The analysis here streamlines the connection of secondary metabolite to producing bacterium, enabling further biological and

  8. Species specificity of symbiosis and secondary metabolism in ascidians.

    PubMed

    Tianero, Ma Diarey B; Kwan, Jason C; Wyche, Thomas P; Presson, Angela P; Koch, Michael; Barrows, Louis R; Bugni, Tim S; Schmidt, Eric W

    2015-03-01

    Ascidians contain abundant, diverse secondary metabolites, which are thought to serve a defensive role and which have been applied to drug discovery. It is known that bacteria in symbiosis with ascidians produce several of these metabolites, but very little is known about factors governing these 'chemical symbioses'. To examine this phenomenon across a wide geographical and species scale, we performed bacterial and chemical analyses of 32 different ascidians, mostly from the didemnid family from Florida, Southern California and a broad expanse of the tropical Pacific Ocean. Bacterial diversity analysis showed that ascidian microbiomes are highly diverse, and this diversity does not correlate with geographical location or latitude. Within a subset of species, ascidian microbiomes are also stable over time (R=-0.037, P-value=0.499). Ascidian microbiomes and metabolomes contain species-specific and location-specific components. Location-specific bacteria are found in low abundance in the ascidians and mostly represent strains that are widespread. Location-specific metabolites consist largely of lipids, which may reflect differences in water temperature. By contrast, species-specific bacteria are mostly abundant sequenced components of the microbiomes and include secondary metabolite producers as major components. Species-specific chemicals are dominated by secondary metabolites. Together with previous analyses that focused on single ascidian species or symbiont type, these results reveal fundamental properties of secondary metabolic symbiosis. Different ascidian species have established associations with many different bacterial symbionts, including those known to produce toxic chemicals. This implies a strong selection for this property and the independent origin of secondary metabolite-based associations in different ascidian species. The analysis here streamlines the connection of secondary metabolite to producing bacterium, enabling further biological and

  9. On the evolutionary significance and metal-binding characteristics of a monolobal transferrin from Ciona intestinalis

    PubMed Central

    Tinoco, Arthur D.; Peterson, Cynthia W.; Lucchese, Baldo; Doyle, Robert P.; Valentine, Ann M.

    2008-01-01

    Transferrins are a family of proteins that bind and transport Fe(III). Modern transferrins are typically bilobal and are believed to have evolved from an ancient gene duplication of a monolobal form. A novel monolobal transferrin, nicatransferrin (nicaTf), was identified in the primitive ascidian species Ciona intestinalis that possesses the characteristic features of the proposed ancestral Tf protein. In this work, nicaTf was expressed in Pichia pastoris. Extensive solution studies were performed on nicaTf, including UV-vis, fluorescence, CD, EPR and NMR spectroscopies, and electrospray time-of-flight mass spectrometry. The expressed protein is nonglycosylated, unlike the protein isolated from the organism. This property does not affect its ability to bind Fe(III). However, Fe(III)-bound nicaTf displays important spectral differences from other Fe(III)-bound transferrins, which are likely the consequence of differences in metal coordination. Coordination differences could also account for the weaker affinity of nicaTf for Fe(III) (log K = 18.5) compared with bilobal human serum transferrin (HsTf) (log K = 22.5 and 21.4). The Fe–nicaTf complex is not labile, as indicated by slow metal removal kinetics by the high-affinity chelator tiron at pH 7.4. The protein alternatively binds up to one equivalent of Ti(IV) or V(V), which suggests that it may transport nonferric metals. These solution studies provide insight into the structure and function of the primitive monolobal transferrin of C. intestinalis for comparison with higher order bilobal transferrins. They suggest that a major advantage for the evolution of modern transferrins, dominantly of bilobal form, is stronger Fe(III) affinity because of cooperativity. PMID:18287008

  10. Genetic and Genomic Toolbox of the Chordate Ciona intestinalis

    PubMed Central

    Stolfi, Alberto; Christiaen, Lionel

    2012-01-01

    The experimental malleability and unique phylogenetic position of the sea squirt Ciona intestinalis as part of the sister group to the vertebrates have helped establish these marine chordates as model organisms for the study of developmental genetics and evolution. Here we summarize the tools, techniques, and resources available to the Ciona geneticist, citing examples of studies that employed such strategies in the elucidation of gene function in Ciona. Genetic screens, germline transgenesis, electroporation of plasmid DNA, and microinjection of morpholinos are all routinely employed, and in the near future we expect these to be complemented by targeted mutagenesis, homologous recombination, and RNAi. The genomic resources available will continue to support the design and interpretation of genetic experiments and allow for increasingly sophisticated approaches on a high-throughput, whole-genome scale. PMID:22964837

  11. Formation of the digestive tract in Ciona intestinalis includes two distinct morphogenic processes between its anterior and posterior parts.

    PubMed

    Nakazawa, Keiichi; Yamazawa, Takumi; Moriyama, Yuuta; Ogura, Yosuke; Kawai, Narudo; Sasakura, Yasunori; Saiga, Hidetoshi

    2013-10-01

    In the ascidian Ciona intestinalis, the digestive tract, an essential system for animals, develops during metamorphosis from the two primordial tissues, the endoderm and endodermal strand, located in the larval trunk and tail, respectively. However, it has been largely unknown how the digestive tract develops from these primordial tissues. We examined the metamorphosing larvae for the tubular formation of the digestive tract, focusing on the epithelial organization of the endoderm, by combined confocal microscopy and computational rendering. The tubular structure of the esophagus to the stomach was formed through the folding and closure of the endodermal epithelia in the central-to-right posterior trunk. By contrast, the intestine was formed in the left posterior trunk through the accumulation and rearrangement of the cells originated from the endodermal strand. This was confirmed by the cell-tracing experiment using Kaede expression construct driven in the endodermal strand. Thus, the tubular formation of the digestive tract in C. intestinalis includes distinct morphogenetic processes and cell lineages between its anterior and posterior parts. This study provides the first detailed description of the digestive tract morphogenesis in C. intestinalis and serves as an important basis toward thorough understanding of its digestive tract development. Copyright © 2013 Wiley Periodicals, Inc.

  12. Ephrin-mediated restriction of ERK1/2 activity delimits the number of pigment cells in the Ciona CNS

    PubMed Central

    Haupaix, Nicolas; Abitua, Philip B.; Sirour, Cathy; Yasuo, Hitoyoshi; Levine, Michael; Hudson, Clare

    2014-01-01

    Recent evidence suggests that ascidian pigment cells are related to neural crest-derived melanocytes of vertebrates. Using live-imaging, we determine a revised cell lineage of the pigment cells in Ciona intestinalis embryos. The neural precursors undergo successive rounds of anterior-posterior (A-P) oriented cell divisions, starting at the blastula 64-cell stage. A previously unrecognized fourth A-P oriented cell division in the pigment cell lineage leads to the generation of the post-mitotic pigment cell precursors. We provide evidence that MEK/ERK signals are required for pigment cell specification until approximately 30 minutes after the final cell division has taken place. Following each of the four A-P oriented cell divisions, ERK1/2 is differentially activated in the posterior sister cells, into which the pigment cell lineage segregates. Eph/ephrin signals are critical during the third A-P oriented cell division to spatially restrict ERK1/2 activation to the posterior daughter cell. Targeted inhibition of Eph/ephrin signals results in, at neurula stages, anterior expansion of both ERK1/2 activation and a pigment cell lineage marker and subsequently, at larval stages, supernumerary pigment cells. We discuss the implications of these findings with respect to the evolution of the vertebrate neural crest. PMID:25062608

  13. The expression of an immune-related phenoloxidase gene is modulated in Ciona intestinalis ovary, test cells, embryos and larva.

    PubMed

    Parrinello, Daniela; Sanfratello, Maria A; Vizzini, Aiti; Cammarata, Matteo

    2015-03-01

    Two distinct Ciona intestinalis phenoloxidases (CinPO1, 2) had previously been cloned and sequenced. The CinPO2 is involved in innate immunity and is expressed by inflammatory hemocytes that populate the tunic and pharynx vessels as a response to LPS inoculation. In situ hybridization and immunohistochemistry assays on histological section, showed that the expression of this gene and the produced protein are shared with oogenesis, embryogenesis and larval morphogenesis. Intriguingly, upregulation of gene transcription was found in the test cell layer that envelopes the ovary follicle, ovulated egg, and gastrula, as well as it was modulated in the zygotic nucleus of outer balstomers of 32-cell embryo, neurula presumptive epidermis tissue and larval mesenchyme. The anti-CinPO2 antibodies, specific for adult inflammatory cells, recognize epitopes in the cytoplasm of ovarian oocytes, ovulated eggs, development stages and larval mesenchyme. The overall findings disclose the precocious activation of the CinPO2 immunity-related gene, and show a developmentally programmed expression of this phenoloxidase. Furthermore, these findings support the multifunctional roles of immunity-related genes and allows us to explore new perspectives on ascidian development and immunity.

  14. Analysis of the Transcription Regulatory Mechanism of Otx During the Development of the Sensory Vesicle in Ciona intestinalis.

    PubMed

    Oonuma, Kouhei; Hirose, Dan; Takatori, Naohito; Saiga, Hidetoshi

    2014-09-01

    Establishment of the anterior-posterior axis is an important event in the development of bilateral animals. A homeodomain transcription factor, Otx, is important for the formation of the anterior part of the embryo, and its mRNA is expressed in a continuous manner in a wide range of animals. This pattern of expression is thought to be important for the formation of anterior neural structures, but the mechanism that regulates Otx expression remains largely unknown. Towards understanding how the transcription of Otx is maintained in the cells of anterior neural structure, the sensory vesicle, during embryogenesis, we examined transcription regulatory mechanisms of Otx, using embryos of the ascidian, Ciona intestinalis, from the gastrula to tailbud stages, which have not been studied previously. We identified two genomic regions capable of mimicking the Otx expression pattern from the gastrula to tailbud stages. Putative transcription factor binding sites required for this activity were identified. Notably, distinct sets of transcription factor binding sites were required at different developmental stages for the expression of Otx, suggesting that the continuity of Otx is supported by distinct transcriptional mechanisms in the gastrula and neurula stages. Along with previous studies using Halocynthia roretzi, the present results provide insight into the evolution of transcriptional regulatory mechanism of Otx.

  15. Ciona intestinalis peroxinectin is a novel component of the peroxidase-cyclooxygenase gene superfamily upregulated by LPS.

    PubMed

    Vizzini, Aiti; Parrinello, Daniela; Sanfratello, Maria Antonietta; Mangano, Valentina; Parrinello, Nicolò; Cammarata, Matteo

    2013-09-01

    Peroxinectins function as hemoperoxidase and cell adhesion factor involved in invertebrate immune reaction. In this study, the ascidian (Ciona intestinalis) peroxinectin gene (CiPxt) and its expression during the inflammatory response have been examined. CiPxt is a new member of the peroxidase-cyclooxygenase gene superfamily that contains both the peroxidase domain and the integrin KGD (Lys-Gly-Asp) binding motif. A phylogenetic tree showed that CiPxt is very close to the chordate group and appears to be the outgroup of mammalian MPO, EPO and TPO clades. The CiPxt molecular structure model resulted superimposable to the human myeloperoxidase. The CiPxt mRNA expression is upregulated by LPS inoculation suggesting it is involved in C. intestinalis inflammatory response. The CiPxt was expressed in hemocytes (compartment/morula cells), vessel epithelium, and unilocular refractile granulocytes populating the inflamed tunic matrix and in the zones 7, 8 and 9 of the endostyle, a special pharynx organs homolog to the vertebrate thyroid gland.

  16. Transforming growth factor β (CiTGF-β) gene expression is induced in the inflammatory reaction of Ciona intestinalis.

    PubMed

    Vizzini, Aiti; Di Falco, Felicia; Parrinello, Daniela; Sanfratello, Maria Antonietta; Cammarata, Matteo

    2016-02-01

    Transforming growth factor (TGF-β) is a well-known component of a regulatory cytokines superfamily that has pleiotropic functions in a broad range of cell types and is involved, in vertebrates, in numerous physiological and pathological processes. In the current study, we report on Ciona intestinalis molecular characterisation and expression of a transforming growth factor β homologue (CiTGF-β). The gene organisation, phylogenetic tree and modelling supported the close relationship with the mammalian TGF suggesting that the C. intestinalis TGF-β gene shares a common ancestor in the chordate lineages. Functionally, real-time PCR analysis showed that CiTGF-β was transcriptionally upregulated in the inflammatory process induced by LPS inoculation, suggesting that is involved in the first phase and significant in the secondary phase of the inflammatory response in which cell differentiation occurs. In situ hybridisation assays revealed that the genes transcription was upregulated in the pharynx, the main organ of the ascidian immune system, and expressed by cluster of hemocytes inside the pharynx vessels. These data supported the view that CiTGF-β is a potential molecule in immune defence systems against bacterial infection.

  17. Ephrin-mediated restriction of ERK1/2 activity delimits the number of pigment cells in the Ciona CNS.

    PubMed

    Haupaix, Nicolas; Abitua, Philip B; Sirour, Cathy; Yasuo, Hitoyoshi; Levine, Michael; Hudson, Clare

    2014-10-01

    Recent evidence suggests that ascidian pigment cells are related to neural crest-derived melanocytes of vertebrates. Using live-imaging, we determine a revised cell lineage of the pigment cells in Ciona intestinalis embryos. The neural precursors undergo successive rounds of anterior-posterior (A-P) oriented cell divisions, starting at the blastula 64-cell stage. A previously unrecognized fourth A-P oriented cell division in the pigment cell lineage leads to the generation of the post-mitotic pigment cell precursors. We provide evidence that MEK/ERK signals are required for pigment cell specification until approximately 30min after the final cell division has taken place. Following each of the four A-P oriented cell divisions, ERK1/2 is differentially activated in the posterior sister cells, into which the pigment cell lineage segregates. Eph/ephrin signals are critical during the third A-P oriented cell division to spatially restrict ERK1/2 activation to the posterior daughter cell. Targeted inhibition of Eph/ephrin signals results in, at neurula stages, anterior expansion of both ERK1/2 activation and a pigment cell lineage marker and subsequently, at larval stages, supernumerary pigment cells. We discuss the implications of these findings with respect to the evolution of the vertebrate neural crest.

  18. The CNS connectome of a tadpole larva of Ciona intestinalis (L.) highlights sidedness in the brain of a chordate sibling

    PubMed Central

    Ryan, Kerrianne; Lu, Zhiyuan; Meinertzhagen, Ian A

    2016-01-01

    Left-right asymmetries in brains are usually minor or cryptic. We report brain asymmetries in the tiny, dorsal tubular nervous system of the ascidian tadpole larva, Ciona intestinalis. Chordate in body plan and development, the larva provides an outstanding example of brain asymmetry. Although early neural development is well studied, detailed cellular organization of the swimming larva’s CNS remains unreported. Using serial-section EM we document the synaptic connectome of the larva’s 177 CNS neurons. These formed 6618 synapses including 1772 neuromuscular junctions, augmented by 1206 gap junctions. Neurons are unipolar with at most a single dendrite, and few synapses. Some synapses are unpolarised, others form reciprocal or serial motifs; 922 were polyadic. Axo-axonal synapses predominate. Most neurons have ciliary organelles, and many features lack structural specialization. Despite equal cell numbers on both sides, neuron identities and pathways differ left/right. Brain vesicle asymmetries include a right ocellus and left coronet cells. DOI: http://dx.doi.org/10.7554/eLife.16962.001 PMID:27921996

  19. Invasion genetics of the Ciona intestinalis species complex: from regional endemism to global homogeneity.

    PubMed

    Zhan, Aibin; Macisaac, Hugh J; Cristescu, Melania E

    2010-11-01

    Determining the degree of population connectivity and investigating factors driving genetic exchange at various geographical scales are essential to understanding population dynamics and spread potential of invasive species. Here, we explore these issues in the highly invasive vase tunicate, Ciona intestinalis, a species whose invasion history has been obscured by its poorly understood taxonomy and population genetics. Recent phylogenetic and comparative genomic studies suggest that C. intestinalis is a cryptic species complex consisting of at least three species. We reconstructed phylogenies based on both mitochondrial (cytochrome c oxidase subunit 3--NADH dehydrogenase subunit 1 region and NADH dehydrogenase subunit 4 gene) and nuclear (internal transcribed spacer 1) sequences, results of which support four major phylogroups corresponding to the previously reported spA, spB and Ciona spp. (spC) as well as an undescribed cryptic species (spD). While spC and spD remain restricted to their native ranges in the Mediterranean Sea and Black Sea, respectively, the highly invasive species (spA and spB) have disjunct global distributions. Despite extensive interspecific divergences, we identified low phylogeographical structure within these two invasive species. Haplotype network analyses revealed comparatively limited mutation steps among haplotypes within each species. Population genetic analyses based on two mtDNA fragments and eight unlinked microsatellites illustrated relatively low population differentiation and high population connectivity at both regional and continental scales in the two invasive species. Human-mediated dispersal coupled with a high potential for natural dispersal is probably responsible for the observed genetic homogeneity.

  20. Origin and Dispersal History of Two Colonial Ascidian Clades in the Botryllus schlosseri Species Complex

    PubMed Central

    Nydam, Marie L.

    2017-01-01

    Human-induced global warming and species introductions are rapidly altering the composition and functioning of Earth’s marine ecosystems. Ascidians (Phylum Chordata, Subphylum Tunicata, Class Ascidiacea) are likely to play an increasingly greater role in marine communities. The colonial ascidian B. schlosseri is a cryptic species complex comprising five genetically divergent clades (A-E). Clade A is a global species, and Clade E has so far been identified in European waters only. Using the largest mitochondrial cytochrome oxidase I datasets yet assembled, we determine the origin and dispersal history of these species. Nucleotide diversity and Approximate Bayesian Computation analyses support a Pacific origin for Clade A, with two likely dispersal scenarios that both show the northwestern Atlantic populations establishing early in the history of the species. Both Discrete Phylogeographic Analysis and Approximate Bayesian Computation support an origin of Clade E on the French side of the English Channel. An unsampled lineage evolved from the French lineage, which reflects the conclusion from the median joining network that not all Clade E lineages have been sampled. This unsampled lineage gave rise to the haplotypes on the English side of the English Channel, which were the ancestors to the Mediterranean and Bay of Biscay populations. Clade E has a wider geographic range than previously thought, and shows evidence of recent range expansion. Both Clade A and Clade E should be considered widespread species: Clade A globally and Clade E within Europe. PMID:28107476

  1. Amylibacter cionae sp. nov., isolated from the sea squirt Ciona savignyi.

    PubMed

    Wang, Dandan; Wei, Yuxi; Cui, Qiu; Li, Wenli

    2017-09-04

    A Gram-stain-negative, non-motile, aerobic and rod-shaped bacterial strain, designated H-12T, was isolated from a sea squirt (Ciona savignyi) collected from Tsingtao Port, Jiaozhou Bay, China, and its taxonomic position was investigated. Strain H-12T grew optimally at 25-30 °C, at pH 7.0-8.0 and in the presence of 3.0-4.0 % (w/v) NaCl. The 16S rRNA gene sequence of strain H-12T exhibited the highest similarity to that of the type strain of Amylibacter marinus (95.3 %). A neighbour-joining phylogenetic tree based on 16S rRNA gene sequences revealed that strain H-12T clustered with the type strain of A. marinus. The predominant ubiquinone in strain H-12T was identified as Q-10. The major fatty acids of strain H-12T were C18 : 1ω7c and C18 : 1ω7c 11-methyl. The major polar lipids detected in strain H-12T were phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, one unidentified aminolipid, two unidentified phospholipids and five unidentified lipids. The DNA G+C content of strain H-12T was 52.7 mol%. On the basis of phylogenetic, chemotaxonomic and phenotypic properties, strain H-12T is considered to represent a novel species within the genus Amylibacter, for which the name Amylibacter cionae sp. nov. is proposed. The type strain is H-12T (=KCTC 52581T=CGMCC 1.15880T).

  2. Embryo Microinjection and Electroporation in the Chordate Ciona intestinalis.

    PubMed

    Kari, Willi; Zeng, Fan; Zitzelsberger, Lena; Will, Johannes; Rothbächer, Ute

    2016-10-16

    Simple model organisms are instrumental for in vivo studies of developmental and cellular differentiation processes. Currently, the evolutionary distance to man of conventional invertebrate model systems and the complexity of genomes in vertebrates are critical challenges to modeling human normal and pathological conditions. The chordate Ciona intestinalis is an invertebrate chordate that emerged from a common ancestor with the vertebrates and may represent features at the interface between invertebrates and vertebrates. A common body plan with much simpler cellular and genomic composition should unveil gene regulatory network (GRN) links and functional genomics readouts explaining phenomena in the vertebrate condition. The compact genome of Ciona, a fixed embryonic lineage with few divisions and large cells, combined with versatile community tools foster efficient gene functional analyses in this organism. Here, we present several crucial methods for this promising model organism, which belongs to the closest sister group to vertebrates. We present protocols for transient transgenesis by electroporation, along with microinjection-mediated gene knockdown, which together provide the means to study gene function and genomic regulatory elements. We extend our protocols to provide information on how community databases are utilized for in silico design of gene regulatory or gene functional experiments. An example study demonstrates how novel information can be gained on the interplay, and its quantification, of selected neural factors conserved between Ciona and man. Furthermore, we show examples of differential subcellular localization in embryonic cells, following DNA electroporation in Ciona zygotes. Finally, we discuss the potential of these protocols to be adapted for tissue specific gene interference with emerging gene editing methods. The in vivo approaches in Ciona overcome major shortcomings of classical model organisms in the quest of unraveling conserved

  3. Embryo Microinjection and Electroporation in the Chordate Ciona intestinalis

    PubMed Central

    Kari, Willi; Zeng, Fan; Zitzelsberger, Lena; Will, Johannes; Rothbächer, Ute

    2016-01-01

    Simple model organisms are instrumental for in vivo studies of developmental and cellular differentiation processes. Currently, the evolutionary distance to man of conventional invertebrate model systems and the complexity of genomes in vertebrates are critical challenges to modeling human normal and pathological conditions. The chordate Ciona intestinalis is an invertebrate chordate that emerged from a common ancestor with the vertebrates and may represent features at the interface between invertebrates and vertebrates. A common body plan with much simpler cellular and genomic composition should unveil gene regulatory network (GRN) links and functional genomics readouts explaining phenomena in the vertebrate condition. The compact genome of Ciona, a fixed embryonic lineage with few divisions and large cells, combined with versatile community tools foster efficient gene functional analyses in this organism. Here, we present several crucial methods for this promising model organism, which belongs to the closest sister group to vertebrates. We present protocols for transient transgenesis by electroporation, along with microinjection-mediated gene knockdown, which together provide the means to study gene function and genomic regulatory elements. We extend our protocols to provide information on how community databases are utilized for in silico design of gene regulatory or gene functional experiments. An example study demonstrates how novel information can be gained on the interplay, and its quantification, of selected neural factors conserved between Ciona and man. Furthermore, we show examples of differential subcellular localization in embryonic cells, following DNA electroporation in Ciona zygotes. Finally, we discuss the potential of these protocols to be adapted for tissue specific gene interference with emerging gene editing methods. The in vivo approaches in Ciona overcome major shortcomings of classical model organisms in the quest of unraveling conserved

  4. Diversity of Actinobacteria Associated with the Marine Ascidian Eudistoma toealensis.

    PubMed

    Steinert, Georg; Taylor, Michael W; Schupp, Peter J

    2015-08-01

    Ascidians have yielded a wide variety of bioactive natural products. The colonial ascidian Eudistoma toealensis from Micronesia has been identified as the source of a series of staurosporine derivatives, though the exact origin of these derivatives is still unknown. To identify known staurosporine-producing microbes associated with E. toealensis, we analyzed with 16S rRNA gene tag pyrosequencing the overall bacterial community and focused on potential symbiotic bacteria already known from other ascidians or other marine hosts, such as sponges. The described microbiota was one of very high diversity, comprising 43 phyla: two from archaea, 34 described bacterial phyla, and seven candidate bacterial phyla. Many bacteria, which are renowned community members of other ascidians and marine holobionts, such as sponges and corals, were also part of the E. toealensis microbial community. Furthermore, two known producers of indolocarbazoles, Salinispora and Verrucosispora, were found with high abundance exclusively in the ascidian tissue, suggesting that microbial symbionts and not the organism itself may be the true producers of the staurosporines in E. toealensis.

  5. Morphological Differences between Larvae of the Ciona intestinalis Species Complex: Hints for a Valid Taxonomic Definition of Distinct Species

    PubMed Central

    Pennati, Roberta; Ficetola, Gentile Francesco; Brunetti, Riccardo; Caicci, Federico; Gasparini, Fabio; Griggio, Francesca; Sato, Atsuko; Stach, Thomas; Kaul-Strehlow, Sabrina; Gissi, Carmela; Manni, Lucia

    2015-01-01

    The cosmopolitan ascidian Ciona intestinalis is the most common model species of Tunicata, the sister-group of Vertebrata, and widely used in developmental biology, genomics and evolutionary studies. Recently, molecular studies suggested the presence of cryptic species hidden within the C. intestinalis species, namely C. intestinalis type A and type B. So far, no substantial morphological differences have been identified between individuals belonging to the two types. Here we present morphometric, immunohistochemical, and histological analyses, as well as 3-D reconstructions, of late larvae obtained by cross-fertilization experiments of molecularly determined type A and type B adults, sampled in different seasons and in four different localities. Our data point to quantitative and qualitative differences in the trunk shape of larvae belonging to the two types. In particular, type B larvae exhibit a longer pre-oral lobe, longer and relatively narrower total body length, and a shorter ocellus-tail distance than type A larvae. All these differences were found to be statistically significant in a Discriminant Analysis. Depending on the number of analyzed parameters, the obtained discriminant function was able to correctly classify > 93% of the larvae, with the remaining misclassified larvae attributable to the existence of intra-type seasonal variability. No larval differences were observed at the level of histology and immunohistochemical localization of peripheral sensory neurons. We conclude that type A and type B are two distinct species that can be distinguished on the basis of larval morphology and molecular data. Since the identified larval differences appear to be valid diagnostic characters, we suggest to raise both types to the rank of species and to assign them distinct names. PMID:25955391

  6. Evolution of Ciona intestinalis Tumor necrosis factor alpha (CiTNFα): Polymorphism, tissues expression, and 3D modeling.

    PubMed

    Vizzini, Aiti; Giovanna, Parisi Maria; Cardinale, Laura; Testasecca, Lelia; Cammarata, Matteo

    2017-02-01

    Although the Tumor necrosis factor gene superfamily seems to be very conserved in vertebrates, phylogeny, tissue expression, genomic and gene organization, protein domains and polymorphism analyses showed that a strong change has happened mostly in invertebrates in which protochordates were a constraint during the immune-molecules history and evolution. RT PCR was used to investigate differential gene expression in different tissues. The expression shown was greater in the pharynx. Single-nucleotide polymorphism has been investigated in Ciona intestinalis Tumor necrosis factor alpha (CiTNFα) mRNA isolated from the pharynx of 30 ascidians collected from Licata, Sicily (Italy), by denaturing gradient gel electrophoresis (DGGE). For this analysis, CiTNFα nucleotide sequence was separated into two fragments, TNF-1 and -2, respectively, of 630 and 540 bp. We defined 23 individual DGGE patterns (named 1 to 10 for TNF-1 and 1 to 13 for TNF-2). Five patterns for TNF-1 accounted for <10% of the individuals, whereas the pattern 13 of TNF-2 accounted for >20% of the individuals. All the patterns were verified by direct sequencing. Single base-pair mutations were observed mainly within COOH-terminus, leading to 30 nucleotide sequence variants and 30 different coding sequences segregating in two main different clusters. Although most of the base mutations were silent, four propeptide variants were detected and six amino acid replacements occurred within COOH-terminus. Statistical tests for neutrality indicated negative selection pressure on signal and mature peptide domains, but possible positive selection pressure on COOH-terminus domain. Lastly we displayed the in silico 3D structure analysis including the CiTNFα variable region.

  7. Morphological Differences between Larvae of the Ciona intestinalis Species Complex: Hints for a Valid Taxonomic Definition of Distinct Species.

    PubMed

    Pennati, Roberta; Ficetola, Gentile Francesco; Brunetti, Riccardo; Caicci, Federico; Gasparini, Fabio; Griggio, Francesca; Sato, Atsuko; Stach, Thomas; Kaul-Strehlow, Sabrina; Gissi, Carmela; Manni, Lucia

    2015-01-01

    The cosmopolitan ascidian Ciona intestinalis is the most common model species of Tunicata, the sister-group of Vertebrata, and widely used in developmental biology, genomics and evolutionary studies. Recently, molecular studies suggested the presence of cryptic species hidden within the C. intestinalis species, namely C. intestinalis type A and type B. So far, no substantial morphological differences have been identified between individuals belonging to the two types. Here we present morphometric, immunohistochemical, and histological analyses, as well as 3-D reconstructions, of late larvae obtained by cross-fertilization experiments of molecularly determined type A and type B adults, sampled in different seasons and in four different localities. Our data point to quantitative and qualitative differences in the trunk shape of larvae belonging to the two types. In particular, type B larvae exhibit a longer pre-oral lobe, longer and relatively narrower total body length, and a shorter ocellus-tail distance than type A larvae. All these differences were found to be statistically significant in a Discriminant Analysis. Depending on the number of analyzed parameters, the obtained discriminant function was able to correctly classify > 93% of the larvae, with the remaining misclassified larvae attributable to the existence of intra-type seasonal variability. No larval differences were observed at the level of histology and immunohistochemical localization of peripheral sensory neurons. We conclude that type A and type B are two distinct species that can be distinguished on the basis of larval morphology and molecular data. Since the identified larval differences appear to be valid diagnostic characters, we suggest to raise both types to the rank of species and to assign them distinct names.

  8. The ascidian pigmented sensory organs: structures and developmental programs.

    PubMed

    Esposito, R; Racioppi, C; Pezzotti, M R; Branno, M; Locascio, A; Ristoratore, F; Spagnuolo, A

    2015-01-01

    The recent advances on ascidian pigment sensory organ development and function represent a fascinating platform to get insight on the basic programs of chordate eye formation. This review aims to summarize current knowledge, at the structural and molecular levels, on the two main building blocks of ascidian light sensory organ, i.e. pigment cells and photoreceptor cells. The unique features of these structures (e.g., simplicity and well characterized cell lineage) are indeed making it possible to dissect the developmental programs at single cell resolution and will soon provide a panel of molecular tools to be exploited for a deep developmental and comparative-evolutionary analysis.

  9. Evidence of a Native Northwest Atlantic COI Haplotype Clade in the Cryptogenic Colonial Ascidian Botryllus schlosseri.

    PubMed

    Yund, Philip O; Collins, Catherine; Johnson, Sheri L

    2015-06-01

    The colonial ascidian Botryllus schlosseri should be considered cryptogenic (i.e., not definitively classified as either native or introduced) in the Northwest Atlantic. Although all the evidence is quite circumstantial, over the last 15 years most research groups have accepted the scenario of human-mediated dispersal and classified B. schlosseri as introduced; others have continued to consider it native or cryptogenic. We address the invasion status of this species by adding 174 sequences to the growing worldwide database for the mitochondrial gene cytochrome c oxidase subunit I (COI) and analyzing 1077 sequences to compare genetic diversity of one clade of haplotypes in the Northwest Atlantic with two hypothesized source regions (the Northeast Atlantic and Mediterranean). Our results lead us to reject the prevailing view of the directionality of transport across the Atlantic. We argue that the genetic diversity patterns at COI are far more consistent with the existence of at least one haplotype clade in the Northwest Atlantic (and possibly a second) that substantially pre-dates human colonization from Europe, with this native North American clade subsequently introduced to three sites in Northeast Atlantic and Mediterranean waters. However, we agree with past researchers that some sites in the Northwest Atlantic have more recently been invaded by alien haplotypes, so that some populations are currently composed of a mixture of native and invader haplotypes.

  10. Toxicity of Hg, Cu, Cd, and Cr on early developmental stages of Ciona intestinalis (Chordata, Ascidiacea) with potential application in marine water quality assessment.

    PubMed

    Bellas, J; Vázquez, E; Beiras, R

    2001-08-01

    The toxicity of mercury, copper, cadmium and chromium on sperm viability, fertilisation, embryogenesis and larval attachment of Ciona intestinalis was examined. Fertilisation rate (FR) showed a small decrease even at the highest metal concentration tested. The median effective concentrations (EC50) reducing rates of embryogenesis and larval attachment by 50% were 54 microg Hg/l (0.27 microM), 46 microg Cu/l (0.72 microM), 838 microg Cd/l (7.46 microM), 10,318 microg Cr/l (198 microM), and 35 microg Hg/l (0.18 microM), 34 microg Cu/l (0.54 microM) and 11,755 microg Cr/l (226 microM), respectively. Therefore, Hg is three times more toxic than Cu (on a molar basis), ca. 30 times more toxic than Cd and ca. 1000 times more toxic than Cr to early stages of C. intestinalis. Rates of larval attachment and embryogenesis were the most sensitive endpoints, although the latter is more advisable for routine assessment of seawater quality because of its greater simplicity. In addition to bivalves and sea-urchins, ascidian embryos can provide biological criteria for seawater quality standards taking into account the sensitivity of a chordate and contributing to the detection of harmful chemicals with no marked effect on the species currently in use in seawater quality bioassays.

  11. LPS injection reprograms the expression and the 3' UTR of a CAP gene by alternative polyadenylation and the formation of a GAIT element in Ciona intestinalis.

    PubMed

    Vizzini, Aiti; Bonura, Angela; Longo, Valeria; Sanfratello, Maria Antonietta; Parrinello, Daniela; Cammarata, Matteo; Colombo, Paolo

    2016-09-01

    The diversification of cellular functions is one of the major characteristics of multicellular organisms which allow cells to modulate their gene expression, leading to the formation of transcripts and proteins with different functions and concentrations in response to different stimuli. CAP genes represent a widespread family of proteins belonging to the cysteine-rich secretory protein, antigen 5 and pathogenesis-related 1 superfamily which, it has been proposed, play key roles in the infection process and the modulation of immune responses in host animals. The ascidian Ciona intestinalis represents a group of proto-chordates with an exclusively innate immune system that has been widely studied in the field of comparative and developmental immunology. Using this biological system, we describe the identification of a novel APA mechanism by which an intronic polyadenylation signal is activated by LPS injection, leading to the formation of a shorter CAP mRNA capable of expressing the first CAP exon plus 19 amino acid residues whose sequence is contained within the first intron of the annotated gene. Furthermore, such an APA event causes the expression of a translational controlling cis-acting GAIT element which is not present in the previously isolated CAP isoform and identified in the 3'-UTR of other immune-related genes, suggesting an intriguing scenario in which both transcriptional and post-transcriptional control mechanisms are involved in the activation of the CAP gene during inflammatory response in C. intestinalis.

  12. Localization of antimicrobial peptides in the tunic of Ciona intestinalis (Ascidiacea, Tunicata) and their involvement in local inflammatory-like reactions

    PubMed Central

    Di Bella, M.A.; Fedders, H.; De Leo, G.; Leippe, M.

    2011-01-01

    Tunicates comprising a wide variety of different species synthesize antimicrobial peptides as important effector molecules of the innate immune system. Recently, two putative gene families coding for antimicrobial peptides were identified in the expressed sequence tag database of the tunicate Ciona intestinalis. Two synthetic peptides representing the cationic core region of one member of each of the families displayed potent antibacterial and antifungal activities. Moreover, the natural peptides were demonstrated to be synthesized and stored in distinct hemocyte types. Here, we investigated the presence of these natural peptides, namely Ci-MAM-A and Ci-PAP-A, in the tunic of C. intestinalis considering that the ascidian tunic is a body surface barrier exposed to constant microbial assault. Furthermore, as the tunic may represent a major route of entry for pathogen invasion after its damage we monitored the location of these peptides upon a local inflammatory-like reaction induced by injection of foreign cells. Using immunocytochemistry and electron microscopy both peptides were localized to the tunic and were massively present in granulocytes of inflamed tissue. Conclusively, antimicrobial peptides may constitute a chemical barrier within the tunic of urochordates. PMID:24371555

  13. A cDNA resource from the basal chordate Ciona intestinalis.

    PubMed

    Satou, Yutaka; Yamada, Lixy; Mochizuki, Yasuaki; Takatori, Naohito; Kawashima, Takeshi; Sasaki, Akane; Hamaguchi, Makoto; Awazu, Satoko; Yagi, Kasumi; Sasakura, Yasunori; Nakayama, Akie; Ishikawa, Hisayoshi; Inaba, Kazuo; Satoh, Nori

    2002-08-01

    The genome of the basal choradate Ciona intestinalis contains a basic set of genes with less redundancy compared to the vertebrate genome. Extensive EST analyses, cDNA sequencing, and clustering yielded "Ciona intestinalis Gene Collection Release 1," which contains cDNA clones for 13,464 genes, covering nearly 85% of the Ciona mRNA species. This release is ready for use in cDNA cloning, micro/macroarray analysis, and other comprehensive genome-wide analyses for further molecular studies of basal chordates.

  14. The secret to a successful relationship: lasting chemistry between ascidians and their symbiotic bacteria.

    PubMed

    Schmidt, Eric W

    2015-03-01

    Bioactive secondary metabolites are common components of marine animals. In many cases, symbiotic bacteria, and not the animals themselves, synthesize the compounds. Among marine animals, ascidians are good models for understanding these symbioses. Ascidians often contain potently bioactive secondary metabolites as their major extractable components. Strong evidence shows that ~8% of the known secondary metabolites from ascidians are made by symbiotic bacteria, and indirect evidence implicates bacteria in the synthesis of many more. Far from being "secondary" to the animals, secondary metabolites are essential components of the interaction between host animals and their symbiotic bacteria. These interactions have complex underlying biology, but the chemistry is clearly ascidian-species specific. The chemical interactions are ancient in at least some cases, and they are widespread among ascidians. Ascidians maintain secondary metabolic symbioses with bacteria that are phylogenetically diverse, indicating a convergent solution to obtaining secondary metabolites and reinforcing the importance of secondary metabolism in animal survival.

  15. The Comparative Organismal Approach in Evolutionary Developmental Biology: Insights from Ascidians and Cavefish.

    PubMed

    Jeffery, William R

    2016-01-01

    Important contributions to evolutionary developmental biology have been made using the comparative organismal approach. As examples, I describe insights obtained from studies of Molgula ascidians and Astyanax cavefish.

  16. *Arsenic (+3 oxidation state) methyltransferase and the methylation of arsenicals in the invertebrate chordate ciona intestinalis

    EPA Science Inventory

    Biotransformation of inorganic arsenic (iAs) involves methylation catalyzed by arsenic (+3 oxidation state) methyltransferase (As3mt) , yielding mono-, di-, and trimethylated arsenicals. A comparative genomic approach focused on Ciona intestinaJis, an invertebrate chordate, was u...

  17. *Arsenic (+3 oxidation state) methyltransferase and the methylation of arsenicals in the invertebrate chordate ciona intestinalis

    EPA Science Inventory

    Biotransformation of inorganic arsenic (iAs) involves methylation catalyzed by arsenic (+3 oxidation state) methyltransferase (As3mt) , yielding mono-, di-, and trimethylated arsenicals. A comparative genomic approach focused on Ciona intestinaJis, an invertebrate chordate, was u...

  18. The Gut of Geographically Disparate Ciona intestinalis Harbors a Core Microbiota

    SciTech Connect

    Dishaw, Larry J.; Flores-Torres, Jaime; Lax, Simon; Gemayel, Kristina; Leigh, Brittany; Melillo, Daniela; Mueller, M. Gail; Natale, Lenina; Zucchetti, Ivana; De Santis, Rosaria; Pinto, Maria Rosaria; Litman, Gary W.; Gilbert, Jack A.

    2014-04-02

    It is now widely understood that all animals engage in complex interactions with bacteria (or microbes) throughout their various life stages. This ancient exchange can involve cooperation and has resulted in a wide range of evolved host-microbial interdependencies, including those observed in the gut. Ciona intestinalis, a filter-feeding basal chordate and classic developmental model that can be experimentally manipulated, is being employed to help define these relationships. Ciona larvae are first exposed internally to microbes upon the initiation of feeding in metamorphosed individuals; however, whether or not these microbes subsequently colonize the gut and whether or not Ciona forms relationships with specific bacteria in the gut remains unknown. Here in this report, we show that the Ciona gut not only is colonized by a complex community of bacteria, but also that samples from three geographically isolated populations reveal striking similarity in abundant operational taxonomic units (OTUs) consistent with the selection of a core community by the gut ecosystem.

  19. The gut of geographically disparate Ciona intestinalis harbors a core microbiota.

    PubMed

    Dishaw, Larry J; Flores-Torres, Jaime; Lax, Simon; Gemayel, Kristina; Leigh, Brittany; Melillo, Daniela; Mueller, M Gail; Natale, Lenina; Zucchetti, Ivana; De Santis, Rosaria; Pinto, Maria Rosaria; Litman, Gary W; Gilbert, Jack A

    2014-01-01

    It is now widely understood that all animals engage in complex interactions with bacteria (or microbes) throughout their various life stages. This ancient exchange can involve cooperation and has resulted in a wide range of evolved host-microbial interdependencies, including those observed in the gut. Ciona intestinalis, a filter-feeding basal chordate and classic developmental model that can be experimentally manipulated, is being employed to help define these relationships. Ciona larvae are first exposed internally to microbes upon the initiation of feeding in metamorphosed individuals; however, whether or not these microbes subsequently colonize the gut and whether or not Ciona forms relationships with specific bacteria in the gut remains unknown. In this report, we show that the Ciona gut not only is colonized by a complex community of bacteria, but also that samples from three geographically isolated populations reveal striking similarity in abundant operational taxonomic units (OTUs) consistent with the selection of a core community by the gut ecosystem.

  20. Differential gene expression along the animal-vegetal axis in the ascidian embryo is maintained by a dual functional protein Foxd.

    PubMed

    Tokuhiro, Shin-Ichi; Tokuoka, Miki; Kobayashi, Kenji; Kubo, Atsushi; Oda-Ishii, Izumi; Satou, Yutaka

    2017-05-01

    In many animal embryos, a specific gene expression pattern is established along the animal-vegetal axis soon after zygotic transcription begins. In the embryo of the ascidian Ciona intestinalis, soon after the division that separates animal and vegetal hemispheres into distinct blastomeres, maternal Gata.a and β-catenin activate specific genes in the animal and vegetal blastomeres, respectively. On the basis of these initial distinct gene expression patterns, gene regulatory networks promote animal cells to become ectodermal tissues and vegetal cells to become endomesodermal tissues and a part of the nerve cord. In the vegetal hemisphere, β-catenin directly activates Foxd, an essential transcription factor gene for specifying endomesodermal fates. In the present study, we found that Foxd also represses the expression of genes that are activated specifically in the animal hemisphere, including Dmrt1, Prdm1-r.a (Bz1), Prdm1-r.b (Bz2), and Otx. A reporter assay showed that Dmrt1 expression was directly repressed by Foxd, and a chromatin immunoprecipitation assay showed that Foxd was bound to the upstream regions of Dmrt1, Prdm1-r.a, Prdm1-r.b, and Otx. Thus, Foxd has a dual function of activating specific gene expression in the vegetal hemisphere and of repressing the expression of genes that are normally expressed in the animal hemisphere. This dual function stabilizes the initial patterning along the animal-vegetal axis by β-catenin and Gata.a.

  1. Valosin-containing protein/p97 interacts with sperm-activating and sperm-attracting factor (SAAF) in the ascidian egg and modulates sperm-attracting activity.

    PubMed

    Kondoh, Eri; Konno, Aru; Inaba, Kazuo; Oishi, Tohru; Murata, Michio; Yoshida, Manabu

    2008-10-01

    Sperm chemotaxis toward an egg is observed in many animals, and the control of sperm-attracting activity is thought to play an important role in ensuring fertilization. However, the mechanism underlying the release of a sperm attractant from an egg is still obscure. In this study, we examined the systems involved in the release of sperm-activating and sperm-attracting factor (SAAF), which is the sperm attractant of the ascidian Ciona intestinalis. Here, we show that the egg acquires sperm-attracting activity after germinal vesicle breakdown. Further, since the cytoplasmic extracts of immature oocytes exhibit no sperm-attracting activity, the SAAF in oocytes may be activated after germinal vesicle breakdown. We found 13 SAAF-binding proteins in an egg plasma membrane extract and identified five proteins by proteomic analysis: valosin-containing protein (VCP)/p97, proteasome alpha 2 subunit, MGC97756 protein, proteasome subunit Y, and beta-tubulin. In particular, the interaction between VCP/p97 and SAAF was confirmed by a pull-down assay. VCP/p97 is initially localized in the germinal vesicle, and during oocyte maturation, it shifts to the endoplasmic reticulum in the cortical regions. Thus, VCP/p97 is a potential modulator of SAAF release from the egg.

  2. Time lapse confocal microscopy of mitochondrial movements in ascidian embryos

    NASA Astrophysics Data System (ADS)

    Sardet, C.; Rouvière, C.; Flannery, B.; Davoust, J.

    1991-05-01

    We have analysed the complex movements of vitally stained mitochondria in living eggs of the ascidian (prochordate) embryo soon after fertilization by time lapse confocal (Laser scanning) Microscopy. Our studies clearly show that it is possible to understand how the subcortical mitochondria rich layer contracts, tears and folds to reach its final location in the future posterior pole of the embryo that gives rise to the muscle cell lineage.

  3. The evolution of larval morphology and swimming performance in ascidians.

    PubMed

    McHenry, Matthew J; Patek, Sheila N

    2004-06-01

    The complexity of organismal function challenges our ability to understand the evolution of animal locomotion. To meet this challenge, we used a combination of biomechanics, phylogenetic comparative analyses, and theoretical morphology to examine evolutionary changes in body shape and how those changes affected swimming performance in ascidian larvae. Results of phylogenetic comparative analyses suggest that coloniality evolved at least three times among ascidians and that colonial species have a convergent larval morphology characterized by a large trunk volume and shorter tail length in proportion to the trunk. To explore the functional significance of this evolutionary change, we first verified the accuracy of a mathematical model of swimming biomechanics in a solitary (C. intestinalis) and a colonial (D. occidentalis) species and then ran numerous simulations of the model that varied in tail length and trunk volume. The results of these simulations were used to construct landscapes of speed and cost of transport predictions within a trunk volume/tail length morphospace. Our results suggest that the reduction of proportionate tail length in colonial species resulted in improved energetic economy of swimming. The increase in the size of larvae with the origin of coloniality facilitated faster swimming with negligible energetic cost, but may have required a reduction in adult fecundity. Therefore, the evolution of ascidians appears to be influenced by a trade-off between the fecundity of the adult stage and the swimming performance of larvae.

  4. Vanadium biochemistry: the unknown role of vanadium-containing cells in ascidians (sea squirts).

    PubMed

    Smith, M J

    1989-05-15

    This article reviews several new developments in vanadium biochemistry, as elucidated from studies of ascidians. A hypothesis correlating ascidian blood cell function to anaerobiosis, via two prominent redox constituents, namely vanadium(III) and the tunichromes, a family of metal ion complexing/reducing hydroquinonoid peptides, is presented.

  5. Commensal Leucothoidae (Crustacea, Amphipoda) of the Ryukyu Archipelago, Japan. Part I: ascidian-dwellers

    PubMed Central

    White, Kristine N.; Reimer, James Davis

    2012-01-01

    Abstract Commensal leucothoid amphipods have been collected from the branchial chambers of their ascidian hosts throughout the Ryukyu Archipelago, Japan. Seven new species are described in two genera with valuable location data and host records. An identification key to ascidian-dwelling Leucothoidae of the Ryukyu Archipelago is provided. PMID:22303128

  6. New distributional data on ascidian fauna (Tunicata: Ascidiacea) from Mandapam coast, Gulf of Mannar, India

    PubMed Central

    Akram, Soban A; Arshan, Kaleem ML

    2016-01-01

    Abstract Background Ascidians play a key role in the ecology and biodiversity of marine ecosystem. Ascidians can be transported in ship ballast water and while attached to ship and boat hulls. Heavy traffic by domestic and international ships as well as cargo vessels between the major and minor ports warrants continuous monitoring for new introductions of ascidians. The Mandapam coast is situated in the Gulf of Mannar, India, a marine hot spot area in the Indian Ocean which provides an environment suitable for the settlement of ascidians. New information A total of 30 species of ascidians were reported from Mandapam coastal waters, of which 26 species were new to the study area and five species: Ecteinascidia turbinata, Eudistoma carnosum, Trididemnum caelatum, T. vermiforme and Didemnum spadix, were new to India. PMID:27099557

  7. Exploiting the extraordinary genetic polymorphism of ciona for developmental genetics with whole genome sequencing.

    PubMed

    Abdul-Wajid, Sarah; Veeman, Michael T; Chiba, Shota; Turner, Thomas L; Smith, William C

    2014-05-01

    Studies in tunicates such as Ciona have revealed new insights into the evolutionary origins of chordate development. Ciona populations are characterized by high levels of natural genetic variation, between 1 and 5%. This variation has provided abundant material for forward genetic studies. In the current study, we make use of deep sequencing and homozygosity mapping to map spontaneous mutations in outbred populations. With this method we have mapped two spontaneous developmental mutants. In Ciona intestinalis we mapped a short-tail mutation with strong phenotypic similarity to a previously identified mutant in the related species Ciona savignyi. Our bioinformatic approach mapped the mutation to a narrow interval containing a single mutated gene, α-laminin3,4,5, which is the gene previously implicated in C. savignyi. In addition, we mapped a novel genetic mutation disrupting neural tube closure in C. savignyi to a T-type Ca(2+) channel gene. The high efficiency and unprecedented mapping resolution of our study is a powerful advantage for developmental genetics in Ciona, and may find application in other outbred species.

  8. Evolution and development of brain sensory organs in molgulid ascidians.

    PubMed

    Jeffery, William R

    2004-01-01

    The ascidian tadpole larva has two brain sensory organs containing melanocytes: the otolith, a gravity receptor, and the ocellus, part of a photoreceptor. One or both of these sensory organs are absent in molgulid ascidians. We show here that developmental changes leading to the loss of sensory pigment cells occur by different mechanisms in closely related molgulid species. Sensory pigment cells are formed through a bilateral determination pathway in which two or more precursor cells are specified as an equivalence group on each side of the embryo. The precursor cells subsequently converge at the midline after neurulation and undergo cell interactions that decide the fates of the otolith and ocellus. Molgula occidentalis and M. oculata, which exhibit a tadpole larva with an otolith but lacking an ocellus, have conserved the bilateral pigment cell determination pathway. Programmed cell death (PCD) is superimposed on this pathway late in development to eliminate the ocellus precursor and supernumerary pigment cells, which do not differentiate into either an otolith or ocellus. In contrast to molgulids with tadpole larvae, no pigment cell precursors are specified on either side of the M. occulta embryo, which forms a tailless (anural) larva lacking both sensory organs, suggesting that the bilateral pigment cell determination pathway has been lost. The bilateral pigment cell determination pathway and superimposed PCD can be restored in hybrids obtained by fertilizing M. occulta eggs with M. oculata sperm, indicating control by a zygotic process. We conclude that PCD plays an important role in the evolution and development of brain sensory organs in molgulid ascidians.

  9. Prochloron-ascidian symbioses: Photosynthetic potential and productivity

    NASA Technical Reports Server (NTRS)

    Lewin, R. A.; Cheng, L.; Alberte, R. S.

    1983-01-01

    The chlorophyll content of didemnid asidians with symbiotic algae (Prochloron) from oligotropic tropical marine waters around Palau, Western Carolin Islands is discussed. Several species contain as much chlorophyll per unit dry weight as many herbaceous crop plants and more than do other symbiotic associations such as lichens, green Hydra, etc. Their chlorphyllA/B ratios (3-9) were generally much lighter than those of angiosperms (2-4). Where they abound, Prochloron - ascidian symbiosis could make a major contribution to the productivity, especially in localized areas of tropical marine waters characterized by low nutrient levels and high irradiance.

  10. Photosymbiotic ascidians from Pari Island (Thousand Islands, Indonesia).

    PubMed

    Hirose, Euichi; Iskandar, Budhi Hascaryo; Wardiatno, Yusli

    2014-01-01

    Photosymbiotic ascidian fauna were surveyed in the subtidal zone off Pari Island in the Thousand Islands (Java Sea, Indonesia). Nine species were recorded: Didemnum molle, Trididemnum miniatum, Lissoclinum patella, L. punctatum, L. timorense, Diplosoma gumavirens, D. simile, D. simileguwa, and D. virens. All of these species have been previously recorded in the Ryukyu Archipelago, Japan. Diplosoma gumavirens and D. simileguwa were originally described from the Ryukyu Archipelago in 2009 and 2005, respectively, and all of the observed species are potentially widely distributed in Indo-West Pacific coral reefs.

  11. Prochloron-ascidian symbioses: Photosynthetic potential and productivity

    NASA Technical Reports Server (NTRS)

    Lewin, R. A.; Cheng, L.; Alberte, R. S.

    1983-01-01

    The chlorophyll content of didemnid asidians with symbiotic algae (Prochloron) from oligotropic tropical marine waters around Palau, Western Carolin Islands is discussed. Several species contain as much chlorophyll per unit dry weight as many herbaceous crop plants and more than do other symbiotic associations such as lichens, green Hydra, etc. Their chlorphyllA/B ratios (3-9) were generally much lighter than those of angiosperms (2-4). Where they abound, Prochloron - ascidian symbiosis could make a major contribution to the productivity, especially in localized areas of tropical marine waters characterized by low nutrient levels and high irradiance.

  12. Photosymbiotic ascidians from Pari Island (Thousand Islands, Indonesia)

    PubMed Central

    Hirose, Euichi; Iskandar, Budhi Hascaryo; Wardiatno, Yusli

    2014-01-01

    Abstract Photosymbiotic ascidian fauna were surveyed in the subtidal zone off Pari Island in the Thousand Islands (Java Sea, Indonesia). Nine species were recorded: Didemnum molle, Trididemnum miniatum, Lissoclinum patella, L. punctatum, L. timorense, Diplosoma gumavirens, D. simile, D. simileguwa, and D. virens. All of these species have been previously recorded in the Ryukyu Archipelago, Japan. Diplosoma gumavirens and D. simileguwa were originally described from the Ryukyu Archipelago in 2009 and 2005, respectively, and all of the observed species are potentially widely distributed in Indo–West Pacific coral reefs. PMID:25061385

  13. Ascidians as excellent models for studying cellular events in the chordate body plan.

    PubMed

    Ogura, Yosuke; Sasakura, Yasunori

    2013-08-01

    The larvae of non-vertebrate chordate ascidians consist of countable numbers of cells. With this feature, ascidians provide us with excellent models for studying cellular events in the construction of the chordate body. This review discusses the recent observations of morphogenetic movements and cell cycles and divisions along with tissue specifications during ascidian embryogenesis. Unequal cleavages take place at the posterior blastomeres during the early cleavage stages of ascidians, and the structure named the centrosome-attracting body restricts the position of the nuclei near the posterior pole to achieve the unequal cleavages. The most-posterior cells differentiate into the primordial germ cells. The gastrulation of ascidians starts as early as the 110-cell stage. During gastrulation, the endodermal cells show two-step changes in cell shape that are crucial for gastrulation. The ascidian notochord is composed of only 40 cells. The 40 cells align to form a single row by an event named the convergent extension, and then the notochord cells undergo vacuolation to transform the notochord into a single hollowed tube. The strictly restricted number of notochord cells is achieved by the regulated number of cell divisions coupled with the differentiation of the cells conducted by a key transcription factor, Brachyury. The dorsally located neural tube is a characteristic of chordates. During the closure of the ascidian neural tube, the epidermis surrounding the neural plate moves toward the midline to close the neural fold. This morphogenetic movement is allowed by an elongation of interphase in the epidermal cell cycles.

  14. Ascidian tunicate extracts attenuate rheumatoid arthritis in a collagen-induced murine model.

    PubMed

    Hong, Seong-Ho; Kwone, Jung-Taek; Lee, Jae-Ho; Lee, Somin; Lee, Ah Young; Cho, Won-Young; Bat-Erdene, Munkhjargal; Choi, Byeong-Dae; Cho, Myung-Haing

    2014-06-01

    Murine rheumatoid arthritis models are often used to investigate the potential therapeutic effects of candidate drugs. The present study has been conducted in order to investigate the therapeutic efficacy of ascidian tunicate extracts in a collagen-induced arthritis DBA1/J mice model. Four types of formulas, ascidian tunicate extracts (ATE), crude ascidian tunicate glycans (ATEC), ascidian tunicate extracts with licorice extracts (ATEL), and crude ascidian tunicate glycans with licorice extracts (ATECL) were orally administered into DBA/1J mice for 3 weeks and paw edema and thickness were evaluated. Changes in inflammatory proteins and cytokines levels were monitored in hind leg tissues by Western blot and quantitative PCR analysis. The oral administration of ascidian tunicate extracts alleviated paw edema and improved the histological hind leg cartilage status. The extracts also reduced the matrix metalloproteinase-9 (MMP-9) protein and prostaglandin E synthase (PGES) levels. In addition, the extracts-treated groups showed increased interleukin-10 (IL-10) levels compared with the non-treated group. These findings suggest that orally administered ascidian tunicate extracts might have potential therapeutic effects for the treatment of rheumatoid arthritis.

  15. Sensing charges of the Ciona intestinalis voltage-sensing phosphatase.

    PubMed

    Villalba-Galea, Carlos A; Frezza, Ludivine; Sandtner, Walter; Bezanilla, Francisco

    2013-11-01

    Voltage control over enzymatic activity in voltage-sensitive phosphatases (VSPs) is conferred by a voltage-sensing domain (VSD) located in the N terminus. These VSDs are constituted by four putative transmembrane segments (S1 to S4) resembling those found in voltage-gated ion channels. The putative fourth segment (S4) of the VSD contains positive residues that likely function as voltage-sensing elements. To study in detail how these residues sense the plasma membrane potential, we have focused on five arginines in the S4 segment of the Ciona intestinalis VSP (Ci-VSP). After implementing a histidine scan, here we show that four arginine-to-histidine mutants, namely R223H to R232H, mediate voltage-dependent proton translocation across the membrane, indicating that these residues transit through the hydrophobic core of Ci-VSP as a function of the membrane potential. These observations indicate that the charges carried by these residues are sensing charges. Furthermore, our results also show that the electrical field in VSPs is focused in a narrow hydrophobic region that separates the extracellular and intracellular space and constitutes the energy barrier for charge crossing.

  16. Three new species of Enterocola van Beneden, 1860 (Crustacea: Copepoda: Cyclopoida) from ascidian hosts.

    PubMed

    Marchenkov, Andrey; Boxshall, Geoff A

    2005-07-01

    Three new species of Enterocola are described: E. dicaudatus n. sp., from the ascidian host Polysyncraton rostrum Monniot & Monniot collected in Tanzania, E. monnioti n. sp., from the ascidian host Polyclinum constellatum Savigny collected in Bahrain, and E. parapterophorus n. sp., from the ascidian host Didemnum granulatum Tokioka taken in Djibouti. In all three species, the mandibles are interpreted as being represented by a pair of setulose seta-like elements that are largely concealed beneath similar paired setulose structures, the labral palps, carried on the labrum. The paired elements interpreted as mandibles are transversely orientated and lie in a groove, the anterior margin of which forms a defined ridge ornamented with spinules.

  17. Bioadhesion in ascidians: a developmental and functional genomics perspective

    PubMed Central

    Pennati, Roberta; Rothbächer, Ute

    2015-01-01

    The development of bioadhesives inspired from marine animals is a promising approach to generate new tissue-compatible medical components. A number of marine species, through their adhesive properties, also represent significant foulers that become increasingly problematic to aquaculture, shipping or local biodiversity. In order to develop more sophisticated man-made glues and/or efficient fouling resistant surfaces, it is important to understand the mechanical, structural and molecular properties of adhesive organs in selected species. Ascidians are marine invertebrates with larvae that opportunistically attach to almost any type of submerged surface to undergo metamorphosis into permanently sessile adults. Not only do they represent a globally important fouling organism, but they are becoming increasingly popular as model organisms for developmental biology. The latter is due to their phylogenetic position as the sister group to the vertebrates and their cellular and molecular accessibility for experimentation. In this paper, we review the mechanisms of larval adhesion in ascidians and draw conclusions from comparative analyses of selected species. We further discuss how knowledge from a developmental and functional genomics point of view can advance our understanding of cellular and molecular signatures and their hierarchical usage in animal adhesive organs. PMID:25657840

  18. Description of Endozoicomonas ascidiicola sp. nov., isolated from Scandinavian ascidians.

    PubMed

    Schreiber, Lars; Kjeldsen, Kasper Urup; Obst, Matthias; Funch, Peter; Schramm, Andreas

    2016-07-01

    Two gram-negative, facultative anaerobic, chemoorganoheterotrophic, motile and rod-shaped bacteria, strains AVMART05(T) and KASP37, were isolated from ascidians (Tunicata, Ascidiaceae) of the genus Ascidiella collected at Gullmarsfjord, Sweden. The strains are the first cultured representatives of an ascidian-specific lineage within the genus Endozoicomonas (Gammaproteobacteria, Oceanospirillales, Hahellaceae). Both strains feature three distinct 16S rRNA gene paralogs, with identities of 98.9-99.1% (AVMART05(T)) and 97.7-98.8% (KASP37) between paralogs. The strains are closely related to Endozoicomonas atrinae and Endozoicomonas elysicola, with which they share 97.3-98.0% 16S rRNA gene sequence identity. Digital DNA-DNA hybridization, average nucleotide identity, and tetra-nucleotide correlation analysis indicate that both strains belong to a single species distinct from their closest relatives. Both strains feature similar DNA G+C contents of 46.70mol% (AVMART05(T)) and 44.64mol% (KASP37). The fatty acid patterns of AVMART05(T) and KASP37 are most similar to those of Endozoicomonas euniceicola and Endozoicomonas gorgoniicola. Based on the polyphasic approach, we propose the species Endozoicomonas ascidiicola sp. nov. to accommodate the newly isolated strains. E. ascidiicola sp. nov. is represented by the type strain AVMART05(T) (=DSM 100913(T)=LMG 29095(T)) and strain KASP37 (=DSM 100914=LMG 29096). Copyright © 2016 Elsevier GmbH. All rights reserved.

  19. Repression of Rx gene on the left side of the sensory vesicle by Nodal signaling is crucial for right-sided formation of the ocellus photoreceptor in the development of Ciona intestinalis.

    PubMed

    Yoshida, Keita; Saiga, Hidetoshi

    2011-06-01

    Nodal signaling plays an essential role in the establishment of left-right asymmetry in various animals. However, it is largely unknown how Nodal signaling is involved in the establishment of the left-right asymmetric morphology. In this study, the role of Nodal signaling in the left-right asymmetric ocellus formation in the ascidian, Ciona intestinalis was dealt with. During the development of C. intestinalis, the ocellus pigment cell forms on the midline and moves to the right side of the midline. Then, the photoreceptor cells form on the right side of the sensory vesicle (SV). Ci-Nodal is expressed on the left side of the SV in the developing tail bud embryo. When Nodal signaling is inhibited, the ocellus pigment cell form but remain on the midline, and expression of marker genes of the ocellus photoreceptor cells is ectopically detected on the left side as well as on the right side of the SV in the larva. Furthermore, Ci-Rx, which is essential for the ocellus differentiation, turns out to be negatively regulated by the Nodal signaling on the left side of the SV, even though it is required for the right-sided photoreceptor formation. These results indicate that Nodal signaling controls the left-right asymmetric ocellus formation in the development of C. intestinalis.

  20. The Whereabouts of an Ancient Wanderer: Global Phylogeography of the Solitary Ascidian Styela plicata

    PubMed Central

    Pineda, Mari Carmen; López-Legentil, Susanna; Turon, Xavier

    2011-01-01

    Genetic tools have greatly aided in tracing the sources and colonization history of introduced species. However, recurrent introductions and repeated shuffling of populations may have blurred some of the genetic signals left by ancient introductions. Styela plicata is a solitary ascidian distributed worldwide. Although its origin remains unclear, this species is believed to have spread worldwide by travelling on ship's hulls. The goals of this study were to infer the genetic structure and global phylogeography of S. plicata and to look for present-day and historical genetic patterns. Two genetic markers were used: a fragment of the mitochondrial gene Cytochrome Oxidase subunit I (COI) and a fragment of the nuclear gene Adenine Nucleotide Transporter/ADP-ATP Translocase (ANT). A total of 368 individuals for COI and 315 for ANT were sequenced from 17 locations worldwide. The levels of gene diversity were moderate for COI to high for ANT. The Mediterranean populations showed the least diversity and allelic richness for both markers, while the Indian, Atlantic and Pacific Oceans had the highest gene and nucleotide diversities. Network and phylogenetic analyses with COI and ANT revealed two groups of alleles separated by 15 and 4 mutational steps, respectively. The existence of different lineages suggested an ancient population split. However, the geographic distributions of these groups did not show any consistent pattern, indicating different phylogeographic histories for each gene. Genetic divergence was significant for many population-pairs irrespective of the geographic distance among them. Stochastic introduction events are reflected in the uneven distribution of COI and ANT allele frequencies and groups among many populations. Our results confirmed that S. plicata has been present in all studied oceans for a long time, and that recurrent colonization events and occasional shuffling among populations have determined the actual genetic structure of this species

  1. Photosymbiotic ascidians in Singapore: turbid waters may reduce living space

    PubMed Central

    Su, Shih-Wei; Hirose, Euichi; Chen, Serina Lee Siew; Mok, Michael Hin-Kiu

    2013-01-01

    Abstract The photosymbiotic ascidian fauna at Changi Beach, Pulau Semakau, Sentosa and St. John’s Island, Singapore were surveyed. A total of five species, Diplosoma simile, Lissoclinum bistratum, Lissoclinum punctatum, Lissoclinum timorense and Trididemnum cyclops, were recorded, with Lissoclinum timorense and Trididemnum cyclops being newly recorded in Singapore. However, no photosymbiotic species were found at Changi Beach probably due to the polluted waters in the region. Coastal development has caused Singapore waters to become turbid, leading to decrease in suitable habitats for photosymbiotic ascidians. Clean waters in Pulau Semakau probably provide a better environment for the growth of photosymbiotic ascidians and this area has a greater variety of these ascidians than the other areas in Singapore. Each of the five species has also been recorded in the Ryukyu Archipelago (Japan) and three species (Diplosoma simile, Lissoclinum bistratum and Trididemnum cyclops) have also been recorded in Taiwan. PMID:23794913

  2. Retroduplication and loss of parental genes is a mechanism for the generation of intronless genes in Ciona intestinalis and Ciona savignyi.

    PubMed

    Kim, Dong Seon; Wang, Yao; Oh, Hye Ji; Choi, Dongjin; Lee, Kangseok; Hahn, Yoonsoo

    2014-12-01

    Tunicates, the sister clade of vertebrates, have miniature genomes and numerous intronless genes compared to other animals. It is still unclear how the tunicates acquired such a large number of intronless genes. Here, we analyzed sequences and intron-exon organizations of homologous genes from two closely related tunicates, Ciona intestinalis and Ciona savignyi. We found seven cases in which ancestral introns of a gene were completely lost in a species after their divergence. In four cases, both the intronless copy and the intron-containing copy were present in the genome, indicating that the intronless copy was generated by retroduplication. In the other three cases, the intron-containing copy was absent, implying it was lost after retroduplication. This result suggests that retroduplication and loss of parental genes is a major mechanism for the accumulation of intronless genes in tunicates.

  3. Ascidian depth zonation on sublittoral hard substrates off deer island, New Brunswick, Canada

    NASA Astrophysics Data System (ADS)

    Hatfield, C.; Logan, A.; Thomas, M. L. H.

    1992-02-01

    The upper surfaces of sublittoral hard substrates in the Deer Island region of the Bay of Fundy, New Brunswick, support diverse, depth-zoned epibenthic communities of which ascidians form a minor part. Their population density was quantitatively studied from photo-transects taken between mean low water (MLW) and 30 m depth at four sites off the Deer Island coast and from 30-140 m depth along two photo-transects in Head Harbour Passage. All photo-analyses were aided by collections from transect survey sites, wharf pilings and salmon cage floats, to yield a total of 15 ascidian species encountered. Ascidians were found at all depths at the four shallow sites. Halocynthia pyriformis and Boltenia ovifera are most common at depths of less than 20 m, while Aplidium pallidum, Didemnum albidum and other species exhibit a marked increase in abundance below this depth. Cluster analysis of ascidians shows an association between B. echinata and B. ovifera, which may reflect resource partitioning, and between A. pallidum-D. albidum and Molgula sp.— A. stellatum, the ecological significance of which are as yet unknown. The community in Head Harbour Passage is animal-dominated and in its deeper sections often shows three-dimensional bottom relief from horse mussel shells. D. albidum, the commonest ascidian, shows a close association with Modiolus modiolus, to which it is normally attached, suggesting that mussel beds may minimize the possibility of dislodgement and even confer a feeding advantage on this ascidian.

  4. The challenges of trafficking hydrolysis prone metals and ascidians as an archetype.

    PubMed

    Gaffney, Jean P; Valentine, Ann M

    2011-06-14

    Some of the metal ions that are required, exploited, or simply managed in biological systems are susceptible to hydrolysis and to hydrolytic precipitation in the aqueous, aerobic environment of much of biology. Organisms have evolved exquisite mechanisms for handling these metal ions, offering striking examples of biological control over inorganic coordination chemistry. This year marks the one hundredth anniversary of the discovery of remarkably high vanadium concentrations in the blood cells of the ascidian. In the ensuing years, these marine invertebrates were established as masters of the biological chemistry of very hydrolysis-prone metals, with various ascidian species accumulating high concentrations of iron, vanadium, and titanium, among others. These three metals have very different histories of biological relevance, and many questions remain about how, and ultimately why, these organisms sequester them. This Perspective addresses the aqueous coordination chemistry that organisms like ascidians must control if they are to manipulate hydrolysis-prone metal ions, and describes some of the ascidian biomolecules that have been implicated in this phenomenon. The recently available genome sequence for one ascidian species offers a glimpse into its metal-management arsenal. It offers the opportunity to map the relatively well-studied paradigm of iron management onto the genome of an organism that is intermediate in evolution between invertebrates and vertebrates. The ascidians have much to teach us about how to manage metals like iron, titanium, and vanadium and how that ability evolved.

  5. The Gut of Geographically Disparate Ciona intestinalis Harbors a Core Microbiota

    DOE PAGES

    Dishaw, Larry J.; Flores-Torres, Jaime; Lax, Simon; ...

    2014-04-02

    It is now widely understood that all animals engage in complex interactions with bacteria (or microbes) throughout their various life stages. This ancient exchange can involve cooperation and has resulted in a wide range of evolved host-microbial interdependencies, including those observed in the gut. Ciona intestinalis, a filter-feeding basal chordate and classic developmental model that can be experimentally manipulated, is being employed to help define these relationships. Ciona larvae are first exposed internally to microbes upon the initiation of feeding in metamorphosed individuals; however, whether or not these microbes subsequently colonize the gut and whether or not Ciona forms relationshipsmore » with specific bacteria in the gut remains unknown. Here in this report, we show that the Ciona gut not only is colonized by a complex community of bacteria, but also that samples from three geographically isolated populations reveal striking similarity in abundant operational taxonomic units (OTUs) consistent with the selection of a core community by the gut ecosystem.« less

  6. Pseudovibrio ascidiaceicola sp. nov., isolated from ascidians (sea squirts).

    PubMed

    Fukunaga, Yukiyo; Kurahashi, Midori; Tanaka, Kenji; Yanagi, Kensuke; Yokota, Akira; Harayama, Shigeaki

    2006-02-01

    Two bacterial strains, F423T and F10102, were isolated from two ascidians, Polycitor proliferus and Botryllidae sp., respectively, which were collected from a beach on the Boso peninsula in Japan. Cells of both isolates were motile, rod-shaped and formed star-shaped aggregates in the early stage of exponential growth, but were coccoid in stationary growth phase. The results of 16S rRNA gene sequence analysis, fatty acid analysis, DNA-DNA hybridization experiments and physiological and biochemical tests indicated that the two strains were members of a novel species of the genus Pseudovibrio for which the name Pseudovibrio ascidiaceicola sp. nov. is proposed. The type strain is F423T (=NBRC 100514T=IAM 15084T=DSM 16392T=KCTC 12308T).

  7. Genetic Distances in Three Ascidian Species determined by PCR Technique

    PubMed Central

    Yoon, Jong-Man

    2016-01-01

    ABSTRACT Seven oligonucleotides primers were shown to generate the shared loci, specific loci, unique shared loci to each species and shared loci by the three species which could be obviously scored. In the present study, 7 oligonucleotides primers produced 401 total loci in the Styela clava (SC) species, 390 in the Halocynthia roretzi (HR) and 434 in the Styela plicata (SP), respectively. Seven oligonucleotides primers generated 275 specific loci in the SC, 341 in the HR and 364 in the SP species, respectively. The oligonucleotides primer BION-23 generated 28 unique loci to each species in the SP species. Especially, the oligonucleotides primer BION-25 produced 7 unique loci to each species, which were identifying each species in the SP species. BION-17 distinguished 21 shared loci by the three ascidian species, major and/or minor fragments of sizes, which were identical in almost all of the samples. Based on the average bandsharing values of all samples, the similarity matrix ranged from 0.519 to 0.774 in the SC species, from 0.261 to 0.683 in the HR species and from 0.346 to 0.730 in the SP species. As regards average bandsharing value (BS) results, individuals from SC species (0.661±0.081) exhibited higher bandsharing values than did individuals from HR species (0.555±0.074) (P<0.05). The dendrogram obtained by the seven oligonucleotides primers indicates three genetic groups. In three ascidian species, the shortest genetic distance (0.071) exhibiting significant molecular difference was also between individual no. 20 and no. 21 within the SP species. PMID:28144642

  8. Proteome map of the neural complex of the tunicate Ciona intestinalis, the closest living relative to vertebrates.

    PubMed

    Saxena, Sandeep; Dupont, Sam; Meghah, Vuppalapaty; Lakshmi, Mula G Meena; Singh, Sachin K; Swamy, Cherukuvada V Brahmendra; Idris, Mohammed M

    2013-03-01

    Ciona intestinalis (the common sea squirt) is the closest living chordate relative to vertebrates with cosmopolitan presence worldwide. It has a relatively simple nervous system and development, making it a widely studied alternative model system in neuroscience and developmental biology. The use of Ciona as a model organism has increased significantly after the draft genome was published. In this study, we describe the first proteome map of the neural complex of C. intestinalis. A total of 544 proteins were identified based on 1DE and 2DE FTMS/ITMSMS analyses. Proteins were annotated against the Ciona database and analyzed to predict their molecular functions, roles in biological processes, and position in constructed network pathways. The identified Ciona neural complex proteome was found to map onto vertebrate nervous system pathways, including cytoskeleton remodeling neurofilaments, cell adhesion through the histamine receptor signaling pathway, γ-aminobutyric acid-A receptor life cycle neurophysiological process, glycolysis, and amino acid metabolism. The proteome map of the Ciona neural complex is the first step toward a better understanding of several important processes, including the evolution and regeneration capacity of the Ciona nervous system.

  9. Chloride channel in vanadocytes of a vanadium-rich ascidian Ascidia sydneiensis samea.

    PubMed

    Ueki, Tatsuya; Yamaguchi, Nobuo; Michibata, Hitoshi

    2003-09-01

    Ascidians, so-called sea squirts, can accumulate high levels of vanadium in the vacuoles of signet ring cells, which are one type of ascidian blood cell and are also called vanadocytes. In addition to containing high concentrations of vanadium in the +3 oxidation state, the proton concentrations in vanadocyte vacuoles are extremely high. In order to elucidate the entire mechanism of the accumulation and reduction of vanadium by ascidian vanadocytes, it is necessary to clarify the participation of anions, which might be involved as counter ions in the active accumulation of both vanadium and protons. We examined the chloride channel, since chloride ions are necessary for the acidification of intracellular vesicles and coexist with H(+)-ATPase. We cloned a cDNA encoding a chloride channel from blood cells of a vanadium-rich ascidian, Ascidia sydneiensis samea. It encoded a 787-amino-acid protein, which showed striking similarity to mammalian ClC3/4/5-type chloride channels. Using a whole-mount in situ hybridization method that we developed for ascidian blood cells, the chloride channel was revealed to be transcribed in vanadocytes, suggesting its participation in the process of vanadium accumulation.

  10. Sunlight Damage To The Solitary Ascidian Chelyosoma productum

    NASA Astrophysics Data System (ADS)

    Flores, E.

    2004-12-01

    Chelyosoma productum (Stimpson) is a temperate solitary ascidian commonly found in Puget Sound and the San Juan Archipelago, Washington, USA. Adult populations are restricted to deeper subtidal regions or shaded shallow-water habitats, such as docks in shaded marinas. C. productum adults have a thin translucent outer tunic that may provide very little if any protection from solar damage. I hypothesized that sunlight may be setting limits on the distribution of this species. Since adult ascidians are sessile and rely on earlier life stages for their distribution, all life stages of Chelyosoma productum were tested. In this study, I examined the effects of sunlight exposure in embryos, larvae, juveniles and adults of Chelyosoma productum. I isolated the PAR, UVA and UVB portions of the spectrum and exposed all life stages using natural sunlight. I also sampled shallow-water dock habitats to see how adult distributions were related to light exposure. The embryonic development in C. productum was negatively affected by any solar exposure. Most embryos exposed to UV light failed to develop normally and those that did could not subsequently settle. This species produces embryos of different colors; two (purple and brown) were observed in my experiments. Damage from light exposure differed between the color morphs. Overall, the brown morph was more tolerant of light exposure than was the purple morph across all life stages. The only exception to this general pattern was that purple embryos were remarkably resistant to light damage. The distribution of C. productum is restricted to areas with no direct solar exposure. However, even within shaded environments where they were abundant, subpopulations related to the color dimorphism were observed. The significance of the brown and purple pigments in embryos and larvae remains largely unknown. However, adults with brown eggs were found to be more prevalent in edge environments where there was higher light exposure. Purple

  11. Down under the tunic: bacterial biodiversity hotspots and widespread ammonia-oxidizing archaea in coral reef ascidians

    PubMed Central

    Erwin, Patrick M; Pineda, Mari Carmen; Webster, Nicole; Turon, Xavier; López-Legentil, Susanna

    2014-01-01

    Ascidians are ecologically important components of marine ecosystems yet the ascidian microbiota remains largely unexplored beyond a few model species. We used 16S rRNA gene tag pyrosequencing to provide a comprehensive characterization of microbial symbionts in the tunic of 42 Great Barrier Reef ascidian samples representing 25 species. Results revealed high bacterial biodiversity (3 217 unique operational taxonomic units (OTU0.03) from 19 described and 14 candidate phyla) and the widespread occurrence of ammonia-oxidizing Thaumarchaeota in coral reef ascidians (24 of 25 host species). The ascidian microbiota was clearly differentiated from seawater microbial communities and included symbiont lineages shared with other invertebrate hosts as well as unique, ascidian-specific phylotypes. Several rare seawater microbes were markedly enriched (200–700 fold) in the ascidian tunic, suggesting that the rare biosphere of seawater may act as a conduit for horizontal symbiont transfer. However, most OTUs (71%) were rare and specific to single hosts and a significant correlation between host relatedness and symbiont community similarity was detected, indicating a high degree of host-specificity and potential role of vertical transmission in structuring these communities. We hypothesize that the complex ascidian microbiota revealed herein is maintained by the dynamic microenvironments within the ascidian tunic, offering optimal conditions for different metabolic pathways such as ample chemical substrate (ammonia-rich host waste) and physical habitat (high oxygen, low irradiance) for nitrification. Thus, ascidian hosts provide unique and fertile niches for diverse microorganisms and may represent an important and previously unrecognized habitat for nitrite/nitrate regeneration in coral reef ecosystems. PMID:24152714

  12. Evidence for dynamic and multiple roles for huntingtin in Ciona intestinalis.

    PubMed

    Idris, Mohammed M; Thorndyke, Michael C; Brown, Euan R

    2013-12-01

    Although mutations in the huntingtin gene (HTT) due to poly-Q expansion cause neuropathology in humans (Huntington’s disease; HD), the normal function(s) of the gene and its protein (HTT) remain obscure. With new information from recently sequenced invertebrate genomes, the study of new animal models opens the possibility of a better understanding of HTT function and its evolution. To these ends, we studied huntingtin expression pattern and dynamics in the invertebrate chordate Ciona intestinalis. Ciona huntingtin (Ci-HTT) shows a biphasic expression pattern during larval development and prior to metamorphosis. A single form of huntingtin protein is present until the early larval stages, at which time two different mass proteins become evident in the metamorphically competent larva. An antibody against Ci-HTT labeled 50 cells in the trunk mesenchyme regions in pre-hatching and hatched larvae and probably represents the distribution of the light form of the protein. Dual labeling with anti-Ci-HTT and anti-aldoketoreductase confirmed the presence of Ci-HTT in mesenchyme cells. Suppression of Ci-HTT RNA by a morpholino oligonucleotide reduced the number and apparent mobility of Ci-HTT positive cells. In Ciona, HTT expression has a dynamic temporal and spatial expression pattern that in ontogeny precedes metamorphosis. Although our results may reflect a derived function for the protein in pre- and post-metamorphic events in Ciona, we also note that as in vertebrates, there is evidence for multiple differential temporal expression, indicating that this protein probably has multiple roles in ontogeny and cell migration.

  13. Sexual and asexual reproduction in the colonial ascidian Botryllus schlosseri.

    PubMed

    Gasparini, Fabio; Manni, Lucia; Cima, Francesca; Zaniolo, Giovanna; Burighel, Paolo; Caicci, Federico; Franchi, Nicola; Schiavon, Filippo; Rigon, Francesca; Campagna, Davide; Ballarin, Loriano

    2015-01-01

    The colonial tunicate Botryllus schlosseri is a widespread filter-feeding ascidian that lives in shallow waters and is easily reared in aquaria. Its peculiar blastogenetic cycle, characterized by the presence of three blastogenetic generations (filtering adults, buds, and budlets) and by recurrent generation changes, has resulted in over 60 years of studies aimed at understanding how sexual and asexual reproduction are coordinated and regulated in the colony. The possibility of using different methodological approaches, from classical genetics to cell transplantation, contributed to the development of this species as a valuable model organism for the study of a variety of biological processes. Here, we review the main studies detailing rearing, staging methods, reproduction and colony growth of this species, emphasizing the asymmetry in sexual and asexual reproduction potential, sexual reproduction in the field and the laboratory, and self- and cross-fertilization. These data, opportunely matched with recent tanscriptomic and genomic outcomes, can give a valuable help to the elucidation of some important steps in chordate evolution. © 2014 Wiley Periodicals, Inc.

  14. Delayed insemination results in embryo mortality in a brooding ascidian.

    PubMed

    Stewart-Savage, J; Phillippi, A; Yund, P O

    2001-08-01

    We explored the effects of temporal variation in sperm availability on fertilization and subsequent larval development in the colonial ascidian Botryllus schlosseri, a brooding hermaphrodite that has a sexual cycle linked to an asexual zooid replacement cycle. We developed a method to quantify the timing of events early in this cycle, and then isolated colonies before the start of the cycle and inseminated them at various times. Colony-wide fertilization levels (assayed by early cleavage) increased from zero to 100% during the period when the siphons of a new generation of zooids were first opening, and remained high for 24 h before slowly declining over the next 48 h. Because embryos are brooded until just before the zooids degenerate at the end of a cycle, delayed fertilization might also affect whether embryos can complete development within the cycle. Consequently, we also determined the effect of delayed insemination on successful embryo development through larval release and metamorphosis. When fertilization was delayed beyond the completion of siphon opening, there was an exponential decline in the percentage of eggs that ultimately produced a metamorphosed larva at the end of the cycle. Thus, even though the majority of oocytes can be fertilized when insemination is delayed for up to 48 h, the resulting embryos cannot complete development before the brooding zooids degenerate.

  15. The larval ascidian nervous system: the chordate brain from its small beginnings.

    PubMed

    Meinertzhagen, I A; Okamura, Y

    2001-07-01

    The body plan of the tadpole larva of ascidians, or sea-squirts, is widely presumed to be close to that of the hypothetical ancestor of all chordate animal groups, including vertebrates. This is nowhere more obvious than in the organization and development of the dorsal tubular nervous system. Several recent developments advocate this model neural system for studies on neurobiology and neurogenesis. These include advances in our understanding of development in ascidian embryos and of differentiation among the cellular progeny of its neural plate; the application of transgenic and mutant approaches to studies on ascidian larval neurones; and the prospect of advances in genomic analyses. In addition to providing ways to study a working chordate brain in miniature, all these offer insights into the ancestral condition of the developing vertebrate brain.

  16. Gut-spilling in chordates: evisceration in the tropical ascidian Polycarpa mytiligera.

    PubMed

    Shenkar, Noa; Gordon, Tal

    2015-04-16

    The ejection of internal organs, i.e., evisceration, is a well-known phenomenon in sea-cucumbers. We report the ability of a member of the Chordate phyla, the tropical ascidian Polycarpa mytiligera, to eviscerate and regenerate its gut within 12 days, and to rebuild its branchial sac within 19 days. Evisceration occurred within 4-43 seconds of gentle mechanical pressure exerted on the tunic in 47% of the tested P. mytiligera. Individuals were able to discard up to 3/4 of their digestive tract via the incurrent siphon by rupture of the branchial sac in this area. Although chemical analysis revealed no significant levels of toxic compounds, the eviscerated guts were unpalatable to the triggerfish and pufferfish on which they were tested, suggesting evisceration as a defense mechanism. Given the close affinity of ascidians to vertebrates, the regeneration pathway of the viscera and branchial sac of ascidians suggests its potential beneficial application in soft tissue regeneration research.

  17. Ascidian recruitment patterns on an artificial reef in Eilat (Red Sea).

    PubMed

    Shenkar, Noa; Zeldman, Yael; Loya, Yossi

    2008-01-01

    Although ascidians are conspicuous members of the fouling community not much is known regarding their recruitment patterns in coral reefs. A 1-year study was carried out along the Red Sea coast of Israel to examine the effects of season and spatial distribution on ascidian recruitment to artificial marine structures. In general, autumn and spring were characterized by higher coverage with a significantly higher percentage of cover of Didemnum granulatum in autumn and higher numbers of Herdmania momus in spring. These species contributed the most to similarity between treatments consequently setting the pattern for each group (colonial and solitary). Halocynthia spinosa had significantly higher numbers during winter and Phallusia nigra was absent in spring and winter. H. momus showed a preference for horizontal surfaces. P. nigra and Ascidia cannelata showed a preference for floating units. It is concluded that the ascidian recruitment patterns are species-specific and vary between seasons, orientation and position on the substrata and in the water column.

  18. Contrasting global genetic patterns in two biologically similar, widespread and invasive Ciona species (Tunicata, Ascidiacea)

    PubMed Central

    Bouchemousse, Sarah; Bishop, John D. D.; Viard, Frédérique

    2016-01-01

    Human-mediated dispersal interplays with natural processes and complicates understanding of the biogeographical history of species. This is exemplified by two invasive tunicates, Ciona robusta (formerly Ciona intestinalis type A) and C. intestinalis (formerly Ciona intestinalis type B), globally distributed and sympatric in Europe. By gathering new mitochondrial sequences that were merged with published datasets, we analysed genetic patterns in different regions, with a focus on 1) their sympatric range and 2) allopatric populations in N and S America and southern Europe. In the sympatric range, the two species display contrasting genetic diversity patterns, with low polymorphism in C. robusta supporting the prevalent view of its recent introduction. In the E Pacific, several genetic traits support the non-native status of C. robusta. However, in the NE Pacific, this appraisal requires a complex scenario of introduction and should be further examined supported by extensive sampling efforts in the NW Pacific (putative native range). For C. intestinalis, Bayesian analysis suggested a natural amphi-North Atlantic distribution, casting doubt on its non-native status in the NW Atlantic. This study shows that both natural and human-mediated dispersal have influenced genetic patterns at broad scales; this interaction lessens our ability to confidently ascertain native vs. non-native status of populations, particularly of those species that are globally distributed. PMID:27137892

  19. Initial deployment of the cardiogenic gene regulatory network in the basal chordate, Ciona intestinalis.

    PubMed

    Woznica, Arielle; Haeussler, Maximilian; Starobinska, Ella; Jemmett, Jessica; Li, Younan; Mount, David; Davidson, Brad

    2012-08-01

    The complex, partially redundant gene regulatory architecture underlying vertebrate heart formation has been difficult to characterize. Here, we dissect the primary cardiac gene regulatory network in the invertebrate chordate, Ciona intestinalis. The Ciona heart progenitor lineage is first specified by Fibroblast Growth Factor/Map Kinase (FGF/MapK) activation of the transcription factor Ets1/2 (Ets). Through microarray analysis of sorted heart progenitor cells, we identified the complete set of primary genes upregulated by FGF/Ets shortly after heart progenitor emergence. Combinatorial sequence analysis of these co-regulated genes generated a hypothetical regulatory code consisting of Ets binding sites associated with a specific co-motif, ATTA. Through extensive reporter analysis, we confirmed the functional importance of the ATTA co-motif in primary heart progenitor gene regulation. We then used the Ets/ATTA combination motif to successfully predict a number of additional heart progenitor gene regulatory elements, including an intronic element driving expression of the core conserved cardiac transcription factor, GATAa. This work significantly advances our understanding of the Ciona heart gene network. Furthermore, this work has begun to elucidate the precise regulatory architecture underlying the conserved, primary role of FGF/Ets in chordate heart lineage specification.

  20. Generation of Germ-Free Ciona intestinalis for Studies of Gut-Microbe Interactions.

    PubMed

    Leigh, Brittany A; Liberti, Assunta; Dishaw, Larry J

    2016-01-01

    Microbes associate with animal hosts, often providing shelter in a nutrient-rich environment. The gut, however, can be a harsh environment with members of the microbiome settling in distinct niches resulting in more stable, adherent biofilms. These diverse communities can provide orders of magnitude more gene products than the host genome; selection and maintenance of a functionally relevant and useful microbiome is now recognized to be an essential component of homeostasis. Germ-free (GF) model systems allow dissection of host-microbe interactions in a simple and direct way where each member of the symbiosis can be studied in isolation. In addition, because immune defenses in the gut are often naïve in GF animals, host immune recognition and responses during the process of colonization can be studied. Ciona intestinalis, a basal chordate, is a well-characterized developmental model system and holds promise for addressing some of these important questions. With transparent juveniles, Ciona can be exposed to distinct bacterial isolates by inoculating GF artificial seawater; concentrated bacteria can subsequently be visualized in vivo if fluorescent stains are utilized. Rearing GF Ciona is a first step in untangling the complex dialogue between bacteria and innate immunity during colonization.

  1. Generation of Germ-Free Ciona intestinalis for Studies of Gut-Microbe Interactions

    PubMed Central

    Leigh, Brittany A.; Liberti, Assunta; Dishaw, Larry J.

    2016-01-01

    Microbes associate with animal hosts, often providing shelter in a nutrient-rich environment. The gut, however, can be a harsh environment with members of the microbiome settling in distinct niches resulting in more stable, adherent biofilms. These diverse communities can provide orders of magnitude more gene products than the host genome; selection and maintenance of a functionally relevant and useful microbiome is now recognized to be an essential component of homeostasis. Germ-free (GF) model systems allow dissection of host-microbe interactions in a simple and direct way where each member of the symbiosis can be studied in isolation. In addition, because immune defenses in the gut are often naïve in GF animals, host immune recognition and responses during the process of colonization can be studied. Ciona intestinalis, a basal chordate, is a well-characterized developmental model system and holds promise for addressing some of these important questions. With transparent juveniles, Ciona can be exposed to distinct bacterial isolates by inoculating GF artificial seawater; concentrated bacteria can subsequently be visualized in vivo if fluorescent stains are utilized. Rearing GF Ciona is a first step in untangling the complex dialogue between bacteria and innate immunity during colonization. PMID:28082961

  2. Ascidians as excellent chordate models for studying the development of the nervous system during embryogenesis and metamorphosis.

    PubMed

    Sasakura, Yasunori; Mita, Kaoru; Ogura, Yosuke; Horie, Takeo

    2012-04-01

    The swimming larvae of the chordate ascidians possess a dorsal hollowed central nervous system (CNS), which is homologous to that of vertebrates. Despite the homology, the ascidian CNS consists of a countable number of cells. The simple nervous system of ascidians provides an excellent experimental system to study the developmental mechanisms of the chordate nervous system. The neural fate of the cells consisting of the ascidian CNS is determined in both autonomous and non-autonomous fashion during the cleavage stage. The ascidian neural plate performs the morphogenetic movement of neural tube closure that resembles that in vertebrate neural tube formation. Following neurulation, the CNS is separated into five distinct regions, whose homology with the regions of vertebrate CNS has been discussed. Following their larval stage, ascidians undergo a metamorphosis and become sessile adults. The metamorphosis is completed quickly, and therefore the metamorphosis of ascidians is a good experimental system to observe the reorganization of the CNS during metamorphosis. A recent study has shown that the major parts of the larval CNS remain after the metamorphosis to form the adult CNS. In contrast to such a conserved manner of CNS reorganization, most larval neurons disappear during metamorphosis. The larval glial cells in the CNS are the major source for the formation of the adult CNS, and some of the glial cells produce adult neurons. © 2012 The Authors Development, Growth & Differentiation © 2012 Japanese Society of Developmental Biologists.

  3. Tunic morphology and viral surveillance in diseased Korean ascidians: Soft tunic syndrome in the edible ascidian, Halocynthia roretzi (Drasche), in aquaculture.

    PubMed

    Kitamura, S-I; Ohtake, S-I; Song, J-Y; Jung, S-J; Oh, M-J; Choi, B-D; Azumi, K; Hirose, E

    2010-02-01

    'Soft tunic syndrome' causes mass mortality in the edible ascidian Halocynthia roretzi in Korean and Japanese aquaculture. In histopathological comparison, there were no specific differences between diseased specimens from Korea and Japan, indicating that soft tunic syndrome occurring in Korea and Japan is the same disease. No bacterial or protozoan cells were microscopically detected in either healthy or diseased tunics suggesting they are not the direct causes of soft tunic syndrome. Attempts were made to isolate virus from affected ascidians taking into account temperature conditions in which soft tunic syndrome is most prevalent in the field. However, no viruses were isolated from diseased or non-diseased specimens using chinook salmon embryo (CHSE-214), flounder fin (FFN) or epithelioma papillosum cyprini (EPC) cell lines.

  4. Multiple origins of the ascidian-Prochloron symbiosis: molecular phylogeny of photosymbiotic and non-symbiotic colonial ascidians inferred from 18S rDNA sequences.

    PubMed

    Yokobori, Shin-Ichi; Kurabayashi, Atsushi; Neilan, Brett A; Maruyama, Tadashi; Hirose, Euichi

    2006-07-01

    In the tropics, certain didemnid ascidians harbor the prokaryotic photosymbiont Prochloron. To date, this photosymbiosis has been found in four didemnid genera that include non-symbiotic species. Here, we report the molecular phylogeny of symbiotic and non-symbiotic didemnids based on their 18S rDNA sequences. The data cover all four genera containing symbiotic species and one other genus comprised of only non-symbiotic species. Near-complete nucleotide sequences of 18S rDNAs were determined for four non-didemnid species and 52 didemnid samples (five genera), including 48 photosymbiotic samples collected from the Ryukyu Archipelago, the Great Barrier Reef, Hawaii, and Bali. Our phylogenetic trees indicated a monophyletic origin of the family Didemnidae, as well as each of the didemnid genera. The results strongly support the hypothesis that establishment of the ascidian-Prochloron symbiosis occurred independently in the Didemnidae lineage at least once in each of the genera that possess symbiotic species.

  5. Blood circulation in the ascidian tunicate Corella inflata (Corellidae)

    PubMed Central

    2016-01-01

    The body of the ascidian tunicate Corella inflata is relatively transparent. Thus, the circulatory system can be visualized by injecting high molecular weight fluorescein labeled dextran into the heart or the large vessels at the ends of the heart without surgery to remove the body wall. In addition, after staining with neutral red, the movement of blood cells can be easily followed to further characterize the circulatory system. The heart is two gently curved concentric tubes extending across the width of the animal. The inner myocardial tube has a partial constriction approximately in the middle. As in other tunicates, the heart is peristaltic and periodically reverses direction. During the branchial phase blood leaves the anterior end of the heart by two asymmetric vessels that connect to the two sides of the branchial basket. Blood then flows in both transverse directions through a complex system of ducts in the basket into large ventral and dorsal vessels which carry blood back to the visceral organs in the posterior of the animal. During the visceral phase blood leaves the posterior end of the heart in two vessels that repeatedly bifurcate and fan into the stomach and gonads. Blood velocity, determined by following individual cells in video frames, is high and pulsatory near the heart. A double peak in velocity at the maximum may be due to the constriction in the middle of the heart tube. Blood velocity progressively decreases with distance from the heart. In peripheral regions with vessels of small diameter blood cells frequently collide with vessel walls and cell motion is erratic. The estimated volume of blood flow during each directional phase is greater than the total volume of the animal. Circulating blood cells are confined to vessels or ducts in the visible parts of the animal and retention of high molecular weight dextran in the vessels is comparable to that seen in vertebrates. These are characteristics of a closed circulatory system. PMID:27994977

  6. Blood circulation in the ascidian tunicate Corella inflata (Corellidae).

    PubMed

    Konrad, Michael W

    2016-01-01

    The body of the ascidian tunicate Corella inflata is relatively transparent. Thus, the circulatory system can be visualized by injecting high molecular weight fluorescein labeled dextran into the heart or the large vessels at the ends of the heart without surgery to remove the body wall. In addition, after staining with neutral red, the movement of blood cells can be easily followed to further characterize the circulatory system. The heart is two gently curved concentric tubes extending across the width of the animal. The inner myocardial tube has a partial constriction approximately in the middle. As in other tunicates, the heart is peristaltic and periodically reverses direction. During the branchial phase blood leaves the anterior end of the heart by two asymmetric vessels that connect to the two sides of the branchial basket. Blood then flows in both transverse directions through a complex system of ducts in the basket into large ventral and dorsal vessels which carry blood back to the visceral organs in the posterior of the animal. During the visceral phase blood leaves the posterior end of the heart in two vessels that repeatedly bifurcate and fan into the stomach and gonads. Blood velocity, determined by following individual cells in video frames, is high and pulsatory near the heart. A double peak in velocity at the maximum may be due to the constriction in the middle of the heart tube. Blood velocity progressively decreases with distance from the heart. In peripheral regions with vessels of small diameter blood cells frequently collide with vessel walls and cell motion is erratic. The estimated volume of blood flow during each directional phase is greater than the total volume of the animal. Circulating blood cells are confined to vessels or ducts in the visible parts of the animal and retention of high molecular weight dextran in the vessels is comparable to that seen in vertebrates. These are characteristics of a closed circulatory system.

  7. Ascidian (Chordata-Tunicata) glycosaminoglycans: extraction, purification, biochemical, and spectroscopic analysis.

    PubMed

    Pavão, Mauro S G

    2015-01-01

    Sulfated polysaccharides with unique structures of the chondroitin/dermatan and heparin/heparan families of sulfated glycosaminoglycans have been described in several species of ascidians (Chordata-Tunicata). These unique sulfated glycans have been isolated from-ascidians and characterized by biochemical and spectroscopic methods. The ascidian glycans can be extracted by different tissues or cells by proteolytic digestion followed by cetylpyridinium chloride/ethanol precipitation. The total glycans are then fractionated by ion-exchange chromatography on DEAE-cellulose and/or Mono Q (HR 5/5) columns. Alternatively, precipitation with different ethanol concentrations can be employed. An initial analysis of the purified ascidian glycans is carried out by agarose gel electrophoresis on diaminopropane/acetate buffer, before or after digestion with specific glycosaminoglycan lyases or deaminative cleavage with nitrous acid. The disaccharides formed by exhaustive degradation of the glycans is purified by gel-filtration chromatography on a Superdex-peptide column and analyzed by HPLC on a strong ion exchange Sax-Spherisorb column. 1H or 13C nuclear magnetic resonance spectroscopy in one or two dimensions is used to confirm the structure of the intact glycans.

  8. Muscle differentiation in a colonial ascidian: organisation, gene expression and evolutionary considerations

    PubMed Central

    Degasperi, Valentina; Gasparini, Fabio; Shimeld, Sebastian M; Sinigaglia, Chiara; Burighel, Paolo; Manni, Lucia

    2009-01-01

    Background Ascidians are tunicates, the taxon recently proposed as sister group to the vertebrates. They possess a chordate-like swimming larva, which metamorphoses into a sessile adult. Several ascidian species form colonies of clonal individuals by asexual reproduction. During their life cycle, ascidians present three muscle types: striated in larval tail, striated in the heart, and unstriated in the adult body-wall. Results In the colonial ascidian Botryllus schlosseri, we investigated organisation, differentiation and gene expression of muscle beginning from early buds to adults and during zooid regression. We characterised transcripts for troponin T (BsTnT-c), adult muscle-type (BsMA2) and cytoplasmic-type (BsCA1) actins, followed by in situ hybridisation (ISH) on sections to establish the spatio-temporal expression of BsTnT-c and BsMA2 during asexual reproduction and in the larva. Moreover, we characterised actin genomic sequences, which by comparison with other metazoans revealed conserved intron patterns. Conclusion Integration of data from ISH, phalloidin staining and TEM allowed us to follow the phases of differentiation of the three muscle kinds, which differ in expression pattern of the two transcripts. Moreover, phylogenetic analyses provided evidence for the close relationship between tunicate and vertebrate muscle genes. The characteristics and plasticity of muscles in tunicates are discussed. PMID:19737381

  9. Natural Products from Antarctic Colonial Ascidians of the Genera Aplidium and Synoicum: Variability and Defensive Role

    PubMed Central

    Núñez-Pons, Laura; Carbone, Marianna; Vázquez, Jennifer; Rodríguez, Jaime; Nieto, Rosa María; Varela, María Mercedes; Gavagnin, Margherita; Avila, Conxita

    2012-01-01

    Ascidians have developed multiple defensive strategies mostly related to physical, nutritional or chemical properties of the tunic. One of such is chemical defense based on secondary metabolites. We analyzed a series of colonial Antarctic ascidians from deep-water collections belonging to the genera Aplidium and Synoicum to evaluate the incidence of organic deterrents and their variability. The ether fractions from 15 samples including specimens of the species A. falklandicum, A. fuegiense, A. meridianum, A. millari and S. adareanum were subjected to feeding assays towards two relevant sympatric predators: the starfish Odontaster validus, and the amphipod Cheirimedon femoratus. All samples revealed repellency. Nonetheless, some colonies concentrated defensive chemicals in internal body-regions rather than in the tunic. Four ascidian-derived meroterpenoids, rossinones B and the three derivatives 2,3-epoxy-rossinone B, 3-epi-rossinone B, 5,6-epoxy-rossinone B, and the indole alkaloids meridianins A–G, along with other minoritary meridianin compounds were isolated from several samples. Some purified metabolites were tested in feeding assays exhibiting potent unpalatabilities, thus revealing their role in predation avoidance. Ascidian extracts and purified compound-fractions were further assessed in antibacterial tests against a marine Antarctic bacterium. Only the meridianins showed inhibition activity, demonstrating a multifunctional defensive role. According to their occurrence in nature and within our colonial specimens, the possible origin of both types of metabolites is discussed. PMID:23015772

  10. Cell type and function of neurons in the ascidian nervous system.

    PubMed

    Horie, Takeo; Nakagawa, Masashi; Sasakura, Yasunori; Kusakabe, Takehiro G

    2009-04-01

    Ascidians, or sea squirts, are primitive chordates, and their tadpole larvae share a basic body plan with vertebrates, including a notochord and a dorsal tubular central nervous system (CNS). The CNS of the ascidian larva is formed through a process similar to vertebrate neurulation, while the ascidian CNS is remarkably simple, consisting of about 100 neurons. Recent identification of genes that are specifically expressed in a particular subtype of neurons has enabled us to reveal neuronal networks at single-cell resolution. Based on the information on neuron subtype-specific genes, different populations of neurons have been visualized by whole-mount in situ hybridization, immunohistochemical staining using specific antibodies, and fluorescence labeling of cell bodies and neurites by a fluorescence protein reporter driven by neuron-specific promoters. Neuronal populations that have been successfully visualized include glutamatergic, cholinergic, gamma-aminobutyric acid/glycinergic, and dopaminergic neurons, which have allowed us to propose functional regionalization of the CNS and a neural circuit for locomotion. Thus, the simple nervous system of the ascidian larva can serve as an attractive model system for studying the development, function, and evolution of the chordate nervous system.

  11. Patellamide E: a new cyclic peptide from the ascidian Lissoclinum patella.

    PubMed

    McDonald, L A; Ireland, C M

    1992-03-01

    A new cyclic peptide, patellamide E [1], was isolated from the ascidian Lissoclinum patella collected at Pulau Salu, Singapore. Its structure was determined by nmr spectroscopy, and its absolute configuration by acid hydrolysis and analysis of the derivatized constituent amino acids by hplc. Patellamide E was mildly cytotoxic against human colon tumor cells in vitro.

  12. The evolution of the vertebrate metzincins; insights from Ciona intestinalis and Danio rerio

    PubMed Central

    Huxley-Jones, Julie; Clarke, Toni-Kim; Beck, Christine; Toubaris, George; Robertson, David L; Boot-Handford, Raymond P

    2007-01-01

    Background The metzincins are a large gene superfamily of proteases characterized by the presence of a zinc protease domain, and include the ADAM, ADAMTS, BMP1/TLL, meprin and MMP genes. Metzincins are involved in the proteolysis of a wide variety of proteins, including those of the extracellular matrix. The metzincin gene superfamily comprises eighty proteins in the human genome and ninety-three in the mouse. When and how the level of complexity apparent in the vertebrate metzincin gene superfamily arose has not been determined in detail. Here we present a comprehensive analysis of vertebrate metzincins using genes from both Ciona intestinalis and Danio rerio to provide new insights into the complex evolution of this gene superfamily. Results We have identified 19 metzincin genes in the ciona genome and 83 in the zebrafish genome. Phylogenetic analyses reveal that the expansion of the metzincin gene superfamily in vertebrates has occurred predominantly by the simple duplication of pre-existing genes rather than by the appearance and subsequent expansion of new metzincin subtypes (the only example of which is the meprin gene family). Despite the number of zebrafish metzincin genes being relatively similar to that of tetrapods (e.g. man and mouse), the pattern of gene retention and loss within these lineages is markedly different. In addition, we have studied the evolution of the related TIMP gene family and identify a single ciona and four zebrafish TIMP genes. Conclusion The complexity seen in the vertebrate metzincin gene families was mainly acquired during vertebrate evolution. The metzincin gene repertoire in protostomes and invertebrate deuterostomes has remained relatively stable. The expanded metzincin gene repertoire of extant tetrapods, such as man, has resulted largely from duplication events associated with early vertebrate evolution, prior to the sarcopterygian-actinopterygian split. The teleost repertoire of metzincin genes in part parallels that of

  13. Dissection of a Ciona regulatory element reveals complexity of cross-species enhancer activity

    PubMed Central

    Chen, Wei-Chung; Pauls, Stefan; Bacha, Jamil; Elgar, Greg; Loose, Matthew; Shimeld, Sebastian M.

    2014-01-01

    Vertebrate genomes share numerous conserved non-coding elements, many of which function as enhancer elements and are hypothesised to be under evolutionary constraint due to a need to be bound by combinations of sequence-specific transcription factors. In contrast, few such conserved elements can be detected between vertebrates and their closest invertebrate relatives. Despite this lack of sequence identity, cross-species transgenesis has identified some cases where non-coding DNA from invertebrates drives reporter gene expression in transgenic vertebrates in patterns reminiscent of the expression of vertebrate orthologues. Such instances are presumed to reflect the presence of conserved suites of binding sites in the regulatory regions of invertebrate and vertebrate orthologues, such that both regulatory elements can correctly interpret the trans-activating environment. Shuffling of binding sites has been suggested to lie behind loss of sequence conservation; however this has not been experimentally tested. Here we examine the underlying basis of enhancer activity for the Ciona intestinalis βγ-crystallin gene, which drives expression in the lens of transgenic vertebrates despite the Ciona lineage predating the evolution of the lens. We construct an interactive gene regulatory network (GRN) for vertebrate lens development, allowing network interactions to be robustly catalogued and conserved network components and features to be identified. We show that a small number of binding motifs are necessary for Ciona βγ-crystallin expression, and narrow down the likely factors that bind to these motifs. Several of these overlap with the conserved core of the vertebrate lens GRN, implicating these sites in cross species function. However when we test these motifs in a transgenic vertebrate they prove to be dispensable for reporter expression in the lens. These results show that current models depicting cross species enhancer function as dependent on conserved binding

  14. Stochasticity in space, persistence in time: genetic heterogeneity in harbour populations of the introduced ascidian Styela plicata

    PubMed Central

    Pineda, Mari-Carmen; Lorente, Beatriz; López-Legentil, Susanna; Palacín, Creu

    2016-01-01

    Spatio-temporal changes in genetic structure among populations provide crucial information on the dynamics of secondary spread for introduced marine species. However, temporal components have rarely been taken into consideration when studying the population genetics of non-indigenous species. This study analysed the genetic structure of Styela plicata, a solitary ascidian introduced in harbours and marinas of tropical and temperate waters, across spatial and temporal scales. A fragment of the mitochondrial gene Cytochrome Oxidase subunit I (COI) was sequenced from 395 individuals collected at 9 harbours along the NW Mediterranean coast and adjacent Atlantic waters (> 1,200 km range) at two time points 5 years apart (2009 and 2014). The levels of gene diversity were relatively low for all 9 locations in both years. Analyses of genetic differentiation and distribution of molecular variance revealed strong genetic structure, with significant differences among many populations, but no significant differences among years. A weak and marginally significant correlation between geographic distance and gene differentiation was found. Our results revealed spatial structure and temporal genetic homogeneity in S. plicata, suggesting a limited role of recurrent, vessel-mediated transport of organisms among small to medium-size harbours. Our study area is representative of many highly urbanized coasts with dense harbours. In these environments, the episodic chance arrival of colonisers appears to determine the genetic structure of harbour populations and the genetic composition of these early colonising individuals persists in the respective harbours, at least over moderate time frames (five years) that encompass ca. 20 generations of S. plicata. PMID:27366653

  15. Stochasticity in space, persistence in time: genetic heterogeneity in harbour populations of the introduced ascidian Styela plicata.

    PubMed

    Pineda, Mari-Carmen; Lorente, Beatriz; López-Legentil, Susanna; Palacín, Creu; Turon, Xavier

    2016-01-01

    Spatio-temporal changes in genetic structure among populations provide crucial information on the dynamics of secondary spread for introduced marine species. However, temporal components have rarely been taken into consideration when studying the population genetics of non-indigenous species. This study analysed the genetic structure of Styela plicata, a solitary ascidian introduced in harbours and marinas of tropical and temperate waters, across spatial and temporal scales. A fragment of the mitochondrial gene Cytochrome Oxidase subunit I (COI) was sequenced from 395 individuals collected at 9 harbours along the NW Mediterranean coast and adjacent Atlantic waters (> 1,200 km range) at two time points 5 years apart (2009 and 2014). The levels of gene diversity were relatively low for all 9 locations in both years. Analyses of genetic differentiation and distribution of molecular variance revealed strong genetic structure, with significant differences among many populations, but no significant differences among years. A weak and marginally significant correlation between geographic distance and gene differentiation was found. Our results revealed spatial structure and temporal genetic homogeneity in S. plicata, suggesting a limited role of recurrent, vessel-mediated transport of organisms among small to medium-size harbours. Our study area is representative of many highly urbanized coasts with dense harbours. In these environments, the episodic chance arrival of colonisers appears to determine the genetic structure of harbour populations and the genetic composition of these early colonising individuals persists in the respective harbours, at least over moderate time frames (five years) that encompass ca. 20 generations of S. plicata.

  16. Amphioxus and ascidian Dmbx homeobox genes give clues to the vertebrate origins of midbrain development.

    PubMed

    Takahashi, Tokiharu; Holland, Peter W H

    2004-07-01

    The ancestral chordate neural tube had a tripartite structure, comprising anterior, midbrain-hindbrain boundary (MHB) and posterior regions. The most anterior region encompasses both forebrain and midbrain in vertebrates. It is not clear when or how the distinction between these two functionally and developmentally distinct regions arose in evolution. Recently, we reported a mouse PRD-class homeobox gene, Dmbx1, expressed in the presumptive midbrain at early developmental stages, and the hindbrain at later stages, with exclusion from the MHB. This gene provides a route to investigate the evolution of midbrain development. We report the cloning, genomic structure, phylogeny and embryonic expression of Dmbx genes from amphioxus and from Ciona, representing the two most closely related lineages to the vertebrates. Our analyses show that Dmbx genes form a distinct, ancient, homeobox gene family, with highly conserved sequence and genomic organisation, albeit more divergent in Ciona. In amphioxus, no Dmbx expression is observed in the neural tube, supporting previous arguments that the MHB equivalent region has been secondarily modified in evolution. In Ciona, the CiDmbx gene is detected in neural cells caudal to Pax2/5/8-positive cells (MHB homologue), in the Hox-positive region, but, interestingly, not in any cells rostral to them. These results suggest that a midbrain homologue is missing in Ciona, and argue that midbrain development is a novelty that evolved specifically on the vertebrate lineage. We discuss the evolution of midbrain development in relation to the ancestry of the tripartite neural ground plan and the origin of the MHB organiser.

  17. Mediterranean diet

    MedlinePlus

    ... provider if you should take a calcium supplement. Wine is a common part of a Mediterranean eating ... but some people should not drink alcohol. Avoid wine if you are prone to alcohol abuse, pregnant, ...

  18. Endozoicomonas Are Specific, Facultative Symbionts of Sea Squirts

    PubMed Central

    Schreiber, Lars; Kjeldsen, Kasper U.; Funch, Peter; Jensen, Jeppe; Obst, Matthias; López-Legentil, Susanna; Schramm, Andreas

    2016-01-01

    Ascidians are marine filter feeders and harbor diverse microbiota that can exhibit a high degree of host-specificity. Pharyngeal samples of Scandinavian and Mediterranean ascidians were screened for consistently associated bacteria by culture-dependent and -independent approaches. Representatives of the Endozoicomonas (Gammaproteobacteria, Hahellaceae) clade were detected in the ascidian species Ascidiella aspersa, Ascidiella scabra, Botryllus schlosseri, Ciona intestinalis, Styela clava, and multiple Ascidia/Ascidiella spp. In total, Endozoicomonas was detected in more than half of all specimens screened, and in 25–100% of the specimens for each species. The retrieved Endozoicomonas 16S rRNA gene sequences formed an ascidian-specific subclade, whose members were detected by fluorescence in situ hybridization (FISH) as extracellular microcolonies in the pharynx. Two strains of the ascidian-specific Endozoicomonas subclade were isolated in pure culture and characterized. Both strains are chemoorganoheterotrophs and grow on mucin (a mucus glycoprotein). The strains tested negative for cytotoxic or antibacterial activity. Based on these observations, we propose ascidian-associated Endozoicomonas to be commensals, living off the mucus continuously secreted into the pharynx. Members of the ascidian-specific Endozoicomonas subclade were also detected in seawater from the Scandinavian sampling site, which suggests acquisition of the symbionts by horizontal transmission. The combined results indicate a host-specific, yet facultative symbiosis between ascidians and Endozoicomonas. PMID:27462299

  19. Endozoicomonas Are Specific, Facultative Symbionts of Sea Squirts.

    PubMed

    Schreiber, Lars; Kjeldsen, Kasper U; Funch, Peter; Jensen, Jeppe; Obst, Matthias; López-Legentil, Susanna; Schramm, Andreas

    2016-01-01

    Ascidians are marine filter feeders and harbor diverse microbiota that can exhibit a high degree of host-specificity. Pharyngeal samples of Scandinavian and Mediterranean ascidians were screened for consistently associated bacteria by culture-dependent and -independent approaches. Representatives of the Endozoicomonas (Gammaproteobacteria, Hahellaceae) clade were detected in the ascidian species Ascidiella aspersa, Ascidiella scabra, Botryllus schlosseri, Ciona intestinalis, Styela clava, and multiple Ascidia/Ascidiella spp. In total, Endozoicomonas was detected in more than half of all specimens screened, and in 25-100% of the specimens for each species. The retrieved Endozoicomonas 16S rRNA gene sequences formed an ascidian-specific subclade, whose members were detected by fluorescence in situ hybridization (FISH) as extracellular microcolonies in the pharynx. Two strains of the ascidian-specific Endozoicomonas subclade were isolated in pure culture and characterized. Both strains are chemoorganoheterotrophs and grow on mucin (a mucus glycoprotein). The strains tested negative for cytotoxic or antibacterial activity. Based on these observations, we propose ascidian-associated Endozoicomonas to be commensals, living off the mucus continuously secreted into the pharynx. Members of the ascidian-specific Endozoicomonas subclade were also detected in seawater from the Scandinavian sampling site, which suggests acquisition of the symbionts by horizontal transmission. The combined results indicate a host-specific, yet facultative symbiosis between ascidians and Endozoicomonas.

  20. Matrix adhesion polarizes heart progenitor induction in the invertebrate chordate Ciona intestinalis

    PubMed Central

    Norton, Jennifer; Cooley, James; Islam, A. F. M. Tariqul; Cota, Christina D.; Davidson, Brad

    2013-01-01

    Cell-matrix adhesion strongly influences developmental signaling. Resulting impacts on cell migration and tissue morphogenesis are well characterized. However, the in vivo impact of adhesion on fate induction remains ambiguous. Here, we employ the invertebrate chordate Ciona intestinalis to delineate an essential in vivo role for matrix adhesion in heart progenitor induction. In Ciona pre-cardiac founder cells, invasion of the underlying epidermis promotes localized induction of the heart progenitor lineage. We found that these epidermal invasions are associated with matrix adhesion along the pre-cardiac cell/epidermal boundary. Through targeted manipulations of RAP GTPase activity, we were able to manipulate pre-cardiac cell-matrix adhesion. Targeted disruption of pre-cardiac cell-matrix adhesion blocked heart progenitor induction. Conversely, increased matrix adhesion generated expanded induction. We were also able to selectively restore cell-matrix adhesion and heart progenitor induction through targeted expression of Ci-Integrin β2. These results indicate that matrix adhesion functions as a necessary and sufficient extrinsic cue for regional heart progenitor induction. Furthermore, time-lapse imaging suggests that cytokinesis acts as an intrinsic temporal regulator of heart progenitor adhesion and induction. Our findings highlight a potentially conserved role for matrix adhesion in early steps of vertebrate heart progenitor specification. PMID:23444358

  1. Matrix adhesion polarizes heart progenitor induction in the invertebrate chordate Ciona intestinalis.

    PubMed

    Norton, Jennifer; Cooley, James; Islam, A F M Tariqul; Cota, Christina D; Davidson, Brad

    2013-03-01

    Cell-matrix adhesion strongly influences developmental signaling. Resulting impacts on cell migration and tissue morphogenesis are well characterized. However, the in vivo impact of adhesion on fate induction remains ambiguous. Here, we employ the invertebrate chordate Ciona intestinalis to delineate an essential in vivo role for matrix adhesion in heart progenitor induction. In Ciona pre-cardiac founder cells, invasion of the underlying epidermis promotes localized induction of the heart progenitor lineage. We found that these epidermal invasions are associated with matrix adhesion along the pre-cardiac cell/epidermal boundary. Through targeted manipulations of RAP GTPase activity, we were able to manipulate pre-cardiac cell-matrix adhesion. Targeted disruption of pre-cardiac cell-matrix adhesion blocked heart progenitor induction. Conversely, increased matrix adhesion generated expanded induction. We were also able to selectively restore cell-matrix adhesion and heart progenitor induction through targeted expression of Ci-Integrin β2. These results indicate that matrix adhesion functions as a necessary and sufficient extrinsic cue for regional heart progenitor induction. Furthermore, time-lapse imaging suggests that cytokinesis acts as an intrinsic temporal regulator of heart progenitor adhesion and induction. Our findings highlight a potentially conserved role for matrix adhesion in early steps of vertebrate heart progenitor specification.

  2. Expression of neuropeptide- and hormone-encoding genes in the Ciona intestinalis larval brain.

    PubMed

    Hamada, Mayuko; Shimozono, Naoki; Ohta, Naoyuki; Satou, Yutaka; Horie, Takeo; Kawada, Tsuyoshi; Satake, Honoo; Sasakura, Yasunori; Satoh, Nori

    2011-04-15

    Despite containing only approximately 330 cells, the central nervous system (CNS) of Ciona intestinalis larvae has an architecture that is similar to the vertebrate CNS. Although only vertebrates have a distinct hypothalamus-the source of numerous neurohormone peptides that play pivotal roles in the development, function, and maintenance of various neuronal and endocrine systems, it is suggested that the Ciona brain contains a region that corresponds to the vertebrate hypothalamus. To identify genes expressed in the brain, we isolated brain vesicles using transgenic embryos carrying Ci-β-tubulin(promoter)::Kaede, which resulted in robust Kaede expression in the larval CNS. The associated transcriptome was investigated using microarray analysis. We identified 565 genes that were preferentially expressed in the larval brain. Among these genes, 11 encoded neurohormone peptides including such hypothalamic peptides as gonadotropin-releasing hormone and oxytocin/vasopressin. Six of the identified peptide genes had not been previously described. We also found that genes encoding receptors for some of the peptides were expressed in the brain. Interestingly, whole-mount in situ hybridization showed that most of the peptide genes were expressed in the ventral brain. This catalog of the genes expressed in the larval brain should help elucidate the evolution, development, and functioning of the chordate brain.

  3. Immune competence of the Ciona intestinalis pharynx: complement system-mediated activity.

    PubMed

    Giacomelli, Stefano; Melillo, Daniela; Lambris, John D; Pinto, Maria Rosaria

    2012-10-01

    In the tunicate Ciona intestinalis, the ciliated pharynx, which connects the external environment to a highly developed and compartmentalized gastrointestinal system, represents the natural portal of entry for a vast and diverse, potentially pathogenic microbial community. To address the role of the pharynx in immune surveillance in Ciona, we asked whether C3, the key component of the complement system, was expressed in this organ and whether the encoded protein was functionally active. We found by real-time PCR that C3, constitutively expressed in the pharynx, is up-regulated by LPS injection. Using two specific anti-CiC3 and anti-CiC3a polyclonal antibodies in immunohistochemical staining of pharynx sections, we found that the gene product was localized to hemocytes of the pharyngeal bars (identified as granular amoebocytes) and in stigmata ciliated cells. Use of the same antibodies in Western blot analysis indicated that CiC3 and its activation products CiC3b and CiC3a are present in pharynx homogenates. Our observation that the amount of the bioactive fragment CiC3a increased in the pharynx of LPS-treated animals provides the first molecular and functional evidence for complement-mediated immunological activity in the tunicate pharynx. Copyright © 2012 Elsevier Ltd. All rights reserved.

  4. Life in cellulose houses: symbiotic bacterial biosynthesis of ascidian drugs and drug leads.

    PubMed

    Schmidt, Eric W; Donia, Mohamed S

    2010-12-01

    Ascidians (tunicates; sea squirts) are sources of diverse, bioactive natural products, one of which is an approved drug and many of which are potent drug leads. It has been shown that symbiotic bacteria living with ascidians produce some of the bioactive compounds isolated from whole animals, and indirect evidence strongly implicates symbiotic bacteria in the synthesis of many others. However, for the majority the producing organism has not been identified. In cases where a symbiotic origin has been definitively assigned, the resulting data lead to improved paths to drug discovery and development from marine animals. This review traces evidence for symbiotic production where such evidence exists and describes the strengths and limitations of that evidence. Copyright © 2010 Elsevier Ltd. All rights reserved.

  5. Ammonia-oxidizing Crenarchaeota and nitrification inside the tissue of a colonial ascidian.

    PubMed

    Martínez-García, Manuel; Stief, Peter; Díaz-Valdés, Marta; Wanner, Gerhard; Ramos-Esplá, Alfonso; Dubilier, Nicole; Antón, Josefa

    2008-11-01

    Marine Crenarchaeota represent an abundant component of the oceanic microbiota that play an important role in the global nitrogen cycle. Here we report the association of the colonial ascidian Cystodytes dellechiajei with putative ammonia-oxidizing Crenarchaeota that could actively be involved in nitrification inside the animal tissue. As shown by 16S rRNA gene analysis, the ascidian-associated Crenarchaeota were phylogenetically related to Nitrosopumilus maritimus, the first marine archaeon isolated in pure culture that grows chemolithoautotrophically oxidizing ammonia to nitrite aerobically. Catalysed reporter deposition (CARD)-FISH revealed that the Crenarchaeota were specifically located inside the tunic tissue of the colony, where moreover the expression of amoA gene was detected. The amoA gene encodes the alpha-subunit of ammonia monooxygenase, which is involved in the first step of nitrification, the oxidation of ammonia to nitrite. Sequencing of amoA gene showed that they were phylogenetically related to amoA genes of N. maritimus and other putative ammonia-oxidizing marine Crenarchaeota. In order to track the suspected nitrification activity inside the ascidian colony under in vivo conditions, microsensor profiles were measured through the tunic tissue. Net NO(x) production was detected in the tunic layer 1200-1800 microm with rates of 58-90 nmol cm(-3) h(-1). Oxygen and pH microsensor profiles showed that the layer of net NO(x) production coincided with O(2) concentrations of 103-116 microM and pH value of 5.2. Together, molecular and microsensor data indicate that Crenarchaeota could oxidize ammonia to nitrite aerobically, and thus be involved in nitrification inside the ascidian tissue.

  6. When shape matters: strategies of different Antarctic ascidians morphotypes to deal with sedimentation.

    PubMed

    Torre, Luciana; Abele, Doris; Lagger, Cristian; Momo, Fernando; Sahade, Ricardo

    2014-08-01

    Climate change leads to increased melting of tidewater glaciers in the Western Antarctic Peninsula region and sediment bearing glacial melt waters negatively affects filter feeding species as solitary ascidians. In previous work the erect-forms Molgula pedunculata and Cnemidocarpa verrucosa (Order Stolidobranchiata) appeared more sensitive than the flat form Ascidia challengeri (Order Phlebobranchiata). Sedimentation exposure is expected to induce up-regulation of anaerobic metabolism by obstructing the organs of gas exchange (environmental hypoxia) or causes enhanced squirting activity (functional hypoxia). In this study we evaluated the possible relationship between ascidian morphotype and their physiological response to sedimentation. Together with some behavioural observations, we analysed the response of anaerobic metabolic parameters (lactate formation and glycogen consumption) in different tissues of three Antarctic ascidians, exposed to high sediment concentrations (200 mgL(-1)). The results were compared to experimental hypoxia (10% pO2) and exercise (induced muscular contraction) effects, in order to discriminate the effect of sediment on each species and morpho-type (erect vs. flat forms). Our results suggest that the styled (erect) C. verrucosa increases muscular squirting activity in order to expulse excessive material, while the flat-form A. challengeri reacts more passively by down-regulating its aerobic metabolism under sediment exposure. Contrary, the erect ascidian M. pedunculata did not show any measurable response to the treatments, indicating that filtration and ingestion activities were not reduced or altered even under high sedimentation (low energetic material) which could be disadvantageous on the long-term and could explain why M. pedunculata densities decline in the study area. Copyright © 2014 Elsevier Ltd. All rights reserved.

  7. Synoxazolidinones A and B: novel bioactive alkaloids from the ascidian Synoicum pulmonaria.

    PubMed

    Tadesse, Margey; Strøm, Morten B; Svenson, Johan; Jaspars, Marcel; Milne, Bruce F; Tørfoss, Veronika; Andersen, Jeanette H; Hansen, Espen; Stensvåg, Klara; Haug, Tor

    2010-11-05

    Bioassay-guided fractionation of the sub-Arctic ascidian Synoicum pulmonaria collected off the Norwegian coast led to the isolation of a novel family of brominated guanidinium oxazolidinones named synoxazolidinones A and B (1 and 2). The backbone of the compounds contains a 4-oxazolidinone ring rarely seen in natural products. The structure of the compounds was determined by spectroscopic methods. The synoxazolidinones exhibited antibacterial and antifungal activities.

  8. Distribution and Localised Effects of the Invasive Ascidian Didemnum perlucidum (Monniot 1983) in an Urban Estuary.

    PubMed

    Simpson, Tiffany Schenk; Wernberg, Thomas; McDonald, Justin I

    2016-01-01

    Didemnid ascidians are notorious marine invaders, fouling infrastructure in many ecosystems globally. However, there have been few reports of direct interactions with native species in their natural environment. The invasive colonial ascidian Didemnum perlucidum was discovered in the Swan River estuary (Western Australia) growing on the native seagrass Halophila ovalis. Given the known effects of other related Didemnum species it was expected that D. perlucidum could adversely affect the seagrass, with possible flow on effects to the rest of the ecosystem. This study aimed to document the distribution and abundance of D. perlucidum in the estuary, and to determine whether this species had a negative impact on H. ovalis or associated flora and fauna. D. perlucidum was largely present near areas of infrastructure, particularly mooring buoys, suggesting these were the source of D. perlucidum recruits on the seagrasses. It showed a clear seasonal pattern in abundance, with highly variable cover and colony size. D. perlucidum had a measurable effect on H. ovalis, with colonies enveloping all plant tissue, likely restricting the photosynthetic ability of individual leaves and total plant biomass. There were also significantly less seagrass-associated mud snails (Batillaria australis) where D. perlucidum cover was high. These results demonstrate the ability of invasive ascidians to colonise and affect native seagrasses and associated biota. Seagrasses are pivotal to the ecological function of many urban estuaries world-wide. Biodiversity in these systems is already vulnerable to multiple stressors from human activities but the potential stress of fouling ascidians may pose an additional and increasing threat in the future.

  9. Shallow-water Ascidians from Matua Island (central Kuril Islands, NW Pacific).

    PubMed

    Sanamyan, Karen; Sanamyan, Nadya

    2017-02-16

    Fifteen species of ascidians were identified in the material collected at Matua Island. Two species are new, Botryllus flavus n. sp. and Distaplia matua n. sp. The first species occurs also at Kamchatka waters, while the second is probably an endemic of Kuril Islands. The genus Macrenteron Redikorzev, 1927 is synonymized with Aplidium Savigny, 1816 and a new name Aplidium macrenteron nom. nov. is proposed for its type species.

  10. Quinone and Hydroquinone Metabolites from the Ascidians of the Genus Aplidium

    PubMed Central

    Bertanha, Camila Spereta; Januário, Ana Helena; Alvarenga, Tavane Aparecida; Pimenta, Letícia Pereira; e Silva, Márcio Luis Andrade; Cunha, Wilson Roberto; Pauletti, Patrícia Mendonça

    2014-01-01

    Ascidians of the genus Aplidium are recognized as an important source of chemical diversity and bioactive natural products. Among the compounds produced by this genus are non-nitrogenous metabolites, mainly prenylated quinones and hydroquinones. This review discusses the isolation, structural elucidation, and biological activities of quinones, hydroquinones, rossinones, longithorones, longithorols, floresolides, scabellones, conicaquinones, aplidinones, thiaplidiaquinones, and conithiaquinones. A compilation of the 13C-NMR spectral data of these compounds is also presented. PMID:24927227

  11. The alien ascidian Styela clava now invading the Sea of Marmara (Tunicata: Ascidiacea).

    PubMed

    Çinar, Melih Ertan

    2016-01-01

    During the implementation of a large project aimed to investigate the benthic community structures of the Sea of Marmara, specimens of the invasive ascidian species Styela clava were collected on natural substrata (rocks) at 10 m depth at one locality (Karamürsel) in İzmit Bay. The specimens were mature, containing gametes, indicating that the species had become established in the area. The Sea of Marmara seems to provide suitable conditions for this species to survive and form proliferating populations.

  12. Distribution and Localised Effects of the Invasive Ascidian Didemnum perlucidum (Monniot 1983) in an Urban Estuary

    PubMed Central

    Wernberg, Thomas; McDonald, Justin I.

    2016-01-01

    Didemnid ascidians are notorious marine invaders, fouling infrastructure in many ecosystems globally. However, there have been few reports of direct interactions with native species in their natural environment. The invasive colonial ascidian Didemnum perlucidum was discovered in the Swan River estuary (Western Australia) growing on the native seagrass Halophila ovalis. Given the known effects of other related Didemnum species it was expected that D. perlucidum could adversely affect the seagrass, with possible flow on effects to the rest of the ecosystem. This study aimed to document the distribution and abundance of D. perlucidum in the estuary, and to determine whether this species had a negative impact on H. ovalis or associated flora and fauna. D. perlucidum was largely present near areas of infrastructure, particularly mooring buoys, suggesting these were the source of D. perlucidum recruits on the seagrasses. It showed a clear seasonal pattern in abundance, with highly variable cover and colony size. D. perlucidum had a measurable effect on H. ovalis, with colonies enveloping all plant tissue, likely restricting the photosynthetic ability of individual leaves and total plant biomass. There were also significantly less seagrass-associated mud snails (Batillaria australis) where D. perlucidum cover was high. These results demonstrate the ability of invasive ascidians to colonise and affect native seagrasses and associated biota. Seagrasses are pivotal to the ecological function of many urban estuaries world-wide. Biodiversity in these systems is already vulnerable to multiple stressors from human activities but the potential stress of fouling ascidians may pose an additional and increasing threat in the future. PMID:27144600

  13. Gut-spilling in chordates: Evisceration in the tropical ascidian Polycarpa mytiligera

    PubMed Central

    Shenkar, Noa; Gordon, Tal

    2015-01-01

    The ejection of internal organs, i.e., evisceration, is a well-known phenomenon in sea-cucumbers. We report the ability of a member of the Chordate phyla, the tropical ascidian Polycarpa mytiligera, to eviscerate and regenerate its gut within 12 days, and to rebuild its branchial sac within 19 days. Evisceration occurred within 4–43 seconds of gentle mechanical pressure exerted on the tunic in 47% of the tested P. mytiligera. Individuals were able to discard up to 3/4 of their digestive tract via the incurrent siphon by rupture of the branchial sac in this area. Although chemical analysis revealed no significant levels of toxic compounds, the eviscerated guts were unpalatable to the triggerfish and pufferfish on which they were tested, suggesting evisceration as a defense mechanism. Given the close affinity of ascidians to vertebrates, the regeneration pathway of the viscera and branchial sac of ascidians suggests its potential beneficial application in soft tissue regeneration research. PMID:25880620

  14. Regulation of nitric-oxide production in hemocytes of the ascidian Phallusia nigra.

    PubMed

    de Barros, Cintia M; Emrich, Laura C; de A Mello, Andressa; da Fonseca, Rodrigo N; Allodi, Silvana

    2014-04-30

    Nitric oxide (NO) production in ascidians is related to immune responses of blood cells, and also to events such as egg fertilization and notochord regression. However, the signaling pathway for NO generation has been little investigated in this animal model. The present contribution identifies the cells involved in NO production and provides new information about a pathway for NO signaling. We were able to identify eight types of blood cells in the hemolymph of the ascidian Phallusia nigra, of which signet ring cells, univacuolar refractile granulocytes, and morula cells were involved in NO production. Zymosan A and lipopolysaccharide (LPS) enhanced NO production by blood cells, and the compound N-nitro-L-arginine-methyl ester (L-NAME) reduced NO production. Finally, the application of protein kinase A (PKA) and protein kinase C (PKC) inhibitors revealed that these molecules participate, together with NFκB, in the regulation of NO production by blood cells of P. nigra. This is the first report to show that PKA and PKC are involved in a signaling pathway that leads to NO production in ascidian blood cells. Copyright © 2014 Elsevier Inc. All rights reserved.

  15. Cockles, barnacles and ascidians compose a subtidal facilitation cascade with multiple hierarchical levels of foundation species.

    PubMed

    Yakovis, Eugeniy; Artemieva, Anna

    2017-03-22

    Facilitation cascades occur when multiple foundation species in a community are involved in a hierarchy of positive interactions, and consist of a primary facilitator which positively affects secondary facilitators, each supporting a suit of dependent species. There is no theoretical limit to the number of levels in a facilitation cascade, yet the existence of more than two has rarely been examined. We manipulated biogenic substrate produced by a primary facilitator (cockle shells) and a secondary facilitator (barnacles and their empty tests) in a space-limited subtidal community to test the hypothesis that solitary ascidians would be the third-level facilitator. In the field, most ascidians were found on barnacles, and most barnacles occupied cockle shells. To produce this pattern, barnacles could nurse ascidians (a longer 'facilitation chain') or outcompete them from cockle shells (a shorter chain). Experimental results clearly supported the nursing hypothesis providing evidence for a facilitation cascade with three hierarchical levels of foundation species. Our findings confirm that like predation and competition, positive interspecific interactions nest into multi-tier hierarchies with numerous levels. While the number of foundation species should increase community stability and resilience as it increases diversity and reduces environmental stress, facilitation chain length may have the opposite effect.

  16. Advances in research on the accumulation, redox behavior, and function of vanadium in ascidians.

    PubMed

    Michibata, Hitoshi; Ueki, Tatsuya

    2010-05-01

    The discovery of high levels of vanadium-containing compounds in ascidian blood cells goes back to 1911. Ascidians, which are also known as tunicates or sea squirts, belong to a subphylum of the Chordata, between the vertebrates and invertebrates. This discovery attracted the attention of an interdisciplinary group of chemists, physiologists, and biochemists, in part because of interest in the possible role of vanadium in oxygen transport as a prosthetic group in respiratory pigments, which was later shown not to be such a role, and in part because of the fact that high levels of vanadium were unknown in other organisms. The intracellular concentration of vanadium in some ascidian species can be as high as 350 mm, which is 107 times that in seawater. Vanadium ions, which are thought to be present in the +5 oxidation state in seawater, are reduced to the +3 oxidation state via the +4 oxidation state and are stored in the vacuoles of vanadium-containing cells called vanadocytes, where high levels of protons and sulfate ions are also found. Recently, many proteins and genes that might be involved in the accumulation and reduction of vanadium have been isolated. In this review, we not only trace the history of vanadium research but also describe recent advances in our understanding of the field from several viewpoints: (i) vanadium-accumulating blood cells, (ii) the energetics of vanadium accumulation, (iii) the redox mechanism of vanadium, (iv) the possible role of sulfate, and (v) the physiological roles of vanadium.

  17. The postbranchial digestive tract of the ascidian, Polyandrocarpa misakiensis (Tunicata: Ascidiacea). 1. Oesophagus.

    PubMed

    Koyama, Hiromichi

    2011-02-01

    The organization of the oesophagus in the budding styelid ascidian, Polyandrocarpa misakiensis, is described. The oesophagus consists of external and internal epithelium, and there are loose connective tissue, blood sinuses, and a muscular layer between them. The internal epithelium is simple columnar, except for the bottom of three folds. The external epithelium is simple squamous. The internal epithelium contains four cell types, i.e., ciliated mucous cells, band cells, endocrine cells, and undifferentiated cells. The ciliated mucous cells have apical cilia and microvilli, and two types of mucous vesicle. The band cells also have apical cilia and electron-dense granules in the apical cytoplasm. The endocrine cells are bottle-shaped, and have electron-dense granules both above and below the nucleus. The undifferentiated cells form pseudostratified epithelium at the bottom of each fold, and they have nuclei with prominent nucleoli. One type of coelomic cell, which has retractile cytoplasm, often migrates in the internal epithelium. Near the stomach, there are many darkly stained round cells clustered around the posterior end of the oesophagus. These two types of coelomic cells may be involved in the defense mechanism against the invasion of foreign organisms. The basic organization of the oesophagus of P. misakiensis is similar to those of other ascidians. However, the presence of three folds is a characteristic of a solitary species, rather than of a colonial species. Although ascidians are chordate invertebrates, the organization of their oesophagus is not very complex, which might reflect their life style.

  18. PSP toxins profile in ascidian Microcosmus vulgaris (Heller, 1877) after human poisoning in Croatia (Adriatic Sea).

    PubMed

    Roje-Busatto, Romana; Ujević, Ivana

    2014-03-01

    Toxins known to cause Paralytic Shellfish Poisoning (PSP) syndrome in humans that can have serious economic consequences for aquaculture were determined in ascidians of the genus Microcosmus. Significant concentrations of toxins were confirmed in all tested samples collected from the western coast of Istria Peninsula (Adriatic Sea, Croatia) when six people were poisoned following the consumption of fresh ascidians. Several species of bivalves that were under continuous monitoring had not accumulated PSP toxins although they were exposed to the same environmental conditions over the survey period. In the present study, HPLC-FLD with pre-column oxidation of PSP toxins has been carried out to provide evidence for the first human intoxication due to consumption of PSP toxic ascidians (Microcosmus vulgaris, Heller, 1877) harvested from the Adriatic Sea. Qualitative analysis established the presence of six PSP toxins: saxitoxin (STX), decarbamoylsaxitoxin (dcSTX), gonyautoxins 2 and 3 (GTX2,3), decarbamoylgonyautoxins 2 and 3 (dcGTX2,3), gonyautoxin 5 (GTX5) and N-sulfocarbamoylgonyautoxins 1 and 2 (C1,2), while quantitative analysis suggested STX and GTX2,3 as dominant toxin types and the ones that contribute the most to the overall toxicity of these samples with concentrations near the regulatory limit.

  19. The Mediterranean

    NASA Astrophysics Data System (ADS)

    Suryawanshi, Vandana

    2017-04-01

    Learning is always a joyful experience for any human being and must always remain so. Children are happiest when they learn through play. The philosophy of my life is to keep encouraging children to think beyond they could achieve easily. I understand children are adaptive to change and take things with an open mind. They are ready to experiment new things and dare to dream big. I am fortunate to be a teacher by profession and thus I always attempt experimenting, observing and participating with other children and adults. Education is not about moulding children the way you think they should be. It is about organizing the natural longing in a human being to know. From birth children are active participants in building their own understanding. I always prepare the environment to help each child build on what they already know. It is such a great pleasure to observe every young kid become excited and curious to know when we teach them. Std 8 Geography the students are very excited to learn about this continent, with the help of Videos and a wall map the Political map of Europe with its countries shown I introduced the topic by asking 'If given a chance which place they would like to visit in Europe' , students are familiar with the countries of their favourite football players and happily pointed out their destination. The Mediterranean Region is a paradise the scenic beauty, the climate, the food along with a variety of fruits which are totally different from Asia increased the curiosity among the students. With the help of case study of the Mediterranean Sea the students were able to research and present the history, the adventure sports the aquatic life and the twenty three beautiful islands located in the Mediterranean Sea. Photos and videos helped me to explain the Mediterranean Sea The Formation of the Mediterranean Sea ( Youtube Video) which is otherwise completely enclosed by land. (The evaporating Mediterranean Sea - BBC (Video) Gibraltar Breach.mov . The

  20. Dynamics of cell polarity in tissue morphogenesis: a comparative view from Drosophila and Ciona.

    PubMed

    Veeman, Michael T; McDonald, Jocelyn A

    2016-01-01

    Tissues in developing embryos exhibit complex and dynamic rearrangements that shape forming organs, limbs, and body axes. Directed migration, mediolateral intercalation, lumen formation, and other rearrangements influence the topology and topography of developing tissues. These collective cell behaviors are distinct phenomena but all involve the fine-grained control of cell polarity. Here we review recent findings in the dynamics of polarized cell behavior in both the Drosophila ovarian border cells and the Ciona notochord. These studies reveal the remarkable reorganization of cell polarity during organ formation and underscore conserved mechanisms of developmental cell polarity including the Par/atypical protein kinase C (aPKC) and planar cell polarity pathways. These two very different model systems demonstrate important commonalities but also key differences in how cell polarity is controlled in tissue morphogenesis. Together, these systems raise important, broader questions on how the developmental control of cell polarity contributes to morphogenesis of diverse tissues across the metazoa.

  1. Investigating sperm cryopreservation in a model tunicate, Ciona intestinalis sp. A.

    PubMed

    Sorrenti, Gerarda; Bagnoli, Anna; Miraglia, Valentina; Crocetta, Fabio; Vitiello, Valentina; Ristoratore, Filomena; Cirino, Paola; Sansone, Giovanni; Sordino, Paolo

    2014-02-01

    In cryopreservation procedures, the capacity to protect the cells from freezing and thawing processes is sensitive to the choice of the cryoprotective agent (CPA) and to its optimal concentration. The advancement of research on Tunicate model species has raised interest in liquid nitrogen cryopreservation for the storage and distribution of genetic resources. Ciona intestinalis (Linnè, 1767) consists of a complex of cryptic taxa that are central to several areas of investigation, from comparative genomics to invasive biology. Here we investigated how five CPAs, three chilling rates and two freezing rates influence semen cryopreservation in C. intestinalis sp. A. By using larval morphology and motility as endpoints, we estimated that long term semen storage requires 10% dimethyl sulfoxide as a protective agent, -1°C/min chilling rate (18°C to 5°C) and -13°C/min freezing rate (5°C to -80°C), followed by immersion in liquid nitrogen.

  2. Dynamic changes in developmental gene expression in the basal chordate Ciona intestinalis.

    PubMed

    Kawashima, Takeshi; Satou, Yutaka; Murakami, Seiko D; Satoh, Nori

    2005-04-01

    Large-scale expressed sequence tags (EST) analysis was used to demonstrate a number of dynamic changes in the global gene expression profile of the basal chordate Ciona intestinalis over the course of its development. The fertilized egg was found to store a great variety of maternal transcripts and, as development proceeds, the organism expresses a progressively smaller repertoire of genes. In addition, a significant portion of genes involved in embryogenesis were observed to be downregulated during metamorphosis, at which point the adult appears to utilize a different set of genes to form its body. At least 25% of the genes involved in development were found to be used multiple times. This kind of information is essential to form a comprehensive understanding of the overarching expression-control plan by which the basic chordate body is formed.

  3. A saturation screen for cis-acting regulatory DNA in the Hox genes of Ciona intestinalis

    SciTech Connect

    Keys, David N.; Lee, Byung-in; Di Gregorio, Anna; Harafuji, Naoe; Detter, Chris; Wang, Mei; Kahsai, Orsalem; Ahn, Sylvia; Arellano, Andre; Zhang, Quin; Trong, Stephan; Doyle, Sharon A.; Satoh, Noriyuki; Satou, Yutaka; Saiga, Hidetoshi; Christian, Allen; Rokhsar, Dan; Hawkins, Trevor L.; Levine, Mike; Richardson, Paul

    2005-01-05

    A screen for the systematic identification of cis-regulatory elements within large (>100 kb) genomic domains containing Hox genes was performed by using the basal chordate Ciona intestinalis. Randomly generated DNA fragments from bacterial artificial chromosomes containing two clusters of Hox genes were inserted into a vector upstream of a minimal promoter and lacZ reporter gene. A total of 222 resultant fusion genes were separately electroporated into fertilized eggs, and their regulatory activities were monitored in larvae. In sum, 21 separable cis-regulatory elements were found. These include eight Hox linked domains that drive expression in nested anterior-posterior domains of ectodermally derived tissues. In addition to vertebrate-like CNS regulation, the discovery of cis-regulatory domains that drive epidermal transcription suggests that C. intestinalis has arthropod-like Hox patterning in the epidermis.

  4. Arsenic (+ 3 oxidation state) methyltransferase and the methylation of arsenicals in the invertebrate chordate Ciona intestinalis.

    PubMed

    Thomas, David J; Nava, Gerardo M; Cai, Shi-Ying; Boyer, James L; Hernández-Zavala, Araceli; Gaskins, H Rex

    2010-01-01

    Biotransformation of inorganic arsenic (iAs) involves methylation catalyzed by arsenic (+ 3 oxidation state) methyltransferase (As3mt) yielding mono-, di-, and trimethylated arsenicals. To investigate the evolution of molecular mechanisms that mediate arsenic biotransformation, a comparative genomic approach focusing on the invertebrate chordate Ciona intestinalis was used. Bioinformatic analyses identified an As3mt gene in the C. intestinalis genome. Constitutive As3mt RNA expression was observed in heart, branchial sac, and gastrointestinal tract. Adult animals were exposed to 0 or 1 ppm of iAs for 1 or 5 days. Steady-state As3mt RNA expression in the gastrointestinal tract was not modulated significantly by 5 days of exposure to iAs. Tissue levels of iAs and its methylated metabolites were determined by hydride generation-cryotrapping-gas chromatography-atomic absorption spectrometry. At either time point, exposure to iAs significantly increased concentrations of iAs and its methylated metabolites in tissues. After 5 days of exposure, total speciated arsenic concentrations were highest in branchial sac (3705 ng/g), followed by heart (1019 ng/g) and gastrointestinal tract (835 ng/g). At this time point, the sum of the speciated arsenical concentrations in gastrointestinal tract and heart equaled or exceeded that of iAs; in branchial sac, iAs was the predominant species present. Ciona intestinalis metabolizes iAs to its methylated metabolites, which are retained in tissues. This metabolic pattern is consistent with the presence of an As3mt ortholog in its genome and constitutive expression of the gene in prominent organs, making this basal chordate a useful model to examine the evolution of arsenic detoxification.

  5. Genome-wide identification and characterization of transcription start sites and promoters in the tunicate Ciona intestinalis

    PubMed Central

    Yokomori, Rui; Shimai, Kotaro; Nishitsuji, Koki; Suzuki, Yutaka; Kusakabe, Takehiro G.; Nakai, Kenta

    2016-01-01

    The tunicate Ciona intestinalis, an invertebrate chordate, has recently emerged as a powerful model organism for gene regulation analysis. However, few studies have been conducted to identify and characterize its transcription start sites (TSSs) and promoters at the genome-wide level. Here, using TSS-seq, we identified TSSs at the genome-wide scale and characterized promoters in C. intestinalis. Specifically, we identified TSS clusters (TSCs), high-density regions of TSS-seq tags, each of which appears to originate from an identical promoter. TSCs were found not only at known TSSs but also in other regions, suggesting the existence of many unknown transcription units in the genome. We also identified candidate promoters of 79 ribosomal protein (RP) genes, each of which had the major TSS in a polypyrimidine tract and showed a sharp TSS distribution like human RP gene promoters. Ciona RP gene promoters, however, did not appear to have typical TATA boxes, unlike human RP gene promoters. In Ciona non-RP promoters, two pyrimidine-purine dinucleotides, CA and TA, were frequently used as TSSs. Despite the absence of CpG islands, Ciona TATA-less promoters showed low expression specificity like CpG-associated human TATA-less promoters. By using TSS-seq, we also predicted trans-spliced gene TSSs and found that their downstream regions had higher G+T content than those of non-trans-spliced gene TSSs. Furthermore, we identified many putative alternative promoters, some of which were regulated in a tissue-specific manner. Our results provide valuable information about TSSs and promoter characteristics in C. intestinalis and will be helpful in future analysis of transcriptional regulation in chordates. PMID:26668163

  6. Genome-wide identification and characterization of transcription start sites and promoters in the tunicate Ciona intestinalis.

    PubMed

    Yokomori, Rui; Shimai, Kotaro; Nishitsuji, Koki; Suzuki, Yutaka; Kusakabe, Takehiro G; Nakai, Kenta

    2016-01-01

    The tunicate Ciona intestinalis, an invertebrate chordate, has recently emerged as a powerful model organism for gene regulation analysis. However, few studies have been conducted to identify and characterize its transcription start sites (TSSs) and promoters at the genome-wide level. Here, using TSS-seq, we identified TSSs at the genome-wide scale and characterized promoters in C. intestinalis. Specifically, we identified TSS clusters (TSCs), high-density regions of TSS-seq tags, each of which appears to originate from an identical promoter. TSCs were found not only at known TSSs but also in other regions, suggesting the existence of many unknown transcription units in the genome. We also identified candidate promoters of 79 ribosomal protein (RP) genes, each of which had the major TSS in a polypyrimidine tract and showed a sharp TSS distribution like human RP gene promoters. Ciona RP gene promoters, however, did not appear to have typical TATA boxes, unlike human RP gene promoters. In Ciona non-RP promoters, two pyrimidine-purine dinucleotides, CA and TA, were frequently used as TSSs. Despite the absence of CpG islands, Ciona TATA-less promoters showed low expression specificity like CpG-associated human TATA-less promoters. By using TSS-seq, we also predicted trans-spliced gene TSSs and found that their downstream regions had higher G+T content than those of non-trans-spliced gene TSSs. Furthermore, we identified many putative alternative promoters, some of which were regulated in a tissue-specific manner. Our results provide valuable information about TSSs and promoter characteristics in C. intestinalis and will be helpful in future analysis of transcriptional regulation in chordates.

  7. Nitric Oxide Acts as a Positive Regulator to Induce Metamorphosis of the Ascidian Herdmania momus

    PubMed Central

    Ueda, Nobuo; Degnan, Sandie M.

    2013-01-01

    Marine invertebrates commonly have a biphasic life cycle in which the metamorphic transition from a pelagic larva to a benthic post-larva is mediated by the nitric oxide signalling pathway. Nitric oxide (NO) is synthesised by nitric oxide synthase (NOS), which is a client protein of the molecular chaperon heat shock protein 90 (HSP90). It is notable, then, that both NO and HSP90 have been implicated in regulating metamorphosis in marine invertebrates as diverse as urochordates, echinoderms, molluscs, annelids, and crustaceans. Specifically, the suppression of NOS activity by the application of either NOS- or HSP90-inhibiting pharmacological agents has been shown consistently to induce the initiation of metamorphosis, leading to the hypothesis that a negative regulatory role of NO is widely conserved in biphasic life cycles. Further, the induction of metamorphosis by heat-shock has been demonstrated for multiple species. Here, we investigate the regulatory role of NO in induction of metamorphosis of the solitary tropical ascidian, Herdmania momus. By coupling pharmacological treatments with analysis of HmNOS and HmHSP90 gene expression, we present compelling evidence of a positive regulatory role for NO in metamorphosis of this species, in contrast to all existing ascidian data that supports the hypothesis of NO as a conserved negative regulator of metamorphosis. The exposure of competent H. momus larvae to a NOS inhibitor or an NO donor results in an up-regulation of NOS and HSP90 genes. Heat shock of competent larvae induces metamorphosis in a temperature dependent manner, up to a thermal tolerance that approaches 35°C. Both larval/post-larval survival and the appearance of abnormal morphologies in H. momus post-larvae reflect the magnitude of up-regulation of the HSP90 gene in response to heat-shock. The demonstrated role of NO as a positive metamorphic regulator in H. momus suggests the existence of inter-specific adaptations of NO regulation in ascidian

  8. Purification of Mitochondrial Proteins HSP60 and ATP Synthase from Ascidian Eggs: Implications for Antibody Specificity

    PubMed Central

    Chenevert, Janet; Pruliere, Gerard; Ishii, Hirokazu; Sardet, Christian; Nishikata, Takahito

    2013-01-01

    Use of antibodies is a cornerstone of biological studies and it is important to identify the recognized protein with certainty. Generally an antibody is considered specific if it labels a single band of the expected size in the tissue of interest, or has a strong affinity for the antigen produced in a heterologous system. The identity of the antibody target protein is rarely confirmed by purification and sequencing, however in many cases this may be necessary. In this study we sought to characterize the myoplasm, a mitochondria-rich domain present in eggs and segregated into tadpole muscle cells of ascidians (urochordates). The targeted proteins of two antibodies that label the myoplasm were purified using both classic immunoaffinity methods and a novel protein purification scheme based on sequential ion exchange chromatography followed by two-dimensional gel electrophoresis. Surprisingly, mass spectrometry sequencing revealed that in both cases the proteins recognized are unrelated to the original antigens. NN18, a monoclonal antibody which was raised against porcine spinal cord and recognizes the NF-M neurofilament subunit in vertebrates, in fact labels mitochondrial ATP synthase in the ascidian embryo. PMF-C13, an antibody we raised to and purified against PmMRF, which is the MyoD homolog of the ascidian Phallusia mammillata, in fact recognizes mitochondrial HSP60. High resolution immunolabeling on whole embryos and isolated cortices demonstrates localization to the inner mitochondrial membrane for both ATP synthase and HSP60. We discuss the general implications of our results for antibody specificity and the verification methods which can be used to determine unequivocally an antibody's target. PMID:23326373

  9. Nitric oxide acts as a positive regulator to induce metamorphosis of the ascidian Herdmania momus.

    PubMed

    Ueda, Nobuo; Degnan, Sandie M

    2013-01-01

    Marine invertebrates commonly have a biphasic life cycle in which the metamorphic transition from a pelagic larva to a benthic post-larva is mediated by the nitric oxide signalling pathway. Nitric oxide (NO) is synthesised by nitric oxide synthase (NOS), which is a client protein of the molecular chaperon heat shock protein 90 (HSP90). It is notable, then, that both NO and HSP90 have been implicated in regulating metamorphosis in marine invertebrates as diverse as urochordates, echinoderms, molluscs, annelids, and crustaceans. Specifically, the suppression of NOS activity by the application of either NOS- or HSP90-inhibiting pharmacological agents has been shown consistently to induce the initiation of metamorphosis, leading to the hypothesis that a negative regulatory role of NO is widely conserved in biphasic life cycles. Further, the induction of metamorphosis by heat-shock has been demonstrated for multiple species. Here, we investigate the regulatory role of NO in induction of metamorphosis of the solitary tropical ascidian, Herdmania momus. By coupling pharmacological treatments with analysis of HmNOS and HmHSP90 gene expression, we present compelling evidence of a positive regulatory role for NO in metamorphosis of this species, in contrast to all existing ascidian data that supports the hypothesis of NO as a conserved negative regulator of metamorphosis. The exposure of competent H. momus larvae to a NOS inhibitor or an NO donor results in an up-regulation of NOS and HSP90 genes. Heat shock of competent larvae induces metamorphosis in a temperature dependent manner, up to a thermal tolerance that approaches 35°C. Both larval/post-larval survival and the appearance of abnormal morphologies in H. momus post-larvae reflect the magnitude of up-regulation of the HSP90 gene in response to heat-shock. The demonstrated role of NO as a positive metamorphic regulator in H. momus suggests the existence of inter-specific adaptations of NO regulation in ascidian

  10. The alien ascidian Styela clava now invading the Sea of Marmara (Tunicata: Ascidiacea)

    PubMed Central

    Çinar, Melih Ertan

    2016-01-01

    Abstract During the implementation of a large project aimed to investigate the benthic community structures of the Sea of Marmara, specimens of the invasive ascidian species Styela clava were collected on natural substrata (rocks) at 10 m depth at one locality (Karamürsel) in İzmit Bay. The specimens were mature, containing gametes, indicating that the species had become established in the area. The Sea of Marmara seems to provide suitable conditions for this species to survive and form proliferating populations. PMID:27047235

  11. Evidence for the heparin-binding ability of the ascidian Xlink domain and insight into the evolution of the Xlink domain in chordates.

    PubMed

    Yoneda, Masahiko; Nakamura, Toshiya; Murai, Miho; Wada, Hiroshi

    2010-07-01

    The vertebrate Xlink domain is found in two types of genes: lecticans and their associated hyaluronan-and-proteoglycan-binding-link-proteins (HAPLNs), which are components of the extracellular matrix, and those represented by CD44 and stabilins, which are expressed on the surface of lymphocytes. In both types of genes, Xlink functions as a hyaluronan binding domain. We have already reported that protochordate ascidians possess only the latter type of gene. The present analysis of the expression of ascidian Xlink domain genes revealed that these genes function in blood cell migration and apoptosis. While the Xlink domain is found in various metazoans, including ascidians and nematodes, hyaluronan is believed to be specific for vertebrates. In comprehensive genome surveys for hyaluronan synthase (HAS), we found no HAS gene in ascidians. We also established that hyaluronan is absent from the ascidian body biochemically. Therefore, ascidians possess the Xlink domain, but they lack HA. We recovered one ascidian Xlink domain gene that encoded a heparin-binding protein, although it shows no affinity for hyaluronan. Based on these findings, we conclude that in invertebrates, the Xlink domain serves as heparin-binding protein domain and functions in blood cell migration and apoptosis. Its binding affinity for HA might have been acquired in the vertebrate lineage.

  12. THE GENERAL ECOLOGY AND GROWTH OF A SOLITARY ASCIDIAN, CORELLA WILLMERIANA.

    PubMed

    Lambert, Gretchen

    1968-10-01

    1. A one-year field study of the ecology of the solitary ascidian Corella willmeriana Herdman was conducted between April, 1966, and April, 1967, at the Bremerton Yacht Club, Bremerton, Washington, where two polyvinyl chloride frames containing glass plates were examined at monthly intervals. 2. The results indicate that Corella is a primary colonizer, preferring to settle on clean surfaces. Growth is rapid during the summer, when sexual maturity, corresponding to a size of 12 mm., may be attained in three months and life span is approximately five months. Individuals grow at a slower rate and live longer during the winter; the life span then is seven or eight months. 3. Very young specimens of Corella are frequently overgrown during the winter by the colonial ascidian Diplosoma macdonaldi. The causes of death of adult Corella are not completely known, although a small percentage of them are eaten by the polyclad flatworm Eurylepta leoparda. A luxuriant spring growth of filamentous diatoms may cause death of adult Corella by smothering them.

  13. Evolution and development of budding by stem cells: ascidian coloniality as a case study.

    PubMed

    Brown, Federico D; Swalla, Billie J

    2012-09-15

    The evolution of budding in metazoans is not well understood on a mechanistic level, but is an important developmental process. We examine the evolution of coloniality in ascidians, contrasting the life histories of solitary and colonial forms with a focus on the cellular and developmental basis of the evolution of budding. Tunicates are an excellent group to study colonial transitions, as all solitary larvae develop with determinant and invariant cleavage patterns, but colonial species show robust developmental flexibility during larval development. We propose that acquiring new stem cell lineages in the larvae may be a preadaptation necessary for the evolution of budding. Brooding in colonial ascidians allows increased egg size, which in turn allows greater flexibility in the specification of cells and cell numbers in late embryonic and pre-metamorphic larval stages. We review hypotheses for changes in stem cell lineages in colonial species, describe what the current data suggest about the evolution of budding, and discuss where we believe further studies will be most fruitful.

  14. Differentiation of papillae and rostral sensory neurons in the larva of the ascidian Botryllus schlosseri (Tunicata).

    PubMed

    Caicci, Federico; Zaniolo, Giovanna; Burighel, Paolo; Degasperi, Valentina; Gasparini, Fabio; Manni, Lucia

    2010-02-15

    During the metamorphosis of tunicate ascidians, the swimming larva uses its three anterior papillae to detect the substrate for settlement, reabsorbs its chordate-like tail, and becomes a sessile oozooid. In view of the crucial role played by the anterior structures and their nerve relations, we applied electron microscopy and immunocytochemistry to study the larva of the colonial ascidian Botryllus schlosseri, following differentiation of the anterior epidermis during late embryogenesis, the larval stage, and the onset of metamorphosis. Rudiments of the papillae appear in the early tail-bud stage as ectodermic protrusions, the apexes of which differentiate into central and peripheral bipolar neurons. Axons fasciculate into two nerves direct to the brain. Distally, the long, rod-like dendritic terminations extend during the larval stage, becoming exposed to sea water. After the larva selects and adheres to the substrate, these neurons retract and regress. Adjacent to the papillae, other scattered neurons insinuate dendrites into the tunic and form the net of rostral trunk epidermal neurons (RTENs) which fasciculate together with the papillary neurons. Our data indicate that the papillae are simple and coniform, the papillary neurons are mechanoreceptors, and the RTENs are chemoreceptors. The interpapillary epidermal area, by means of an apocrine secretion, provides sticky material for temporary adhesion of the larva to the substrate.

  15. Population dynamics of the solitary ascidian Herdmania momus (Savignyi, 1816) in Jeju Island, Korea

    NASA Astrophysics Data System (ADS)

    Yi, Chang Ho; Kim, Jeong Ha

    2016-09-01

    In 2007-2008, a population outbreak of the solitary ascidian Herdmania momus was observed along the coasts of Jeju Island and the ascidians became the top dominant species in macrobenthic communities. We conducted field experiments on population dynamics and demographic parameters including seasonal density fluctuation, vertical distribution, recruitment patterns, mortality, and growth rate. Recruitment continued year-round except in April following the lowest water temperature season in February. Population density was highest in August-October, concomitantly with the peak in recruitment, related to the highest water temperature. The lowest density was observed in February-April when the water temperature was lowest. In April all individuals disappeared from all permanent plots, resulting in a maximum life span of 10-12 months. Individuals of newly recruited H. momus grew to have an adult size of 4-6 cm in only 3-4 months. Higher densities were observed in the shallower areas (5 m) compared to the deeper zones. More recruitment occurred on vertical slopes than horizontal substrates, but substrate pre-occupancy caused no difference in recruitment. This is the first report on H. momus population-level investigation in Korea, which provides valuable baseline information on the current status of population demography and their potential community-level influence on Jeju coasts.

  16. Neurula rotation determines left-right asymmetry in ascidian tadpole larvae.

    PubMed

    Nishide, Kazuhiko; Mugitani, Michio; Kumano, Gaku; Nishida, Hiroki

    2012-04-01

    Tadpole larvae of the ascidian Halocynthia roretzi show morphological left-right asymmetry. The tail invariably bends towards the left side within the vitelline membrane. The structure of the larval brain is remarkably asymmetric. nodal, a conserved gene that shows left-sided expression, is also expressed on the left side in H. roretzi but in the epidermis unlike in vertebrates. We show that nodal signaling at the late neurula stage is required for stereotypic morphological left-right asymmetry at later stages. We uncover a novel mechanism to break embryonic symmetry, in which rotation of whole embryos provides the initial cue for left-sided expression of nodal. Two hours prior to the onset of nodal expression, the neurula embryo rotates along the anterior-posterior axis in a counterclockwise direction when seen in posterior view, and then this rotation stops when the left side of the embryo is oriented downwards. It is likely that epidermis monocilia, which appear at the neurula rotation stage, generate the driving force for the rotation. When the embryo lies on the left side, protrusion of the neural fold physically prevents it from rotating further. Experiments in which neurula rotation is perturbed by various means, including centrifugation and sandwiching between glass, indicate that contact of the left epidermis with the vitelline membrane as a consequence of neurula rotation promotes nodal expression in the left epidermis. We suggest that chemical, and not mechanical, signals from the vitelline membrane promote nodal expression. Neurula rotation is also conserved in other ascidian species.

  17. The recently-described ascidian species Molgula tectiformis is a direct developer.

    PubMed

    Tagawa, K; Jeffery, W R; Satoh, N

    1997-04-01

    Molgula tectiformis is a new ascidian species recently described by Nishikawa (1991). In Otsuchi Bay, Iwate, Japan, they are easily obtainable from cages for culturing scallops. We report here that M. tectiformis is another example of a direct developer: their embryonic development is lacking the tadpole larva. The fertilized egg is orange and about 150 microns in diameter. At 18 degrees C, the egg cleaves at about 20 min intervals and gastrulation occurs about 5 hr after fertilization. In contrast to conventionally-developing ascidians, M. tectiformis does not form a tadpole larva. Immediately before hatching, three stolons or ampullae begin to extend from the tailless embryo. After hatching the stolons mediate the attachment of the juvenile body to the substratum. Histochemistry for tissue-specific enzyme activity did not detect muscle-specific acetyl-cholinesterase, endoderm-specific alkaline phosphatase, and pigment cell-specific tyrosinase. In addition, in situ hybridization could not prove the presence of muscle actin gene transcripts in the embryo. These results suggest that these larval tissues do not differentiate in M. tectiformis embryos. Because M. tectiformis is common and gravid year-around in Otsuchi Bay, this direct developer provides the opportunity for further analysis of molecular changes during evolution that cause an alternative mode of development.

  18. Preparation and Antitumor Activity of CS5931, A Novel Polypeptide from Sea Squirt Ciona Savignyi

    PubMed Central

    Chen, Xiaoshuang; Xu, Huanli; Li, Bo; Wang, Feng; Chen, Xiaoliang; Kong, Dexin; Lin, Xiukun

    2016-01-01

    CS5931 is a novel anticancer agent isolated from the sea squirt Ciona savignyi. However, its content in the species is very low, and developing a novel approach for production of the polypeptide is promising. In the present study, we expressed and purified the polypeptide from E. coli, and the fermentation conditions were studied using response surface methodology. The yield of CS5931 was increased from 2.0 to 7.5 mg/L. The denaturing and renaturation conditions were also studied. Using the optimized renaturation condition, the anticancer activity of refolding CS5931 was increased significantly; the value of IC50 was decreased from 23.2 to 11.6 μM. In vivo study using xenograft nude mice bearing HCT116 cancer cells revealed that CS5931 was able to inhibit the growth of tumor significantly. The study provides a useful approach for obtaining enough amount of CS5931 for further study. This study is also important for developing the polypeptide as a novel anticancer agent. PMID:27007382

  19. Regeneration, Stem Cells, and Aging in the Tunicate Ciona: Insights from the Oral Siphon.

    PubMed

    Jeffery, William R

    2015-01-01

    Regeneration studies in the tunicate Ciona intestinalis have recently been focused on the potential of adult stem cells to replace injured tissues and organs during the adult life cycle using the oral siphon (OS) as a model. The OS has oral siphon pigment organs (OPOs) along its rim and an underlying network of muscle fibers in its tube. Different regeneration processes are triggered by OS amputation at the tip, along the tube, or at the base. One process involves the replacement of OPOs without new cell division by direct differentiation of locally deployed stem cells or stem cells that migrate from the branchial sac. Another process involves blastema formation by the migration of progenitor cells produced from branchial sac stem cells. The capacity for complete and accurate OS regeneration declines continuously during the adult life cycle. Finally, after an age threshold is reached, OS regeneration ceases in old animals. The loss of regeneration capacity in old animals involves the depletion of stem cells in the branchial sac, the inability of branchial sac progenitor cells to migrate to the sites of regeneration, and defective oral pigment organ replacement. The significance of the OS model for studying regeneration, stem cells, and aging will be enhanced by the application of molecular methods.

  20. Bacterial diversity associated with the tunic of the model chordate Ciona intestinalis.

    PubMed

    Blasiak, Leah C; Zinder, Stephen H; Buckley, Daniel H; Hill, Russell T

    2014-02-01

    The sea squirt Ciona intestinalis is a well-studied model organism in developmental biology, yet little is known about its associated bacterial community. In this study, a combination of 454 pyrosequencing of 16S ribosomal RNA genes, catalyzed reporter deposition-fluorescence in situ hybridization and bacterial culture were used to characterize the bacteria living inside and on the exterior coating, or tunic, of C. intestinalis adults. The 454 sequencing data set demonstrated that the tunic bacterial community structure is different from that of the surrounding seawater. The observed tunic bacterial consortium contained a shared community of <10 abundant bacterial phylotypes across three individuals. Culture experiments yielded four bacterial strains that were also dominant groups in the 454 sequencing data set, including novel representatives of the classes Alphaproteobacteria and Flavobacteria. The relatively simple bacterial community and availability of dominant community members in culture make C. intestinalis a promising system in which to investigate functional interactions between host-associated microbiota and the development of host innate immunity.

  1. Molecular characterisation, evolution and expression analysis of g-type lysozymes in Ciona intestinalis.

    PubMed

    Di Falco, Felicia; Cammarata, Matteo; Vizzini, Aiti

    2017-02-01

    Lysozyme is an important defense molecule of the innate immune system. Known for its bactericidal properties, lysozyme catalyzes the hydrolysis of b-(1,4)-glycosidic bonds between the N-acetyl glucosamine and N-acetyl muramic acid in the peptidoglycan layer of bacterial cell walls. In this study, the complete coding sequence of four g-type lysozymes were identified in Ciona intestinalis. Phylogenetic analysis and modelling supported the hypothesis of a close relationship with the vertebrate g-type lysozymes suggesting that the C. intestinalis g-type lysozyme genes (CiLys-g1, Cilys-g2, CiLys-g3, CiLys-g4) share a common ancestor in the chordate lineage. Protein motif searches indicated that C. intestinalis g-type lysozymes contain a GEWL domain with a GXXQ signature, typical of goose lysozymes. Quantitative Real-Time PCR analysis results showed that transcripts are expressed in various tissues from C. intestinalis. In order to determine the involvement of C. intestinalis g-type lysozymes in immunity, their expression was analyzed in the pharynx, showing that transcripts were significantly up-regulated in response to a challenge with lipopolysaccharide (LPS). These data support the view that CiLys g-type are molecules with potential for immune defense system against bacterial infection.

  2. Isolation of a novel LPS-induced component of the ML superfamily in Ciona intestinalis.

    PubMed

    Vizzini, Aiti; Bonura, Angela; Longo, Valeria; Sanfratello, Maria Antonietta; Parrinello, Daniela; Cammarata, Matteo; Colombo, Paolo

    2015-11-01

    ML superfamily represents a group of proteins playing important roles in lipid metabolism and innate immune response. In this study, we report the identification of the first component of the ML superfamily in the invertebrate Ciona intestinalis by means of a subtractive hybridization strategy. Sequence homology and phylogenetic analysis showed that this protein forms a specific clade with vertebrate components of the Niemann-Pick type C2 protein and, for this reason, it has been named Ci-NPC2. The putative Ci-NPC2 is a 150 amino acids long protein with a short signal peptide, seven cysteine residues, three putative lipid binding site and a three-dimensional model showing a characteristic β-strand structure. Gene expression analysis demonstrated that the Ci-NPC2 protein is positively upregulated after LPS inoculum with a peak of expression 1 h after challenge. Finally, in-situ hybridization demonstrated that the Ci-NPC2 protein is preferentially expressed in hemocytes inside the vessel lumen.

  3. Mining antimicrobial peptides from small open reading frames in Ciona intestinalis.

    PubMed

    Lu, Yongzhong; Zhuang, Yu; Liu, Jie

    2014-01-01

    Though being able to encode various kinds of bioactive peptides, small open reading frames (sORFs) are poorly annotated in many genomic data. The present study was conducted to evaluate the potential of sORFs in encoding antimicrobial peptides (AMPs) in the basal chordate model Ciona intestinalis. About 4.8 m genomic sequence was first retrieved for sORFs mining by the program sORF finder, then the sORFs were translated into amino acid sequences for AMP prediction via CAMP server, and thereafter, ten putative AMPs were selected for expression and antimicrobial activity validation. In total, over 180 peptides deduced from the sORFs were predicted to be AMPs. Among the ten tested peptides, six were found to have significant expressed sequence tag matches, providing strong evidence for gene expression; five were proved to be active against the bacterial strains. These results indicate that many sORFs in C. intestinalis genome contain AMP information. This work can serve as an important initial step to investigate the role of sORFs in the innate defense of C. intestinalis.

  4. Raman spectroscopic imaging of the whole Ciona intestinalis embryo during development.

    PubMed

    Nakamura, Mitsuru J; Hotta, Kohji; Oka, Kotaro

    2013-01-01

    Intracellular composition and the distribution of bio-molecules play central roles in the specification of cell fates and morphogenesis during embryogenesis. Consequently, investigation of changes in the expression and distribution of bio-molecules, especially mRNAs and proteins, is an important challenge in developmental biology. Raman spectroscopic imaging, a non-invasive and label-free technique, allows simultaneous imaging of the intracellular composition and distribution of multiple bio-molecules. In this study, we explored the application of Raman spectroscopic imaging in the whole Ciona intestinalis embryo during development. Analysis of Raman spectra scattered from C. intestinalis embryos revealed a number of localized patterns of high Raman intensity within the embryo. Based on the observed distribution of bio-molecules, we succeeded in identifying the location and structure of differentiated muscle and endoderm within the whole embryo, up to the tailbud stage, in a label-free manner. Furthermore, during cell differentiation, we detected significant differences in cell state between muscle/endoderm daughter cells and daughter cells with other fates that had divided from the same mother cells; this was achieved by focusing on the Raman intensity of single Raman bands at 1002 or 1526 cm(-1), respectively. This study reports the first application of Raman spectroscopic imaging to the study of identifying and characterizing differentiating tissues in a whole chordate embryo. Our results suggest that Raman spectroscopic imaging is a feasible label-free technique for investigating the developmental process of the whole embryo of C. intestinalis.

  5. Differential gene expression in notochord and nerve cord fate segregation in the Ciona intestinalis embryo.

    PubMed

    Kobayashi, Kenji; Yamada, Lixy; Satou, Yutaka; Satoh, Nori

    2013-09-01

    During early embryogenesis, embryonic cells gradually restrict their developmental potential and are eventually destined to give rise to one type of cells. Molecular mechanisms underlying developmental fate restriction are one of the major research subjects within developmental biology. In this article, this subject was addressed by combining blastomere isolation with microarray analysis. During the 6th cleavage of the Ciona intestinalis embryo, from the 32-cell to the 64-cell stage, four mother cells divide into daughter cells with two distinct fates, one giving rise to notochord precursor cells and the other to nerve cord precursors. Approximately 2,200 each of notochord and nerve cord precursor cells were isolated, and their mRNA expression profiles were compared by microarray. This analysis identified 106 and 68 genes, respectively, that are differentially expressed in notochord and nerve cord precursor cells. These included not only genes for transcription factors and signaling molecules but also those with generalized functions observed in many types of cells. In addition, whole-mount in situ hybridization showed dynamic spatial expression profiles of these genes during segregation of the two fates: partitioning of transcripts present in the mother cells into either type of daughter cells, and initiation of preferential gene expression in either type of cells.

  6. Raman Spectroscopic Imaging of the Whole Ciona intestinalis Embryo during Development

    PubMed Central

    Nakamura, Mitsuru J.; Hotta, Kohji; Oka, Kotaro

    2013-01-01

    Intracellular composition and the distribution of bio-molecules play central roles in the specification of cell fates and morphogenesis during embryogenesis. Consequently, investigation of changes in the expression and distribution of bio-molecules, especially mRNAs and proteins, is an important challenge in developmental biology. Raman spectroscopic imaging, a non-invasive and label-free technique, allows simultaneous imaging of the intracellular composition and distribution of multiple bio-molecules. In this study, we explored the application of Raman spectroscopic imaging in the whole Ciona intestinalis embryo during development. Analysis of Raman spectra scattered from C. intestinalis embryos revealed a number of localized patterns of high Raman intensity within the embryo. Based on the observed distribution of bio-molecules, we succeeded in identifying the location and structure of differentiated muscle and endoderm within the whole embryo, up to the tailbud stage, in a label-free manner. Furthermore, during cell differentiation, we detected significant differences in cell state between muscle/endoderm daughter cells and daughter cells with other fates that had divided from the same mother cells; this was achieved by focusing on the Raman intensity of single Raman bands at 1002 or 1526 cm−1, respectively. This study reports the first application of Raman spectroscopic imaging to the study of identifying and characterizing differentiating tissues in a whole chordate embryo. Our results suggest that Raman spectroscopic imaging is a feasible label-free technique for investigating the developmental process of the whole embryo of C. intestinalis. PMID:23977129

  7. Scanning x-ray microscopy of living and freeze-dried blood cells in two vanadium-rich ascidian species, Phallusia mammillata and Ascidia sydneiensis samea.

    PubMed

    Ueki, Tatsuya; Takemoto, Kuniko; Fayard, Barbara; Salomé, Murielle; Yamamoto, Akitsugu; Kihara, Hiroshi; Susini, Jean; Scippa, Silvia; Uyama, Taro; Michibata, Hitoshi

    2002-01-01

    Some ascidians (sea squirts) accumulate the transitional metal vanadium in their blood cells at concentrations of up to 350 mM, about 10(7) times its concentration found in seawater. There are approximately 10 different types of blood cell in ascidians. The identity of the true vanadium-containing blood cell (vanadocyte) is controversial and little is known about the subcellular distribution of vanadium. A scanning x-ray microscope installed at the ID21 beamline of the European Synchroton Radiation Facility to visualize vanadium in ascidian blood cells. Without fixation, freezing or staining realized the visualization of vanadium localized in living signet ring cells and vacuolated amoebocytes of two vanadium-rich ascidian species, Phallusia mammillata and Ascidia sydneiensis samea. A combination of transmission and fluorescence images of signet ring cells suggested that in both species the vacuoles contain vanadium.

  8. Urochordate ascidians possess a single isoform of Aurora kinase that localizes to the midbody via TPX2 in eggs and cleavage stage embryos.

    PubMed

    Hebras, Celine; McDougall, Alex

    2012-01-01

    Aurora kinases are key proteins found throughout the eukaryotes that control mitotic progression. Vertebrate Aurora-A and B kinases are thought to have evolved from a single Aurora-kinase isoform closest to that found in present day urochordates. In urochordate ascidians Aurora binds both TPX2 (a vertebrate AURKA partner) and INCENP (a vertebrate AURKB partner) and localizes to centrosomes and spindle microtubules as well as chromosomes and midbody during both meiosis and mitosis. Ascidian Aurora also displays this localization pattern during mitosis in echinoderms, strengthening the idea that non-vertebrate deuterostomes such as the urochordates and echinoderms possess a single form of Aurora kinase that has properties of vertebrate Aurora-kinase A and B. In the ascidian, TPX2 localizes to the centrosome and the spindle poles also as in vertebrates. However, we were surprised to find that TPX2 also localized strongly to the midbody in ascidian eggs and embryos. We thus examined more closely Aurora localization to the midbody by creating two separate point mutations of ascidian Aurora predicted to perturb binding to TPX2. Both forms of mutated Aurora behaved as predicted: neither localized to spindle poles where TPX2 is enriched. Interestingly, neither form of mutated Aurora localized to the midbody where TPX2 is also enriched, suggesting that ascidian Aurora midbody localization required TPX2 binding in ascidians. Functional analysis revealed that inhibition of Aurora kinase with a pharmacological inhibitor or with a dominant negative kinase dead form of Aurora caused cytokinesis failure and perturbed midbody formation during polar body extrusion. Our data support the view that vertebrate Aurora-A and B kinases evolved from a single non-vertebrate deuterostome ancestor. Moreover, since TPX2 localizes to the midbody in ascidian eggs and cleavage stage embryos it may be worthwhile re-assessing whether Aurora A kinase or TPX2 localize to the midbody in eggs and

  9. Urochordate Ascidians Possess a Single Isoform of Aurora Kinase That Localizes to the Midbody via TPX2 in Eggs and Cleavage Stage Embryos

    PubMed Central

    Hebras, Celine; McDougall, Alex

    2012-01-01

    Aurora kinases are key proteins found throughout the eukaryotes that control mitotic progression. Vertebrate Aurora-A and B kinases are thought to have evolved from a single Aurora-kinase isoform closest to that found in present day urochordates. In urochordate ascidians Aurora binds both TPX2 (a vertebrate AURKA partner) and INCENP (a vertebrate AURKB partner) and localizes to centrosomes and spindle microtubules as well as chromosomes and midbody during both meiosis and mitosis. Ascidian Aurora also displays this localization pattern during mitosis in echinoderms, strengthening the idea that non-vertebrate deuterostomes such as the urochordates and echinoderms possess a single form of Aurora kinase that has properties of vertebrate Aurora-kinase A and B. In the ascidian, TPX2 localizes to the centrosome and the spindle poles also as in vertebrates. However, we were surprised to find that TPX2 also localized strongly to the midbody in ascidian eggs and embryos. We thus examined more closely Aurora localization to the midbody by creating two separate point mutations of ascidian Aurora predicted to perturb binding to TPX2. Both forms of mutated Aurora behaved as predicted: neither localized to spindle poles where TPX2 is enriched. Interestingly, neither form of mutated Aurora localized to the midbody where TPX2 is also enriched, suggesting that ascidian Aurora midbody localization required TPX2 binding in ascidians. Functional analysis revealed that inhibition of Aurora kinase with a pharmacological inhibitor or with a dominant negative kinase dead form of Aurora caused cytokinesis failure and perturbed midbody formation during polar body extrusion. Our data support the view that vertebrate Aurora-A and B kinases evolved from a single non-vertebrate deuterostome ancestor. Moreover, since TPX2 localizes to the midbody in ascidian eggs and cleavage stage embryos it may be worthwhile re-assessing whether Aurora A kinase or TPX2 localize to the midbody in eggs and

  10. Antagonizing Retinoic Acid and FGF/MAPK Pathways Control Posterior Body Patterning in the Invertebrate Chordate Ciona intestinalis

    PubMed Central

    Pasini, Andrea; Manenti, Raoul; Rothbächer, Ute; Lemaire, Patrick

    2012-01-01

    Vertebrate embryos exploit the mutual inhibition between the RA and FGF signalling pathways to coordinate the proliferative elongation of the main body axis with the progressive patterning and differentiation of its neuroectodermal and paraxial mesodermal structures. The evolutionary history of this patterning system is still poorly understood. Here, we investigate the role played by the RA and FGF/MAPK signals during the development of the tail structures in the tunicate Ciona intestinalis, an invertebrate chordate belonging to the sister clade of vertebrates, in which the prototypical chordate body plan is established through very derived morphogenetic processes. Ciona embryos are constituted of few cells and develop according to a fixed lineage; elongation of the tail occurs largely by rearrangement of postmitotic cells; mesoderm segmentation and somitogenesis are absent. We show that in the Ciona embryo, the antagonism of the RA and FGF/MAPK signals is required to control the anteroposterior patterning of the tail epidermis. We also demonstrate that the RA, FGF/MAPK and canonical Wnt pathways control the anteroposterior patterning of the tail peripheral nervous system, and reveal the existence of distinct subpopulations of caudal epidermal neurons with different responsiveness to the RA, FGF/MAPK and canonical Wnt signals. Our data provide the first demonstration that the use of the antagonism between the RA and FGF signals to pattern the main body axis predates the emergence of vertebrates and highlight the evolutionary plasticity of this patterning strategy, showing that in different chordates it can be used to pattern different tissues within the same homologous body region. PMID:23049976

  11. Antagonizing retinoic acid and FGF/MAPK pathways control posterior body patterning in the invertebrate chordate Ciona intestinalis.

    PubMed

    Pasini, Andrea; Manenti, Raoul; Rothbächer, Ute; Lemaire, Patrick

    2012-01-01

    Vertebrate embryos exploit the mutual inhibition between the RA and FGF signalling pathways to coordinate the proliferative elongation of the main body axis with the progressive patterning and differentiation of its neuroectodermal and paraxial mesodermal structures. The evolutionary history of this patterning system is still poorly understood. Here, we investigate the role played by the RA and FGF/MAPK signals during the development of the tail structures in the tunicate Ciona intestinalis, an invertebrate chordate belonging to the sister clade of vertebrates, in which the prototypical chordate body plan is established through very derived morphogenetic processes. Ciona embryos are constituted of few cells and develop according to a fixed lineage; elongation of the tail occurs largely by rearrangement of postmitotic cells; mesoderm segmentation and somitogenesis are absent. We show that in the Ciona embryo, the antagonism of the RA and FGF/MAPK signals is required to control the anteroposterior patterning of the tail epidermis. We also demonstrate that the RA, FGF/MAPK and canonical Wnt pathways control the anteroposterior patterning of the tail peripheral nervous system, and reveal the existence of distinct subpopulations of caudal epidermal neurons with different responsiveness to the RA, FGF/MAPK and canonical Wnt signals. Our data provide the first demonstration that the use of the antagonism between the RA and FGF signals to pattern the main body axis predates the emergence of vertebrates and highlight the evolutionary plasticity of this patterning strategy, showing that in different chordates it can be used to pattern different tissues within the same homologous body region.

  12. Evolution of the ascidian anural larva: evidence from embryos and molecules.

    PubMed

    Jeffery, W R; Swalla, B J; Ewing, N; Kusakabe, T

    1999-05-01

    Most ascidians pass through a tadpole (urodele) larval stage, although some species have derived a tailless (anural) larva. New insights into the evolution of anural larvae in the Roscovita clade of molgulid ascidians were obtained from studing embryonic development of the transitional anural species Molgula bleizi and from phylogenetic analysis based on muscle and cytoskeletal actin gene sequences. By observing in vitro fertilized eggs, we found that M. bleizi, previously described as a typical anural developer, actually forms a short immotile tail during embryogenesis. The short tail contains notochord lineage cells, which undergo abbreviated morphogenetic movements but eventually arrest in development. Molgula bleizi tail muscle lineage cells produce the muscle enzyme acetylcholinesterase (AChE) but do not express muscle actin genes. The presence of a short tail, a vestigial notochord, and AChE-positive muscle cells suggest that M. bleizi is a recently derived anural species. An M. bleizi larval muscle actin gene (MbMA1) was isolated, sequenced, and shown to be a pseudogene based on critical deletions in its coding region that would result in a nonfunctional actin protein. The mutations in MbMA1 are distinct from and have evolved independent of the larval muscle actin pseudogenes MoccMA1a and MoccMA1b in Molgula occulta, another anural developer in the Roscovita clade. Pseudogene formation explains the absence of muscle actin mRNA in M. bleizi embryos. The 3' untranslated region of an M. bleizi cytoskeletal actin gene was also isolated and sequenced. Phylogenetic trees reconstructed using muscle and cytoskeletal actin sequences suggest that the anural developer M. bleizi evolved prior to the divergence of the urodele developer Molgula oculata and the anural developer M. occulta in the Roscovita clade. Since M. bleizi lives attached to hard substrata in the tidal zone, whereas M. oculata and M. occulta live buried in subtidal sand flats, our results suggest that

  13. Wnt5 is required for notochord cell intercalation in the ascidian Halocynthia roretzi

    PubMed Central

    Niwano, Tomoko; Takatori, Naohito; Kumano, Gaku; Nishida, Hiroki

    2009-01-01

    Background information. In the embryos of various animals, the body elongates after gastrulation by morphogenetic movements involving convergent extension. The Wnt/PCP (planar cell polarity) pathway plays roles in this process, particularly mediolateral polarization and intercalation of the embryonic cells. In ascidians, several factors in this pathway, including Wnt5, have been identified and found to be involved in the intercalation process of notochord cells. Results. In the present study, the role of the Wnt5 genes, Hr-Wnt5α (Halocynthia roretzi Wnt5α) and Hr-Wnt5β, in convergent extension was investigated in the ascidian H. roretzi by injecting antisense oligonucleotides and mRNAs into single precursor blastomeres of various tissues, including notochord, at the 64-cell stage. Hr-Wnt5α is expressed in developing notochord and was essential for notochord morphogenesis. Precise quantitative control of its expression level was crucial for proper cell intercalation. Overexpression of Wnt5 proteins in notochord and other tissues that surround the notochord indicated that Wnt5α plays a role within the notochord, and is unlikely to be the source of polarizing cues arising outside the notochord. Detailed mosaic analysis of the behaviour of individual notochord cells overexpressing Wnt5α indicated that a Wnt5α-manipulated cell does not affect the behaviour of neighbouring notochord cells, suggesting that Wnt5α works in a cell-autonomous manner. This is further supported by comparison of the results of Wnt5α and Dsh (Dishevelled) knockdown experiments. In addition, our results suggest that the Wnt/PCP pathway is also involved in mediolateral intercalation of cells of the ventral row of the nerve cord (floor plate) and the endodermal strand. Conclusion. The present study highlights the role of the Wnt5α signal in notochord convergent extension movements in ascidian embryos. Our results raise the novel possibility that Wnt5α functions in a cell-autonomous manner

  14. Diiron centre mutations in Ciona intestinalis alternative oxidase abolish enzymatic activity and prevent rescue of cytochrome oxidase deficiency in flies.

    PubMed

    Andjelković, Ana; Oliveira, Marcos T; Cannino, Giuseppe; Yalgin, Cagri; Dhandapani, Praveen K; Dufour, Eric; Rustin, Pierre; Szibor, Marten; Jacobs, Howard T

    2015-12-17

    The mitochondrial alternative oxidase, AOX, carries out the non proton-motive re-oxidation of ubiquinol by oxygen in lower eukaryotes, plants and some animals. Here we created a modified version of AOX from Ciona instestinalis, carrying mutations at conserved residues predicted to be required for chelation of the diiron prosthetic group. The modified protein was stably expressed in mammalian cells or flies, but lacked enzymatic activity and was unable to rescue the phenotypes of flies knocked down for a subunit of cytochrome oxidase. The mutated AOX transgene is thus a potentially useful tool in studies of the physiological effects of AOX expression.

  15. Cytotoxicity of the Ascidian Cystodytes dellechiajei Against Tumor Cells and Study of the Involvement of Associated Microbiota in the Production of Cytotoxic Compounds

    PubMed Central

    Martinez-García, Manuel; Diaz-Valdés, Marta; Ramos-Esplá, Alfonso; Salvador, Nélida; Lopez, Patricia; Larriba, Eduardo; Antón, Josefa

    2007-01-01

    Many cytotoxic compounds of therapeutic interest have been isolated from marine invertebrates, and some of them have been reported to be of microbial origin. Pyridoacridine alkaloids are the main compounds extracted from the ascidian Cystodytes dellechiajei. Here we describe the in vitro antiproliferative activity against different tumor cell lines of the ascidian extracts and provide some insights on the role of the microbial community associated with the tunicate in the production of these compounds. C. dellechiajei extracts showed remarkably high antiproliferative activity (IC50 ≤5 μg/mL) in human lung carcinoma A-549, colon adenocarcinoma H-116, pancreatic adenocarcinoma PSN-1 and breast carcinoma SKBR3 cell lines. Moreover, we found that the maximum activity was located in the tunic tissue of the colony, which harbours a microbial community. In order to ascertain the involvement of this community in the synthesis of the bioactive compounds different approachs that included culture and culture independent methods were carried out. We undertook a screening for antiproliferative activities of the bacterial isolates from the ascidian, as well as a comprative analysis of the cytotoxic activities and the microbial communities from two color morphs of the ascidian, green and blue. In addition, the changes of the antiproliferative activities and the composition of the microbial communities were studied from ascidians kept in aquaria and treated with antibiotics for one month. Our data obtained from the different experiments did not point out to bacteria as the source of the cytotoxic compounds, suggesting thus an ascidian origin. PMID:18463720

  16. Ultrastructures and classification of circulating hemocytes in 9 botryllid ascidians (chordata: ascidiacea).

    PubMed

    Hirose, Euichi; Shirae, Maki; Saito, Yasunori

    2003-05-01

    Ultrastructures of circulating hemocytes were studied in 9 botryllid ascidians. The hemocytes are classified into five types: hemoblasts, phagocytes, granulocytes, morula cells, and pigment cells. These five types are always found in the 9 species. They should represent the major hemocyte types of the circulating cells in the blood. Hemoblasts are small hemocytes having a high nucleus/cytoplasm ratio. There are few granular or vacuolar inclusions in the cytoplasm. Phagocytes have phagocytic activity and their shape is variable depending on the amount of engulfed materials. In granulocytes, shape and size of granules are different among the species. Morula cells are characterized by several vacuoles filled with electron dense materials. In pigment cells, the bulk of the cytoplasm is occupied by one or a few vacuoles containing pigment granules. We also described some other hemocyte types found in particular species. Furthermore, we encountered free oocytes circulating in the blood in two species, Botryllus primigenus and Botrylloides lentus.

  17. Experimental allometry: effect of size manipulation on metabolic rate of colonial ascidians

    PubMed Central

    Nakaya, Fumio; Saito, Yasunori; Motokawa, Tatsuo

    2005-01-01

    The allometric scaling of metabolic rate of organisms, the three-quarters power rule, has led to a questioning of the basis for the relation. We attacked this problem experimentally for the first time by employing the modular organism, the ascidian that forms a single layered flat colony, as a model system. The metabolic rate and colony size followed the three-quarters power relation, which held even after the colony size was experimentally manipulated. Our results established that the three-quarters power relation is a real continuous function, not an imaginary statistical regression. The fact that all the hypotheses failed to explain why the two-dimensional organism adhered to the three-quarters power relation led us to propose a new hypothesis, in which the allometric relation derives from the self-organized criticality based on local interaction between modulus-comprising organisms. PMID:16191604

  18. [Distribution and abundance of the ascidian Ecteinascidia turbinata (Ascidiacea: Perophoridae) in Cuba].

    PubMed

    Hernández-Zanuy, Aida; Carballo, José Luis; García-Cagide, Alida; Naranjo, Santiago; Esquivel, Macario

    2007-03-01

    Permanently submerged mangrove roots (Rhizophora mangle) are the main habitat of the ascidian Ecteinascidia turbinata in Cuba. It was occasionally found on black coral (Antiphates caribeana) between 22 and 38 meters deep. This species exhibits a wide distribution in all the mangrove keys surrounding the Island of Cuba but does not occur in riparian or fringing mangroves. Populations of this species are abundant in Cuba: in 75% of the 58 localities sampled the species was present and in 57% more than 50% of the roots held at least one colony. The highest colony densities were found in the northern coast of Pinar del Rio province with values near one colony per lineal meter of mangrove root. We found the highest density (1.46 col/m) and greatest biomass at Jutías Key, with values between 25 and 660 g/m. The average of wet biomass in the studied mangroves was 73.63 g/m.

  19. Structure of ascidiacyclamide as the ethanol water solvate, a cytotoxic cyclic peptide from Ascidian.

    PubMed

    In, Y; Doi, M; Inoue, M; Ishida, T; Hamada, Y; Shioiri, T

    1994-12-15

    The X-ray crystal structure determination of the C2H5OH.H2O solvate of ascidiacyclamide (C36H52N8O6S2), a cytotoxic cyclic peptide from marine tunicate Ascidian, revealed a C2-symmetric saddle-shaped rectangular conformation of the molecule. The water and ethanol molecules are located on the crystallographic diad axis and are held by hydrogen bonds and van der Waals contacts with the polar ring N atoms and nonpolar D-Val side-chain atoms, respectively. The molecular conformation and the interaction with solvent molecules are nearly the same as those of the compound with C2H5OH.2H2O [Ishida, In, Doi, Inoue, Hamada & Shioiri (1992). Biopolymers, 32, 131-143].

  20. Unfractionated Heparin and New Heparin Analogues from Ascidians (Chordate-Tunicate) Ameliorate Colitis in Rats*

    PubMed Central

    Belmiro, Celso L. R.; Castelo-Branco, Morgana T. L.; Melim, Leandra M. C.; Schanaider, Alberto; Elia, Celeste; Madi, Kalil; Pavão, Mauro S. G.; de Souza, Heitor S. P.

    2009-01-01

    The anti-inflammatory effect of mammalian heparin analogues, named dermatan sulfate and heparin, isolated from the ascidian Styela plicata was accessed in a TNBS-induced colitis model in rats. Subcutaneous administration of the invertebrate compounds during a 7-day period drastically reduced inflammation as observed by the normalization of the macroscopic and histological characteristics of the colon. At the molecular level, a decrease in the production of TNF-α, TGF-β, and VEGF was observed, as well as a reduction of NF-κB and MAPK kinase activation. At the cellular level, the heparin analogues attenuated lymphocyte and macrophage recruitment and epithelial cell apoptosis. A drastic reduction in collagen-mediated fibrosis was also observed. No hemorrhagic events were observed after glycan treatment. These results strongly indicate the potential therapeutic use of these compounds for the treatment of colonic inflammation with a lower risk of hemorrhage when compared with mammalian heparin. PMID:19258310

  1. 3-Bromohomofascaplysin A, a fascaplysin analogue from a Fijian Didemnum sp. ascidian

    PubMed Central

    Lu, Zhenyu; Ding, Yuanqing; Li, Xing-Cong; Djigbenou, Daignon R.; Grimberg, Brian T.; Ferreira, Daneel; Ireland, Chris M.; Van Wagoner, Ryan M.

    2011-01-01

    A new fascaplysin analogue, 3-bromohomofascaplysin A (1), along with two known analogues, homofascaplysin A (2) and fascaplysin (3), were isolated from a Fijian Didemnum sp. ascidian. The absolute configurations of 3-bromohomofascaplysin A (1) and homofascaplysin A (2) were determined via experimental and theoretically calculated ECD spectra. The differential activities of 1–3 against different blood-borne life stages of the malaria pathogen Plasmodium falciparum were assessed. Homofascaplysin A (2) displayed an IC50 of 0.55 ± 0.11 nM against ring stage parasites and 105 ± 38 nM against all live parasites. Given the stronger resistance of ring stage parasites against most current antimalarials relative to the other blood stages, homofascaplysin A (2) represents a promising agent for treatment of drug resistant malaria. PMID:21696970

  2. [Diet of the ascidian Ecteinascidia turbinata (Ascidiacea: Perophoridae) in two mangrove areas of Cuba].

    PubMed

    Hernández-Zanuy, Aida; García-Cagide, Alida; Borrero, Nilda; Carballo, José Luis

    2007-06-01

    Diet of the ascidian Ecteinascidia turbinata (Ascidiacea: Perophoridae) in two mangrove areas of Cuba. Stomach contents of 88 zooids ofEcteinascidia turbinata Herdman 1880 and the qualitative and quantitative composition of phytoplankton in the water column were studied in Santa Fe (North Coast of Havana) and Punta del Este (SW of Cuba). We identified 59 microalgal species and four tintinnids in the stomachs. Cell size was 75-165 microm in length and 2-105 microm in width. There were not significant differences in microalgal diversity in the water column in the two locations. In both locations, the diatoms had the largest number of species and individuals in stomachs and water. In Santa Fe, dinoflagellate biomass was larger in water and stomach contents, while in Punta del Este the contribution of each group to the stomach content was similar to that of the water column. This species filters in a constant and irregular way during the day, independently of food availability.

  3. Efficient targeted mutagenesis of the chordate Ciona intestinalis genome with zinc-finger nucleases.

    PubMed

    Kawai, Narudo; Ochiai, Hiroshi; Sakuma, Tetsushi; Yamada, Lixy; Sawada, Hitoshi; Yamamoto, Takashi; Sasakura, Yasunori

    2012-06-01

    Zinc-finger nucleases (ZFNs) are engineered nucleases that induce DNA double-strand breaks (DSBs) at target sequences. They have been used as tools for generating targeted mutations in the genomes of multiple organisms in both animals and plants. The DSB induced by ZFNs is repaired by non-homologous end joining (NHEJ) or by homologous recombination (HR) mechanisms. Non-homologous end joining induces some errors because it is independent of a reference DNA sequence. Through the NHEJ mechanism, ZFNs generate insertional or deletional mutations at the target sequence. We examined the usability, specificity and toxicity of ZFNs in the basal chordate Ciona intestinalis. As the target of ZFNs, we chose an enhanced green fluorescent protein (EGFP) gene artificially inserted in the C. intestinalis genome because this locus is neutral for the development and growth of C. intestinalis, and the efficiency of mutagenesis with ZFNs can thus be determined without any bias. We introduced EGFP -ZFN mRNAs into the embryos of an EGFP -transgenic line and observed the mutation frequency in the target site of EGFP . We also examined the effects of the EGFP -ZFNs at off-target sites resembling the EGFP target sequence in the C. intestinalis genome in order to examine the specificity of ZFNs. We further investigated the influence of ZFNs on embryogenesis, and showed that adequate amounts of ZFNs, which do not disrupt embryogenesis, can efficiently induce mutations on the on-target site with less effect on the off-target sites. This suggests that target mutagenesis with ZFNs will be a powerful technique in C. intestinalis.

  4. Crossing the species barrier: genomic hotspots of introgression between two highly divergent Ciona intestinalis species.

    PubMed

    Roux, Camille; Tsagkogeorga, Georgia; Bierne, Nicolas; Galtier, Nicolas

    2013-07-01

    Inferring a realistic demographic model from genetic data is an important challenge to gain insights into the historical events during the speciation process and to detect molecular signatures of selection along genomes. Recent advances in divergence population genetics have reported that speciation in face of gene flow occurred more frequently than theoretically expected, but the approaches used did not account for genome-wide heterogeneity (GWH) in introgression rates. Here, we investigate the impact of GWH on the inference of divergence with gene flow between two cryptic species of the marine model Ciona intestinalis by analyzing polymorphism and divergence patterns in 852 protein-coding sequence loci. These morphologically similar entities are highly diverged molecular-wise, but evidence of hybridization has been reported in both laboratory and field studies. We compare various speciation models and test for GWH under the approximate Bayesian computation framework. Our results demonstrate the presence of significant extents of gene flow resulting from a recent secondary contact after >3 My of divergence in isolation. The inferred rates of introgression are relatively low, highly variable across loci and mostly unidirectional, which is consistent with the idea that numerous genetic incompatibilities have accumulated over time throughout the genomes of these highly diverged species. A genomic map of the level of gene flow identified two hotspots of introgression, that is, large genome regions of unidirectional introgression. This study clarifies the history and degree of isolation of two cryptic and partially sympatric model species and provides a methodological framework to investigate GWH at various stages of speciation process.

  5. The Simple Chordate Ciona intestinalis Has a Reduced Complement of Genes Associated with Fanconi Anemia.

    PubMed

    Stanley, Edward C; Azzinaro, Paul A; Vierra, David A; Howlett, Niall G; Irvine, Steven Q

    2016-01-01

    Fanconi anemia (FA) is a human genetic disease characterized by congenital defects, bone marrow failure, and increased cancer risk. FA is associated with mutation in one of 24 genes. The protein products of these genes function cooperatively in the FA pathway to orchestrate the repair of DNA interstrand cross-links. Few model organisms exist for the study of FA. Seeking a model organism with a simpler version of the FA pathway, we searched the genome of the simple chordate Ciona intestinalis for homologs of the human FA-associated proteins. BLAST searches, sequence alignments, hydropathy comparisons, maximum likelihood phylogenetic analysis, and structural modeling were used to infer the likelihood of homology between C. intestinalis and human FA proteins. Our analysis indicates that C. intestinalis indeed has a simpler and potentially functional FA pathway. The C. intestinalis genome was searched for candidates for homology to 24 human FA and FA-associated proteins. Support was found for the existence of homologs for 13 of these 24 human genes in C. intestinalis. Members of each of the three commonly recognized FA gene functional groups were found. In group I, we identified homologs of FANCE, FANCL, FANCM, and UBE2T/FANCT. Both members of group II, FANCD2 and FANCI, have homologs in C. intestinalis. In group III, we found evidence for homologs of FANCJ, FANCO, FANCQ/ERCC4, FANCR/RAD51, and FANCS/BRCA1, as well as the FA-associated proteins ERCC1 and FAN1. Evidence was very weak for the existence of homologs in C. intestinalis for any other recognized FA genes. This work supports the notion that C. intestinalis, as a close relative of vertebrates, but having a much reduced complement of FA genes, offers a means of studying the function of certain FA proteins in a simpler pathway than that of vertebrate cells.

  6. The Simple Chordate Ciona intestinalis Has a Reduced Complement of Genes Associated with Fanconi Anemia

    PubMed Central

    Stanley, Edward C.; Azzinaro, Paul A.; Vierra, David A.; Howlett, Niall G.; Irvine, Steven Q.

    2016-01-01

    Fanconi anemia (FA) is a human genetic disease characterized by congenital defects, bone marrow failure, and increased cancer risk. FA is associated with mutation in one of 24 genes. The protein products of these genes function cooperatively in the FA pathway to orchestrate the repair of DNA interstrand cross-links. Few model organisms exist for the study of FA. Seeking a model organism with a simpler version of the FA pathway, we searched the genome of the simple chordate Ciona intestinalis for homologs of the human FA-associated proteins. BLAST searches, sequence alignments, hydropathy comparisons, maximum likelihood phylogenetic analysis, and structural modeling were used to infer the likelihood of homology between C. intestinalis and human FA proteins. Our analysis indicates that C. intestinalis indeed has a simpler and potentially functional FA pathway. The C. intestinalis genome was searched for candidates for homology to 24 human FA and FA-associated proteins. Support was found for the existence of homologs for 13 of these 24 human genes in C. intestinalis. Members of each of the three commonly recognized FA gene functional groups were found. In group I, we identified homologs of FANCE, FANCL, FANCM, and UBE2T/FANCT. Both members of group II, FANCD2 and FANCI, have homologs in C. intestinalis. In group III, we found evidence for homologs of FANCJ, FANCO, FANCQ/ERCC4, FANCR/RAD51, and FANCS/BRCA1, as well as the FA-associated proteins ERCC1 and FAN1. Evidence was very weak for the existence of homologs in C. intestinalis for any other recognized FA genes. This work supports the notion that C. intestinalis, as a close relative of vertebrates, but having a much reduced complement of FA genes, offers a means of studying the function of certain FA proteins in a simpler pathway than that of vertebrate cells. PMID:27279728

  7. The development of three identified motor neurons in the larva of an ascidian, Halocynthia roretzi.

    PubMed

    Okada, Toshiaki; Katsuyama, You; Ono, Fumihito; Okamura, Yasushi

    2002-04-15

    The generation of distinct classes of motor neurons underlies the development of complex motile behavior in all animals and is well characterized in chordates. Recent molecular studies indicate that the ascidian larval central nervous system (CNS) exhibits anteroposterior regionalization similar to that seen in the vertebrate CNS. To extend the understanding about the diversity of motor neurons in the ascidian larva, we have identified the number, position, and projection of individual motor neurons in Halocynthia roretzi, using a green fluorescent protein under the control of a neuron-specific promoter. Three pairs of motor neurons, each with a distinct shape and innervation pattern, were identified along the anteroposterior axis of the neural tube: the anterior and posterior pairs extend their axons toward dorsal muscle cells, whereas the middle pair project their axons toward ventral muscle. Overexpression of a dominant-negative form of a potassium channel in these cells resulted in paralysis on the injected side, thus these cells must constitute the major population of motor neurons responsible for swimming behavior. Lim class homeobox genes have been known as candidate genes that determine subtypes of motor neurons. Therefore, the expression pattern of Hrlim, which is a Lim class homeobox gene, was examined in the motor neuron precursors. All three motor neurons expressed Hrlim at the tailbud stage, although each down-regulated Hrlim at a different time. Misexpression of Hrlim in the epidermal lineage led to ectopic expression of TuNa2, a putative voltage-gated channel gene normally expressed predominantly in the three pairs of motor neurons. Hrlim may control membrane excitability of motor neurons by regulating ion channel gene expression.

  8. Ciona intestinalis as a marine model system to study some key developmental genes targeted by the diatom-derived aldehyde decadienal.

    PubMed

    Lettieri, Anna; Esposito, Rosaria; Ianora, Adrianna; Spagnuolo, Antonietta

    2015-03-17

    The anti-proliferative effects of diatoms, described for the first time in copepods, have also been demonstrated in benthic invertebrates such as polychaetes, sea urchins and tunicates. In these organisms PUAs (polyunsaturated aldehydes) induce the disruption of gametogenesis, gamete functionality, fertilization, embryonic mitosis, and larval fitness and competence. These inhibitory effects are due to the PUAs, produced by diatoms in response to physical damage as occurs during copepod grazing. The cell targets of these compounds remain largely unknown. Here we identify some of the genes targeted by the diatom PUA 2-trans-4-trans-decadienal (DD) using the tunicate Ciona intestinalis. The tools, techniques and genomic resources available for Ciona, as well as the suitability of Ciona embryos for medium-to high-throughput strategies, are key to their employment as model organisms in different fields, including the investigation of toxic agents that could interfere with developmental processes. We demonstrate that DD can induce developmental aberrations in Ciona larvae in a dose-dependent manner. Moreover, through a preliminary analysis, DD is shown to affect the expression level of genes involved in stress response and developmental processes.

  9. Characterization and transcription studies of a phytochelatin synthase gene from the solitary tunicate Ciona intestinalis exposed to cadmium.

    PubMed

    Franchi, Nicola; Piccinni, Ester; Ferro, Diana; Basso, Giuseppe; Spolaore, Barbara; Santovito, Gianfranco; Ballarin, Loriano

    2014-07-01

    The major thiol-containing molecules involved in controlling the level of intracellular ROS in eukaryotes, acting as a nonenzymatic detoxification system, are metallothioneins (MTs), glutathione (GSH) and phytochelatins (PCs). Both MTs and GSH are well-known in the animal kingdom. PC was considered a prerogative of the plant kingdom but, in 2001, a phytochelatin synthase (PCS) gene was described in the nematode Caenorhabditis elegans; additional genes encoding this enzyme were later described in the earthworm Eisenia fetida and in the parasitic nematode Schistosoma mansoni but scanty data are available, up to now, for Deuterostomes. Here, we describe the molecular characteristics and transcription pattern, in the presence of Cd, of a PCS gene from the invertebrate chordate Ciona intestinalis, a ubiquitous solitary tunicate and demonstrate the presence of PCs in tissue extracts. We also studied mRNA localization by in situ hybridization. In addition, we analyzed the behavior of hemocytes and tunic cells consequent to Cd exposure as well as the transcription pattern of the Ciona orthologous for proliferating cell nuclear antigen (PCNA), usually considered a proliferation marker, and observed that cell proliferation occurs after 96h of Cd treatment. This matches the hypothesis of Cd-induced cell proliferation, as already suggested by previous data on the expression of a metallothionein gene in the same animal.

  10. Molecular and morphological discrimination between an invasive ascidian, Ascidiella aspersa, and its congener A. scabra (Urochordata: Ascidiacea).

    PubMed

    Nishikawa, Teruaki; Oohara, Ichiro; Saitoh, Kenji; Shigenobu, Yuya; Hasegawa, Natsuki; Kanamori, Makoto; Baba, Katsuhisa; Turon, Xavier; Bishop, John D D

    2014-03-01

    The solitary ascidian Ascidiella aspersa (Müller, 1776) has sometimes been regarded as conspecific with A. scabra (Müller, 1776), although previous detailed morphological comparisons have indicated that the two are distinguishable by internal structures. Resolution of this taxonomic issue is important because A. aspersa has been known as a notoriously invasive ascidian, doing much damage to aquaculture e.g. in Hokkaido, Japan. We collected many specimens from European waters (including the Swedish coast, near the type localities of these two species) and Hokkaido, Japan (as an alien population) and made molecular phylogenetic analyses using the mitochondrial cytochrome c oxidase subunit I (COI) gene, and found that in terms of COI sequences all the analyzed specimens were clustered into two distinct groups, one of which is morphologically referable to A. aspersa and the other to A. scabra. Thus, these two species should be regarded as distinct from each other.

  11. A quantitative study of elements in the blood cells of the tropical ascidian Phallusia philippinensis using the nuclear microscope

    NASA Astrophysics Data System (ADS)

    Thong, P. S. P.; Law, N. C.; Orlic, I.; Lane, D. J. W.; Watt, F.

    1995-09-01

    Two types of blood cells of the tropical ascidian Phallusia philippinensis have been studied using the NUS nuclear microscope. Morula cells were found to contain 1-2% (by weight) of vanadium and 0.1-0.85% of bromine and signet ring cells about 0.16% vanadium and much reduced levels of bromine (about 50 ppm or less). The signet ring cells were also found to contain very high levels of sulphur (20 wt%), calcium (16 wt%) and oxygen (40 wt% of the cell matrix). The occurrence of sulphur and oxygen in the approximate ratios of 1:4 adds weight to the suggestion by several authors of the presence of sulphates in the ascidian blood cells. Such high levels of sulphur and oxygen were not seen in the morula cells.

  12. Microenvironmental Ecology of the Chlorophyll b-Containing Symbiotic Cyanobacterium Prochloron in the Didemnid Ascidian Lissoclinum patella

    PubMed Central

    Kühl, Michael; Behrendt, Lars; Trampe, Erik; Qvortrup, Klaus; Schreiber, Ulrich; Borisov, Sergey M.; Klimant, Ingo; Larkum, Anthony W. D.

    2012-01-01

    The discovery of the cyanobacterium Prochloron was the first finding of a bacterial oxyphototroph with chlorophyll (Chl) b, in addition to Chl a. It was first described as Prochloron didemni but a number of clades have since been described. Prochloron is a conspicuously large (7–25 μm) unicellular cyanobacterium living in a symbiotic relationship, primarily with (sub-) tropical didemnid ascidians; it has resisted numerous cultivation attempts and appears truly obligatory symbiotic. Recently, a Prochloron draft genome was published, revealing no lack of metabolic genes that could explain the apparent inability to reproduce and sustain photosynthesis in a free-living stage. Possibly, the unsuccessful cultivation is partly due to a lack of knowledge about the microenvironmental conditions and ecophysiology of Prochloron in its natural habitat. We used microsensors, variable chlorophyll fluorescence imaging and imaging of O2 and pH to obtain a detailed insight to the microenvironmental ecology and photobiology of Prochloron in hospite in the didemnid ascidian Lissoclinum patella. The microenvironment within ascidians is characterized by steep gradients of light and chemical parameters that change rapidly with varying irradiances. The interior zone of the ascidians harboring Prochloron thus became anoxic and acidic within a few minutes of darkness, while the same zone exhibited O2 super-saturation and strongly alkaline pH after a few minutes of illumination. Photosynthesis showed lack of photoinhibition even at high irradiances equivalent to full sunlight, and photosynthesis recovered rapidly after periods of anoxia. We discuss these new insights on the ecological niche of Prochloron and possible interactions with its host and other microbes in light of its recently published genome and a recent study of the overall microbial diversity and metagenome of L. patella. PMID:23226144

  13. Microenvironmental Ecology of the Chlorophyll b-Containing Symbiotic Cyanobacterium Prochloron in the Didemnid Ascidian Lissoclinum patella.

    PubMed

    Kühl, Michael; Behrendt, Lars; Trampe, Erik; Qvortrup, Klaus; Schreiber, Ulrich; Borisov, Sergey M; Klimant, Ingo; Larkum, Anthony W D

    2012-01-01

    The discovery of the cyanobacterium Prochloron was the first finding of a bacterial oxyphototroph with chlorophyll (Chl) b, in addition to Chl a. It was first described as Prochloron didemni but a number of clades have since been described. Prochloron is a conspicuously large (7-25 μm) unicellular cyanobacterium living in a symbiotic relationship, primarily with (sub-) tropical didemnid ascidians; it has resisted numerous cultivation attempts and appears truly obligatory symbiotic. Recently, a Prochloron draft genome was published, revealing no lack of metabolic genes that could explain the apparent inability to reproduce and sustain photosynthesis in a free-living stage. Possibly, the unsuccessful cultivation is partly due to a lack of knowledge about the microenvironmental conditions and ecophysiology of Prochloron in its natural habitat. We used microsensors, variable chlorophyll fluorescence imaging and imaging of O(2) and pH to obtain a detailed insight to the microenvironmental ecology and photobiology of Prochloron in hospite in the didemnid ascidian Lissoclinum patella. The microenvironment within ascidians is characterized by steep gradients of light and chemical parameters that change rapidly with varying irradiances. The interior zone of the ascidians harboring Prochloron thus became anoxic and acidic within a few minutes of darkness, while the same zone exhibited O(2) super-saturation and strongly alkaline pH after a few minutes of illumination. Photosynthesis showed lack of photoinhibition even at high irradiances equivalent to full sunlight, and photosynthesis recovered rapidly after periods of anoxia. We discuss these new insights on the ecological niche of Prochloron and possible interactions with its host and other microbes in light of its recently published genome and a recent study of the overall microbial diversity and metagenome of L. patella.

  14. The proteomic response of sea squirts (genus Ciona) to acute heat stress: a global perspective on the thermal stability of proteins.

    PubMed

    Serafini, Loredana; Hann, Jay B; Kültz, Dietmar; Tomanek, Lars

    2011-09-01

    Congeners belonging to the genus Ciona have disparate distributions limited by temperature. Ciona intestinalis is more widespread with a cosmopolitan distribution ranging from tropical to sub-arctic zones, while Ciona savignyi is limited to temperate-latitudes of the northern Pacific Ocean. To compare the heat stress response between congeners, we quantified changes in protein expression using proteomics. Animals were exposed to 22°C, 25°C, and 28°C for 6h, then recovered at a control temperature (13°C) for 16h (high heat stress experiment). In a second experiment we exposed animals to lower levels of heat stress at 18°C, 20°C, and 23°C, with a 16°C control. A quantitative analysis, using 2D gel electrophoresis and MALDI-TOF/TOF mass spectrometry (with a 69% and 93% identification rate for Ciona intestinalis and Ciona savignyi, respectively), showed changes in a number of protein functional groups, including molecular chaperones, extracellular matrix proteins, calcium-binding proteins, cytoskeletal proteins and proteins involved in energy metabolism. Our results indicate that C. intestinalis maintains higher constitutive levels of molecular chaperones than C. savignyi, suggesting that it is prepared to respond faster to thermal stress. Systematic discrepancies between estimated versus predicted molecular masses of identified proteins differed between protein families and were more pronounced under high heat conditions, suggesting that thermal sensitivities are lower for cytoskeletal proteins and ATP-synthase than for any other protein group represented on 2D gels. Copyright © 2011 Elsevier Inc. All rights reserved.

  15. Calmodulin and immunophilin are required as functional partners of a ryanodine receptor in ascidian oocytes at fertilization.

    PubMed

    Albrieux, M; Moutin, M J; Grunwald, D; Villaz, M

    2000-09-01

    Fertilization of oocytes incites numerous changes relying on Ca(2+) signaling. In inseminated ascidian eggs, an increase in the egg surface membrane, monitored by a change in electrical capacitance, is recorded at the onset of meiosis resumption. This membrane addition to the cell surface is controlled by calcium release through a ryanodine receptor (RyR), sensitive to cyclic ADP-ribose. Using confocal microscopy analysis of ascidian oocytes immunostained with anti-RyR antibody, we show here that this calcium channel is asymmetrically located in the vegetal cortical zone. Interestingly, the increase in cell capacitance occurring at fertilization is correlated with a fluorescent signal, imaged by the marker of vesicle trafficking FM 1-43, located close to the RyR region. Two putative partners of RyR, namely an FKBP-like protein and a calmodulin, are identified in these oocyte extracts by detection of enzyme activity and PCR amplification. Both are necessary to sustain ryanodine receptor activity in these oocytes since the membrane insertion triggered by fertilization is inhibited by the FKBP ligand rapamycin and by a calmodulin antagonist peptide. These findings suggest that exocytosis in ascidian eggs is triggered at fertilization by a functional Ca(2+) release unit operating as a complex of several proteins, including a calmodulin and an immunophilin, around the intracellular calcium channel itself. Copyright 2000 Academic Press.

  16. Isolation of an early neural maker gene abundantly expressed in the nervous system of the ascidian, Halocynthia roretzi.

    PubMed

    Yagi, K; Makabe, K W

    2001-01-01

    Ascidian tadpole larvae possess a primitive nervous system, which is a prospective prototype of the chordate nervous system. It is composed of relatively few cells but sufficient for complex larval behavior. Here we report on HrETR-1, a gene zygotically expressed in a large proportion of the developing neural cells of the ascidian, Halocynthia roretzi. HrETR-1 is an early neural marker which can be used for analyzing neural differentiation. HrETR-1 expression intensified in most neural cells of genes isolated to date, in both central and peripheral nervous systems including palps as early as the 110-cell stage. Using this gene as a probe, we characterized neural cells in the nervous system as well as confirming their origins. Also, we recognized three types of peripheral epidermal neurons which presumably correlate to the larval neurons previously reported for another ascidian. Among these, five bilateral neurons located in the anterior region of the trunk appeared to be derived from a8.26 blastomeres.

  17. Activation of a tunicate (Ciona intestinalis) xenobiotic receptor orthologue by both natural toxins and synthetic toxicants.

    PubMed

    Fidler, Andrew E; Holland, Patrick T; Reschly, Erica J; Ekins, Sean; Krasowski, Matthew D

    2012-02-01

    Vertebrate xenobiotic receptors are ligand-activated nuclear receptors (NRs) that bind exogenous biologically active chemicals before activating the transcription of genes involved in xenobiotic metabolism and excretion. Typically, xenobiotic receptors have ligand binding domains (LBDs) that can accommodate a structurally diverse array of molecules and in addition display high levels of inter-taxa sequence diversity suggestive of positive selection. Pursuing the idea that xenobiotic receptors may adaptively evolve to bind toxic chemicals commonly present in an organism's environment/diet, we examined ligand binding by a xenobiotic receptor orthologue of a marine filter-feeding organism. The solitary tunicate Ciona intestinalis (Phylum Chordata) genome encodes an orthologue of the vertebrate pregnane X receptor (PXR) and vitamin D receptor (VDR), here denoted CiVDR/PXRα. In a luciferase reporter assay the CiVDR/PXRα was activated, at nanomolar concentrations, by two of four natural marine microalgal biotoxins tested (okadaic acid, EC50 = 18.2 ± 0.9 nM and pectenotoxin-2, EC50 = 37.0 ± 3.5 nM) along with 1 of 11 synthetic toxicants (esfenvalerate: EC50 = 0.59 ± 0.7 μM). Two related C. intestinalis NRs, orthologous to vertebrate farnesoid X receptor and liver X receptors, respectively, along with the PXR of a freshwater fish (zebrafish, Danio rerio), were not activated by any of the 15 chemicals tested. In contrast, human PXR was activated by okadaic acid at similar concentrations to CiVDR/PXRα (EC50 = 7.2 ± 1.1 nM) but not by pectenotoxin-2. A common features pharmacophore developed for the CiVDR/PXRα ligand consisted of an off-center hydrogen bond acceptor flanked by two hydrophobic regions. The results of this study are consistent with the original hypothesis that natural toxins, present in the diet of filter-feeding marine invertebrates, may have acted as selective agents in the molecular evolution of tunicate xenobiotic receptors. Bioassays based on

  18. Exploring the mechanism of action of the sperm-triggered calcium-wave pacemaker in ascidian zygotes.

    PubMed

    Carroll, Michael; Levasseur, Mark; Wood, Chris; Whitaker, Michael; Jones, Keith T; McDougall, Alex

    2003-12-15

    In ascidians, as in mammals, sperm trigger repetitive Ca2+-waves that originate from cortical pacemakers situated in the vegetal hemisphere of the zygotes. In ascidians, a vegetal protrusion termed the contraction pole (CP) acts as the Ca2+-wave pacemaker, but the mechanism that underlies the generation of a Ca2+-wave pacemaker is not known. Here, we tested four hypotheses to determine which factors at the CP are involved in setting the pace of the ascidian Ca2+-wave pacemaker: (1) localized Ca2+ influx; (2) accumulation of phosphatidylinositol (4,5)bisphosphate [PtdIns(4,5)P2]; (3) accumulation of cortical endoplasmic reticulum (cER); and (4) enrichment of the sperm activating factor. We developed a method of dynamically monitoring the location of the CP during fertilization using a plekstrin homology (PH) domain from phospholipase Cdelta1 coupled to green fluorescent protein (GFP) that binds PtdIns(4,5)P2. We found that eggs in Ca2+-free sea water displayed Ca2+ waves that originated from the CP, showing that enhanced CP Ca2+ influx does not determine the origin of the pacemaker. Also, disruption of the PH::GFP-labelled CP once it had formed did not dislodge the Ca2+-wave pacemaker from that site. Next, when we prevented the accumulation of cER at the CP, all of the Ca2+ waves came from the site of sperm-egg fusion and the frequency of Ca2+ oscillations was unaltered. These data show that local Ca2+ influx, the accumulation of PtdIns(4,5)P2 and cER at the CP are not required for Ca2+-wave pacemaker function and instead suggest that a factor associated with the sperm determines the site of the Ca2+-wave pacemaker. Finally, when we injected ascidian sperm extract into the centre of unfertilized ascidian eggs that had been treated with microfilament- and microtubule-disrupting drugs, all the Ca2+ waves still originated from near the plasma membrane, showing that the sperm factor does not require an intact cortex if it is enriched near the plasma membrane (PM). We

  19. From oocyte to 16-cell stage: cytoplasmic and cortical reorganizations that pattern the ascidian embryo.

    PubMed

    Sardet, Christian; Paix, Alexandre; Prodon, François; Dru, Philippe; Chenevert, Janet

    2007-07-01

    The dorsoventral and anteroposterior axes of the ascidian embryo are defined before first cleavage by means of a series of reorganizations that reposition cytoplasmic and cortical domains established during oogenesis. These domains situated in the periphery of the oocyte contain developmental determinants and a population of maternal postplasmic/PEM RNAs. One of these RNAs (macho-1) is a determinant for the muscle cells of the tadpole embryo. Oocytes acquire a primary animal-vegetal (a-v) axis during meiotic maturation, when a subcortical mitochondria-rich domain (myoplasm) and a domain rich in cortical endoplasmic reticulum (cER) and maternal postplasmic/PEM RNAs (cER-mRNA domain) become polarized and asymmetrically enriched in the vegetal hemisphere. Fertilization at metaphase of meiosis I initiates a series of dramatic cytoplasmic and cortical reorganizations of the zygote, which occur in two major phases. The first major phase depends on sperm entry which triggers a calcium wave leading in turn to an actomyosin-driven contraction wave. The contraction concentrates the cER-mRNA domain and myoplasm in and around a vegetal/contraction pole. The precise localization of the vegetal/contraction pole depends on both the a-v axis and the location of sperm entry and prefigures the future site of gastrulation and dorsal side of the embryo. The second major phase of reorganization occurs between meiosis completion and first cleavage. Sperm aster microtubules and then cortical microfilaments cause the cER-mRNA domain and myoplasm to reposition toward the posterior of the zygote. The location of the posterior pole depends on the localization of the sperm centrosome/aster attained during the first major phase of reorganization. Both cER-mRNA and myoplasm domains localized in the posterior region are partitioned equally between the first two blastomeres and then asymmetrically over the next two cleavages. At the eight-cell stage the cER-mRNA domain compacts and gives rise to

  20. Molecular basis of canalization in an ascidian species complex adapted to different thermal conditions

    PubMed Central

    Sato, Atsuko; Kawashima, Takeshi; Fujie, Manabu; Hughes, Samantha; Satoh, Noriyuki; Shimeld, Sebastian M.

    2015-01-01

    Canalization is a result of intrinsic developmental buffering that ensures phenotypic robustness under genetic variation and environmental perturbation. As a consequence, animal phenotypes are remarkably consistent within a species under a wide range of conditions, a property that seems contradictory to evolutionary change. Study of laboratory model species has uncovered several possible canalization mechanisms, however, we still do not understand how the level of buffering is controlled in natural populations. We exploit wild populations of the marine chordate Ciona intestinalis to show that levels of buffering are maternally inherited. Comparative transcriptomics show expression levels of genes encoding canonical chaperones such as Hsp70 and Hsp90 do not correlate with buffering. However the expression of genes encoding endoplasmic reticulum (ER) chaperones does correlate. We also show that ER chaperone genes are widely conserved amongst animals. Contrary to previous beliefs that expression level of Heat Shock Proteins (HSPs) can be used as a measurement of buffering levels, we propose that ER associated chaperones comprise a cellular basis for canalization. ER chaperones have been neglected by the fields of development, evolution and ecology, but their study will enhance understanding of both our evolutionary past and the impact of global environmental change. PMID:26577490

  1. A Maternal System Initiating the Zygotic Developmental Program through Combinatorial Repression in the Ascidian Embryo

    PubMed Central

    Oda-Ishii, Izumi; Kubo, Atsushi; Kari, Willi; Suzuki, Nobuhiro; Rothbächer, Ute

    2016-01-01

    Maternal factors initiate the zygotic developmental program in animal embryos. In embryos of the chordate, Ciona intestinalis, three maternal factors—Gata.a, β-catenin, and Zic-r.a—are required to establish three domains of gene expression at the 16-cell stage; the animal hemisphere, vegetal hemisphere, and posterior vegetal domains. Here, we show how the maternal factors establish these domains. First, only β-catenin and its effector transcription factor, Tcf7, are required to establish the vegetal hemisphere domain. Second, genes specifically expressed in the posterior vegetal domain have additional repressive cis-elements that antagonize the activity of β-catenin/Tcf7. This antagonizing activity is suppressed by Zic-r.a, which is specifically localized in the posterior vegetal domain and binds to DNA indirectly through the interaction with Tcf7. Third, Gata.a directs specific gene expression in the animal hemisphere domain, because β-catenin/Tcf7 weakens the Gata.a-binding activity for target sites through a physical interaction in the vegetal cells. Thus, repressive regulation through protein-protein interactions among the maternal transcription factors is essential to establish the first distinct domains of gene expression in the chordate embryo. PMID:27152625

  2. Vascular budding in Symplegma brakenhielmi and the evolution of coloniality in styelid ascidians.

    PubMed

    Gutierrez, Stefania; Brown, Federico D

    2017-03-15

    Individuals of colonial animals (e.g. zooids) are in continuous turnover. In ascidians colonial or solitary species have evolved by convergence multiple times. Colonial Botryllus and Botrylloides are well-studied genera that exhibit colony-wide developmental mechanisms that regulate synchronous and orchestrated cycles of budding and turnover of zooids. The origins of modular developmental mechanisms that facilitated the evolution of coloniality in this group remain unclear. To reconstruct ancestral states of coloniality we studied Symplegma brakenhielmi, a sister taxon of the botryllids. S. brakenhielmi zooids are embedded in a common tunic and present a similar vascular system as the botrylloides, however development and turnover of zooids occurs asynchronously and in a more independent manner. We generated a table of common stages of budding in Symplegma and Botryllus for comparative studies of asexual development. We tested dependent processes of budding among individuals of the colony by systemic bud or zooid removals. Although our results showed a higher degree of independence in bud development in S. brakenhielmi, we found a subtle colony-wide regulatory mechanism of modular development, i.e. new buds expedited development after the removal of all buds in the colony. Next, we characterized external morphology, ultrastructure, and abundance of circulatory blood cells in the vascular system of S. brakenhielmi. Macrophage-like cells (MLCs) are involved in zooid resorption and turnover. Proportions of MLCs in the blood of S. brakenhielmi corresponded to the peak of occurrence of this cell type during the budding cycle of B. schlosseri. We found several new blood cell types in S. brakenhielmi, including two cell types that resemble circulatory progenitor stem cells of other botryllid colonial ascidians. These cells showed features of undifferentiated cells and expressed mitotic marker Phospho-histone H3. Comparative studies of S. brakenhielmi and B. schlosseri

  3. Ecological observations on the colonial ascidian Didemnum sp. in a New England tide pool habitat

    USGS Publications Warehouse

    Valentine, P.C.; Carman, M.R.; Blackwood, D.S.; Heffron, E.J.

    2007-01-01

    The colonial ascidian Didemnum sp. has colonized northwestern Atlantic coastal habitats from southern Long Island, New York, to Eastport, Maine. It is also present in offshore habitats of the Georges Bank fishing grounds. It threatens to alter fisheries habitats and shellfish aquacultures. Observations in a tide pool at Sandwich, MA from December 2003 to February 2006 show that Didemnum sp. tolerates water temperatures ranging from ≤ 1 to > 24 °C, with daily changes of up to 11 °C. It attaches to pebbles, cobbles, and boulders, and it overgrows other tunicates, seaweeds, sponges, and bivalves. From May to mid July, colonies appear as small patches on the bottoms of rocks. Colonies grow rapidly from July to September, with some growth into December, and they range in color from pink to pale yellow to pale orange. Colony health declines from October through April, presumably in response to changes in water temperatures, and this degenerative process is manifested by color changes, by the appearance of small dark brown spots that represent clumps of fecal pellets in the colony, by scavenging by periwinkles, and by a peeling-away of colonies from the sides of cobbles and boulders. At Sandwich, colonies died that were exposed to air at low tide. The species does not exhibit this seasonal cycle of growth and decline in subtidal habitats (40–65 m) on the Georges Bank fishing grounds where the daily climate is relatively stable and annual water temperatures range from 4 to 15 °C. Experiments in the tide pool with small colony fragments (5 to 9 cm2) show they re-attach and grow rapidly by asexual budding, increasing in size 6- to 11-fold in the first 15 days. Didemnum sp. at Sandwich has no known predators except for common periwinkles (Littorina littorea) that graze on degenerating colonies in the October to April time period and whenever colonies are stressed by desiccation. The tendencies of the ascidian (1) to attach to firm substrates, (2) to rapidly overgrow

  4. Isolation and Characterization of a Shewanella Phage–Host System from the Gut of the Tunicate, Ciona intestinalis

    PubMed Central

    Leigh, Brittany; Karrer, Charlotte; Cannon, John P.; Breitbart, Mya; Dishaw, Larry J.

    2017-01-01

    Outnumbering all other biological entities on earth, bacteriophages (phages) play critical roles in structuring microbial communities through bacterial infection and subsequent lysis, as well as through horizontal gene transfer. While numerous studies have examined the effects of phages on free-living bacterial cells, much less is known regarding the role of phage infection in host-associated biofilms, which help to stabilize adherent microbial communities. Here we report the cultivation and characterization of a novel strain of Shewanella fidelis from the gut of the marine tunicate Ciona intestinalis, inducible prophages from the S. fidelis genome, and a strain-specific lytic phage recovered from surrounding seawater. In vitro biofilm assays demonstrated that lytic phage infection affects biofilm formation in a process likely influenced by the accumulation and integration of the extracellular DNA released during cell lysis, similar to the mechanism that has been previously shown for prophage induction. PMID:28327522

  5. Complete DNA sequence of the mitochondrial genome of the ascidian Halocynthia roretzi (Chordata, Urochordata).

    PubMed

    Yokobori, S i; Ueda, T; Feldmaier-Fuchs, G; Pääbo, S; Ueshima, R; Kondow, A; Nishikawa, K; Watanabe, K

    1999-12-01

    The complete nucleotide sequence of the 14,771-bp-long mitochondrial (mt) DNA of a urochordate (Chordata)-the ascidian Halocynthia roretzi-was determined. All the Halocynthia mt-genes were found to be located on a single strand, which is rich in T and G rather than in A and C. Like nematode and Mytilus edulis mtDNAs, that of Halocynthia encodes no ATP synthetase subunit 8 gene. However, it does encode an additional tRNA gene for glycine (anticodon TCT) that enables Halocynthia mitochondria to use AGA and AGG codons for glycine. The mtDNA carries an unusual tRNA(Met) gene with a TAT anticodon instead of the usual tRNA(Met)(CAT) gene. As in other metazoan mtDNAs, there is not any long noncoding region. The gene order of Halocynthia mtDNA is completely different from that of vertebrate mtDNAs except for tRNA(His)-tRNA(Ser)(GCU), suggesting that evolutionary change in the mt-gene structure is much accelerated in the urochordate line compared with that in vertebrates. The amino acid sequences of Halocynthia mt-proteins deduced from their gene sequences are quite different from those in other metazoans, indicating that the substitution rate in Halocynthia mt-protein genes is also accelerated.

  6. Fertilization in an egg-brooding colonial ascidian does not vary with population density.

    PubMed

    Phillippi, Aimee; Hamann, Ellen; Yund, Philip O

    2004-06-01

    The possibility that free-spawning marine organisms may be subject to fertilization failure at low population density (due to the effects of sperm dilution) has sparked much interest, but these effects have been demonstrated only in a few species that broadcast their eggs. Some egg-brooding species may overcome dilution effects by filtering low concentrations of sperm from seawater and fertilizing eggs throughout an extended period of time. We examined the effects of population density and size on fertilization in Botryllus schlosseri, a hermaphroditic colonial ascidian that free-spawns sperm, but broods eggs. We experimentally manipulated the size and density of mating groups and surveyed fertilization levels in natural populations that varied in density. Fertilization was not affected by variation in population size or density in either the experimental or natural populations. Near the end of the reproductive season, some eggs may have been fertilized too late to complete development, suggesting a temporal form of sperm limitation that has not been considered in other systems. We also detected greater variability in fertilization levels at lower population density. Nevertheless, these results suggest that caution must be used in extrapolating reported density effects on fertilization to all taxa of free-spawners; density effects may be reduced in brooders that have efficient sperm collection mechanisms.

  7. Global Phylogeography of the Widely Introduced North West Pacific Ascidian Styela clava

    PubMed Central

    Goldstien, Sharyn J.; Dupont, Lise; Viard, Frédérique; Hallas, Paul J.; Nishikawa, Teruaki; Schiel, David R.; Gemmell, Neil J.; Bishop, John D. D.

    2011-01-01

    The solitary ascidian Styela clava Herdman, 1882 is considered to be native to Japan, Korea, northern China and the Russian Federation in the NW Pacific, but it has spread globally over the last 80 years and is now established as an introduced species on the east and west coasts of North America, Europe, Australia and New Zealand. In eastern Canada it reaches sufficient density to be a serious pest to aquaculture concerns. We sequenced a fragment of the cytochrome oxidase subunit I mitochondrial gene (COI) from a total of 554 individuals to examine the genetic relationships of 20 S. clava populations sampled throughout the introduced and native ranges, in order to investigate invasive population characteristics. The data presented here show a moderate level of genetic diversity throughout the northern hemisphere. The southern hemisphere (particularly New Zealand) displays a greater amount of haplotype and nucleotide diversity in comparison. This species, like many other invasive species, shows a range of genetic diversities among introduced populations independent of the age of incursion. The successful establishment of this species appears to be associated with multiple incursions in many locations, while other locations appear to have experienced rapid expansion from a potentially small population with reduced genetic diversity. These contrasting patterns create difficulties when attempting to manage and mitigate a species that continues to spread among ports and marinas around the world. PMID:21364988

  8. Immunotoxicity in ascidians: antifouling compounds alternative to organotins-IV. The case of zinc pyrithione.

    PubMed

    Cima, Francesca; Ballarin, Loriano

    2015-03-01

    New biocides such as the organometallic compound zinc pyrithione (ZnP) have been massively introduced by many countries in formulations of antifouling paints following the ban on tributyltin (TBT). The effects of sublethal concentrations (LC50=82.5 μM, i.e., 26.2 mg/l) on cultured haemocytes of the ascidian Botryllus schlosseri have been investigated and compared with TBT. The percentage of haemocytes with amoeboid morphology and containing phagocytised yeast cells were significantly (p<0.05) reduced after exposure to 0.1 (31.7 μg/l) and 0.5 μM (158 μg/l), respectively. An antagonistic interaction in inducing cytoskeletal alterations was observed when ZnP and TBT were co-present in the exposure medium. ZnP affected only the actin component. As caused by TBT, ZnP induced apoptosis and inhibited both oxidative phosphorylation and lysosomal activities. In contrast to the case of TBT, a decrement in Ca(2+)-ATPase activity and a decrease in cytosolic Ca(2+) were detected after incubation at the highest concentration (1 μM, i.e., 317.7 μg/l) used. In comparison with other antifouling compounds, ZnP shows as much toxicity as TBT to cultured haemocytes at extremely low concentrations interfering with fundamental cell activities.

  9. Global phylogeography of the widely introduced North West Pacific ascidian Styela clava.

    PubMed

    Goldstien, Sharyn J; Dupont, Lise; Viard, Frédérique; Hallas, Paul J; Nishikawa, Teruaki; Schiel, David R; Gemmell, Neil J; Bishop, John D D

    2011-02-22

    The solitary ascidian Styela clava Herdman, 1882 is considered to be native to Japan, Korea, northern China and the Russian Federation in the NW Pacific, but it has spread globally over the last 80 years and is now established as an introduced species on the east and west coasts of North America, Europe, Australia and New Zealand. In eastern Canada it reaches sufficient density to be a serious pest to aquaculture concerns. We sequenced a fragment of the cytochrome oxidase subunit I mitochondrial gene (COI) from a total of 554 individuals to examine the genetic relationships of 20 S. clava populations sampled throughout the introduced and native ranges, in order to investigate invasive population characteristics. The data presented here show a moderate level of genetic diversity throughout the northern hemisphere. The southern hemisphere (particularly New Zealand) displays a greater amount of haplotype and nucleotide diversity in comparison. This species, like many other invasive species, shows a range of genetic diversities among introduced populations independent of the age of incursion. The successful establishment of this species appears to be associated with multiple incursions in many locations, while other locations appear to have experienced rapid expansion from a potentially small population with reduced genetic diversity. These contrasting patterns create difficulties when attempting to manage and mitigate a species that continues to spread among ports and marinas around the world.

  10. Nuclear microscope analysis of blood cells from the tropical ascidian Phallusia philippinensis

    NASA Astrophysics Data System (ADS)

    Hogarth, A. N.; Thong, P. S. P.; Lane, D. J. W.; Watt, F.

    1997-07-01

    The present study examines the concentrations of vanadium, bromine and sulphur contained within cryofixed/freeze dried blood cells of the ascidian Phallusia philippinensis (Millar, 1975). Elemental profiles of seven cell types were obtained using the National University of Singapore nuclear microscope. Morula cells were found to contain the following mean values; 0.8% vanadium, 3.5% bromine and 6.1% sulphur. Signet ring cells contained 0.5% vanadium, 2.4% bromine and 1.5% sulphur. Compartment cells had 0.1% vanadium, 2.1% bromine and 2.4% sulphur. Other less abundant cell types such as lymphocytes, macrogranular amoebocytes, carotenoid pigment cells and granular amoebocytes were also analysed and found to contain 0.4%, 0.7%, 0.2% and 1.0% vanadium, 2.0%, 1.6%, 0.6% and 1.2% bromine and 1.3%, 1.5%, 0.3% and 4.3% sulphur respectively. Sulphur occurred in high levels in all cell types, which could indicate its involvement in the vanadium concentration process, while bromine, incorporated into complexes, may be utilised for anti-fouling rather than as a deterrent to predators.

  11. Complete DNA sequence of the mitochondrial genome of the ascidian Halocynthia roretzi (Chordata, Urochordata).

    PubMed Central

    Yokobori, S i; Ueda, T; Feldmaier-Fuchs, G; Pääbo, S; Ueshima, R; Kondow, A; Nishikawa, K; Watanabe, K

    1999-01-01

    The complete nucleotide sequence of the 14,771-bp-long mitochondrial (mt) DNA of a urochordate (Chordata)-the ascidian Halocynthia roretzi-was determined. All the Halocynthia mt-genes were found to be located on a single strand, which is rich in T and G rather than in A and C. Like nematode and Mytilus edulis mtDNAs, that of Halocynthia encodes no ATP synthetase subunit 8 gene. However, it does encode an additional tRNA gene for glycine (anticodon TCT) that enables Halocynthia mitochondria to use AGA and AGG codons for glycine. The mtDNA carries an unusual tRNA(Met) gene with a TAT anticodon instead of the usual tRNA(Met)(CAT) gene. As in other metazoan mtDNAs, there is not any long noncoding region. The gene order of Halocynthia mtDNA is completely different from that of vertebrate mtDNAs except for tRNA(His)-tRNA(Ser)(GCU), suggesting that evolutionary change in the mt-gene structure is much accelerated in the urochordate line compared with that in vertebrates. The amino acid sequences of Halocynthia mt-proteins deduced from their gene sequences are quite different from those in other metazoans, indicating that the substitution rate in Halocynthia mt-protein genes is also accelerated. PMID:10581290

  12. Multilocus genetic analyses differentiate between widespread and spatially restricted cryptic species in a model ascidian.

    PubMed

    Bock, Dan G; MacIsaac, Hugh J; Cristescu, Melania E

    2012-06-22

    Elucidating the factors that shape species distributions has long been a fundamental goal in ecology and evolutionary biology. In spite of significant theoretical advancements, empirical studies of range limits have lagged behind. Specifically, little is known about how the attributes that allow species to expand their ranges and become widespread vary across phylogenies. Here, we studied the ascidian Botryllus schlosseri, a worldwide invasive species that is also characterized by marked genetic subdivision. Our study includes phylogenetic and population genetic data based on mitochondrial and nuclear genes, as well as polymorphic microsatellites for B. schlosseri colonies sampled from the southern and northern coasts of Europe and the eastern and western coasts of North America. We demonstrate that this well-known model organism comprises three highly divergent and probably reproductively isolated cryptic species (A, D and E), with two more (B and C) being suggested by data retrieved from GenBank. Among these, species A, recovered in all of the surveyed regions, is by far the most common and widespread. By contrast, species B-E, occurring mostly in sites from northern Europe, are considerably more geographically restricted. These findings, along with inferences made on transport opportunity, suggest that divergent evolutionary histories promoted differences in invasive potential between B. schlosseri sibling species, indicating that attributes that facilitate dramatic shifts in range limits can evolve more easily and frequently than previously thought. We propose environmental disturbance as a selective force that could have shaped the evolution of invasiveness in the B. schlosseri complex.

  13. Localization of CiCBR in the invertebrate chordate Ciona intestinalis: evidence of an ancient role for cannabinoid receptors as axonal regulators of neuronal signalling.

    PubMed

    Egertová, Michaela; Elphick, Maurice R

    2007-06-01

    CiCBR is a G-protein-coupled receptor in the sea-squirt Ciona intestinalis and the first ortholog of vertebrate CB(1) and CB(2) cannabinoid receptors to be identified in an invertebrate (Elphick et al. [2003] Gene 302:95-101). Here we have used Western blotting and immunocytochemistry to examine expression of CiCBR in adult Ciona, employing novel antibodies to the C-terminal tail of CiCBR. Consistent with the expected mass for CiCBR, a approximately 47-kDa band was detected in Ciona membranes, and immunocytochemical analysis of serial sections of Ciona revealed intense immunoreactivity in the cerebral ganglion localised in a dense meshwork of fibers in the neuropile. Accordingly, Western blot analysis of neural complex homogenates revealed the presence of a approximately 47-kDa band. CiCBR immunoreactivity was also observed in axons exiting the ganglion in the anterior and posterior nerves, and analysis of whole-mount preparations revealed that these axons project over the interior surface of the oral and atrial siphons. Isolated CiCBR-immunoreactive axons not associated with the anterior and posterior nerves were observed projecting through the cortical layer of the cerebral ganglion. Central and peripheral CiCBR-immunoreactive fibers were studded with intensely stained varicosities, indicative of a role for CiCBR in regulation of axonal release of neurotransmitters, neuromodulators, or neurohormones. Collectively, our data suggest that the well-established role that the CB(1) receptor has as an axonal regulator of neurotransmitter release in mammals may have originated with ancestral-type cannabinoid receptors in invertebrate chordates before the emergence of CB(1)- and CB(2)-type receptors in vertebrates.

  14. Mechanisms of helical swimming: asymmetries in the morphology, movement and mechanics of larvae of the ascidian Distaplia occidentalis.

    PubMed

    McHenry, M J

    2001-09-01

    A great diversity of unicellular and invertebrate organisms swim along a helical path, but it is not well understood how asymmetries in the body shape or the movement of propulsive structures affect a swimmer's ability to perform the body rotation necessary to move helically. The present study found no significant asymmetries in the body shape of ascidian larvae (Distaplia occidentalis) that could operate to rotate the body during swimming. By recording the three-dimensional movement of free-swimming larvae, it was found that the tail possessed two bends, each with constant curvature along their length. As these bends traveled posteriorly, the amplitude of curvature changes was significantly greater in the concave-left direction than in the concave-right direction. In addition to this asymmetry, the tail oscillated at an oblique angle to the midline of the trunk. These asymmetries generated a yawing moment that rotated the body in the counterclockwise direction from a dorsal view, according to calculations from hydrodynamic theory. The tails of resting larvae were bent in the concave-left direction with a curvature statistically indistinguishable from the median value for tail curvature during swimming. The flexural stiffness of the tails of larvae, measured in three-point bending, may be great enough to allow the resting curvature of the tail to have an effect on the symmetry of kinematics. This work suggests that asymmetrical tail motion is an important mechanism for generating a yawing moment during swimming in ascidian larvae and that these asymmetries may be caused by the tail's bent shape. Since helical motion requires that moments also be generated in the pitching or rolling directions, other mechanisms are required to explain fully how ascidian larvae generate and control helical swimming.

  15. Origin of the Henze solution/precipitate from morula cells of the blood of the ascidian Phallusia mammillata

    NASA Astrophysics Data System (ADS)

    Nette, Geoffrey; Scippa, S.; de Vincentiis, Mario

    The "Henze solution", derived originally from the aqueous extraction of pelleted whole blood from the ascidian Phallusia mammillata, was examined using spectral studies. The aqueous extraction of fractionated blood cells including compartment cells, signet ring cells, and morula cells obtained using cell separation techniques were also examined. It was found that this Henze solution, and the Henze precipitate itself derived from this solution, emanated solely from the morula cells. Furthermore, it was found that this solution is formed independently of the vanadium metal ions otherwise associated with the vanadocytes. Observation of the Henze precipitate by light microscopy shows that this material partially forms crystallites or microglasses.

  16. Biology of the invasive ascidian Ascidiella aspersa in its native habitat: Reproductive patterns and parasite load

    NASA Astrophysics Data System (ADS)

    Lynch, Sharon A.; Darmody, Grainne; O'Dwyer, Katie; Gallagher, Mary Catherine; Nolan, Sinead; McAllen, Rob; Culloty, Sarah C.

    2016-11-01

    The European sea squirt Ascidiella aspersa is a solitary tunicate native to the northeastern Atlantic, commonly found in shallow and sheltered marine ecosystems where it is capable of forming large clumps and outcompeting other invertebrate fauna at settlement. To date, there have been relatively few studies looking at the reproductive biology and health status of this invasive species. Between 2006 and 2010 sampling of a native population took place to investigate gametogenesis and reproductive cycle and to determine the impact of settlement depth on reproduction. In addition, parasite diversity and impact was assessed. A staging system to assess reproductive development was determined. The study highlighted that from year to year the tunicate could change its reproductive strategy from single sex to hermaphrodite, with spawning possible throughout the year. Depth did not impact on sex determination, however, gonad maturation and spawning occurred earlier in individuals in deeper waters compared to shallow depth and it also occurred later in A. aspersa at sites further away from the open sea. Four significant parasite groups including eugregarines, ciliates, trematodes and turbellarians were detected and prevalence of parasite infections increased in A. aspersa at sites with a reduced water flow rate. This study demonstrates the high biotic potential of this ascidian bioinvader to have a negative impact on native fauna in an introduced ecosystem, due to its highly efficient reproductive and resource allocation strategies. Artificial structures such as mooring lines can harbour large aggregations of A. aspersa, however, these manmade habitats may facilitate the colonisation and establishment of this invasive species in the benthos. Additionally, the parasite communities that A. aspersa harbour may also exacerbate its negative impact, both ecologically and economically, in an introduced area by possibly leading to the emergence of new disease in native species i

  17. The candidate Fu/HC gene in Botryllusschlosseri (Urochordata) and ascidians' historecognition--an oxymoron?

    PubMed

    Rinkevich, Buki; Douek, Jacob; Rabinowitz, Claudette; Paz, Guy

    2012-04-01

    Allorecognition, distinguishing self from non-self allogeneic tissues is the underlying basis of innate immunity. In the colonial tunicate Botryllus schlosseri this historecognition is governed at a single genetic locus, Fu/HC (for fusibility/histocompatibility), with hundreds of co-dominantly expressed alleles. Several years ago, De Tomaso et al. (2005) have revolutionized the discipline of invertebrate allorecognition by describing a novel form of immune recognition in B. schlosseri, a non-vertebrate candidate histocompatibility gene (cFu/HC), revealing that allorecognition machinery in urochordates has nothing in common with the vertebrates' MHC-based histocompatibility. The authors reported absolute concordance of fusibility and cFu/HC genotype, predicted fusion/rejection outcomes in allorecognition settings from allelic polymorphism at the cFu/HC, also claiming cFu/HC gene expressions only in tissues directly engaged in histocompatibility. Here, we raise queries for the validity of the results and conclusions of De Tomaso et al. (2005) publication. Our reservations include discrepancies in the paper's results, including the perplexing absence of key sequencing material from public domains and above all, our own impugning outcomes. These include cloning efforts, in situ hybridization results, semi quantitative PCR outcomes, and the incongruence emerged between fusion/rejection profiles and cFu/HC segregated polymorphism that separately and cumulatively contradict the original publication. We conclude that Botryllus histocompatibility properties are not signaled in the claimed cFu/HC and that cFu/HC gene is unlikely the allodeterminant for Botryllus histocompatibility locus. Hence, the molecular nature of the Fu/HC locus in botryllid ascidians is still awaiting elucidation.

  18. The hydrodynamics of locomotion at intermediate Reynolds numbers: undulatory swimming in ascidian larvae (Botrylloides sp.).

    PubMed

    McHenry, Matthew J; Azizi, Emanuel; Strother, James A

    2003-01-01

    Understanding how the shape and motion of an aquatic animal affects the performance of swimming requires knowledge of the fluid forces that generate thrust and drag. These forces are poorly understood for the large diversity of animals that swim at Reynolds numbers (Re) between 10(0) and 10(2). We experimentally tested quasi-steady and unsteady blade-element models of the hydrodynamics of undulatory swimming in the larvae of the ascidian Botrylloides sp. by comparing the forces predicted by these models with measured forces generated by tethered larvae and by comparing the swimming speeds predicted with measurements of the speed of freely swimming larvae. Although both models predicted mean forces that were statistically indistinguishable from measurements, the quasi-steady model predicted the timing of force production and mean swimming speed more accurately than the unsteady model. This suggests that unsteady force (i.e. the acceleration reaction) does not play a role in the dynamics of steady undulatory swimming at Re approximately 10(2). We explored the relative contribution of viscous and inertial force to the generation of thrust and drag at 10(0)10(2)) and low (<10(0)) Re, the fluid forces that generate thrust cannot be assumed to be the same as those that generate drag at intermediate Re.

  19. The invariant cleavage pattern displayed by ascidian embryos depends on spindle positioning along the cell's longest axis in the apical plane and relies on asynchronous cell divisions

    PubMed Central

    Dumollard, Rémi; Minc, Nicolas; Salez, Gregory; Aicha, Sameh Ben; Bekkouche, Faisal; Hebras, Céline; Besnardeau, Lydia; McDougall, Alex

    2017-01-01

    The ascidian embryo is an ideal system to investigate how cell position is determined during embryogenesis. Using 3D timelapse imaging and computational methods we analyzed the planar cell divisions in ascidian early embryos and found that spindles in every cell tend to align at metaphase in the long length of the apical surface except in cells undergoing unequal cleavage. Furthermore, the invariant and conserved cleavage pattern of ascidian embryos was found to consist in alternate planar cell divisions between ectoderm and endomesoderm. In order to test the importance of alternate cell divisions we manipulated zygotic transcription induced by β-catenin or downregulated wee1 activity, both of which abolish this cell cycle asynchrony. Crucially, abolishing cell cycle asynchrony consistently disrupted the spindle orienting mechanism underpinning the invariant cleavage pattern. Our results demonstrate how an evolutionary conserved cell cycle asynchrony maintains the invariant cleavage pattern driving morphogenesis of the ascidian blastula. DOI: http://dx.doi.org/10.7554/eLife.19290.001 PMID:28121291

  20. Transmission of cyanobacterial symbionts during embryogenesis in the coral reef ascidians Trididemnum nubilum and T. clinides (Didemnidae, Ascidiacea, Chordata).

    PubMed

    Kojima, Aoi; Hirose, Euichi

    2012-02-01

    Vertical transmission of cyanobacterial symbionts occurs in didemnid ascidians harboring Prochloron as an obligate symbiont; the photosymbionts are transferred from the parental ascidian colony to the offspring in various ways depending on host species. Although several didemnids harbor non-Prochloron cyanobacteria in their tunics, few studies have reported the processes of vertical transmission in these didemnids. Here we describe the histological processes of the transmission of cyanobacteria in two didemnids, Trididemnum nubilum harboring Synechocystis and T. clinides harboring three cyanobacterial species. In both species, the photosymbionts in the tunic of the parent colony were apparently captured by the tunic cells of the host and transferred to the embryos brooded in the tunic. The symbiont cells were then incorporated into the inner tunic of the embryo. This mode of transmission is essentially the same as that of T. miniatum harboring Prochloron in the tunic, although there are some differences among species in the timing of the release of the symbionts from the tunic cells. We suggest that the similar modes of vertical transmission are an example of convergent evolution caused by constraints in the distribution patterns of symbiont cells in the host colony.

  1. Isolation of cDNA clones for mRNAs transcribed zygotically during cleavage in the ascidian, Halocynthia roretzi.

    PubMed

    Miya, Takahito; Nishida, Hiroki

    2002-02-01

    The ascidian larva consists of a relatively small number of different cell types, and the cell lineages during embryogenesis have been well described. The clonal restriction of developmental fate takes place considerably early in development. The fates of most of the blastomeres become tissue-restricted by the 110-cell stage, just before the onset of gastrulation. To elucidate the molecular basis of the early events of fate determination in the ascidian Halocynthia roretzi, we isolated the genes for which zygotic expression is initiated during the early cleavage stages. Here we report 18 genes isolated by subtractive hybridization screening between 110-cell embryos and fertilized eggs. The expression of most (13) of the genes was initiated at the 32-cell stage. The genes were subdivided into three groups according to their spatial expression patterns. The first group included clones expressed throughout almost the entire embryo. The second and third groups represented clones expressed mainly in the animal hemisphere and in a subset of vegetal blastomeres, respectively. One of the genes, HrHesl1, encoded a polypeptide containing the bHLH domain that is similar to those of the Hairy/Enhancer of split/Deadpan family of transcriptional repressors. HrHesl1 was expressed exclusively in epidermal precursor cells during cleavage. Another gene named HrWnt-5 beta was expressed in muscle precursors.

  2. Trypsin-like enzyme from eggs of the ascidian (protochordate), Halocynthia roretzi. Purification, properties, and physiological role.

    PubMed

    Sawada, H; Kawahigashi, M; Yokosawa, H; Ishii, S

    1985-12-15

    A trypsin-like enzyme has been purified to apparent homogeneity from eggs of the ascidian, Halocynthia roretzi, by a procedure including column chromatography on diethylaminoethyl-cellulose, phenyl-Sepharose, and soybean trypsin inhibitor-immobilized Sepharose 4B. The molecular weight of the enzyme was estimated to be 31,000 and 33,000 by gel electrophoresis in sodium dodecyl sulfate under the reducing and the nonreducing conditions, respectively. The isoelectric point of the enzyme was 4.8. The pH optimum of the activity was 8.4. The enzyme was stable between pH 6 and 9 in the presence of 0.005% Brij 35 as a stabilizer. Substrate specificity of the purified enzyme was broad toward various peptidyl-arginine (or -lysine) 4-methylcoumaryl-7-amides and was similar to that of a trypsin-like enzyme found in the fertilization product. The purified enzyme was inhibited by diisopropyl fluorophosphate and a variety of trypsin inhibitors including leupeptin, but not, or scarcely, inhibited by p-chloromercuribenzoic acid, pepstatin, chymostatin, bestatin, elastatinal, and tosyl-phenylalanyl-chloromethane. The rankings in the potencies of leupeptin and its six analogs as the inhibitors of the purified enzyme were well correlated with those found in their inhibitory effects on the expansion of perivitelline space. Thus, the trypsin-like enzyme possibly present in the fertilization product participates in the expansion of perivitelline space of the egg during fertilization of the ascidian.

  3. Morphological characterization of the tunic in the edible ascidian, Halocynthia roretzi (Drasche), with remarks on 'soft tunic syndrome' in aquaculture.

    PubMed

    Hirose, E; Ohtake, S-I; Azumi, K

    2009-05-01

    'Soft tunic syndrome' is a serious problem in the aquaculture of the edible ascidian, Halocynthia roretzi (Drasche), and often leads to mass mortality. Here, we describe the tunic morphology of intact and diseased ascidians to reveal structural differences between them. Morphologically, diseased tunics are not very different from intact tunics, although the former are thinner and softer than the latter. While several types of cells are distributed in the tunic, the cell types and their cytomorphologies were almost identical in both groups. As bacterial/protozoan cells were not found in either intact or diseased tunics, they are not the direct cause of soft tunic syndrome. The most remarkable difference was in the bundles of tunic fibres that compose the tunic matrix; in intact tunics, the thick bundles interlace to form a firm matrix, whereas in soft tunics, the tunic fibres do not form thick bundles. Furthermore, areas of low fibre density were found in diseased tunics. Therefore, soft tunic syndrome probably causes inhibition of bundle formation and degradation of tunic bundles, creating areas of low fibre density, although the causes remain unknown.

  4. Deep Sequencing of Mixed Total DNA without Barcodes Allows Efficient Assembly of Highly Plastic Ascidian Mitochondrial Genomes

    PubMed Central

    Rubinstein, Nimrod D.; Feldstein, Tamar; Shenkar, Noa; Botero-Castro, Fidel; Griggio, Francesca; Mastrototaro, Francesco; Delsuc, Frédéric; Douzery, Emmanuel J.P.; Gissi, Carmela; Huchon, Dorothée

    2013-01-01

    Ascidians or sea squirts form a diverse group within chordates, which includes a few thousand members of marine sessile filter-feeding animals. Their mitochondrial genomes are characterized by particularly high evolutionary rates and rampant gene rearrangements. This extreme variability complicates standard polymerase chain reaction (PCR) based techniques for molecular characterization studies, and consequently only a few complete Ascidian mitochondrial genome sequences are available. Using the standard PCR and Sanger sequencing approach, we produced the mitochondrial genome of Ascidiella aspersa only after a great effort. In contrast, we produced five additional mitogenomes (Botrylloides aff. leachii, Halocynthia spinosa, Polycarpa mytiligera, Pyura gangelion, and Rhodosoma turcicum) with a novel strategy, consisting in sequencing the pooled total DNA samples of these five species using one Illumina HiSeq 2000 flow cell lane. Each mitogenome was efficiently assembled in a single contig using de novo transcriptome assembly, as de novo genome assembly generally performed poorly for this task. Each of the new six mitogenomes presents a different and novel gene order, showing that no syntenic block has been conserved at the ordinal level (in Stolidobranchia and in Phlebobranchia). Phylogenetic analyses support the paraphyly of both Ascidiacea and Phlebobranchia, with Thaliacea nested inside Phlebobranchia, although the deepest nodes of the Phlebobranchia–Thaliacea clade are not well resolved. The strategy described here thus provides a cost-effective approach to obtain complete mitogenomes characterized by a highly plastic gene order and a fast nucleotide/amino acid substitution rate. PMID:23709623

  5. Deep sequencing of mixed total DNA without barcodes allows efficient assembly of highly plastic ascidian mitochondrial genomes.

    PubMed

    Rubinstein, Nimrod D; Feldstein, Tamar; Shenkar, Noa; Botero-Castro, Fidel; Griggio, Francesca; Mastrototaro, Francesco; Delsuc, Frédéric; Douzery, Emmanuel J P; Gissi, Carmela; Huchon, Dorothée

    2013-01-01

    Ascidians or sea squirts form a diverse group within chordates, which includes a few thousand members of marine sessile filter-feeding animals. Their mitochondrial genomes are characterized by particularly high evolutionary rates and rampant gene rearrangements. This extreme variability complicates standard polymerase chain reaction (PCR) based techniques for molecular characterization studies, and consequently only a few complete Ascidian mitochondrial genome sequences are available. Using the standard PCR and Sanger sequencing approach, we produced the mitochondrial genome of Ascidiella aspersa only after a great effort. In contrast, we produced five additional mitogenomes (Botrylloides aff. leachii, Halocynthia spinosa, Polycarpa mytiligera, Pyura gangelion, and Rhodosoma turcicum) with a novel strategy, consisting in sequencing the pooled total DNA samples of these five species using one Illumina HiSeq 2000 flow cell lane. Each mitogenome was efficiently assembled in a single contig using de novo transcriptome assembly, as de novo genome assembly generally performed poorly for this task. Each of the new six mitogenomes presents a different and novel gene order, showing that no syntenic block has been conserved at the ordinal level (in Stolidobranchia and in Phlebobranchia). Phylogenetic analyses support the paraphyly of both Ascidiacea and Phlebobranchia, with Thaliacea nested inside Phlebobranchia, although the deepest nodes of the Phlebobranchia-Thaliacea clade are not well resolved. The strategy described here thus provides a cost-effective approach to obtain complete mitogenomes characterized by a highly plastic gene order and a fast nucleotide/amino acid substitution rate.

  6. Familial Mediterranean Fever.

    PubMed

    Kucuk, Adem; Gezer, Ilknur Albayrak; Ucar, Ramazan; Karahan, Ali Yavuz

    2014-01-01

    Familial Mediterranean Fever is an autosomal recessive inherited disease with a course of autoinflammation, which is characterized by the episodes of fever and serositis. It affects the populations from Mediterranean basin. Genetic mutation of the disease is on MEFV gene located on short arm of Chromosome 16. The disease is diagnosed based on clinical evaluation. Amyloidosis is the most important complication. The only agent that decreases the development of amyloidosis and the frequency and severity of the episodes is colchicine, which has been used for about 40 years. In this review, we aimed to discuss especially the most recent advances about Familial Mediterranean Fever which is commonly seen in our population.

  7. Mediterranean California, Chapter 13

    Treesearch

    M.E. Fenn; E.B. Allen; L.H. Geiser

    2011-01-01

    The Mediterranean California ecoregion (CEC 1997; Fig 2.2) encompasses the greater Central Valley, Sierra foothills, and central coast ranges of California south to Mexico and is bounded by the Pacific Ocean, Sierra Nevada Mountains and Mojave Desert.

  8. Familial Mediterranean Fever

    MedlinePlus

    ... don't use genetic tests as the sole method of diagnosing familial Mediterranean fever. There's no cure ... may be options, though these treatments are considered experimental. Other medications include rilonacept (Arcalyst) and anakinra (Kineret). ...

  9. Tracking the Mediterranean Abyss

    NASA Astrophysics Data System (ADS)

    Aracri, S.; Schroeder, K.; Chiggiato, J.; Bryden, H. L.; McDonagh, E.; Josey, S. A.; Hello, Y.; Borghini, M.

    2016-02-01

    The Mediterranean Sea is well known to be a miniature ocean with small enough timescales to allow the observation of main oceanographic events, e.g. deep water formation and overturning circulation, in a human life time. This renders the Mediterranean Sea the perfect observatory to study and forecast the behaviour of the world ocean. Considering the coherence between NAO (North Atlantic Oscillation), AMO (Atlantic Multidecadal Oscillation) and Mediterranean oscillation and bearing in mind that the Mediterranean outflow at Gibraltar constitutes a constant source of intermediate, warm and saline water, it has been suggested that "the system composed of the North Atlantic, the Mediterranean Sea/Gibraltar Strait and the Arctic Sea/Fram Strait might work as a unique oceanographic entity, with the physical processes within the straits determining the exchange of the fresh and salty waters between the marginal seas and the open ocean".In the light of the present knowledge the Mediterranean might, then, be considered as a key oceanographic observatory site. The deep sea is still challenging to monitor, especially given the latest years lack of fundings and ships availability. Therefore optimizing the existing methods and instrumentation has become a priority. This work is focused on the North-Western Mediterranean basin, where deep water formation events often occur in the Gulf of Lion as well as deep convection in the neighbour Ligurian Sea. A different application of submarine robots - Mermaids- designed to observe underwater seismic waves aiming to improve ocean tomography is presented. In order to improve our knowledge of the North-Western Mediterranean abyssal circulation we track Mermaids extracting their velocity, correcting it and comparing it with the historically estimated values and with the geostrophic velocity extracted from a 40 years long hydrographic datasets.

  10. Neural differentiation in cleavage-arrested ascidian blastomeres induced by a proteolytic enzyme.

    PubMed Central

    Okado, H; Takahashi, K

    1993-01-01

    1. As previously reported, ectodermal a4-2 blastomeres isolated from 8-cell embryos of the ascidian, Halocynthia roretzi or aurantium, and cultured under conditions of cleavage arrest always differentiated into an epidermal phenotype, showing long-lasting Ca(2+)-dependent action potentials and/or tunic on the cell surface. a4-2 blastomeres contacted by a chordamesodermal blastomere, A4-1, differentiated into a neural phenotype, characterized by fast Na(+)-dependent spikes. Differentiation to a similar neural phenotype occurred when isolated a4-2 blastomeres from H. aurantium embryos were treated with > 0.003% subtilisin for 60 min at the 32-cell stage of the control embryo. Comparisons between induction by cell contact and induction by proteolytic enzymes were made and showed them to be similar in several respects. 2. When the serine protease, subtilisin, was used as the neural inducer, neural competence of a4-2 blastomeres, measured as the percentage frequency of the induction of Na+ spikes, increased after the 32-cell stage and decreased during the gastrula stage. The time course of the neural competence was the same as that for contact with the A4-1 blastomere. 3. The neural competence of four different ectodermal blastomeres isolated from the 16-cell embryo was also examined using subtilisin as a neural inducer, and by contact with the A4-1 blastomere from the 8-cell embryo. The competence was higher in anterior blastomeres than in posterior blastomeres for both types of induction. This regional difference in neural competence along the antero-posterior axis paralleled that expected from neural cell lineage during normal development, i.e. blastomeres with more cells of neural lineage among their derivatives showed higher competence. 4. Streptomyces subtilisin inhibitor, SSI (0.1%), a specific protease inhibitor for subtilisin-type serine proteases, significantly suppressed (50%) neural induction of the ectodermal blastomere, a4-2, by contact with the

  11. LPS challenge regulates gene expression and tissue localization of a Ciona intestinalis gene through an alternative polyadenylation mechanism.

    PubMed

    Vizzini, Aiti; Bonura, Angela; Parrinello, Daniela; Sanfratello, Maria Antonietta; Longo, Valeria; Colombo, Paolo

    2013-01-01

    A subtractive hybridization strategy for the identification of differentially expressed genes was performed between LPS-challenged and naive Ciona intestinalis. This strategy allowed the characterization of two transcripts (Ci8short and Ci8long) generated by the use of two Alternative Polyadenylation sites. The Ci8long transcript contains a protein domain with relevant homology to several components of the Receptor Transporting Protein (RTP) family not present in the Ci8short mRNA. By means of Real Time PCR and Northern Blot, the Ci8short and Ci8long transcripts showed a different pattern of gene expression with the Ci8short mRNA being strongly activated after LPS injection in the pharynx. In situ hybridization analysis demonstrated that the activation of the APA site also influenced the tissue localization of the Ci8short transcript. This analysis showed that the Ci8long mRNA was expressed in hemocytes meanwhile the Ci8short mRNA was highly transcribed also in vessel endothelial cells and in the epithelium of pharynx. These findings demonstrated that regulation of gene expression based on different polyadenylation sites is an ancestral powerful strategy influencing both the level of expression and tissue distribution of alternative transcripts.

  12. Aquaculture fouling: Efficacy of potassium monopersulphonate triple salt based disinfectant (Virkon® Aquatic) against Ciona intestinalis.

    PubMed

    Paetzold, S Christine; Davidson, Jeff

    2011-07-01

    With the increasing spread of invasive marine species and their detrimental effects on aquaculture operations globally, mitigation strategies need to be optimized to mitigate economic impacts. The efficacy of a potassium monopersulphonate triple salt based disinfectant used in the aquaculture industry (Virkon® Aquatic at 0.5-5%) was evaluated against the solitary tunicate Ciona intestinalis, as well as the susceptibility of three different age groups of C. intestinalis to the treatment and the effect of the disinfectant on mussel mortality. Younger C. intestinalis were most affected by all treatments, and almost all immersion applications significantly decreased the biomass of C. intestinalis compared to untreated plates. Disinfectant solutions of ≥ 1% reduced biomass below pre-treatment levels. Mussel mortality was low, especially for solutions <3%. C. intestinalis should be treated 4 weeks post-settlement to maximize antifouling treatment effects. Immersion in 3% disinfectant for 30 s reduced the biomass of C. intestinalis by up to 89% and would be feasible in field applications using existing treatment equipment.

  13. Toxic effects of two pesticides, Imazalil and Triadimefon, on the early development of the ascidian Phallusia mammillata (Chordata, Ascidiacea).

    PubMed

    Pennati, Roberta; Groppelli, Silvia; Zega, Giuliana; Biggiogero, Maira; De Bernardi, Fiorenza; Sotgia, Cristina

    2006-09-12

    Azole compounds are fungicides used in agriculture and in clinical area and are suspected to produce craniofacial malformations in vertebrates. Toxicity tests on sperm viability, fertilization and embryogenesis of the solitary ascidian Phallusia mammillata were performed to evaluate the effects of two azole derivatives, Imazalil and Triadimefon. Ascidian (Chordata, Ascidiacea) embryos and larvae could provide biological criteria for seawater quality standards because the larvae are lecitotrophic and have a short pelagic period, allowing to run the larval toxicity tests over a short period of time. Imazalil and Triadimefon proved to have strong consequences on P. mammillata. They could influence the reproductive rate of the animal exerting their effects at different levels: acting as spermiotoxic agents, inhibiting fertilization and impairing embryological development. Fertilization rate significantly decreased after 30min exposure of sperm to 25microM Imazalil (P<0.0001) and after exposure of both gametes to 50microM Imazalil (P<0.05) and 1mM Triadimefon (P<0.0001) as compared to controls. Malformations caused by exposure of embryos to both substances were dose dependent. Imazalil median teratogenic concentration (TC50 concentration, the concentration that resulted in 50% malformed larvae) value was 0.67microM and median lethal concentration (LC50, the concentration that resulted in 50% embryos dead before completing the development) value was 10.23microM while for Triadimefon TC50 value was 29.56 and LC50 value was 173.7microM. Larvae developed from embryos treated with Imazalil and Triadimefon showed alterations of the anterior structures of the trunk: papillary nerves and the anterior central nervous system failed to correctly differentiate, as showed by immunostaining with anti-beta-tubulin antibody. Comparing the anomalies caused by retinoic acid, reported in a previous study, it was possible to hypothesize that malformations induced by Imazalil and

  14. A new genus of Asterocheridae (Copepoda: Siphonostomatoida) ectoassociate of the ascidian Eudistoma vannamei Millar, 1977 (Polycitoridae) from Brazil.

    PubMed

    Johnsson, Rodrigo; Bahia, Cristiano; Neves, Elizabeth

    2016-05-23

    Asterocheres Boeck, 1860 is the largest genus of the siphonostomatoid copepod family Asterocheridae, containing 63 valid species. The genus is known for its symbiotic relationships with many marine invertebrate taxa, especially sponges, cnidarians, bryozoans, and echinoderms. Recent studies have restricted the diagnosis of this genus. Consequently, many species are now considered as species inquirendae. The present paper describes a new species living externally on the tunic of Eudistoma vannamei Millar, 1977, an endemic ascidian from Brazil. As the new species does not fit Asterocheres in the strict sense, a new genus is erected to accommodate it. Setacheres gen. nov. is characterized by its possession of two distal setae on the third endopodal segment of P3, thus differing from the distal seta and spine pattern that is deemed as diagnostic of Asterocheres. A revision and comparison of Asterocheres´ species inquirendae revealed eight species sharing the same generic characteristics and were thus reallocated as members of the new genus.

  15. Verruculides A and B, two new protein tyrosine phosphatase 1B inhibitors from an Indonesian ascidian-derived Penicillium verruculosum.

    PubMed

    Yamazaki, Hiroyuki; Nakayama, Wataru; Takahashi, Ohgi; Kirikoshi, Ryota; Izumikawa, Yuta; Iwasaki, Kohei; Toraiwa, Kengo; Ukai, Kazuyo; Rotinsulu, Henki; Wewengkang, Defny S; Sumilat, Deiske A; Mangindaan, Remy E P; Namikoshi, Michio

    2015-08-15

    Two new merosesquiterpenes, verruculides A (1) and B (2), were isolated from a culture broth of the Indonesian ascidian-derived Penicillium verruculosum TPU1311, together with three known congeners, chrodrimanins A (3), B (4), and H (5). The structures of 1 and 2 were assigned on the basis of their spectroscopic data (1D and 2D NMR, HRMS, UV, CD, and IR). Compound 2 had a linear sesquiterpene moiety and was considered to be the derivative of the biosynthetic precursor for 1 and 3-5. Compounds 1, 3, and 5 inhibited the activity of protein tyrosine phosphatase 1B (PTP1B) with IC50 values of 8.4, 8.5, and 14.9 μM, respectively. Compound 2 showed 40% inhibition at 23.1 μM, while 4 was not active at 20.7 μM. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Interspecific hybridization between an anural and urodele ascidian: differential expression of urodele features suggests multiple mechanisms control anural development.

    PubMed

    Swalla, B J; Jeffery, W R

    1990-12-01

    Anural development in the ascidian Molgula occulta was examined using tissue-specific markers and interspecific hybridization. Unlike most ascidians, which develop into a swimming tadpole larva (urodele development), M. occulta eggs develop into a tailless slug-like larva (anural development) which metamorphoses into an adult. M. occulta embryos show conventional early cleavage patterns, gastrulation, and neurulation, but then diverge from the urodele developmental mode during larval morphogenesis. M. occulta larvae do not contain a pigmented sensory cell in their brain or form a tail with differentiated notochord and muscle cells. As shown by in situ hybridization with cloned probes and analysis of in vitro translation products, M. occulta embryos do not accumulate high levels of alpha actin or myosin heavy chain mRNA. In contrast, acetylcholinesterase is expressed in muscle lineage cells, indicating that various muscle cell features are differentially suppressed. M. occulta embryos also lack tyrosinase activity, suggesting that suppression of brain pigment cell differentiation occurs at an early step in development. M. occulta eggs fertilized with sperm from Molgula oculata (a closely related urodele species) develop into hybrid larvae exhibiting some of the missing urodele features. Some hybrid embryos develop tyrosinase activity and differentiate a brain pigment cell and a short row of notochord cells, and form a short tail. These urodele features appeared together or separately in different hybrid embryos suggesting that they develop by independent mechanisms. In contrast, alpha actin and myosin heavy chain mRNA accumulation was not enhanced in hybrid embryos. These results suggest that multiple mechanisms control anural development.

  17. Anticancer effects of brominated indole alkaloid Eudistomin H from marine ascidian Eudistoma viride against cervical cancer cells (HeLa).

    PubMed

    Rajesh, Rajaian Pushpabai; Annappan, Murugan

    2015-01-01

    Marine invertebrates called ascidians are prolific producers of bioactive substances. The ascidian Eudistoma viride, distributed along the Southeast coast of India, was investigated for its in vitro cytotoxic activity against human cervical carcinoma (HeLa) cells by the MTT assay. The crude methanolic extract of E. viride, with an IC50 of 53 μg/ml, was dose-dependently cytotoxic. It was more potent at 100 μg/ml than cyclohexamide (1 μg/ml), reducing cell viability to 9.2%. Among nine fractions separated by chromatography, ECF-8 exhibited prominent cytoxic activity at 10 μg/ml. The HPLC fraction EHF-21 of ECF-8 was remarkably dose- and time-dependently cytotoxic, with 39.8% viable cells at 1 μg/ml compared to 51% in cyclohexamide-treated cells at the same concentration; the IC50 was 0.49 μg/ml. Hoechst staining of HeLa cells treated with EHF-21 at 0.5 μg/ml revealed apoptotic events such an cell shrinkage, membrane blebbing, chromatin condensation and formation of apoptotic bodies. Cell size and granularity study showed changes in light scatter, indicating the characteristic feature of cells dying by apoptosis. The cell-cycle analysis of HeLa cells treated with fraction EHF-21 at 1 μg/ml showed the marked arrest of cells in G0/G1, S and G2/M phases and an increase in the sub G0/G1 population indicated an increase in the apoptotic cell population. The statistical analysis of the sub-G1 region showed a dose-dependent induction of apoptosis. DNA fragmentation was also observed in HeLa cells treated with EHF-21. The active EHF-21 fraction, a brominated indole alkaloid Eudistomin H, led to apoptotic death of HeLa cells.

  18. Biogeography of Mediterranean Invasions

    NASA Astrophysics Data System (ADS)

    Groves, R. H.; di Castri, F.

    The Mediterranean basin, California, Chile, the western Cape of South Africa, and southern Australia share a Mediterranean climate characterized by cool, wet winters and hot, dry summers. These five regions have differing patterns of human settlement, but similarities in natural vegetation and some faunal assemblages. These likenesses are enhanced with time by an increasing level of biotic exchange among the regions. An initiative of a subcommittee of SCOPE (Scientific Committee on Problems of the Environment), which realized that the integrity of many natural ecosystems is being threatened by the ingress of invasive species, this book uniquely documents the introduced floras and faunas, especially plants, buds, and mammals, in these five regions of Mediterranean climate, and aims to increase our understanding of the ecology of biological invasions. In doing so, it points a way to more effectively manage the biota of these regions.

  19. Mediterranean, our sea

    NASA Astrophysics Data System (ADS)

    Markaki, Foteini

    2017-04-01

    My school (1o EPAL Ymittos -Athens, Greece) is a technical school of secondary education and throughout this school year being drafted a program of environmental education. The main theme is the Mediterranean Sea, the biggest closed sea extending between three continents. Topics studied: 1. Biodiversity and the risks threat. 2. The geophysics that characterize (earthquakes, volcanoes explosions, etc). 3. The Mediterranean Sea as environment anthropogenesis, a mosaic of other cultures and even place current notions of social phenomena (refugees). Pedagogical Objectives: Cognitive/Enviromental: 1. To investigate and understand the biodiversity of the Mediterranean Sea and the risks to threaten and phenomena that characterize. 2. To understand the position of the Mediterranean Sea in the land and the role of the historical, cultural and social human environment. 3. To come in contact with texts literary, social, articles on the Mediterranean. Psychomotor: 1. To work together and collect information for the Mediterranean Sea. 2. Experiential approach to the natural environment. 3. Develop critical thinking. 4. Undertake responsibilities for the presentation of the program. Emotional: 1. To feel joy from participation in the program. 2. Being sensitized and configure attitudes and actions of respect towards the environment. Methodology implementation: Teamwork. Interdisciplinary - holistic to dissemination of program recordings to courses curriculum. Study in the field. Gathering information from newspapers, magazines, internet, maps, and photographs. Experiential method- Project. Assessment methods and self-assessment. Fields of courses: Greek language- History- Biology- Chemistry- Technology Dissemination of results: Make a page of social media (facebook), a blog, enhancing environmental awareness via video, make an electronic poster.

  20. Localization and enzymatic activity profiles of the proteases responsible for tachykinin-directed oocyte growth in the protochordate, Ciona intestinalis.

    PubMed

    Aoyama, Masato; Kawada, Tsuyoshi; Satake, Honoo

    2012-03-01

    We previously substantiated that Ci-TK, a tachykinin of the protochordate, Ciona intestinalis (Ci), triggered oocyte growth from the vitellogenic stage (stage II) to the post-vitellogenic stage (stage III) via up-regulation of the gene expression and enzymatic activity of the proteases: cathepsin D, carboxypeptidase B1, and chymotrypsin. In the present study, we have elucidated the localization, gene expression and activation profile of these proteases. In situ hybridization showed that the Ci-cathepsin D mRNA was present exclusively in test cells of the stage II oocytes, whereas the Ci-carboxypeptidase B1 and Ci-chymotrypsin mRNAs were detected in follicular cells of the stage II and stage III oocytes. Double-immunostaining demonstrated that the immunoreactivity of Ci-cathepsin D was largely colocalized with that of the receptor of Ci-TK, Ci-TK-R, in test cells of the stage II oocytes. Ci-cathepsin D gene expression was detected at 2h after treatment with Ci-TK, and elevated for up to 5h, and then slightly decreased. Gene expression of Ci-carboxypeptidase B1 and Ci-chymotrypsin was observed at 5h after treatment with Ci-TK, and then decreased. The enzymatic activities of Ci-cathepsin D, Ci-carboxypeptidase B1, and Ci-chymotrypsin showed similar alterations with 1-h lags. These gene expression and protease activity profiles verified that Ci-cathepsin D is initially activated, which is followed by the activation of Ci-carboxypeptidase B1 and Ci-chymotrypsin. Collectively, the present data suggest that Ci-TK directly induces Ci-cahtepsin D in test cells expressing Ci-TK receptor, leading to the secondary activation of Ci-chymotrypsin and Ci-carboxypeptidase B1 in the follicle in the tachykininergic oocyte growth pathway.

  1. Fatty Acid and Lipid Profiles with Emphasis on n-3 Fatty Acids and Phospholipids from Ciona intestinalis.

    PubMed

    Zhao, Yadong; Wang, Miao; Lindström, Mikael E; Li, Jiebing

    2015-10-01

    In order to establish Ciona intestinalis as a new bioresource for n-3 fatty acids-rich marine lipids, the animal was fractionated into tunic and inner body tissues prior to lipid extraction. The lipids obtained were further classified into neutral lipids (NL), glycolipids (GL) and phospholipids (PL) followed by qualitative and quantitative analysis using GC-FID, GC-MS, (1)H NMR, 2D NMR, MALDI-TOF-MS and LC-ESI-MS methods. It was found that the tunic and inner body tissues contained 3.42-4.08% and 15.9-23.4% of lipids respectively. PL was the dominant lipid class (42-60%) irrespective of the anatomic fractions. From all lipid fractions and classes, the major fatty acids were 16:0, 18:1n-9, C20:1n-9, C20:5n-3 (EPA) and C22:6n-3 (DHA). The highest amounts of long chain n-3 fatty acids, mainly EPA and DHA, were located in PL from both body fractions. Cholestanol and cholesterol were the dominant sterols together with noticeable amounts of stellasterol, 22 (Z)-dehydrocholesterol and lathosterol. Several other identified and two yet unidentified sterols were observed for the first time from C. intestinalis. Different molecular species of phosphatidylcholine (34 species), sphingomyelin (2 species), phosphatidylethanolamine (2 species), phosphatidylserine (10 species), phosphatidylglycerol (9 species), ceramide (38 species) and lysophospholipid (5 species) were identified, representing the most systematic PL profiling knowledge so far for the animal. It could be concluded that C. intestinalis lipids should be a good alternative for fish oil with high contents of n-3 fatty acids. The lipids would be more bioavailable due to the presence of the fatty acids being mainly in the form of PL.

  2. Distinguishing contemporary hybridization from past introgression with postgenomic ancestry-informative SNPs in strongly differentiated Ciona species.

    PubMed

    Bouchemousse, Sarah; Liautard-Haag, Cathy; Bierne, Nicolas; Viard, Frédérique

    2016-11-01

    Biological introductions bring into contact species that can still hybridize. The evolutionary outcomes of such secondary contacts may be diverse (e.g. adaptive introgression from or into the introduced species) but are not yet well examined in the wild. The recent secondary contact between the non-native sea squirt Ciona robusta (formerly known as C. intestinalis type A) and its native congener C. intestinalis (formerly known as C. intestinalis type B), in the Western English Channel, provides an excellent case study to examine. To examine contemporary hybridization between the two species, we developed a panel of 310 ancestry-informative SNPs from a population transcriptomic study. Hybridization rates were examined on 449 individuals sampled in eight sites from the sympatric range and five sites from allopatric ranges. The results clearly showed an almost complete absence of contemporary hybridization between the two species in syntopic localities, with only one-first-generation hybrid and no other genotype compatible with recent backcrosses. Despite the almost lack of contemporary hybridization, shared polymorphisms were observed in sympatric and allopatric populations of both species. Furthermore, one allopatric population from SE Pacific exhibited a higher rate of shared polymorphisms compared to all other C. robusta populations. Altogether, these results indicate that the observed level of shared polymorphism is more probably the outcome of ancient gene flow spread afterwards at a worldwide scale. They also emphasize efficient reproductive barriers preventing hybridization between introduced and native species, which suggests hybridization should not impede too much the expansion and the establishment of the non-native species in its introduction range.

  3. Characterization of a novel vanadium-binding protein (VBP-129) from blood plasma of the vanadium-rich ascidian Ascidia sydneiensis samea.

    PubMed

    Yoshihara, Masao; Ueki, Tatsuya; Yamaguchi, Nobuo; Kamino, Kei; Michibata, Hitoshi

    2008-02-01

    The ascidians, the so-called sea squirts, accumulate high levels of vanadium, a transition metal. Since Henze first observed this physiologically unusual phenomenon about one hundred years ago, it has attracted interdisciplinary attention from chemists, physiologists, and biochemists. The maximum concentration of vanadium in ascidians can reach 350 mM, and most of the vanadium ions are stored in the +3 oxidation state in the vacuoles of vanadium-accumulating blood cells known as vanadocytes. Many proteins involved in the accumulation and reduction of vanadium in the vanadocytes, blood plasma, and digestive tract have been identified. However, the process by which vanadium is taken in prior to its accumulation in vanadocytes has not been elucidated. In the present study, a novel vanadium-binding protein, designated VBP-129, was identified from blood plasma of the vanadium-rich ascidian Ascidia sydneiensis samea. Although VBP-129 mRNA was transcribed in all A. sydneiensis samea tissues examined, the VBP-129 protein was exclusively localized in blood plasma and muscle cells of this ascidian. It bound not only to VO(2+) but also to Fe(3+), Co(2+), Cu(2+), and Zn(2+); on the other hand, a truncated form of VBP-129, designated VBP-88, bound only to Co(2+), Cu(2+) and Zn(2+). In a pull-down assay, an interaction between VanabinP and VBP-129 occurred both in the presence and the absence of VO(2+). These results suggest that VBP-129 and VanabinP function cooperatively as metallochaperones in blood plasma.

  4. Expression of Ciona intestinalis variable region-containing chitin-binding proteins during development of the gastrointestinal tract and their role in host-microbe interactions.

    PubMed

    Liberti, Assunta; Melillo, Daniela; Zucchetti, Ivana; Natale, Lenina; Dishaw, Larry J; Litman, Gary W; De Santis, Rosaria; Pinto, Maria Rosaria

    2014-01-01

    Variable region-containing chitin-binding proteins (VCBPs) are secreted, immune-type molecules that have been described in both amphioxus, a cephalochordate, and sea squirt, Ciona intestinalis, a urochordate. In adult Ciona, VCBP-A, -B and -C are expressed in hemocytes and the cells of the gastrointestinal tract. VCBP-C binds bacteria in the stomach lumen and functions as an opsonin in vitro. In the present paper the expression of VCBPs has been characterized during development using in situ hybridization, immunohistochemical staining and quantitative polymerase chain reaction (qPCR) technologies. The expression of VCBP-A and -C is detected first in discrete areas of larva endoderm and becomes progressively localized during differentiation in the stomach and intestine, marking the development of gut tracts. In "small adults" (1-2 cm juveniles) expression of VCBP-C persists and VCBP-A gradually diminishes, ultimately replaced by expression of VCBP-B. The expression of VCBP-A and -C in stage 7-8 juveniles, at which point animals have already started feeding, is influenced significantly by challenge with either Gram-positive or -negative bacteria. A potential role for VCBPs in gut-microbiota interactions and homeostasis is indicated.

  5. The population genomics of a fast evolver: high levels of diversity, functional constraint, and molecular adaptation in the tunicate Ciona intestinalis.

    PubMed

    Tsagkogeorga, Georgia; Cahais, Vincent; Galtier, Nicolas

    2012-01-01

    Phylogenomics has revealed the existence of fast-evolving animal phyla in which the amino acid substitution rate, averaged across many proteins, is consistently higher than in other lineages. The reasons for such differences in proteome-wide evolutionary rates are still unknown, largely because only a handful of species offer within-species genomic data from which molecular evolutionary processes can be deduced. In this study, we use next-generation sequencing technologies and individual whole-transcriptome sequencing to gather extensive polymorphism sequence data sets from Ciona intestinalis. Ciona is probably the best-characterized member of the fast-evolving Urochordata group (tunicates), which was recently identified as the sister group of the slow-evolving vertebrates. We introduce and validate a maximum-likelihood framework for single-nucleotide polymorphism and genotype calling, based on high-throughput short-read typing. We report that the C. intestinalis proteome is characterized by a high level of within-species diversity, efficient purifying selection, and a substantial percentage of adaptive amino acid substitutions. We conclude that the increased rate of amino acid sequence evolution in tunicates, when compared with vertebrates, is the consequence of both a 2-6 times higher per-year mutation rate and prevalent adaptive evolution.

  6. Expression of Ciona intestinalis Variable Region-Containing Chitin-Binding Proteins during Development of the Gastrointestinal Tract and Their Role in Host-Microbe Interactions

    PubMed Central

    Liberti, Assunta; Melillo, Daniela; Zucchetti, Ivana; Natale, Lenina; Dishaw, Larry J.; Litman, Gary W.; De Santis, Rosaria; Pinto, Maria Rosaria

    2014-01-01

    Variable region-containing chitin-binding proteins (VCBPs) are secreted, immune-type molecules that have been described in both amphioxus, a cephalochordate, and sea squirt, Ciona intestinalis, a urochordate. In adult Ciona, VCBP-A, -B and -C are expressed in hemocytes and the cells of the gastrointestinal tract. VCBP-C binds bacteria in the stomach lumen and functions as an opsonin in vitro. In the present paper the expression of VCBPs has been characterized during development using in situ hybridization, immunohistochemical staining and quantitative polymerase chain reaction (qPCR) technologies. The expression of VCBP-A and -C is detected first in discrete areas of larva endoderm and becomes progressively localized during differentiation in the stomach and intestine, marking the development of gut tracts. In “small adults” (1–2 cm juveniles) expression of VCBP-C persists and VCBP-A gradually diminishes, ultimately replaced by expression of VCBP-B. The expression of VCBP-A and -C in stage 7–8 juveniles, at which point animals have already started feeding, is influenced significantly by challenge with either Gram-positive or -negative bacteria. A potential role for VCBPs in gut-microbiota interactions and homeostasis is indicated. PMID:24788831

  7. The Mediterranean pine engraver

    Treesearch

    Jana C. Lee; Sheri L. Smith; Steven J. Seybold

    2005-01-01

    In May 2004, a new exotic bark beetle for North America was discovered in baited flight traps in Fresno, California during an annual bark beetle and woodborer survey by the California Department of Food and Agriculture. This bark beetle was identified as Orthotomicus erosus (Wollaston), the Mediterranean pine engraver, a well-documented pest of...

  8. Explore Mediterranean in classroom

    NASA Astrophysics Data System (ADS)

    Balesevic, Ivana

    2017-04-01

    I am a science teacher at a primary school and my students are very interested in science. Through this year I will work with my students, organizing several workshops and or results will be presented on poster. I will work with several groups (4-6) students 8th grade. In this poster all activities will be presented, showing how science is easy to learn even in a classroom. 1. Workshop > Chemical characteristic of sea water Using school laboratory each group of students will analyze the physical and chemical characteristic of sea water and they have to explain the results to younger student's 5th and 6th grade. The final result will be presented on poster. 2. Workshop> Meet the Mediterranean life During this workshop students will work in different groups. The aim of the workshop is to meet lots of species that we can find in Mediterranean using movies, phone applications, internet explorer, science books and school collections of invertebrates … 3. Workshop>Stop the pollution Several groups of students have to debate about causes of pollution and possibilities for prevention. At the end of workshop we will organize a quiz. Student's answers and suggestions will be shown on the poster. 4. Workshop> How we see the Mediterranean During this workshop students will make models of Mediterranean in 2d and 3d perspective, using different materials. They can show on models parts of Mediterranean area, country, sea... After making models students need to visit 5th and 6th grade classes, to show them and explain the final results. Few models will be presented on poster

  9. Is there a Mediterranean bioethics?

    PubMed

    Mallia, Pierre

    2012-11-01

    Is there a special Mediterranean approach to Bioethics and if so what are the roots of this approach? And why not a Bosphorus, or a 'lake Michigan' bioethics? The answer to such a question depends on the focus one takes on defining 'Mediterranean'? On the one hand one can refer to the Mediterranean region which includes the surrounding coasts, having Europe on its northern coast line, northern Africa on its southern coast line (and these will include the north and South West coasts), and in the Eastern region countries which border with Middle-Eastern countries. This approach is the approach currently being taken by European Parliamentarians when they speak about the Mediterranean, namely including countries like France, Italy and Libya. On the other hand there is the look upon the Mediterranean as 'Southern Europe'; this is a more 'traditional' way on how westerners view the Mediterranean. This common approach is often recognized when, for example, we speak of 'Mediterranean diet', or, 'Mediterranean Temperament'. It would include Eastern countries like Greece and Cyprus. This article focuses on these two approaches to Mediterranean ethics after discussing issues pertaining to the region which are important to define in this context. It then analyses the need for having a Mediterranean approach to bioethical issues.

  10. Southeastern Mediterranean Panorama

    NASA Technical Reports Server (NTRS)

    1991-01-01

    This oblique northwestward looking panorama view shows the southeastern Mediterranean (29.0N, 33.0E) in great detail. The Sinai Peninsula, the eastern Arabian Desert, the Nile River Valley and Delta as well as the Qatara Depression in Egypt are all prominently portrayed. Even Mt. Sinai is visible in the lower left center of the view. The dusty atmosphere of the region can be seen as a general haziness in the atmosphere.

  11. The Mediterranean salinity crisis

    SciTech Connect

    Hsue, K.J.

    1988-08-01

    That the Mediterranean Sea underwent a salinity crisis during the Miocene (Messinian) is proven by the 1970 JOIDES deep sea drilling expedition. Subsequent work by ocean drilling and by studies on land have recorded the history of this crisis. Based upon the deep desiccated-basin model, the use of event-stratigraphy, calibrated by strontium-isotope dating and magnetostratigraphy, has enabled them to decipher the following events between 6.0 and 5.1 Ma: (1) deposition of marine diatom-rich sediments in a partially restricted basin, (2) first desiccation of the Mediterranean when Calcare di base was deposited at a time of isolation from the Atlantic because of a glacial eustatic drop of sea level, (3) influx of marine waters through southern Spanish basins to furnish brines for the deposition of the main salt, (4) Intra-Messinian desiccation, as evidenced by the erosional unconformity above the lower evaporite, (5) Intra-Messinian denudation, when reefs grew on Cyprus and marine sediments were deposited in basins, (6) frequency isolations due to oscillating sea level, when the upper evaporite was deposited, (7) Lago mare, formation of freshwater and brackish lakes due to influx of Paratethys water, (8) opening of the Gibraltar and Pliocene inundation of the Mediterranean.

  12. SMED - Sulphur MEditerranean Dispersion

    NASA Astrophysics Data System (ADS)

    Salerno, Giuseppe G.; Sellitto, Pasquale; Corradini, Stefano; Di Sarra, Alcide Giorgio; Merucci, Luca; Caltabiano, Tommaso; La Spina, Alessandro

    2016-04-01

    Emissions of volcanic gases and particles can have profound impacts on terrestrial environment, atmospheric composition, climate forcing, and then on human health at various temporal and spatial scales. Volcanic emissions have been identified as one of the largest sources of uncertainty in our understanding of recent climate change trends. In particular, a primary role is acted by sulphur dioxide emission due to its conversion to volcanic sulphate aerosol via atmospheric oxidation. Aerosols may play a key role in the radiative budget and then in photochemistry and tropospheric composition. Mt. Etna is one of the most prodigious and persistent emitters of gasses and particles on Earth, accounting for about 10% of global average volcanic emission of CO2 and SO2. Its sulphur emissions stand for 0.7 × 106 t S/yr9 and then about 10 times bigger than anthropogenic sulphur emissions in the Mediterranean area. Centrepiece of the SMED project is to advance the understanding of volcanogenic sulphur dioxide and sulphate aerosol particles dispersion and radiative impact on the downwind Mediterranean region by an integrated approach between ground- and space-based observations and modelling. Research is addressed by exploring the potential relationship between proximal SO2 flux and aerosol measured remotely in the volcanic plume of Mt. Etna between 2000 and 2014 and distal aerosol ground-based measurements in Lampedusa, Greece, and Malta from AERONET network. Ground data are combined with satellite multispectral polar and geostationary imagers able to detect and retrieve volcanic ash and SO2. The high repetition time of SEVIRI (15 minutes) will ensure the potential opportunity to follow the entire evolution of the volcanic cloud, while, the higher spatial resolution of MODIS (1x1 km2), are exploited for investigating the probability to retrieve volcanic SO2 abundances from passive degassing. Ground and space observations are complemented with atmospheric Lagrangian model

  13. Lineage divergence, local adaptation across a biogeographic break, and artificial transport, shape the genetic structure in the ascidian Pyura chilensis

    PubMed Central

    Segovia, Nicolás I.; Gallardo-Escárate, Cristian; Poulin, Elie; Haye, Pilar A.

    2017-01-01

    Marine benthic organisms inhabit a heterogeneous environment in which connectivity between populations occurs mainly through dispersive larval stages, while local selective pressures acting on early life history stages lead to non-random mortality, shaping adaptive genetic structure. In order to test the influence of local adaptation and neutral processes in a marine benthic species with low dispersal, in this study we used Genotyping by Sequencing technology to compare the neutral and putatively selected signals (neutral and outlier loci, respectively) in SNPs scattered throughout the genome in six local populations of the commercially exploited ascidian Pyura chilensis along the southeast Pacific coast (24°–42°S). This species is sessile as an adult, has a short-lived larval stage, and may also be dispersed by artificial transport as biofouling. We found that the main signal in neutral loci was a highly divergent lineage present at 39°S, and a subjacent signal that indicated a separation at 30°S (north/south), widely reported in the area. North/south separation was the main signal in outlier loci, and the linage divergence at 39°S was subjacent. We conclude that the geographic structure of the genetic diversity of outlier and neutral loci was established by different strengths of environmental, historical and anthropogenic factors. PMID:28300177

  14. Photoadaptation and protection against active forms of oxygen in the symbiotic procaryote Prochloron sp. and its ascidian host

    SciTech Connect

    Lesser, M.P.; Stochaj, W.R. )

    1990-06-01

    Superoxide dismutase, ascorbate, peroxidase, and catalase activities were studied in the symbiotic photosynthetic procaryote Prochloron sp. and its ascidian host Lissoclinum patella. The protein-specific activities of these antioxidant enzymes in the Prochloron sp. and L. patella collected at different depths from the Great Barrier Reef, Australia, were directly proportional to irradiance, whereas the pigment concentrations in the Prochloron sp. were inversely proportional to irradiance. The presence of a cyanide-sensitive superoxide dismutase, presumably a Cu-An metalloprotein, in the Prochloron sp. extends the possible phylogenetic distribution of this protein. The concentration of UV-absorbing mycosporine-like amino acids in inversely proportional to irradiance in both the host and symbiont, suggesting that these compounds may not provide sufficient protection against UV radiation in high-irradiance environments. The significant differences in the specific activities of these antioxidant enzymes, cellular photosynthetic pigment concentrations, and UV-absorbing compounds from high- and low-irradiance habitats constitute an adaptive response to different photic environments. These photoadaptive responses are essential to prevent inhibition of photosynthesis by high fluxes of visible and UV radiation.

  15. Larval adhesive organs and metamorphosis in ascidians. II. The mechanism of eversion of the papillae of Distaplia occidentalis.

    PubMed

    Cloney, R A

    1979-09-01

    The cup-shaped adhesive papillae of Distaplia occidentalis evert at the onset of metamorphosis and each transforms into a hyperboloidal configuration. The rate of transformation is a function of temperature. At 14 degrees C complete eversion takes about 30 seconds. Myoepithelial cells that extend from the rim to the base on the cup contract. Simultaneously the central part of the papilla advances 60--70 micrometers. During the last phases of eversion, collocytes (cells that secrete adhesives) on the inner wall of the cup and on the sides of the axial protrusion flow outward and form a collar-like structure. The myoepithelial cells contain arrays of thick and thin filaments. These become compacted during contraction. The surfaces of these cells become extensively folded as they shorten to about 1/3 of rest length. According to the proposed model the myoepithelial cells are the driving force in papillary eversion. Immediately after eversion is completed the papillae begin to retract. Eversion of the papillae is not inhibited by cytochalasin B, but the process of retraction is reversibly inhibited. Some histological characteristics of five types of everting papillae in four families of ascidians are compared.

  16. Lineage divergence, local adaptation across a biogeographic break, and artificial transport, shape the genetic structure in the ascidian Pyura chilensis.

    PubMed

    Segovia, Nicolás I; Gallardo-Escárate, Cristian; Poulin, Elie; Haye, Pilar A

    2017-03-16

    Marine benthic organisms inhabit a heterogeneous environment in which connectivity between populations occurs mainly through dispersive larval stages, while local selective pressures acting on early life history stages lead to non-random mortality, shaping adaptive genetic structure. In order to test the influence of local adaptation and neutral processes in a marine benthic species with low dispersal, in this study we used Genotyping by Sequencing technology to compare the neutral and putatively selected signals (neutral and outlier loci, respectively) in SNPs scattered throughout the genome in six local populations of the commercially exploited ascidian Pyura chilensis along the southeast Pacific coast (24°-42°S). This species is sessile as an adult, has a short-lived larval stage, and may also be dispersed by artificial transport as biofouling. We found that the main signal in neutral loci was a highly divergent lineage present at 39°S, and a subjacent signal that indicated a separation at 30°S (north/south), widely reported in the area. North/south separation was the main signal in outlier loci, and the linage divergence at 39°S was subjacent. We conclude that the geographic structure of the genetic diversity of outlier and neutral loci was established by different strengths of environmental, historical and anthropogenic factors.

  17. Msxb is a core component of the genetic circuitry specifying the dorsal and ventral neurogenic midlines in the ascidian embryo.

    PubMed

    Roure, Agnès; Darras, Sébastien

    2016-01-01

    The tail ascidian larval peripheral nervous system is made up of epidermal sensory neurons distributed more or less regularly in ventral and dorsal midlines. Their formation occurs in two-steps: the ventral and dorsal midlines are induced as neurogenic territories by Fgf9/16/20 and Admp respectively. The Delta2/Notch interaction then controls the number of neurons that form. The genetic machinery acting between the inductive processes taking place before gastrulation and neuron specification at tailbud stages are largely unknown. The analysis of seven transcription factors expressed in the forming midlines revealed an unexpected complexity and dynamic of gene expression. Their systematic overexpression confirmed that these genes do not interact following a linear cascade of activation. However, the integration of our data revealed the distinct key roles of the two upstream factors Msxb and Nkx-C that are the earliest expressed genes and the only ones able to induce neurogenic midline and ESN formation. Our data suggest that Msxb would be the primary midline gene integrating inputs from the ventral and dorsal inducers and launching a pan-midline transcriptional program. Nkx-C would be involved in tail tip specification, in maintenance of the pan-midline network and in a posterior to anterior wave controlling differentiation.

  18. Catecholamines are produced by ascidian immune cells: The involvement of PKA and PKC in the adrenergic signaling pathway.

    PubMed

    de Abreu Mello, Andressa; Fernandes de Souza, Jéssica; Nunes da Fonseca, Rodrigo; Allodi, Silvana; Monteiro de Barros, Cintia

    2017-03-01

    The stress response is a complex mechanism, which includes changes in the immune system to enable organisms to maintain homeostasis. The neurohormones dopamine, noradrenaline (NA) and adrenalin are responsible for the physiological modulations that occur during acute stress. In the present study, we analyzed the effects of NA on the immune system specific to nitric-oxide (NO) production by subpopulations of immune cells (hemocytes) of the ascidian Phallusia nigra. We also investigated the capability of immune cells to produce catecholamine (CA). Finally, we tested the involvement of protein kinase A (PKA) and C (PKC) in the NA downstream signaling pathway. The results revealed that NA can reduce NO production by P. nigra hemocytes threefold, and that signet-ring cells, univacuolar refractile granulocytes and morula cells are the cell types most involved in this event. A challenge effected with Zymosan A induced CA production, and co-incubation with both inhibitors of the second messengers PKA and PKC revealed the involvement of these molecules in the adrenergic pathway of P. nigra hemocytes. Taken together, these results suggest that NO production can be down-regulated by NA through α- and β-adrenoceptors via the second messengers PKA and PKC.

  19. T-Type Ca2+ Current Activity during Oocyte Growth and Maturation in the Ascidian Styela plicata

    PubMed Central

    Gallo, Alessandra; Russo, Gian Luigi; Tosti, Elisabetta

    2013-01-01

    Voltage-dependent calcium currents play a fundamental role during oocyte maturation, mostly L-type calcium currents, whereas T-type calcium currents are involved in sperm physiology and cell growth. In this paper, using an electrophysiological and pharmacological approach, we demonstrated, for the first time in oocytes, that T-type calcium currents are present with functional consequences on the plasma membrane of growing immature oocytes of the ascidian Styela plicata. We classified three subtypes of immature oocytes at the germinal vesicle stage on the basis of their size, morphology and accessory cellular structures. These stages were clearly associated with an increased activity of T-type calcium currents and hyperpolarization of the plasma membrane. We also observed that T-type calcium currents oscillate in the post-fertilization embryonic stages, with minimal amplitude of the currents in the zygote and maximal at 8-cell stage. In addition, chemical inhibition of T-type calcium currents, obtained by applying specific antagonists, induced a significant reduction in the rate of cleavage and absence of larval formation. We suggest that calcium entry via T-type calcium channels may act as a potential pacemaker in regulating cytosolic calcium involved in fertilization and early developmental events. PMID:23349937

  20. A statistical approach for optimization of alkaline lipase production by ascidian associated-Halobacillus trueperi RSK CAS9.

    PubMed

    Sathishkumar, Ramamoorthy; Ananthan, Gnanakkan; Iyappan, Kathirvel; Stalin, Chinnathambi

    2015-12-01

    A marine ascidian-associated bacterium, Halobacillus trueperi RSK CAS9, was optimized for lipase production by response surface methodology using marine waste as substrate. The central composite design was employed, and the optimal medium constituents for maximum lipase production (1355.81 U/ml) were determined to be tuna powder (14.58 g/l), olive oil (5.05 ml/l); NaCl (72.42 g/l), temperature (45 °C) and pH 9.0. An alkaline lipase was purified to 8.46 fold with 1193.59 U mg(-1) specific activities with the molecular weight of 44 kDa. The activity was substantially inhibited by EDTA and PMSF, indicating that it was a metalloenzyme serine residue which was essential for catalytic activity. Thus, lipase production by microbial conversion of marine fish wastes in this study suggested its potential utilization for the production of high value products.

  1. Symmetrical reproductive compatibility of two species in the Ciona intestinalis (Ascidiacea) species complex, a model for marine genomics and developmental biology.

    PubMed

    Sato, Atsuko; Shimeld, Sebastian M; Bishop, John D D

    2014-06-01

    The sea squirt Ciona intestinalis species complex is a widely used model system for genomics and developmental biology, as well as ecology. Contrary to previous reports, here we show no difference in the success of development and hatching between hybrid and conspecific crosses between the two species within this complex known as types A and B, from a region in the English Channel where they are sympatric. We grew laboratory hybrids in the field for three months, and successfully obtained reproductive adults. In back-crosses of F1 laboratory hybrids to parental types, normal larvae were obtained. We conclude that hybrid crosses generate viable offspring and the resulting hybrids are interfertile with types A and B. However we also show that introgression in the natural sympatric population remains low. We discuss possible pre-zygotic and post-zygotic mechanisms which reproductively isolate these species.

  2. [Massive mortality of marine invertebrates: an unprecedented event in northwestern Mediterranean].

    PubMed

    Perez, T; Garrabou, J; Sartoretto, S; Harmelin, J G; Francour, P; Vacelet, J

    2000-10-01

    An unprecedented mass mortality event has been observed at the end of the summer 1999 along the coasts of Provence (France) and Ligury (Italy). This event has severely affected a wide array of sessile filter-feeder invertebrates from hard-substratum communities, such as sponges (particularly the keratose sponges Hippospongia and Spongia), cnidarians (particularly the anthozoans Corallium, Paramuricea, Eunicella and Cladocora), bivalves, ascidians and bryozoans. Along the Provence coasts, the outbreak spread from east to west. Exceptionally high and constant temperatures of the whole water column (23-24 degrees C, for over one month, down to 40 m) could have determined an environmental context favourable to the mass mortality event. Like the thermal anomaly, the mortality is limited in depth. However, we cannot ascertain whether temperature had a direct effect on organisms or acted in synergy with a latent and/or waterborne agent (microbiological or chemical). Taking into account the global warming context in the NW-Mediterranean, monitoring programs of physical-chemical parameters and vulnerable populations should rapidly be set up.

  3. Reviving the Mediterranean Olive Community

    ERIC Educational Resources Information Center

    Zaferatos, Nicholas C.

    2011-01-01

    This article presents the findings of a collaborative investigation by six nongovernment organisations (NGOs) from five European-Mediterranean countries to identify a framework for reversing rural marginalisation in Mediterranean communities through sustainable forms of community-based agricultural development. The project brought together…

  4. Reviving the Mediterranean Olive Community

    ERIC Educational Resources Information Center

    Zaferatos, Nicholas C.

    2011-01-01

    This article presents the findings of a collaborative investigation by six nongovernment organisations (NGOs) from five European-Mediterranean countries to identify a framework for reversing rural marginalisation in Mediterranean communities through sustainable forms of community-based agricultural development. The project brought together…

  5. The Hemolymph of the ascidian Styela plicata (Chordata-Tunicata) contains heparin inside basophil-like cells and a unique sulfated galactoglucan in the plasma.

    PubMed

    de Barros, Cintia M; Andrade, Leonardo R; Allodi, Silvana; Viskov, Christian; Mourier, Pierre A; Cavalcante, Moisés C M; Straus, Anita H; Takahashi, Helio K; Pomin, Vitor H; Carvalho, Vinicius F; Martins, Marco A; Pavão, Mauro S G

    2007-01-19

    The hemolymph of ascidians (Chordata-Tunicata) contains different types of hemocytes embedded in a liquid plasma. In the present study, heparin and a sulfated heteropolysaccharide were purified from the hemolymph of the ascidian Styela plicata. The heteropolysaccharide occurs free in the plasma, is composed of glucose ( approximately 60%) and galactose ( approximately 40%), and is highly sulfated. Heparin, on the other hand, occurs in the hemocytes, and high performance liquid chromatography of the products formed by degradation with specific lyases revealed that it is composed mainly by the disaccharides DeltaUA(2SO(4))-1-->4-beta-d-GlcN(SO(4)) (39.7%) and DeltaUA(2SO(4))-1-->4-beta-d-GlcN(SO(4))(6SO(4)) (38.2%). Small amounts of the 3-O-sulfated disaccharides DeltaUA(2SO(4))-1-->4-beta-d-GlcN(SO(4))(3SO(4)) (9.8%) and DeltaUA(2SO(4))-1-->4-beta-d-GlcN(SO(4))(3SO(4))(6SO(4)) (3.8%) were also detected. These 3-O-sulfated disaccharides were demonstrated to be essential for the binding of the hemocyte heparin to antithrombin III. Electron microscopy techniques were used to characterize the ultrastructure of the hemocytes and to localize heparin and histamine in these cells. At least five cell types were recognized and classified as univacuolated and multivacuolated cells, amebocytes, hemoblasts, and granulocytes. Immunocytochemistry showed that heparin and histamine co-localize in intracellular granules of only one type of hemocyte, the granulocyte. These results show for the first time that in ascidians, a sulfated galactoglucan circulates free in the plasma, and heparin occurs as an intracellular product of a circulating basophil-like cell.

  6. A new species of photosymbiotic ascidian from the Ryukyu Archipelago, Japan, with remarks on the stability of stigma number in photosymbiotic Diplosoma species.

    PubMed

    Hirose, Euichi; Oka, Atsushi T

    2008-12-01

    Diplosoma variostigmatum sp. nov. is a photosymbiotic ascidian in the family Didemnidae. This species is characterized by the unique pattern of its stigma number, which is often variable within the same zooid, as well as within the same colony. The total number of stigmata per half branchial sac varies from 22 (five, six, six, and five stigmata from the top to bottom rows) to 27 (seven, seven, seven, and six). In contrast, the stigma patterns are constant in D. ooru, D. simile, D. simileguwa, and D. virens. Thus, the stigma number and its stability (or variability) seem to be important as characters for the taxonomy of the photosymbiotic Diplosoma species.

  7. A new notodelphyid copepod, Paranotodelphys illgi n. sp. (Copepoda: Cyclopoida), parasitic in the ascidian Corynascidia herdmani Ritter in the North Pacific.

    PubMed

    Marchenkov, Andrey; Boxshall, Geoff A

    2003-01-01

    A new species of the Notodelphyidae belonging to Paranotodelphys Schellenberg, 1922 is described from both sexes. The new species, Paranotodelphys illgi, can be distinguished from its congeners by the combination of its 10-segmented female antennule, the bisetose fifth legs and the extreme elongation of the caudal rami of the female, which are more than 10 times longer than wide and 4.4 times longer than the anal somite. The male has non-geniculate antennules. The new species was collected at a depth of 508m in the North Pacific Ocean, from the ascidian Corynascidia herdmani Ritter.

  8. CIPRO 2.5: Ciona intestinalis protein database, a unique integrated repository of large-scale omics data, bioinformatic analyses and curated annotation, with user rating and reviewing functionality.

    PubMed

    Endo, Toshinori; Ueno, Keisuke; Yonezawa, Kouki; Mineta, Katsuhiko; Hotta, Kohji; Satou, Yutaka; Yamada, Lixy; Ogasawara, Michio; Takahashi, Hiroki; Nakajima, Ayako; Nakachi, Mia; Nomura, Mamoru; Yaguchi, Junko; Sasakura, Yasunori; Yamasaki, Chisato; Sera, Miho; Yoshizawa, Akiyasu C; Imanishi, Tadashi; Taniguchi, Hisaaki; Inaba, Kazuo

    2011-01-01

    The Ciona intestinalis protein database (CIPRO) is an integrated protein database for the tunicate species C. intestinalis. The database is unique in two respects: first, because of its phylogenetic position, Ciona is suitable model for understanding vertebrate evolution; and second, the database includes original large-scale transcriptomic and proteomic data. Ciona intestinalis has also been a favorite of developmental biologists. Therefore, large amounts of data exist on its development and morphology, along with a recent genome sequence and gene expression data. The CIPRO database is aimed at collecting those published data as well as providing unique information from unpublished experimental data, such as 3D expression profiling, 2D-PAGE and mass spectrometry-based large-scale analyses at various developmental stages, curated annotation data and various bioinformatic data, to facilitate research in diverse areas, including developmental, comparative and evolutionary biology. For medical and evolutionary research, homologs in humans and major model organisms are intentionally included. The current database is based on a recently developed KH model containing 36,034 unique sequences, but for higher usability it covers 89,683 all known and predicted proteins from all gene models for this species. Of these sequences, more than 10,000 proteins have been manually annotated. Furthermore, to establish a community-supported protein database, these annotations are open to evaluation by users through the CIPRO website. CIPRO 2.5 is freely accessible at http://cipro.ibio.jp/2.5.

  9. NK4 Antagonizes Tbx1/10 to Promote Cardiac versus Pharyngeal Muscle Fate in the Ascidian Second Heart Field

    PubMed Central

    Wang, Wei; Razy-Krajka, Florian; Siu, Eric; Ketcham, Alexandra; Christiaen, Lionel

    2013-01-01

    The heart and head muscles share common developmental origins and genetic underpinnings in vertebrates, including humans. Parts of the heart and cranio-facial musculature derive from common mesodermal progenitors that express NKX2-5, ISL1, and TBX1. This ontogenetic kinship is dramatically reflected in the DiGeorge/Cardio-Velo-Facial syndrome (DGS/CVFS), where mutations of TBX1 cause malformations in the pharyngeal apparatus and cardiac outflow tract. Cardiac progenitors of the first heart field (FHF) do not require TBX1 and segregate precociously from common progenitors of the second heart field (SHF) and pharyngeal muscles. However, the cellular and molecular mechanisms that govern heart versus pharyngeal muscle specification within this lineage remain elusive. Here, we harness the simplicity of the ascidian larva to show that, following asymmetric cell division of common progenitors, NK4/NKX2-5 promotes GATAa/GATA4/5/6 expression and cardiac specification in the second heart precursors by antagonizing Tbx1/10-mediated inhibition of GATAa and activation of Collier/Olf/EBF (COE), the determinant of atrial siphon muscle (ASM) specification. Our results uncover essential regulatory connections between the conserved cardio-pharyngeal factor Tbx1/10 and muscle determinant COE, as well as a mutual antagonism between NK4 and Tbx1/10 activities upstream of GATAa and COE. The latter cross-antagonism underlies a fundamental heart versus pharyngeal muscle fate choice that occurs in a conserved lineage of cardio-pharyngeal progenitors. We propose that this basic ontogenetic motif underlies cardiac and pharyngeal muscle development and evolution in chordates. PMID:24311985

  10. Purification and characterization of a 39,000-Da serine proteinase from the hemolymph of a solitary ascidian, Halocynthia roretzi.

    PubMed

    Shishikura, F; Abe, T; Ohtake, S; Tanaka, K

    1997-09-01

    A new endogenous serine proteinase from the cell-free hemolymph of a solitary ascidian, Halocythia roretzi, was purified by a combination of ammonium sulfate fractionation, hydrophobic interaction chromatography on TSKgel Toyopearl HW 65 F, ion exchange chromatography on TSKgel DEAE-Toyopearl 650 M, affinity chromatography on Arginine-Sepharose 4B, gel filtration on TSKgel Toyopearl HW 65F and hydroxyapatite chromatography on Bio-Gel HT. The serine proteinase is a single polypeptide chain whose molecular weight and isoelectric point are 39 kDa and about 7.6 pI, respectively. The most susceptible substrate was Boc-Leu-Gly-Arg-4-methyl-coumaryl-7-amide (MCA), and activity was optimal at pH 8. The enzyme was relatively stable at high temperatures; about 50% activity was retained even at 60 degrees C for 30 min in 50 mM Tris-HCl, pH 8.0, containing 0.5 M NaCl, and 0.05% Brij-35. The enzyme was characterized by the inhibitory effects of synthetic or natural inhibitors, substrate specificity toward 26 peptidyl-MCAs, proteinase activity toward natural proteins and complex formation with a serine proteinase inhibitor (58 kDa) previously found in H. roretzi hemolymph, indicating that the enzyme was a member of serine proteinases and strongly inhibited by the 58 kDa serine proteinase inhibitor as well as human antithrombin III. We also demonstrated the clotting enzyme activity of the purified serine proteinase toward bovine fibrinogen and Limulus coagulogen, a fibrinogen-like clottable protein of horseshoe crabs.

  11. NK4 antagonizes Tbx1/10 to promote cardiac versus pharyngeal muscle fate in the ascidian second heart field.

    PubMed

    Wang, Wei; Razy-Krajka, Florian; Siu, Eric; Ketcham, Alexandra; Christiaen, Lionel

    2013-12-01

    The heart and head muscles share common developmental origins and genetic underpinnings in vertebrates, including humans. Parts of the heart and cranio-facial musculature derive from common mesodermal progenitors that express NKX2-5, ISL1, and TBX1. This ontogenetic kinship is dramatically reflected in the DiGeorge/Cardio-Velo-Facial syndrome (DGS/CVFS), where mutations of TBX1 cause malformations in the pharyngeal apparatus and cardiac outflow tract. Cardiac progenitors of the first heart field (FHF) do not require TBX1 and segregate precociously from common progenitors of the second heart field (SHF) and pharyngeal muscles. However, the cellular and molecular mechanisms that govern heart versus pharyngeal muscle specification within this lineage remain elusive. Here, we harness the simplicity of the ascidian larva to show that, following asymmetric cell division of common progenitors, NK4/NKX2-5 promotes GATAa/GATA4/5/6 expression and cardiac specification in the second heart precursors by antagonizing Tbx1/10-mediated inhibition of GATAa and activation of Collier/Olf/EBF (COE), the determinant of atrial siphon muscle (ASM) specification. Our results uncover essential regulatory connections between the conserved cardio-pharyngeal factor Tbx1/10 and muscle determinant COE, as well as a mutual antagonism between NK4 and Tbx1/10 activities upstream of GATAa and COE. The latter cross-antagonism underlies a fundamental heart versus pharyngeal muscle fate choice that occurs in a conserved lineage of cardio-pharyngeal progenitors. We propose that this basic ontogenetic motif underlies cardiac and pharyngeal muscle development and evolution in chordates.

  12. Epimerization of D-glucose to L-galactose during the biosynthesis of a sulfated L-galactan in the ascidian tunic

    SciTech Connect

    Mourao, P.A.S. )

    1991-04-09

    The sulfated polysaccharides occurring in the tunic of ascidians are unique among known sulfated polysaccharides in that their major constituent sugar is galactose, which occurs exclusively in the L-enantiomeric form. In vitro incorporation experiments using tunic slices incubated with {sup 14}C-labeled sugars revealed that cells from this tissue epimerize D-isomers of hexose into L-galactose during the biosynthesis of their constituent polysaccharides. Compared with other hexoses, the precursor D-({sup 14}C)glucose has the highest rate of incorporation and produces the highest proportion of L-galactose units. This metabolic pathway is distinct from the epimerization of D-mannose to L-galactose through its guanosine 5{prime}-diphosphate nucleotide, described previously in an alga and in a snail. Therefore, the epimerization of D-glucose to L-galactose in the ascidian tunic occurs through a novel metabolic route, which involves inversion of the configuration of carbon atoms 2, 3, and 5 of the hexosyl moieties.

  13. Vanadium accumulation in ascidian coelomic cells is associated with enhanced pentose phosphate pathway capacity but not overall aerobic or anaerobic metabolism.

    PubMed

    Treberg, Jason R; Stacey, Joy E; Driedzic, William R

    2012-04-01

    Some suborders of ascidians (sea squirts) accumulate remarkable levels of the heavy metal vanadium while others accumulate negligible amounts. The function of this vanadium is unclear, but enhanced pentose phosphate pathway (PPP) has been implicated in its reduction and accumulation. We compared aspects of intermediary metabolism in coelomic cells from ascidian species that have a wide range of vanadium accumulation including non-accumulators. All species appear to have similar aerobic poise with no apparent link to vanadium accumulation. Similarly, all species examined have a limited anaerobic poise that does not seem to relate to vanadium levels. Based on the activities of phosphoglucose isomerase and glucose-6 phosphate dehydrogenase we demonstrate that, relative to the capacity for entry into glycolysis, vanadium-accumulating species have enhanced capacity to metabolize glucose-6 phosphate via the PPP compared to non-accumulators. This finding provides the first comparative support for enhanced PPP capacity linked to vanadium accumulation in tunicates. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. Southeastern Mediterranean Panorama

    NASA Image and Video Library

    1991-06-14

    STS040-152-180 (5-24 June 1991) --- The Sinai Peninsula dominates this north-looking, oblique view. According to NASA photo experts studying the STS 40 imagery, the Red Sea in the foreground is clear of river sediment because of the prevailing dry climate of the Middle East. The great rift of the Gulf of Aqaba extends northward to Turkey (top right) through the Dead Sea. The international boundary between Israel and Egypt, reflecting different rural landscapes, stands out clearly. The Nile River runs through the frame. NASA photo experts believe the haze over the Mediterranean to be wind-borne dust. The photo was taken with an Aero-Linhof large format camera.

  15. Qibla in the Mediterranean

    NASA Astrophysics Data System (ADS)

    Rius-Piniés, Mònica

    Orientation toward Mecca has been compulsory for Muslims in all time periods and in all places. In fact, mosques were built in such a way as to help believers to pray toward the right direction. Nevertheless, the alignment of the sacred buildings was not always exact, and many did not actually face the Kaaba. There are many reasons for this "mistake", the main one being that at the time of the construction of the most important mosques, the astronomical and geographical knowledge needed to make accurate calculations was lacking. In the Mediterranean area, the scholars who were most involved in this task were the fuqahā' (experts in Islamic jurisprudence) who were sometimes well versed in astronomical knowledge or, at least, were skilled in the practice of popular astronomy. The combination of astronomy and religion, mixed with the political and topographical conditions, produces a unique area of study which remains controversial today.

  16. Familial Mediterranean fever.

    PubMed

    Padeh, Shai; Berkun, Yackov

    2016-09-01

    Familial Mediterranean fever (FMF) is the oldest and most common of the hereditary autoinflammatory diseases (AIDs). A large body of information has been accumulated over recent years on the pathophysiology, diagnosis and treatment of these diseases. The purpose of this review is to bring an up-to-date summary of the clinic manifestations, diagnostic criteria and treatment of FMF. An overview of the pathophysiologic basis of FMF as part of the AID is discussed. Over the last year, attempts to establish new criteria for childhood FMF, new guidelines for treatment and follow-up of disease and novel treatment for FMF were made. A comparison of the different disease severity scores for research purposes suggests that a new score is needed. New evidence for antiinterleukin-1 blockade as a new treatment modality is described. New diagnostic criteria, disease severity score, treatment and follow-up guidelines have been proposed, and need validation in the next several years.

  17. Familial Mediterranean Fever

    PubMed Central

    Migita, Kiyoshi; Agematsu, Kazunaga; Yazaki, Masahide; Nonaka, Fumiaki; Nakamura, Akinori; Toma, Tomoko; Kishida, Dai; Uehara, Ritei; Nakamura, Yoshikazu; Jiuchi, Yuka; Masumoto, Junya; Furukawa, Hiroshi; Ida, Hiroaki; Terai, Chihiro; Nakashima, Yoshikazu; Kawakami, Atsushi; Nakamura, Tadashi; Eguchi, Katsumi; Yasunami, Michio; Yachie, Akihiro

    2014-01-01

    Abstract Familial Mediterranean fever (FMF) is an autoinflammatory disease caused by MEditerranean FeVer gene (MEFV) mutations. In Japan, patients with FMF have been previously reported, including a mild or incomplete form. Several factors are presumed to contribute to the variable penetrance and to the phenotypic variability of FMF. We conducted the current study to investigate the correlation of variable clinical presentations and MEFV genotypic distributions in Japanese FMF patients. We analyzed demographic, clinical, and genetic data for 311 FMF patients enrolled in the study. Clinically, we classified FMF into 2 phenotypes: 1) the “typical” form of FMF, and 2) the “atypical” form of FMF according to the Tel Hashomer criteria. Patients with the typical FMF phenotype had a higher frequency of febrile episodes, a shorter duration of febrile attacks, more frequent thoracic pain, abdominal pain, a family history of FMF, and MEFV exon 10 mutations. Conversely, patients with the atypical FMF phenotype had a lower frequency of fever episodes and more frequent arthritis in atypical distribution, myalgia, and MEFV exon 3 mutations. Multivariate analysis showed that the variable associated with typical FMF presentation was the presence of MEFV exon 10 mutations. Typical FMF phenotype frequencies were decreased in patients carrying 2 or a single low-penetrance mutations compared with those carrying 2 or a single high-penetrance mutations (M694I), with an opposite trend for the atypical FMF phenotype. In addition, patients having more than 2 MEFV mutations had a younger disease onset and a higher prevalence of thoracic pain than those carrying a single or no mutations. Thus, MEFV exon 10 mutations are associated with the more typical FMF phenotype. In contrast, more than half of the Japanese FMF patients without MEFV exon 10 mutations presented with an atypical FMF phenotype, indicating that Japanese FMF patients tend to be divided into 2 phenotypes by a variation

  18. Orthodontics in the Mediterranean Area.

    PubMed

    Calò, Claudia; Garino, Francesco; Cozzani, Mauro

    2006-01-01

    The article describes on the results of the first Workshop that SIDO organized in December 2005 in Cairo getting together twenty Countries who discussed Orthodontics in the Mediterranean Area. The aim was to take a picture, for the first time, of the state of Orthodontics in the different Mediterranean Countries using it as basis for a further co-operation. Progress in Orthodontics (published in English, indexed in Medline, free of charge website and downloading) was offered by SIDO as the means for a scientific and organizational sharing, and to spread a Mediterranean Orthodontic culture all over the world. The first step toward mutual understanding was completed and for the first time we collected data and the Mediterranean Orthodontic Integration Project (MOIP), on which we can base a coming meeting and debate, was introduced.

  19. Cancer and Mediterranean dietary traditions.

    PubMed

    Trichopoulou, A; Lagiou, P; Kuper, H; Trichopoulos, D

    2000-09-01

    The incidence of cancer overall in Mediterranean countries is lower than in Scandinavian countries, the United Kingdom, and the United States. This is mostly accounted for by the lower incidence among Mediterranean countries of cancer of the large bowel, breast, endometrium, and prostate. These forms of cancer have been linked to dietary factors, particularly low consumption of vegetables and fruit, and to a certain extent, high consumption of meat. The traditional Mediterranean diet is characterized by high consumption of foods of plant origin, relatively low consumption of red meat, and high consumption of olive oil, which in several studies has been reported to be more beneficial against cancer than other forms of added lipids. By taking into account the established or presumed nutritional causation of major forms of cancer and the composition of the traditional Mediterranean diet, estimates can be derived concerning the fraction of cancer occurrence in highly developed Western countries that could be attributed to their diets in comparison with the healthy traditional Mediterranean diet. Although estimates can only be crude, it can be calculated that up to 25% of the incidence of colorectal cancer, approximately 15% of the incidence of breast cancer, and approximately 10% of the incidence of prostate, pancreas, and endometrial cancer could be prevented if the populations of highly developed Western countries could shift to the traditional healthy Mediterranean diet.

  20. Assessment of coastal marine pollution in Galicia (NW Iberian Peninsula); metal concentrations in seawater, sediments and mussels (Mytilus galloprovincialis) versus embryo-larval bioassays using Paracentrotus lividus and Ciona intestinalis.

    PubMed

    Beiras, R; Bellas, J; Fernández, N; Lorenzo, J I; Cobelo-García, A

    2003-10-01

    Sediments from three Galician Rias were tested for toxicity using sea-urchin and ascidian sediment elutriate embryo-larval bioassays. Trace metal contents in seawater, sediments and mussels were also determined and subjected to multidimensional scaling methods which grouped stations according to chemical contamination. High metal contents were found in seawater, sediments and mussels from the Ria of Pontevedra, and moderate levels were detected in the Ria of Vigo and Ria of Arousa. The results revealed that samples assessed as toxic, according to the sea-urchin and ascidian embryo-larval bioassays, were among the most polluted by trace metals. A good agreement was reported between ordination plots resulting from applying multidimensional scaling to the chemical data, and the results of the biological endpoints tested.

  1. Calcium signaling by cyclic ADP-ribose, NAADP, and inositol trisphosphate are involved in distinct functions in ascidian oocytes.

    PubMed

    Albrieux, M; Lee, H C; Villaz, M

    1998-06-05

    ADP-ribosyl cyclase catalyzes the synthesis of two structurally and functionally different Ca2+ releasing molecules, cyclic ADP-ribose (cADPR) from beta-NAD and nicotinic acid-adenine dinucleotide phosphate (NAADP) from beta-NADP. Their Ca2+-mobilizing effects in ascidian oocytes were characterized in connection with that induced by inositol 1,4,5-trisphosphate (InsP3). Fertilization of the oocyte is accompanied by a decrease in the oocyte Ca2+ current and an increase in membrane capacitance due to the addition of membrane to the cell surface. Both of these electrical changes could be induced by perfusion, through a patch pipette, of nanomolar concentrations of cADPR or its precursor, beta-NAD, into unfertilized oocytes. The changes induced by beta-NAD showed a distinctive delay consistent with its enzymatic conversion to cADPR. The cADPR-induced changes were inhibited by preloading the oocytes with a Ca2+ chelator, indicating the effects were due to Ca2+ release induced by cADPR. Consistently, ryanodine (at high concentration) or 8-amino-cADPR, a specific antagonist of cADPR, but not heparin, inhibited the cADPR-induced changes. Both inhibitors likewise blocked the membrane insertion that normally occurred at fertilization consistent with it being mediated by a ryanodine receptor. The effects of NAADP were different from those of cADPR. Although NAADP induced a similar decrease in the Ca2+ current, no membrane insertion occurred. Moreover, pretreatment of the oocytes with NAADP inhibited the post-fertilization Ca2+ oscillation while cADPR did not. A similar Ca2+ oscillation could be artificially induced by perfusing into the oocytes a high concentration of InsP3 and NAADP could likewise inhibit such an InsP3-induced oscillation. This work shows that three independent Ca2+ signaling pathways are present in the oocytes and that each is involved in mediating distinct changes associated with fertilization. The results are consistent with a hierarchical organization of

  2. Structural investigations of the p53/p73 homologs from the tunicate species Ciona intestinalis reveal the sequence requirements for the formation of a tetramerization domain.

    PubMed

    Heering, Jan; Jonker, Hendrik R A; Löhr, Frank; Schwalbe, Harald; Dötsch, Volker

    2016-02-01

    Most members of the p53 family of transcription factors form tetramers. Responsible for determining the oligomeric state is a short oligomerization domain consisting of one β-strand and one α-helix. With the exception of human p53 all other family members investigated so far contain a second α-helix as part of their tetramerization domain. Here we have used nuclear magnetic resonance spectroscopy to characterize the oligomerization domains of the two p53-like proteins from the tunicate Ciona intestinalis, representing the closest living relative of vertebrates. Structure determination reveals for one of the two proteins a new type of packing of this second α-helix on the core domain that was not predicted based on the sequence, while the other protein does not form a second helix despite the presence of crucial residues that are conserved in all other family members that form a second helix. By mutational analysis, we identify a proline as well as large hydrophobic residues in the hinge region between both helices as the crucial determinant for the formation of a second helix.

  3. Ciona intestinalis galectin (CiLgals-a and CiLgals-b) genes are differentially expressed in endostyle zones and challenged by LPS.

    PubMed

    Parrinello, Daniela; Sanfratello, Maria Antonietta; Vizzini, Aiti; Parrinello, Nicolò; Cammarata, Matteo

    2015-01-01

    Immunohistochemical and in situ hybridization assays were performed to answer the question whether the endostyle, that is the initial gastro-intestinal trait of Ciona intestinalis pharynx, is involved in galectin (CiLgals-a and CiLgals-b) production during the pharynx inflammatory response to LPS inoculation. Specific anti-CiLgal-a and anti-CiLgals-b antibodies, and oligonucleotide probes, that mark inflammatory hemocytes inside the pharynx vessels and vessel epithelium as shown by a previous paper, were assayed on endostyle histological sections. For the first time, we show that galectins are produced by endostyle zones, and both CiLgals-a and -b genes are upregulated by LPS. CiLgals-a and CiLgals-b are constitutively expressed in the endostyle zone 2 and 3, respectively, both genes are upregulated by LPS in the zone 2, and CiLgals-b in the zone 3 and 4. The antibody-reacting material contained in intracellular and extracellular large vesicles suggest an unexpected vesicle-dependent transporting mechanism of galectins not provided with signal peptide. Differential expression and gene upregulation in not-treated and LPS-treated specimens, support the role of endostyle galectins both in filter feeding and defense responses.

  4. A reverse search for antimicrobial peptides in Ciona intestinalis: identification of a gene family expressed in hemocytes and evaluation of activity.

    PubMed

    Fedders, Henning; Leippe, Matthias

    2008-01-01

    In search of antimicrobial peptides in the tunicate Ciona intestinalis, we used the recently completed genome project and the substantial number of expressed sequence tag (EST) data available as a screening matrix. By this means, we identified a putative gene family that exhibits several structural features typical of antimicrobial peptides. We designed and synthesized a peptide corresponding to the core region of a member of this peptide family, which is predicted to adopt an amphipathic alpha-helical structure. The synthetic peptide exerted potent antimicrobial activity against a variety of bacteria and against the yeast Candida albicans but was not cytolytic for mammalian erythrocytes. Moreover, by employing a non-cell-permeable fluorescent dye it became evident that the peptide kills bacteria by permeabilizing their cytoplasmic membranes. Using the synthetic peptide as an antigen, we generated specific antibodies and localized the natural parent molecule to a compartment of a distinct hemocyte type, the univacuolar refractile granulocytes. As C. intestinalis apparently does not possess gene products that resemble well-known antimicrobial peptides of tunicates and of other animals, the aforementioned peptide family may represent a potent armamentarium of the hemocytes to combat microbial infection in sea squirts.

  5. Adaptive evolution of Mediterranean pines.

    PubMed

    Grivet, Delphine; Climent, José; Zabal-Aguirre, Mario; Neale, David B; Vendramin, Giovanni G; González-Martínez, Santiago C

    2013-09-01

    Mediterranean pines represent an extremely heterogeneous assembly. Although they have evolved under similar environmental conditions, they diversified long ago, ca. 10 Mya, and present distinct biogeographic and demographic histories. Therefore, it is of special interest to understand whether and to what extent they have developed specific strategies of adaptive evolution through time and space. To explore evolutionary patterns, the Mediterranean pines' phylogeny was first reconstructed analyzing a new set of 21 low-copy nuclear genes with multilocus Bayesian tree reconstruction methods. Secondly, a phylogenetic approach was used to search for footprints of natural selection and to examine the evolution of multiple phenotypic traits. We identified two genes (involved in pines' defense and stress responses) that have likely played a role in the adaptation of Mediterranean pines to their environment. Moreover, few life-history traits showed historical or evolutionary adaptive convergence in Mediterranean lineages, while patterns of character evolution revealed various evolutionary trade-offs linking growth-development, reproduction and fire-related traits. Assessing the evolutionary path of important life-history traits, as well as the genomic basis of adaptive variation is central to understanding the past evolutionary success of Mediterranean pines and their future response to environmental changes.

  6. Establishment and succession of an epibiotic community on chromated copper arsenate-treated wood in Mediterranean waters.

    PubMed

    Karayanni, Hera; Kormas, Konstantinos Ar; Cragg, Simon; Nicolaidou, Artemis

    2010-01-01

    Colonization and succession of an epibiotic animal community on chromated copper arsenate (CCA)-treated wood were studied for 18 months in the eastern Mediterranean (Saronikos Gulf, Aegean Sea). Pine wood panels, 200 x 100 x 25 mm, impregnated with CCA at retentions of 0, 12, 24, and 48 kg m(-3) were used. The abundance or surface coverage of the most characteristic taxa (polychaetes, mollusca, crustacea bryozoa, sponges, ascidians) was measured in situ, while 12 months after submersion two panels of each retention were removed and examined in the laboratory. A total of 26 taxa were identified, among which polychaetes of the family Serpulidae dominated. The controls carried the largest number of species (17) but the lowest number of individuals. On panels with CCA retentions of 12 and 24 kg m(-3), 14 and 16 species were observed, respectively, while at 48 kg m(-3), only 9 species were found. Only the controls were affected by boring bivalves of the family Teredinidae and started to break up within 3 months of submersion. Statistically significant differences in barnacle and polychaete abundance were found between treated and untreated panels. There were no significant differences among panels treated at the three CCA loadings. Ordination by nonmetric multidimensional scaling showed a seasonal effect on the colonization of the treated panels, with the highest recruitment during the warmer months of the study.

  7. [Endemic zoonosis in Mediterranean area].

    PubMed

    Fenga, Concettina; Pugliese, Michela

    2013-01-01

    The Mediterranean is historically considered an area of high concentration of zoonoses. Mediterranean countries socio-economic features have favoured, over time, the onset of different types of zoonosis. Many of these may affect many occupational categories, first of all farmers, people working in abattoirs and processing products of animal origin. New farming activities and technologies have generated new occupational and zoonotic risks. These changes have influenced zoonosis epidemiology and have led to a gradual decrease in the number of diseases and to a reduction of some biological risks. However, brucellosis, Q fever, bovine tuberculosis cystic echinococcosis remain a strong example of zoonosis and a real risk, in the Mediterranean area especially. Therefore, an interdisciplinary collaboration between Veterinary Service, Public Health and Occupational medicine is necessary in order to plan territorial prevention.

  8. Two new species of photosymbiotic ascidians of the genus Diplosoma from the Ryukyu Archipelago, with partial sequences of the COI gene.

    PubMed

    Hirose, Euichi; Oka, Atsushi T; Hirose, Mamiko

    2009-05-01

    Two new species of Diplosoma are described from coral reefs in the Ryukyu Archipelago, Japan: D. watanabei sp. nov. and D. gumavirens sp. nov. Colonies of both species are green due to algal symbionts of Prochloron, which are distributed in the common cloacal cavity. Both species are characterized by the point of emergence of the retractor muscle and a unique combination of stigma numbers among the four rows of the branchial sac. There are three adhesive papillae in the embryos of D. watanabei, as in most ascidian embryos and larvae, whereas the pre-hatching embryos of D. gumavirens sp. nov. posses 12-16 adhesive papillae. Partial sequences of the cytochrome c oxidase subunit I (COI) gene discriminated the new species from each other and from congeners.

  9. Predicting the Eastern Mediterranean Basin

    NASA Astrophysics Data System (ADS)

    Royden, L.

    2011-12-01

    From ~30 Ma onwards, the evolution of the Mediterranean region has been dominated by the rapid migration of thin-skinned thrust-belts. Thrust belt migration has been accommodated by the opening of "back arc type" basins within the upper plate of the thrust belts. Migration of these thrust belts is associated with the migration of subduction systems where subduction and thrusting are driven largely by negative slab buoyancy. Where the subducting slab has large negative buoyancy, thrust belt migration is commonly rapid. Where buoyant continental lithosphere enters the subduction system, subduction ceases quickly. Hence the large-scale tectonic evolution of the Mediterranean basin is largely pre-conditioned by its paleogeography. This can be quantitatively illustrated for the Hellenic subduction system where the post Eocene evolution of the Hellenic thrust belt can be ascribed to the buoyancy of the lithosphere subducted. Entry of the Ionian oceanic lithosphere into a slow-moving trench at 10-15 Ma explains the increase in subduction rate along the central part of the trench, to ~35 mm/yr at present, while subduction rates along strike to the northeast, where continental/transitional crust is subducted, remain less than ~10 mm/yr. Using quantitative modeling of the Hellenic subduction system in post Eocene time, it is possible to approximate how this thrust belt, and the active subduction belt of the eastern Mediterranean, will evolve over the next 10 m.y. This exercise suggests that the large-scale evolution of the eastern Mediterranean basin will be strikingly similar to that of the western Mediterranean basin from 20-0 Ma. This highlights the common dynamic mechanism that shapes the large scale deformation and morphology of the Mediterranean basin.

  10. Fine- and regional-scale genetic structure of the exotic ascidian Styela clava (Tunicata) in southwest England, 50 years after its introduction.

    PubMed

    Dupont, L; Viard, F; Dowell, M J; Wood, C; Bishop, J D D

    2009-02-01

    Styela clava, an ascidian native to the northwest Pacific, was first recorded in the Atlantic at Plymouth, southwest England, in 1953. It now ranges in the northeast Atlantic from Portugal to northern Denmark, and has colonized the east coast of North America. Within the region of first introduction, we aimed to characterize current genetic diversity in the species, elucidate the respective roles of human-aided vs. natural dispersal, and assess the extent of larval dispersal by looking for genetic differentiation at very small scales. Eight sites, mostly marinas, were studied along c. 200 km of coast in southwest England encompassing Plymouth. Five microsatellite loci were genotyped in 303 individuals to analyse gene flow at regional (among sites) and fine (within sites) scales. F-statistics and assignment tests were used to investigate regional genetic structure. At the fine scale, deviation from mutation-drift equilibrium was tested, and isolation by distance and genetic clustering analyses were undertaken. Significant genetic differentiation existed between sites, unrelated to geographical separation; migration between geographically distant marinas was inferred, highlighting the likely importance of human-mediated dispersal in range expansion and occupancy by S. clava. Fine-scale population structure was present within at least four sites, which may be explained by the limited dispersal ability of this ascidian and recruitment from differentiated pools of larvae. Populations in enclosed marinas had higher self-recruitment rates than those in open sites. Some marinas might therefore function as reservoirs of propagules for subsequent spread, whereas others might be sinks for migrants.

  11. The sexual and mating system of the shrimp Odontonia katoi (Palaemonidae, Pontoniinae), a symbiotic guest of the ascidian Polycarpa aurata in the Coral Triangle.

    PubMed

    Baeza, J Antonio; Hemphill, Carrie A; Ritson-Williams, Raphael

    2015-01-01

    Theory predicts that monogamy is adaptive in symbiotic crustaceans inhabiting relatively small and morphologically simple hosts in tropical environments where predation risk away from hosts is high. We tested this prediction in the shrimp Odontonia katoi, which inhabits the atrial chamber of the ascidian Polycarpa aurata in the Coral Triangle. Preliminary observations in O. katoi indicated that males were smaller than females, which is suggestive of sex change (protandry) in some symbiotic organisms. Thus, we first investigated the sexual system of O. katoi to determine if this shrimp was sequentially hermaphroditic. Morphological identification and size frequency distributions indicated that the population comprised males that, on average, were smaller than females. Gonad dissections demonstrated the absence of transitional individuals. Thus, O. katoi is a gonochoric species with reverse sexual dimorphism. The population distribution of O. katoi in its ascidian host did not differ significantly from a random distribution and shrimps inhabiting the same host individual as pairs were found with a frequency similar to that expected by chance alone. This is in contrast to that reported for other socially monogamous crustaceans in which pairs of heterosexual conspecifics are found in host individuals more frequently than expected by chance alone. Thus, the available information argues against monogamy in O. katoi. Furthermore, that a high frequency of solitary females were found brooding embryos and that the sex ratio was skewed toward females suggests that males might be roaming among hosts in search of receptive females in O. katoi. Symbiotic crustaceans can be used as a model system to understand the adaptive value of sexual and mating systems in marine invertebrates.

  12. An ultraviolet-sensitive maternal mRNA encoding a cytoskeletal protein may be involved in axis formation in the ascidian embryo

    SciTech Connect

    Jeffery, W.R. )

    1990-09-01

    Ultraviolet (uv) irradiation of the vegetal hemisphere of fertilized eggs during ooplasmic segregation inhibits subsequent gastrulation and axis formation in ascidian embryos. The molecular basis of this phenomenon was investigated in by comparing in vivo protein synthesis and in vitro mRNA translation in normal and uv-irradiated embryos of the ascidian Styela clava. Analysis of protein synthesis by (35S)methionine incorporation, two-dimensional (2D) gel electrophoresis, and autoradiography showed that only 21 of 433 labeled polypeptides were missing or decreased in labeling intensity in uv-irradiated embryos. The most prominent of these was a 30,000 molecular weight (pI 6.0) polypeptide (p30). Extraction of gastrulae with the nonionic detergent Triton X-100 showed that p30 is retained in the detergent insoluble residue, suggesting that it is associated with the cytoskeleton. Several lines of evidence suggest that p30 may be involved in axis formation. First, p30 labeling peaks during gastrulation, when the embryonic axis is being established. Second, axis formation and p30 labeling are abolished by the same threshold uv dose, which is distinct from that required to inactivate muscle cell development. Third, the uv sensitivity period for abolishing p30 labeling and axis formation are both restricted to ooplasmic segregation. In vitro translation of egg RNA followed by 2D gel electrophoresis and autoradiography of the protein products showed that p30 is encoded by a maternal mRNA. The translation of p30 mRNA was abolished by uv irradiation of fertilized eggs during ooplasmic segregation suggesting that this message is a uv-sensitive target. The results are consistent with the hypothesis that uv irradiation blocks gastrulation and axis formation by inhibiting the translation of maternal mRNA localized in the vegetal hemisphere of the fertilized egg.

  13. The Sexual and Mating System of the Shrimp Odontonia katoi (Palaemonidae, Pontoniinae), a Symbiotic Guest of the Ascidian Polycarpa aurata in the Coral Triangle

    PubMed Central

    Baeza, J. Antonio; Hemphill, Carrie A.; Ritson-Williams, Raphael

    2015-01-01

    Theory predicts that monogamy is adaptive in symbiotic crustaceans inhabiting relatively small and morphologically simple hosts in tropical environments where predation risk away from hosts is high. We tested this prediction in the shrimp Odontonia katoi, which inhabits the atrial chamber of the ascidian Polycarpa aurata in the Coral Triangle. Preliminary observations in O. katoi indicated that males were smaller than females, which is suggestive of sex change (protandry) in some symbiotic organisms. Thus, we first investigated the sexual system of O. katoi to determine if this shrimp was sequentially hermaphroditic. Morphological identification and size frequency distributions indicated that the population comprised males that, on average, were smaller than females. Gonad dissections demonstrated the absence of transitional individuals. Thus, O. katoi is a gonochoric species with reverse sexual dimorphism. The population distribution of O. katoi in its ascidian host did not differ significantly from a random distribution and shrimps inhabiting the same host individual as pairs were found with a frequency similar to that expected by chance alone. This is in contrast to that reported for other socially monogamous crustaceans in which pairs of heterosexual conspecifics are found in host individuals more frequently than expected by chance alone. Thus, the available information argues against monogamy in O. katoi. Furthermore, that a high frequency of solitary females were found brooding embryos and that the sex ratio was skewed toward females suggests that males might be roaming among hosts in search of receptive females in O. katoi. Symbiotic crustaceans can be used as a model system to understand the adaptive value of sexual and mating systems in marine invertebrates. PMID:25799577

  14. Impact and recovery from a mass mortality event of the gorgonian Paramuricea clavata populations on the french Mediterranean coasts

    NASA Astrophysics Data System (ADS)

    Bonhomme, D.; Garrabou, J.; Perez, T.; Sartoretto, S.; Harmelin, J. G.

    2003-04-01

    An unprecedented mass mortality occurred in the NW Mediterranean in summer 1999. This event affected 30 species of invertebrates from 5 different Phyla (sponges, cnidarians, molluscs, ascidians, bryozoans) on several hundred kilometres of shoreline from the Bay of Genoa in Italy to the Bay of Marseilles in France. The most affected taxa were sponges and cnidarians. This mass mortality took place under an unusual environmental context characterized by high and stable water column temperatures. The impact of the mass mortality and recovery (1999-2002) of populations of the gorgonian Paramuricea clavata from Provence's coasts (France) have been studied. Most surveyed populations displayed 50 % of affected colonies at different degree (dead, severe and low damage), although a great variability in rates of mortality and tissue loss was also evident depending among sites. The mortality impact decreased with depth, a pattern which supports the hypothesis that temperature played a key role in this event. Surveys on populations for which pre-mortality data (density and size structure) was attested that recovery was far from completion three years after the mortality. Considering the growth rate of P. clavata, full recovery of the most affected local populations will take several decades will be necessary to return to pre-mortality values. In the context of the global change and taking into account the correlation with the temperature, one may expect that the frequency of of these kind of events will increase in the next decades. Dramatic consequences can be expected for the conservation of the affected species in shallow habitats, in particular, and for the Mediterranean biodiversity, in general.

  15. Postfire chaparral regeneration under mediterranean and non-mediterranean climates

    USGS Publications Warehouse

    Keeley, Jon E.; Fotheringham, Connie J.; Rundel, Philip W.

    2012-01-01

    This study compares postfire regeneration and diversity patterns in fire-prone chaparral shrublands from mediterranean (California) and non-mediterranean-type climates (Arizona). Vegetation sampling was conducted in tenth hectare plots with nested subplots for the first two years after fire. Floras in the two regions were compared with Jaccard's Index and importance of families and genera compared with dominance-diversity curves. Although there were 44 families in common between the two regions, the dominant families differed; Poaceae and Fabaceae in Arizona and Hydrophyllaceae and Rosaceae in California. Dominance diversity curves indicated in the first year a more equable distribution of families in Arizona than in California. Woody plants were much more dominant in the mediterranean climate and herbaceous plants more dominant in the bimodal rainfall climate. Species diversity was comparable in both regions at the lowest spatial scales but not at the tenth hectare scale. Due to the double growing season in the non-mediterranean region, the diversity for the first year comprised two different herbaceous floras in the fall and spring growing seasons. The Mediterranean climate in California, in contrast, had only a spring growing season and thus the total diversity for the first year was significantly greater in Arizona than in California for both annuals and herbaceous perennials. Chaparral in these two climate regimes share many dominant shrub species but the postfire communities are very different. Arizona chaparral has both a spring and fall growing season and these produce two very different postfire floras. When combined, the total annual diversity was substantially greater in Arizona chaparral.

  16. Mediterranean Diet and Diabetes: Prevention and Treatment

    PubMed Central

    Georgoulis, Michael; Kontogianni, Meropi D.; Yiannakouris, Nikos

    2014-01-01

    The aim of the present review is to examine current scientific knowledge on the association between the Mediterranean diet and diabetes mellitus (mostly type 2 diabetes). A definition of the Mediterranean diet and the tools widely used to evaluate adherence to this traditional diet (Mediterranean diet indices) are briefly presented. The review focuses on epidemiological data linking adherence to the Mediterranean diet with the risk of diabetes development, as well as evidence from interventional studies assessing the effect of the Mediterranean diet on diabetes control and the management of diabetes-related complications. The above mentioned data are explored on the basis of evaluating the Mediterranean diet as a whole dietary pattern, rather than focusing on the effect of its individual components. Possible protective mechanisms of the Mediterranean diet against diabetes are also briefly discussed. PMID:24714352

  17. [Mediterranean diet: not only food].

    PubMed

    da Vico, Letizia; Agostini, Susanna; Brazzo, Silvia; Biffi, Barbara; Masini, Maria Luisa

    2012-09-01

    The proposal of a Mediterranean way of life is much more than advise how to eat. The Mediterranean Diet, a model of Sustainable Diet, is an example of how to combine personal choices, economic, social and cultural rights, protective of human health and the ecosystem. There is in fact fundamental interdependence between dietary requirements, nutritional recommendations, production and consumption of food. In literature studies and nutritional and epidemiological monitoring activities at national and international level have found a lack of adherence to this lifestyle, due to the spread of the economy, lifestyles of the Western type and globalization of the production and consumption. To encourage the spread of a culture and a constant practice of the Mediterranean Diet, there are some tools that are presented in this article. The Mediterranean Diet Pyramid in addition to the recommendations on the frequency and portions of food, focuses on the choice of how to cook and eat food. The "Double Food Pyramid" encourages conscious food choices based on "healthy eating and sustainability. All the nutrition professionals and dietitians in particular should be constantly striving to encourage the adoption of a sustainable and balanced nutrition.

  18. PON1 and Mediterranean Diet.

    PubMed

    Lou-Bonafonte, José M; Gabás-Rivera, Clara; Navarro, María A; Osada, Jesús

    2015-05-27

    The Mediterranean diet has been proven to be highly effective in the prevention of cardiovascular diseases. Paraoxonase 1 (PON1) has been implicated in the development of those conditions, especially atherosclerosis. The present work describes a systematic review of current evidence supporting the influence of Mediterranean diet and its constituents on this enzyme. Despite the differential response of some genetic polymorphisms, the Mediterranean diet has been shown to exert a protective action on this enzyme. Extra virgin olive oil, the main source of fat, has been particularly effective in increasing PON1 activity, an action that could be due to low saturated fatty acid intake, oleic acid enrichment of phospholipids present in high-density lipoproteins that favor the activity, and increasing hepatic PON1 mRNA and protein expressions induced by minor components present in this oil. Other Mediterranean diet constituents, such as nuts, fruits and vegetables, have been effective in modulating the activity of the enzyme, pomegranate and its compounds being the best characterized items. Ongoing research on compounds isolated from all these natural products, mainly phenolic compounds and carotenoids, indicates that some of them are particularly effective, and this may enhance the use of nutraceuticals and functional foods capable of potentiating PON1 activity.

  19. PON1 and Mediterranean Diet

    PubMed Central

    Lou-Bonafonte, José M.; Gabás-Rivera, Clara; Navarro, María A.; Osada, Jesús

    2015-01-01

    The Mediterranean diet has been proven to be highly effective in the prevention of cardiovascular diseases. Paraoxonase 1 (PON1) has been implicated in the development of those conditions, especially atherosclerosis. The present work describes a systematic review of current evidence supporting the influence of Mediterranean diet and its constituents on this enzyme. Despite the differential response of some genetic polymorphisms, the Mediterranean diet has been shown to exert a protective action on this enzyme. Extra virgin olive oil, the main source of fat, has been particularly effective in increasing PON1 activity, an action that could be due to low saturated fatty acid intake, oleic acid enrichment of phospholipids present in high-density lipoproteins that favor the activity, and increasing hepatic PON1 mRNA and protein expressions induced by minor components present in this oil. Other Mediterranean diet constituents, such as nuts, fruits and vegetables, have been effective in modulating the activity of the enzyme, pomegranate and its compounds being the best characterized items. Ongoing research on compounds isolated from all these natural products, mainly phenolic compounds and carotenoids, indicates that some of them are particularly effective, and this may enhance the use of nutraceuticals and functional foods capable of potentiating PON1 activity. PMID:26024295

  20. The formation and positioning of cilia in Ciona intestinalis embryos in relation to the generation and evolution of chordate left-right asymmetry.

    PubMed

    Thompson, Helen; Shaw, Michael K; Dawe, Helen R; Shimeld, Sebastian M

    2012-04-15

    In the early mouse embryo monocilia on the ventral node rotate to generate a leftward flow of fluid. This nodal flow is essential for the left-sided expression of nodal and pitx2, and for subsequent asymmetric organ patterning. Equivalent left fluid flow has been identified in other vertebrates, including Xenopus and zebrafish, indicating it is an ancient vertebrate mechanism. Asymmetric nodal and Pitx expression have also been identified in several invertebrates, including the vertebrates' nearest relatives, the urochordates. However whether cilia regulate this asymmetric gene expression remains unknown, and previous studies in urochordates have not identified any cilia prior to the larval stage, when asymmetry is already long established. Here we use Scanning and Transmission Electron Microscopy and immunofluorescence to investigate cilia in the urochordate Ciona intestinalis. We show that single cilia are transiently present on each ectoderm cell of the late neurula/early tailbud stage embryo, a time point just before onset of asymmetric nodal expression. Mapping the position of each cilium on these cells shows they are posteriorly positioned, something also described for mouse node cilia. The C. intestinalis cilia have a 9+0 ring ultrastructure, however we find no evidence of structures associated with motility such as dynein arms, radial spokes or nexin. Furthermore the 9+0 ring structure becomes disorganised immediately after the cilia have exited the cell, indicative of cilia which are not capable of motility. Our results indicate that although cilia are present prior to molecular asymmetries, they are not motile and hence cannot be operating in the same way as the flow-generating cilia of the vertebrate node. We conclude that the cilia may have a role in the development of C. intestinalis left-right asymmetry but that this would have to be in a sensory capacity, perhaps as mechanosensors as hypothesised in two-cilia physical models of vertebrate cilia

  1. The heart of Ciona intestinalis: eicosanoid-generating capacity and the effects of precursor fatty acids and eicosanoids on heart rate.

    PubMed

    Pope, Edward C; Rowley, Andrew F

    2002-06-01

    Eicosanoids are a group of oxygenated fatty-acid derivatives formed from C20 polyunsaturated fatty acids including arachidonic and eicosapentaenoic acids. In mammals, these compounds have been shown to be key molecules in several physiological processes including regulation of the vascular system. This study determined whether eicosanoids or their precursors are involved in the regulation of heart rate in the sea squirt Ciona intestinalis. Eicosanoid generation by both heart and blood cells was measured. The major lipoxygenase products formed were both derivatives of eicosapentaenoic acid, namely 8- and 12-hydroxyeicosapentaenoic acids (8-HEPE and 12-HEPE). Smaller amounts of 8,15-dihydroxyeicosapentaenoic acid (8,15-diHEPE) were also formed. The cyclo-oxygenase product prostaglandin E was also found in small amounts in the heart. Isolated hearts were exposed either to these fatty acid precursors or to 8-HEPE, 12-HEPE or prostaglandin E3, and the effect on heart rate was recorded. Both eicosapentaenoic and arachidonic acids stimulated the heart rate at concentrations between 50 and 200 micromol l(-1). 12-HEPE (5 micromol l(-1)) and prostaglandin E3 (50 micromoll(-1)) caused a modest increase in heart rate, while 8-HEPE had no significant effects at any of the time periods studied (

  2. Characterization of the teneurin C-terminal associated peptide (TCAP) in the vase tunicate, Ciona intestinalis: A novel peptide system associated with energy metabolism and reproduction.

    PubMed

    Colacci, Michael; De Almeida, Reuben; Chand, Dhan; Lovejoy, Sabine R; Sephton, Dawn; Vercaemer, Benedikte; Lovejoy, David A

    2015-05-15

    The vase tunicate, Ciona intestinalis, is a protochordate and is considered a sister lineage to the chordates. The recent sequencing of its genome has made this species a particularly important model to understand the genetic basis of vertebrate evolution. However, C. intestinalis is also a highly invasive species along the Atlantic coast of North America and other regions of the world which have caused considerable economic stress due to its biofouling actions and, in particular, negative impacts on the mussel- and oyster-based aquaculture industry. Despite this background, little is known about C. intestinalis physiology. The teneurin C-terminal associated peptides (TCAP) are a family of highly conserved peptide hormones found in most metazoans. Moreover, these peptides have been implicated in the inhibition of stress and stimulation of feeding-based metabolism. We have, therefore, identified this peptide using an in silico approach and characterized its immunological expression in tissues using a mouse polyclonal antiserum. These data indicate that its primary structure is more similar to invertebrate TCAPs relative to vertebrate TCAPs. Immunological expression indicates that it is highly expressed in the digestive tract and gonads consistent with findings in vertebrates. Synthetic mouse TCAP-1 administered into the brachial basket significantly increases the incidence of non-stress contractile behaviors. These findings support the hypothesis that TCAP is a bioactive peptide in C. intestinalis. Thus, C. intestinalis and tunicates in general may offer a simple model to investigate peptide interaction while providing information on how to control this invasive species. Crown Copyright © 2015. Published by Elsevier Inc. All rights reserved.

  3. Fire regime in Mediterranean ecosystem

    NASA Astrophysics Data System (ADS)

    Biondi, Guido; Casula, Paolo; D'Andrea, Mirko; Fiorucci, Paolo

    2010-05-01

    The analysis of burnt areas time series in Mediterranean regions suggests that ecosystems characterising this area consist primarily of species highly vulnerable to the fire but highly resilient, as characterized by a significant regenerative capacity after the fire spreading. In a few years the area burnt may once again be covered by the same vegetation present before the fire. Similarly, Mediterranean conifer forests, which often refers to plantations made in order to reforest the areas most severely degraded with high erosion risk, regenerate from seed after the fire resulting in high resilience to the fire as well. Only rarely, and usually with negligible damages, fire affects the areas covered by climax species in relation with altitude and soil types (i.e, quercus, fagus, abies). On the basis of these results, this paper shows how the simple Drossel-Schwabl forest fire model is able to reproduce the forest fire regime in terms of number of fires and burned area, describing whit good accuracy the actual fire perimeters. The original Drossel-Schwabl model has been slightly modified in this work by introducing two parameters (probability of propagation and regrowth) specific for each different class of vegetation cover. Using model selection methods based on AIC, the model with the optimal number of classes with different fire behaviour was selected. Two different case studies are presented in this work: Regione Liguria and Regione Sardegna (Italy). Both regions are situated in the center of the Mediterranean and are characterized by a high number of fires and burned area. However, the two regions have very different fire regimes. Sardinia is affected by the fire phenomenon only in summer whilst Liguria is affected by fires also in winter, with higher number of fires and larger burned area. In addition, the two region are very different in vegetation cover. The presence of Mediterranean conifers, (Pinus Pinaster, Pinus Nigra, Pinus halepensis) is quite spread in

  4. Handbook for Forecasters in the Mediterranean. Part 2. Regional Forecasting Aids for the Mediterranean Basin.

    DTIC Science & Technology

    1980-12-01

    Occurrences.....................11-7 3. Forecasting Rules.........................11-11 III. TYRRHENIAN SEA - CENTRAL MEDITERRANEAN AREA...the Tyrrhenian Sea -Central Mediterranean Area addressed in Section II1. A complex coastal topography characterizes the north and the south coasts of...is only 3 n mi. 11-25 III. TYRRHENIAN SEA - CENTRAL MEDITERRANEAN AREA 1. OVERVIEW 1.1 REGIONAL GEOGRAPHY The Tyrrhenian Sea -Central Mediterranean

  5. Mediterranean outflow mixing and dynamics.

    PubMed

    Price, J F; Baringer, M O; Lueck, R G; Johnson, G C; Ambar, I; Parrilla, G; Cantos, A; Kennelly, M A; Sanford, T B

    1993-02-26

    The Mediterranean Sea produces a salty, dense outflow that is strongly modified by entrainment as it first begins to descend the continental slope in the eastern Gulf of Cadiz. The current accelerates to 1.3 meters per second, which raises the internal Froude number above 1, and is intensely turbulent through its full thickness. The outflow loses about half of its density anomaly and roughly doubles its volume transport as it entrains less saline North Atlantic Central water. Within 100 kilometers downstream, the current is turned by the Coriolis force until it flows nearly parallel to topography in a damped geostrophic balance. The mixed Mediterranean outflow continues westward, slowly descending the continental slope until it becomes neutrally buoyant in the thermocline where it becomes an important water mass.

  6. Mediterranean diet and childhood asthma.

    PubMed

    Calatayud-Sáez, F M; Calatayud Moscoso Del Prado, B; Gallego Fernández-Pacheco, J G; González-Martín, C; Alguacil Merino, L F

    2016-01-01

    The incidence of bronchial hyperreactivity has increased to one-third of the population in developed countries, which requires the adoption of preventive and therapeutic measures. The objective of the present study was to assess the effects of a traditional Mediterranean diet on patients diagnosed with childhood asthma and determine if there is a beneficial effect from this dietary intervention. Prospective before-after comparison study of 50 girls and 54 boys aged 1-5 years, who were enrolled in the 1-year programme "Learning to Eat from the Mediterranean", designed to promote the adoption of a traditional Mediterranean diet. We studied the clinical and therapeutic variables and anthropometric measurements. All studied symptomatic indicators (number and intensity of asthmatic attack, infections and hospital admissions) showed a positive and statistically significant evolution of bronchial hyperreactivity from the first weeks of the intervention onwards. Throughout the treatment, 32.2% of patients remained free of crisis, 35.3% of the patients only had one attack throughout the year and 24.9% had two episodes, compared to 4.73 episodes on average in the previous year. The use of inhaled corticosteroids markedly decreased from 3.92 ± 1.61 to 1.11 ± 1.09 times per patient per year (P<0.001) and that of inhaled bronchodilators decreased from 4.14 ± 1.61 to 1.12 ± 1.40 (P<0.001). As a result, the families involved in the programme reported a high level of satisfaction. The adoption of a traditional Mediterranean diet could contribute significantly to the improvement of patients diagnosed with childhood asthma. Copyright © 2014 SEICAP. Published by Elsevier Espana. All rights reserved.

  7. Treatment of Mediterranean visceral leishmaniasis.

    PubMed Central

    Gradoni, L.; Bryceson, A.; Desjeux, P.

    1995-01-01

    Up-to-date information is given on the epidemiological situation of zoonotic visceral leishmaniasis (ZVL) in nine Mediterranean countries, and on drug regimens adopted in the management of ZVL patients in each country. Results of experimental and clinical trials on the efficacy and tolerability of liposomal amphotericin B in laboratory animals and in patients with ZVL are presented, as well as conclusions and recommendations on drug regimens to be used in the treatment of ZVL. PMID:7743590

  8. Gadolinium, a mechano-sensitive channel blocker, inhibits osmosis-initiated motility of sea- and freshwater fish sperm, but does not affect human or ascidian sperm motility.

    PubMed

    Krasznai, Zoltán; Morisawa, Masaaki; Krasznai, Zoárd Tibor; Morisawa, Sachiko; Inaba, Kazuo; Bazsáné, Zsuzsa Kassai; Rubovszky, Bálint; Bodnár, Béla; Borsos, Antal; Márián, Teréz

    2003-08-01

    Exposure to hypo-osmotic or hyperosmotic environment triggers the initiation of fish sperm motility. In this article, we report that calcium and potassium channel blockers do not influence motility of puffer fish sperm but calmodulin antagonists reversibly decrease it, suggesting that calmodulin-Ca(2+) interactions are prerequisite for the initiation of sperm motility in this species. Gadolinium (a stretch activated ion channel blocker) decreased the motility of puffer fish sperm from 92 +/- 3% to 6 +/- 3% and that of carp sperm from 91 +/- 7% to 3.5 +/- 4.3% in a dose-dependent manner (10-40 micro M). The effect of gadolinium was reversible, suggesting that stretch activated ion channels participate in the initiation of sperm motility of the two species. Gadolinium inhibits changes in the isoelectric point of certain proteins of puffer fish sperm, which occur when sperm motility is initiated in a hypertonic solution. Anisotropy measurements showed that hypo-osmotic treatment, which initiates carp sperm motility, increased membrane fluidity. When hypo-osmotic treatment was given in the presence of gadolinium, the sperm membrane remained as rigid as in quiescent cells, while motility was blocked. By contrast, gadolinium did not influence the motility parameters of Ciona or human sperm. Based on these lines of evidence, we suggest that conformational changes of mechanosensitive membrane proteins are involved in osmolality-dependent but not osmolality-independent sperm.

  9. Mantle dynamics in the Mediterranean

    NASA Astrophysics Data System (ADS)

    Faccenna, Claudio; Becker, Thorsten W.

    2016-04-01

    The Mediterranean offers a unique avenue to study the driving forces of tectonic deformation within a complex mobile belt. Lithospheric dynamics are affected by slab rollback and collision of two large, slowly moving plates, forcing fragments of continental and oceanic lithosphere to interact. Here, we review the rich and growing set of constraints from geological reconstructions, geodetic data, and crustal and upper mantle heterogeneity imaged by structural seismology. We discuss a conceptual and quantitative framework for the causes of surface deformations. Exploring existing and newly developed tectonic and numerical geodynamic models, we illustrate the role of mantle convection on surface geology. A coherent picture emerges which can be outlined by two, almost symmetric, upper mantle convection cells. The down-wellings are found in the centre of the Mediterranean, and are associated with the descent of the Tyrrhenian and the Hellenic slabs. During plate convergence, these slabs migrated, driving return flow of the asthenosphere from the backarc regions. These currents can be found at large distance from the subduction zones, and are at present expressed in two upwellings beneath Anatolia and eastern Iberia. This convection system provides an explanation for the general pattern of seismic anisotropy in the Mediterranean, the first-order Anatolia and Adria microplate kinematics, and the positive dynamic topography of Anatolia and Eastern Iberia. More generally, it is an illustration of upper mantle, small-scale convection leading to intraplate deformation and complex plate boundary reconfiguration at the westernmost terminus of the Tethyan collision.

  10. [Amyloidosis and familial Mediterranean fever].

    PubMed

    Pras, M

    1986-01-01

    Familial Mediterranean Fever (F. M. F.) is an autosomal recessive disorder occurring most commonly in Sepharadi Jews and Armenians. Two phenotypic features characterize the disease: brief episodic febrile attacks of peritonitis, pleuritis or synovitis recurring from childhood or adolescence and the development of systemic amyloidosis. Attacks are accompanied by striking elevations of acute phase proteins, including serum amyloid A protein. The amyloidosis of Familial Mediterranean Fever is of the AA type, and manifest clinically as a nephropathy that passes through proteinuria, nephrotic and uremic stages to renal death. Although there is ethnic variation in the incidence of amyloidosis of F. M. F. in our patient population--predominantly Sepharadi Jews of North African extraction--an amyloidotic death at an early age is their genetic destiny. Since the introduction in 1972 of colchicine to prevent the febrile attacks, the drug has been proven and become the main stay of therapy. Today, colchicine has been shown to be effective in preventing amyloidosis as well as the febrile attacks in Familial Mediterranean Fever. End stage renal disease is not the end of the road for patients with F.M.F. because of improving outlook for dialysis and renal transplantation in these patients.

  11. A Boolean Function for Neural Induction Reveals a Critical Role of Direct Intercellular Interactions in Patterning the Ectoderm of the Ascidian Embryo

    PubMed Central

    Mochizuki, Atsushi; Satou, Yutaka

    2015-01-01

    A complex system of multiple signaling molecules often produce differential gene expression patterns in animal embryos. In the ascidian embryo, four signaling ligands, Ephrin-A.d (Efna.d), Fgf9/16/20, Admp, and Gdf1/3-r, coordinately induce Otx expression in the neural lineage at the 32-cell stage. However, it has not been determined whether differential inputs of all of these signaling pathways are really necessary. It is possible that differential activation of one of these signaling pathways is sufficient and the remaining signaling pathways are activated in all cells at similar levels. To address this question, we developed a parameter-free method for determining a Boolean function for Otx expression in the present study. We treated activities of signaling pathways as Boolean values, and we also took all possible patterns of signaling gradients into consideration. We successfully determined a Boolean function that explains Otx expression in the animal hemisphere of wild-type and morphant embryos at the 32-cell stage. This Boolean function was not inconsistent with three sensing patterns, which represented whether or not individual cells received sufficient amounts of the signaling molecules. These sensing patterns all indicated that differential expression of Otx in the neural lineage is primarily determined by Efna.d, but not by differential inputs of Fgf9/16/20, Admp, and Gdf1/3-r signaling. To confirm this hypothesis experimentally, we simultaneously knocked-down Admp, Gdf1/3-r, and Fgf9/16/20, and treated this triple morphant with recombinant bFGF and BMP4 proteins, which mimic Fgf9/16/20 and Admp/Gdf1/3-r activity, respectively. Although no differential inputs of Admp, Gdf1/3-r and Fgf9/16/20 signaling were expected under this experimental condition, Otx was expressed specifically in the neural lineage. Thus, direct cell–cell interactions through Efna.d play a critical role in patterning the ectoderm of the early ascidian embryo. PMID:26714026

  12. A Boolean Function for Neural Induction Reveals a Critical Role of Direct Intercellular Interactions in Patterning the Ectoderm of the Ascidian Embryo.

    PubMed

    Ohta, Naoyuki; Waki, Kana; Mochizuki, Atsushi; Satou, Yutaka

    2015-12-01

    A complex system of multiple signaling molecules often produce differential gene expression patterns in animal embryos. In the ascidian embryo, four signaling ligands, Ephrin-A.d (Efna.d), Fgf9/16/20, Admp, and Gdf1/3-r, coordinately induce Otx expression in the neural lineage at the 32-cell stage. However, it has not been determined whether differential inputs of all of these signaling pathways are really necessary. It is possible that differential activation of one of these signaling pathways is sufficient and the remaining signaling pathways are activated in all cells at similar levels. To address this question, we developed a parameter-free method for determining a Boolean function for Otx expression in the present study. We treated activities of signaling pathways as Boolean values, and we also took all possible patterns of signaling gradients into consideration. We successfully determined a Boolean function that explains Otx expression in the animal hemisphere of wild-type and morphant embryos at the 32-cell stage. This Boolean function was not inconsistent with three sensing patterns, which represented whether or not individual cells received sufficient amounts of the signaling molecules. These sensing patterns all indicated that differential expression of Otx in the neural lineage is primarily determined by Efna.d, but not by differential inputs of Fgf9/16/20, Admp, and Gdf1/3-r signaling. To confirm this hypothesis experimentally, we simultaneously knocked-down Admp, Gdf1/3-r, and Fgf9/16/20, and treated this triple morphant with recombinant bFGF and BMP4 proteins, which mimic Fgf9/16/20 and Admp/Gdf1/3-r activity, respectively. Although no differential inputs of Admp, Gdf1/3-r and Fgf9/16/20 signaling were expected under this experimental condition, Otx was expressed specifically in the neural lineage. Thus, direct cell-cell interactions through Efna.d play a critical role in patterning the ectoderm of the early ascidian embryo.

  13. Mediterranean Ocean Colour Chlorophyll trend

    NASA Astrophysics Data System (ADS)

    rinaldi, eleonora; colella, simone; santoleri, rosalia

    2014-05-01

    Monitoring chlorophyll (Chl) concentration, seen as a proxy for phytoplankton biomass, is an efficient tool in order to understand the response of marine ecosystem to human pressures. This is particularly important along the coastal regions, in which the strong anthropization and the irrational exploitation of resources represent a persistent threat to the biodiversity. The aim of this work is to assess the effectiveness and feasibility of using Ocean Color (OC) data to monitor the environmental changes in Mediterranean Sea and to develop a method for detecting trend from OC data that can constitute a new indicator of the water quality within the EU Marine Strategy Framework Directive implementation. In this study the Mediterranean merged Case1-Case2 chlorophyll product, produced by CNR-ISAC and distributed in the framework of MyOcean, is analyzed. This product is obtained by using two different bio-optical algorithms for open ocean (Case1) and coastal turbid (Case2) waters; this improves the quality of the Chl satellite estimates, especially near the coast. In order to verify the real capability of the this product for estimating Chl trend and for selecting the most appropriated statistical test to detect trend in the Mediterranean Sea, a comparison between OC and in situ data are carried out. In-situ Chl data are part of the European Environment Information and Observation Network (Eionet) of the European Environmental Agency (EEA). Four different statistical approaches to estimate trend have been selected and used to compare trend values obtained with in-situ and OC data. Results show that the best agreement between in-situ and OC trend is achieved using the Mann- Kendall test. The Mediterranean trend map obtained applying this test to the de-seasonalized OC time series shows that, in accordance with the results of many authors, the case 1 waters of Mediterranean sea are characterized by a negative trend. However, the most intense trend signals, both negative

  14. Combining environmental suitability and population abundances to evaluate the invasive potential of the tunicate Ciona intestinalis along the temperate South American coast

    PubMed Central

    Estay, Sergio A.; Labra, Fabio A.; Lima, Mauricio

    2015-01-01

    The tunicate Ciona intestinalis is an opportunistic invader with high potential for causing economic losses in aquaculture centers. Recent phylogenetic and population genetic analysis support the existence of a genetic complex described as C. intestinalis with two main dominant species (sp A and B) occurring worldwide. In Chile, the species has been observed around 30°S of latitude, but no official reports exist for the presence of C. intestinalis in southern regions (above 40°S), where most of the mollusk aquaculture centers are located. Here, we used occurrences from multiple invaded regions and extensive field sampling to model and validate the environmental conditions that allow the species to persist and to find the geographic areas with the most suitable environmental conditions for the spread of C. intestinalis in the Chilean coast. By studying the potential expansion of C. intestinalis southward in the Chilean Coast, we aimed to provide valuable information that might help the development of control plans before the species becomes a significant problem, especially above 40°S. Our results highlight that, by using portions of the habitat that are apparently distinguishable, the species seem to be not only genetically distinct, but ecologically distinct as well. The two regional models fitted for sp A and for sp B showed disagreement on which sections of Chilean coastline are considered more suitable for these species. While the model for sp A identifies moderately to highly suitable areas between 30° and 40°S, the model for sp B classifies the areas around 45°S as the most appropriate. Data from field sampling show a positive linear relationship between density of C. intestinalis and the index of suitability for sp A in aquaculture centers. Understanding the relation of the distinct species with the surrounding environment provided valuable insights about probable routes of dispersion in Chile, especially into those areas considered suitable for

  15. Combining environmental suitability and population abundances to evaluate the invasive potential of the tunicate Ciona intestinalis along the temperate South American coast.

    PubMed

    Januario, Stella M; Estay, Sergio A; Labra, Fabio A; Lima, Mauricio

    2015-01-01

    The tunicate Ciona intestinalis is an opportunistic invader with high potential for causing economic losses in aquaculture centers. Recent phylogenetic and population genetic analysis support the existence of a genetic complex described as C. intestinalis with two main dominant species (sp A and B) occurring worldwide. In Chile, the species has been observed around 30°S of latitude, but no official reports exist for the presence of C. intestinalis in southern regions (above 40°S), where most of the mollusk aquaculture centers are located. Here, we used occurrences from multiple invaded regions and extensive field sampling to model and validate the environmental conditions that allow the species to persist and to find the geographic areas with the most suitable environmental conditions for the spread of C. intestinalis in the Chilean coast. By studying the potential expansion of C. intestinalis southward in the Chilean Coast, we aimed to provide valuable information that might help the development of control plans before the species becomes a significant problem, especially above 40°S. Our results highlight that, by using portions of the habitat that are apparently distinguishable, the species seem to be not only genetically distinct, but ecologically distinct as well. The two regional models fitted for sp A and for sp B showed disagreement on which sections of Chilean coastline are considered more suitable for these species. While the model for sp A identifies moderately to highly suitable areas between 30° and 40°S, the model for sp B classifies the areas around 45°S as the most appropriate. Data from field sampling show a positive linear relationship between density of C. intestinalis and the index of suitability for sp A in aquaculture centers. Understanding the relation of the distinct species with the surrounding environment provided valuable insights about probable routes of dispersion in Chile, especially into those areas considered suitable for

  16. Mitochondrial inhibitor sodium azide inhibits the reorganization of mitochondria-rich cytoplasm and the establishment of the anteroposterior axis in ascidian embryo.

    PubMed

    Ishii, Hirokazu; Shirai, Takuma; Makino, Chisato; Nishikata, Takahito

    2014-02-01

    In ascidian eggs, cytoplasmic and cortical reorganization, previously called ooplasmic segregation, occurs in two phases during the first cell cycle. In the second phase of reorganization, the mitochondria-rich cytoplasm (myoplasm) moves to the future posterior side, concurrent with sperm aster migration along the egg cortex. Although this reorganization is the critical step for establishing the anteroposterior axis, its molecular mechanism is not fully understood. In this study, we showed that low concentrations of the mitochondrial inhibitor sodium azide (NaN3 ), which showed the low toxicity in sperm, inhibited the second phase of reorganization without the microtubule depolymerization. In the NaN3 -treated embryo, the sperm aster was not attracted to the cortex and altered its migration pathway; therefore, the myoplasm remained at the vegetal pole. Consequently, the anteroposterior axis was not established. Another mitochondrial inhibitor, oligomycin, did not affect these processes. These results suggest that NaN3 inhibits unknown molecules that are important for the second phase of reorganization. Identifying the target molecule of NaN3 will lead to a molecular understanding of cytoplasmic and cortical reorganization.

  17. Molecular conformation of ascidiacyclamide, a cytotoxic cyclic peptide from Ascidian: X-ray analyses of its free form and solvate crystals.

    PubMed

    Ishida, T; In, Y; Doi, M; Inoue, M; Hamada, Y; Shioiri, T

    1992-02-01

    In order to investigate the conformational variation of ascidiacyclamide, a cytotoxic cyclic peptide from marine tunicate Ascidian, single crystals were prepared from ethanol and aqueous ethanol solutions as its free form (crystal I) and H2O/0.5 C2H5OH solvate (crystal II), respectively, and were determined by the x-ray diffraction method. Crystal I showed a pseudo C2-symmetric saddle-shaped rectangular conformation. Similar conformations were also observed in crystal II, where there were two crystallographically independent C2-symmetric molecules (named Mol-A and -B) per asymmetric unit. Mol-A and -B included H2O and H2O/C2H5OH solvents within their ring structures, respectively. These water and ethanol molecules were located on the crystallographic dyad axes, and were stabilized by the van der Waals contacts (including hydrogen bonds) with the polar-ring N atoms and nonpolar D-Val side-chain atoms. The conformational characteristics of ascidiacyclamide and its fluctuation/variation were discussed based on the present and previously reported x-ray results.

  18. Structural analysis of a sulfated galactan from the tunic of the ascidian Microcosmus exasperatus and its inhibitory effect of the intrinsic coagulation pathway.

    PubMed

    Restrepo-Espinosa, Diana C; Román, Yony; Colorado-Ríos, Jhonny; de Santana-Filho, Arquimedes Paixão; Sassaki, Guilherme Lanzi; Cipriani, Thales R; Martínez, Alejandro; Iacomini, Marcello; Pavão, Mauro S G

    2017-09-01

    Several bioactive sulfated galactans have been isolated from the tunic of different species of ascidians. The biological activity of this kind of polysaccharides has been related with the presence and position of sulfate groups, and by the chemical composition of this kind of polysaccharides. A sulfated galactan (1000RS) was isolated from the tunic of the Brazilian ascidia Microcosmus exasperatus through proteolytic digestion, ethanol precipitation, dialysis and freeze-thaw cycles. Homogeneity and molecular weight were estimated by using size exclusion chromatography. Monosaccharide composition and type of linkage were assessed by Gas chromatography coupled to mass spectrometry and the sulfate content was quantified through gelatin/BaCl2 method. These experiments along with NMR and FTIR analysis allowed to claim that the galactan backbone is mainly composed of 4-linked α-l-Galp units. In addition, they permitted to establish that some of the galactose residues are sulfated at the 3-position. This sulfated polysaccharide, which has an average molecular mass of 439.5kDa, presents anticoagulant effect in a dose-dependent manner through the inhibition of the intrinsic coagulation pathway. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Plate tectonics of the Mediterranean region.

    PubMed

    McKenzie, D P

    1970-04-18

    The seismicity and fault plane solutions in the Mediterranean area show that two small rapidly moving plates exist in the Eastern Mediterranean, and such plates may be a common feature of contracting ocean basins. The results show that the concepts of plate tectonics apply to instantaneous motions across continental plate boundaries.

  20. The Mediterranean Plastic Soup: synthetic polymers in Mediterranean surface waters

    NASA Astrophysics Data System (ADS)

    Suaria, Giuseppe; Avio, Carlo G.; Mineo, Annabella; Lattin, Gwendolyn L.; Magaldi, Marcello G.; Belmonte, Genuario; Moore, Charles J.; Regoli, Francesco; Aliani, Stefano

    2016-11-01

    The Mediterranean Sea has been recently proposed as one of the most impacted regions of the world with regards to microplastics, however the polymeric composition of these floating particles is still largely unknown. Here we present the results of a large-scale survey of neustonic micro- and meso-plastics floating in Mediterranean waters, providing the first extensive characterization of their chemical identity as well as detailed information on their abundance and geographical distribution. All particles >700 μm collected in our samples were identified through FT-IR analysis (n = 4050 particles), shedding for the first time light on the polymeric diversity of this emerging pollutant. Sixteen different classes of synthetic materials were identified. Low-density polymers such as polyethylene and polypropylene were the most abundant compounds, followed by polyamides, plastic-based paints, polyvinyl chloride, polystyrene and polyvinyl alcohol. Less frequent polymers included polyethylene terephthalate, polyisoprene, poly(vinyl stearate), ethylene-vinyl acetate, polyepoxide, paraffin wax and polycaprolactone, a biodegradable polyester reported for the first time floating in off-shore waters. Geographical differences in sample composition were also observed, demonstrating sub-basin scale heterogeneity in plastics distribution and likely reflecting a complex interplay between pollution sources, sinks and residence times of different polymers at sea.

  1. The Mediterranean Plastic Soup: synthetic polymers in Mediterranean surface waters

    PubMed Central

    Suaria, Giuseppe; Avio, Carlo G.; Mineo, Annabella; Lattin, Gwendolyn L.; Magaldi, Marcello G.; Belmonte, Genuario; Moore, Charles J.; Regoli, Francesco; Aliani, Stefano

    2016-01-01

    The Mediterranean Sea has been recently proposed as one of the most impacted regions of the world with regards to microplastics, however the polymeric composition of these floating particles is still largely unknown. Here we present the results of a large-scale survey of neustonic micro- and meso-plastics floating in Mediterranean waters, providing the first extensive characterization of their chemical identity as well as detailed information on their abundance and geographical distribution. All particles >700 μm collected in our samples were identified through FT-IR analysis (n = 4050 particles), shedding for the first time light on the polymeric diversity of this emerging pollutant. Sixteen different classes of synthetic materials were identified. Low-density polymers such as polyethylene and polypropylene were the most abundant compounds, followed by polyamides, plastic-based paints, polyvinyl chloride, polystyrene and polyvinyl alcohol. Less frequent polymers included polyethylene terephthalate, polyisoprene, poly(vinyl stearate), ethylene-vinyl acetate, polyepoxide, paraffin wax and polycaprolactone, a biodegradable polyester reported for the first time floating in off-shore waters. Geographical differences in sample composition were also observed, demonstrating sub-basin scale heterogeneity in plastics distribution and likely reflecting a complex interplay between pollution sources, sinks and residence times of different polymers at sea. PMID:27876837

  2. The Mediterranean Plastic Soup: synthetic polymers in Mediterranean surface waters.

    PubMed

    Suaria, Giuseppe; Avio, Carlo G; Mineo, Annabella; Lattin, Gwendolyn L; Magaldi, Marcello G; Belmonte, Genuario; Moore, Charles J; Regoli, Francesco; Aliani, Stefano

    2016-11-23

    The Mediterranean Sea has been recently proposed as one of the most impacted regions of the world with regards to microplastics, however the polymeric composition of these floating particles is still largely unknown. Here we present the results of a large-scale survey of neustonic micro- and meso-plastics floating in Mediterranean waters, providing the first extensive characterization of their chemical identity as well as detailed information on their abundance and geographical distribution. All particles >700 μm collected in our samples were identified through FT-IR analysis (n = 4050 particles), shedding for the first time light on the polymeric diversity of this emerging pollutant. Sixteen different classes of synthetic materials were identified. Low-density polymers such as polyethylene and polypropylene were the most abundant compounds, followed by polyamides, plastic-based paints, polyvinyl chloride, polystyrene and polyvinyl alcohol. Less frequent polymers included polyethylene terephthalate, polyisoprene, poly(vinyl stearate), ethylene-vinyl acetate, polyepoxide, paraffin wax and polycaprolactone, a biodegradable polyester reported for the first time floating in off-shore waters. Geographical differences in sample composition were also observed, demonstrating sub-basin scale heterogeneity in plastics distribution and likely reflecting a complex interplay between pollution sources, sinks and residence times of different polymers at sea.

  3. Importance of United States Naval Forward Presence in Mediterranean Affairs

    DTIC Science & Technology

    2001-06-01

    Mediterranean Region, and beyond. 24 This theme is developed by Fernand Braudel , The Mediterranean and...Relations,” Janes Intelligence Review, v. 13, no. 2, February 2000. Braudel , Fernand , “The Mediterranean and the Mediterranean World in the Age of

  4. The maternal genes Ci-p53/p73-a and Ci-p53/p73-b regulate zygotic ZicL expression and notochord differentiation in Ciona intestinalis embryos.

    PubMed

    Noda, Takeshi

    2011-12-01

    I isolated a Ciona intestinalis homolog of p53, Ci-p53/p73-a, in a microarray screen of rapidly degraded maternal mRNA by comparing the transcriptomes of unfertilized eggs and 32-cell stage embryos. Higher expression of the gene in eggs and lower expression in later embryonic stages were confirmed by whole-mount in situ hybridization (WISH) and quantitative reverse transcription-PCR (qRT-PCR); expression was ubiquitous in eggs and early embryos. Knockdown of Ci-p53/p73-a by injection of antisense morpholino oligonucleotides (MOs) severely perturbed gastrulation cell movements and expression of notochord marker genes. A key regulator of notochord differentiation in Ciona embryos is Brachyury (Ci-Bra), which is directly activated by a zic-like gene (Ci-ZicL). The expression of Ci-ZicL and Ci-Bra in A-line notochord precursors was downregulated in Ci-p53/p73-a knockdown embryos. Maternal expression of Ci-p53/p73-b, a homolog of Ci-p53/p73-a, was also detected. In Ci-p53/p73-b knockdown embryos, gastrulation cell movements, expression of Ci-ZicL and Ci-Bra in A-line notochord precursors, and expression of notochord marker gene at later stages were perturbed. The upstream region of Ci-ZicL contains putative p53-binding sites. Cis-regulatory analysis of Ci-ZicL showed that these sites are involved in expression of Ci-ZicL in A-line notochord precursors at the 32-cell and early gastrula stages. These results suggest that p53 genes are maternal factors that play a crucial role in A-line notochord differentiation in C. intestinalis embryos by regulating Ci-ZicL expression.

  5. Infrequently asked questions about the Mediterranean diet.

    PubMed

    Bellisle, France

    2009-09-01

    Numerous health benefits have been attributed to the 'Mediterranean diet' over the last decades. Selecting foods that were common in the Mediterranean regions (especially Crete) in the 1970s, with a frequent and abundant intake of fruit, vegetables, fish, olive oil and perhaps wine, has been reported to be associated with wide-ranging benefits including improved glucose metabolism and decreased risk of type 2 diabetes, obesity and CVD. While the respective contributions of various types of food have been widely investigated, less attention has been paid to other factors, also characteristic of the Mediterranean lifestyle, which may contribute to the health benefits perhaps as much as specific food choices. Traditionally, the Mediterranean diet was consumed in the context of a particular lifestyle, with a fixed number of daily meals, generally consumed at later hours (compared to North of Europe), and some specific meal-related behaviours such as the post-lunch siesta. In addition, the Mediterranean diet and lifestyle that were so beneficial to health, 40 years ago, were considerably different from present-day practices. The changes are particularly clear in younger individuals and countries of the Mediterranean region presently have the highest child overweight rates in Europe. The present paper will address research about meal and satiety patterns and examine how the recent changes from traditional practices are likely to have an impact on health risks and benefits in Mediterranean populations.

  6. Follicle cell trypsin-like protease HrOvochymase: Its cDNA cloning, localization, and involvement in the late stage of oogenesis in the ascidian Halocynthia roretzi.

    PubMed

    Mino, Masako; Sawada, Hitoshi

    2016-04-01

    We previously reported that the sperm trypsin-like protease HrAcrosin and its precursor HrProacrosin participate in fertilization of the ascidian Halocynthia roretzi. The HrProacrosin gene is annotated in the H. roretzi genome database as Harore.CG.MTP2014.S89.g15383; our previously reported sequence of HrProacrosin gene appeared to include four nucleotides inserted near the 3'-end of HrProacrosin, resulting in a frame-shift mutation and a premature termination codon. The gene architecture of HrProacrosin and Harore.CG.MTP2014.S89.g15383 resembles that of Xenopus laevis ovochymase-1/OVCH1 and ovochymase-2/OVCH2, which encode egg extracellular polyproteases. Considering these new observations, we evaluated the cDNA cloning, expression, localization, and function of Harore.CG.MTP2014.S89.g15383, herein designated as HrOvochymase/HrOVCH. We found that HrOVCH cDNA consists of a single open reading frame of 1,575 amino acids, containing a signal peptide, three trypsin-like protease domains, and six CUB domains. HrOVCH was transcribed by the testis and ovary, but the majority of protein exists in ovarian follicle cells surrounding eggs. An anti-HrOVCH antibody inhibited elevation of the vitelline coat at a late stage of oogenesis, during the period when self-sterility is acquired. As trypsin inhibitors are reported to block the acquisition of self-sterility during oogenesis, whereas trypsin induces the acquisition of self-sterility and elevation of the vitelline coat in defolliculated ovarian eggs, we propose that HrOVCH may play a role in the acquisition of self-sterility by late-stage H. roretzi oocytes.

  7. The occurrence of the colonial ascidian Didemnum sp. on Georges Bank gravel habitat: ecological observations and potential effects on groundfish and scallop fisheries

    USGS Publications Warehouse

    Valentine, P.C.; Collie, J.S.; Reid, R.N.; Asch, R.G.; Guida, V.G.; Blackwood, D.S.

    2007-01-01

    The colonial ascidian Didemnum sp. is present on the Georges Bank fishing grounds in a gravel habitat where the benthic invertebrate fauna has been monitored annually since 1994. The species was not noted before 2002 when large colonies were first observed; and by 2003 and 2004 it covered large areas of the seabed at some locations. The latest survey in 2005 documented the tunicate's presence in two gravel areas that total more than 67 nm2 (230 km2). The affected area is located on the Northern Edge of the bank in United States waters near the U.S./Canada boundary ( Fig. 1). This is the first documented offshore occurrence of a species that has colonized eastern U.S. coastal waters from New York to Maine during the past 15–20 years ( U.S. Geological Survey, 2006). Video imagery shows colonies coalescing to form large mats that cover more than 50% of the seabed along some video/photo transects. The affected area is an immobile pebble and cobble pavement that lies at water depths of 40 to 65 m where strong semidiurnal tidal currents reach speeds of 1 to 2 kt (50–100 cm/s). The water column is mixed year round, ensuring a constant supply of nutrients to the seabed. Annual temperatures range from 4 to 15 °C ( Mountain and Holzwarth, 1989). The gravel areas are bounded by sand ridges whose mobile surfaces are moved daily by the strong tidal currents. Studies commenced here in 1994 to characterize the gravel habitat and to document the effects of fishing disturbance on it ( Collie et al., 2005).

  8. The colonial ascidian Didemnum sp. A: current distribution, basic biology and potential threat to marine communities of the northeast and west coasts of North America

    USGS Publications Warehouse

    Bullard, S.G.; Lambert, G.; Carman, M.R.; Byrnes, J.; Whitlatch, R.B.; Ruiz, G.; Miller, R.J.; Harris, L.; Valentine, P.C.; Collie, J.S.; Pederson, J.; McNaught, D.C.; Cohen, A.N.; Asch, R.G.; Dijkstra, J.; Heinonen, K.

    2007-01-01

    Didemnum sp. A is a colonial ascidian with rapidly expanding populations on the east and west coasts of North America. The origin of Didemum sp. A is unknown. Populations were first observed on the northeast coast of the U.S. in the late 1980s and on the west coast during the 1990s. It is currently undergoing a massive population explosion and is now a dominant member of many subtidal communities on both coasts. To determine Didemnum sp. A's current distribution, we conducted surveys from Maine to Virginia on the east coast and from British Columbia to southern California on the west coast of the U.S. between 1998 and 2005. In nearshore locations Didemnum sp. A currently ranges from Eastport, Maine to Shinnecock Bay, New York on the east coast. On the west coast it has been recorded from Humboldt Bay to Port San Luis in California, several sites in Puget Sound, Washington, including a heavily fouled mussel culture facility, and several sites in southwestern British Columbia on and adjacent to oyster and mussel farms. The species also occurs at deeper subtidal sites (up to 81 m) off New England, including Georges, Stellwagen and Tillies Banks. On Georges Bank numerous sites within a 230 km2 area are 50–90% covered by Didemnum sp. A; large colonies cement the pebble gravel into nearly solid mats that may smother infaunal organisms. These observations suggest that Didemnum sp. A has the potential to alter marine communities and affect economically important activities such as fishing and aquaculture.

  9. Familial Mediterranean Fever and Hypercoagulability

    PubMed Central

    Tayer-Shifman, Oshrat E.; Ben-Chetrit, Eldad

    2011-01-01

    Familial Mediterranean fever (FMF) is an autosomal recessive hereditary disease which is characterized by recurrent attacks of fever and peritonitis, pleuritis, arthritis, or erysipelas-like skin disease. As such, FMF is a prototype of autoinflammatory diseases where genetic changes lead to acute inflammatory episodes. Systemic inflammation – in general - may increase procoagulant factors, and decrease natural anticoagulants and fibrinolytic activity. Therefore, it is anticipated to see more thrombotic events among FMF patients compared with healthy subjects. However, reviewing the current available literature and based upon our personal experience, thrombotic events related purely to FMF are very rare. Possible explanation for this discrepancy is that along with the procoagulant activity during FMF acute attacks, anticoagulant and fibrinolytic changes are also taking place. Colchicine which is the treatment of choice in FMF may also play a role in reducing inflammation thereby decreasing hypercoagulability. PMID:21713077

  10. Metagenomes of Mediterranean Coastal Lagoons

    PubMed Central

    Ghai, Rohit; Hernandez, Claudia Mella; Picazo, Antonio; Mizuno, Carolina Megumi; Ininbergs, Karolina; Díez, Beatriz; Valas, Ruben; DuPont, Christopher L.; McMahon, Katherine D.; Camacho, Antonio; Rodriguez-Valera, Francisco

    2012-01-01

    Coastal lagoons, both hypersaline and freshwater, are common, but still understudied ecosystems. We describe, for the first time, using high throughput sequencing, the extant microbiota of two large and representative Mediterranean coastal lagoons, the hypersaline Mar Menor, and the freshwater Albufera de Valencia, both located on the south eastern coast of Spain. We show there are considerable differences in the microbiota of both lagoons, in comparison to other marine and freshwater habitats. Importantly, a novel uncultured sulfur oxidizing Alphaproteobacteria was found to dominate bacterioplankton in the hypersaline Mar Menor. Also, in the latter prokaryotic cyanobacteria were almost exclusively comprised by Synechococcus and no Prochlorococcus was found. Remarkably, the microbial community in the freshwaters of the hypertrophic Albufera was completely in contrast to known freshwater systems, in that there was a near absence of well known and cosmopolitan groups of ultramicrobacteria namely Low GC Actinobacteria and the LD12 lineage of Alphaproteobacteria. PMID:22778901

  11. Metagenomes of Mediterranean coastal lagoons.

    PubMed

    Ghai, Rohit; Hernandez, Claudia Mella; Picazo, Antonio; Mizuno, Carolina Megumi; Ininbergs, Karolina; Díez, Beatriz; Valas, Ruben; DuPont, Christopher L; McMahon, Katherine D; Camacho, Antonio; Rodriguez-Valera, Francisco

    2012-01-01

    Coastal lagoons, both hypersaline and freshwater, are common, but still understudied ecosystems. We describe, for the first time, using high throughput sequencing, the extant microbiota of two large and representative Mediterranean coastal lagoons, the hypersaline Mar Menor, and the freshwater Albufera de Valencia, both located on the south eastern coast of Spain. We show there are considerable differences in the microbiota of both lagoons, in comparison to other marine and freshwater habitats. Importantly, a novel uncultured sulfur oxidizing Alphaproteobacteria was found to dominate bacterioplankton in the hypersaline Mar Menor. Also, in the latter prokaryotic cyanobacteria were almost exclusively comprised by Synechococcus and no Prochlorococcus was found. Remarkably, the microbial community in the freshwaters of the hypertrophic Albufera was completely in contrast to known freshwater systems, in that there was a near absence of well known and cosmopolitan groups of ultramicrobacteria namely Low GC Actinobacteria and the LD12 lineage of Alphaproteobacteria.

  12. Familial Mediterranean fever: current perspectives

    PubMed Central

    Sönmez, Hafize Emine; Batu, Ezgi Deniz; Özen, Seza

    2016-01-01

    Familial Mediterranean fever (FMF) is the most frequent monogenic autoinflammatory disease, and it is characterized by recurrent attacks of fever and polyserositis. The disease is associated with mutations in the MEFV gene encoding pyrin, which causes exaggerated inflammatory response through uncontrolled production of interleukin 1. The major long-term complication of FMF is amyloidosis. Colchicine remains the principle therapy, and the aim of treatment is to prevent acute attacks and the consequences of chronic inflammation. With the evolution in the concepts about the etiopathogenesis and genetics of the disease, we have understood that FMF is more complicated than an ordinary autosomal recessive monogenic disorder. Recently, recommendation sets have been generated for interpretation of genetic testing and genetic diagnosis of FMF. Here, we have reviewed the current perspectives in FMF in light of recent recommendations. PMID:27051312

  13. Mediterranean Ocean Colour Chlorophyll Trends

    PubMed Central

    Colella, Simone; Falcini, Federico; Rinaldi, Eleonora; Sammartino, Michela; Santoleri, Rosalia

    2016-01-01

    In being at the base of the marine food web, phytoplankton is particularly important for marine ecosystem functioning (e.g., biodiversity). Strong anthropization, over-exploitation of natural resources, and climate change affect the natural amount of phytoplankton and, therefore, represent a continuous threat to the biodiversity in marine waters. In particular, a concerning risks for coastal waters is the increase in nutrient inputs of terrestrial/anthropogenic origin that can lead to undesirable modifications of phytoplankton concentration (i.e., eutrophication). Monitoring chlorophyll (Chl) concentration, which is a proxy of phytoplankton biomass, is an efficient tool for recording and understanding the response of the marine ecosystem to human pressures and thus for detecting eutrophication. Here, we compute Chl trends over the Mediterranean Sea by using satellite data, also highlighting the fact that remote sensing may represent an efficient and reliable solution to synoptically control the “good environmental status” (i.e., the Marine Directive to achieve Good Environmental Status of EU marine waters by 2020) and to assess the application of international regulations and environmental directives. Our methodology includes the use of an ad hoc regional (i.e., Mediterranean) algorithm for Chl concentration retrieval, also accounting for the difference between offshore (i.e., Case I) and coastal (i.e., Case II) waters. We apply the Mann-Kendall test and the Sens’s method for trend estimation to the Chl concentration de-seasonalized monthly time series, as obtained from the X-11 technique. We also provide a preliminary analysis of some particular trends by evaluating their associated inter-annual variability. The high spatial resolution of our approach allows a clear identification of intense trends in those coastal waters that are affected by river outflows. We do not attempt to attribute the observed trends to specific anthropogenic events. However, the

  14. Night Views Over the Mediterranean Sea

    NASA Image and Video Library

    This video over the Mediterranean Sea was taken by the crew of Expedition 29 aboard the International Space Station. This sequence of shots was taken on Oct. 6, 2011, from 22:58:09 to 23:13:15 GMT,...

  15. [Mediterranean diet. Characteristics and health benefits].

    PubMed

    Serra Majem, Lluís; García Alvarez, Alicia; Ngo de la Cruz, Joy

    2004-06-01

    The purpose of this article is to define the concept of Mediterranean Diet or Diets and to describe the associated health benefits recognised by the scientific community. The characteristics of the Mediterranean Diet are described as well as the effects the foods comprising it have on the most common pathologies such as cardiovascular disease and cancer. The Spanish Society of Community Nutrition's consensus based Healthy Diet Pyramid, along with Greece's pyramid for food guidelines for the adult population (from the Greek Ministry of Health), are presented and compared. They are the graphic representation of the food and physical activity guides of two typically Mediterranean countries. Nutritional and sociological trends are also discussed and their impact on the evolution of the Mediterranean Diet.

  16. Genetics Home Reference: familial Mediterranean fever

    MedlinePlus

    ... Epub 2013 Sep 9. Review. Citation on PubMed Lidar M, Kedem R, Berkun Y, Langevitz P, Livneh ... 090401. Epub 2009 Dec 15. Citation on PubMed Lidar M, Livneh A. Familial Mediterranean fever: clinical, molecular ...

  17. New hexactinellid sponges from deep Mediterranean canyons.

    PubMed

    Boury-Esnault, Nicole; Vacelet, Jean; Dubois, Maude; Goujard, Adrien; Fourt, Maïa; Pérez, Thierry; Chevaldonné, Pierre

    2017-02-21

    During the exploration of the NW Mediterranean deep-sea canyons (MedSeaCan and CorSeaCan cruises), several hexactinellid sponges were observed and collected by ROV and manned submersible. Two of them appeared to be new species of Farrea and Tretodictyum. The genus Farrea had so far been reported with doubt from the Mediterranean and was listed as "taxa inquirenda" for two undescribed species. We here provide a proper description for the specimens encountered and sampled. The genus Tretodictyum had been recorded several times in the Mediterranean and in the near Atlantic as T. tubulosum Schulze, 1866, again with doubt, since the type locality is the Japan Sea. We here confirm that the Mediterranean specimens are a distinct new species which we describe. We also provide18S rDNA sequences of the two new species and include them in a phylogenetic tree of related hexactinellids.

  18. Complete nucleotide sequence of the mitochondrial genome of Doliolum nationalis with implications for evolution of urochordates.

    PubMed

    Yokobori, Shin-ichi; Oshima, Tairo; Wada, Hiroshi

    2005-02-01

    The evolutionary history of the diverse lifestyles adopted by urochordates has attracted intense interest because it may effect the evolutionary history of vertebrates. Here, we report the complete mitochondrial (mt) DNA sequence of the pelagic thaliacean doliolid Doliolum nationalis. The doliolid mt genome shares the unusual tRNAs of trnM(uau) and trnG(ucu) with other ascidians, such as Halocynthia and Ciona. On the other hand, the gene order of the doliolid mt genome is significantly different from that of any ascidian species or vertebrate reported to date. Phylogenetic analyses of the amino acid sequences of 12 protein-coding genes strongly support the sister-grouping of doliolids and the Phlebobranch ascidian Ciona, with the Stolidobranch ascidian alocynthia as the outgroup, thereby providing strong support for the paraphyly of ascidians, as has been suggested by 18S rDNA studies. Given the paraphyletic nature of ascidians, it seems likely that the common ancestor of ascidians and thaliaceans was sessile, as are the present-day ascidians, and that the thaliaceans subsequently evolved a pelagic lifestyle.

  19. The Strategic Importance of the Mediterranean

    DTIC Science & Technology

    1990-03-26

    Mediterranean and the straits of Bosphorus. From ancient times, man has struggled to control thebe vital maritime waterways. This geographic analysis shows that...axis to Greece and Italy. From the American’s viewpoint, the sea joins the United * States Central Command (USCENTCOM) and the United States European...separated southern flank nations, Turkey and Greece . An American control over the Mediterranean accomplishes the following: o Maintains maritime control in an

  20. Towards the advancement of the Mediterranean food cultures.

    PubMed

    Dernini, Sandro

    2006-02-01

    The Forum on Mediterranean Food Cultures has the purpose to foster an interdisciplinary dialogue to acknowledge the underestimated role of the Mediterranean diet and of the Mediterranean food cultures for an effective sustainable development in the Mediterranean. It is addressed towards the achievement of food security and a broader nutritional well-being in the entire Mediterranean area. The Forum uses a creative approach for the development of community-based programmes to manage the emerging trend of childhood overweight and obesity, as well as to reduce the increasing erosion of the Mediterranean food cultural heritage.

  1. Mediterranean diet adherence in the Mediterranean healthy eating, aging and lifestyle (MEAL) study cohort.

    PubMed

    Marventano, Stefano; Godos, Justyna; Platania, Alessio; Galvano, Fabio; Mistretta, Antonio; Grosso, Giuseppe

    2017-05-31

    A decline in adherence to the Mediterranean dietary pattern has been observed over the last years. The aim of this study was to assess the level of adherence to the Mediterranean diet and possible determinants in the Mediterranean healthy Eating, Aging and Lifestyle (MEAL) study cohort. Demographic and dietary data of 1937 individuals were collected in 2014-2015 from the general population of Catania, Sicily (Italy). Food frequency questionnaires and the MEDI-LITE score were used to assess adherence to the Mediterranean diet. The score well characterised consumption of major food groups, micro- and macro-nutrients. The cohort had a general good adherence, but only a minority was highly adherent. High adherence was directly associated with education, non-smoking and physical activity and inversely with high occupational status. In conclusions, Mediterranean diet is still followed in Sicily; however, nutrition education campaigns should promote healthy traditional dietary patterns in certain groups of individuals.

  2. The Eratosthenes Seamount - Eastern Mediterranean

    NASA Astrophysics Data System (ADS)

    Ehrhardt, A.; Schnabel, M.; Damm, V.

    2012-04-01

    The Eratosthenes Seamount forms a prominent landmark in the Eastern Mediterranean. It is located south of Cyprus with the Levantine Basin on its eastern side, the Herodotus Basin on its western side and the Nile Cone south of the seamount. The Eratosthenes Seamount rises up to 750 m below sea surface and is about 1200 m higher than the surrounding seafloor of the Levantine Basin and the Nile Cone sediments. The Eratosthenes Seamount is considered as a continental fragment of the former African-Nubian Plate that was rifted to its present position relative to Africa during the formation of the Tethyan Ocean. In 2010 a detailed geophysical survey was carried out in the area of the Eratosthenes Seamount by the Federal Institute for Geosciences and Natural Resources of Germany including multichannel seismic (MCS), refraction seismics, magnetic, gravity and magnetotelluric data acquisition. First results show a highly deformed seamount, with a plateau-like top that is impacted by west-east trending graben formation. The slopes of the seamount are eroded showing deep incised ripple patterns and recent submarine landslides. The Eratosthenes Seamount produces also a prominent magnetic and gravity anomaly, both supporting its uniqueness in the area of the Eastern Mediterranean. Velocity information by refraction seismic modeling, as well as the models of the magnetic and gravity data show evidence for a volcanic core of the seamount with carbonate layers on top of the volcanic core. The slopes of the seamount terminate against a conspicuous rim-like escarpment that forms in addition the northern and western termination of the Messinian Evaporites in the study area. The MCS and refraction seismic data show a very deep Levantine Basin with maximum acoustic basement depths of 12 to 14 km very close to the slope of the Eratosthenes Seamount. The deepest sediments resolved by the MCS data are of Lower Cretaceous to Jurassic age. The refraction seismic model shows a 14 km thick

  3. Atlantic forcing of the Mediterranean oligotrophy

    NASA Astrophysics Data System (ADS)

    Huertas, I. E.; RíOs, A. F.; GarcíA-Lafuente, J.; Navarro, G.; Makaoui, A.; SáNchez-RomáN, A.; Rodriguez-Galvez, S.; Orbi, A.; RuíZ, J.; PéRez, F. F.

    2012-06-01

    The Mediterranean Sea shows a peculiar anomaly in its nutrient pattern compared to the global ocean, as there is decrease in nutrient concentration from west to east. This feature has been attributed to the antiestuarine circulation at the Strait of Gibraltar, where an eastward flow of Atlantic nutrient-poor surface waters is compensated by a westward countercurrent of Mediterranean nutrient-rich deep waters. This water exchange has been suggested as the ultimate cause for the oligotrophy of the Mediterranean basin, even though only a few studies have accurately examined the magnitude of the nutrient flux through the Strait of Gibraltar. In this work, data from the Gibraltar Fixed Time series (GIFT) between 2005 and 2008 were used to assess nutrient distributions. Applying a two-layer model of water mass exchange and using the Mediterranean outflow recorded in situ, the net export of nutrients from the Mediterranean to the Atlantic was calculated as 139 and 4.8 Gmol yr-1 of nitrate and phosphate, respectively. The results also demonstrated that the Atlantic inflow is not nutrient depleted and in particular contains significant levels of phosphate, which is the limiting factor for biological productivity in the eastern Mediterranean. The distribution of the quasi-conservative parameter N* in the western and eastern basins indicated that nitrate-deficient surface waters are transformed into phosphate-deficient bottom waters by internal cycling processes. Therefore, phosphate depletion in the Mediterranean does not have its origin in the entry of a phosphorus-impoverished Atlantic inflow through the Strait of Gibraltar.

  4. Summer fire predictability in a Mediterranean environment

    NASA Astrophysics Data System (ADS)

    Marcos, Raül; Turco, Marco; Bedía, Joaquín; Llasat, Maria Carmen; Provenzale, Antonello

    2015-04-01

    Each year approximately 500000 hectares burn in Europe. Most of them are consequence of Mediterranean summer fires that lead to damages to the natural environment causing important economic and life losses. In order to allow the preparedness of adequate prevention measures in European Mediterranean regions, a better understanding of the summer fire predictability is crucial. Climate is a primary driver of the interannual variability of fires in Mediterranean-type ecosystems, controlling fuel flammability and fuel structure [1, 2]. That is, summer fires are linked to current-year climate values (proxies for the climatic factors that affect fuel flammability) and to antecedent climate variables (proxies for the climatic factors influencing fine fuel availability and connectivity). In our contribution we explore the long-term predictability of wildfires in a Mediterranean region (NE Spain), driving a multiple linear regression model with observed antecedent climate variables and with predicted variables from the ECMWF System-4 seasonal forecast. The approaches are evaluated through a leave-one-out cross-validation over the period 1983-2010. While the ECMWF System-4 proved of limited usefulness due to its limited skill, the model driven with antecedent climate variables alone allowed for satisfactory long-term prediction of above-normal fire activity, suggesting the feasibility of successful seasonal prediction of summer fires in Mediterranean-type regions. *References [1] M. Turco, M. C. Llasat, J. von Hardenberg, and A. Provenzale. Impact of climate variability on summer fires in a mediterranean environment (northeastern iberian peninsula). Climatic Change, 116:665-678, 2013. [2] M. Turco, M. C. Llasat, J. von Hardenberg, and A. Provenzale. Climate change impacts on wildfires in a Mediterranean environment. Climatic Change, 125: 369-380, 2014.

  5. Mercury speciation in the Mediterranean Sea

    NASA Astrophysics Data System (ADS)

    Horvat, M.; Kotnik, J.; Ogrinc, N.; Fajon, V.; Logar, M.; Gibicar, D.; Vaupotic, J.; Pirrone, N.

    2004-12-01

    An interesting feature of mercury biogeochemistry in the Mediterranean is that several fish species from the Mediterranean show higher concentrations of Hg in their tissues than same fish species in the Atlantic ocean. although the concentrations of mercury in the open waters of both oceans are similar. It has been suggested that the higher mercury levels noted in many larger pelagic fish species in the Mediterranean are not related to anthropogenic inputs, but rather are due to the higher than average natural environmental levels of this metal originating from the Mediterranean mercury anomaly. Although elevated Hg levels have been noted in environmental matrices from the Mediterranean regions adjacent to known mercury anomalies, the data do not clearly indicate that the effects of these anomalies have been transmitted to open waters or to lower trophic level species living in these waters. In the present contribution data obtained during three oceanographic cruises carried out in the framework of the MERCYMS project (An integrate approach to assess mercury cycling in the Mediterranean basin) funded by EU in the period between 2000 and 2003 will be presented. Measurements included total mercury measurements and its speciation (reactive Hg, total Hg and monomethylmercury (MMHg) in filtered and non-filtered sea water samples, dissolved gaseous mercury (DGM) and dimethylmercury (DMHg) in open and coastal waters of the Mediterranean Sea. Radon, as a tracer gas of tectonic activity was also measured in depth profiles. The results presented clearly show that hg species distribution in surface and deep oceanic waters is affected by several dynamic processes such as photochemical transformation at the surface, phytoplankton biomass stratification in the photic zone, development of an oxygen depletion zone at intermediate depths and diffusion from deeper layers due to biological and/or tectonic activities.

  6. [Sacroiliitis in familial Mediterranean fever].

    PubMed

    Connemann, B J; Steinhoff, J; Benstein, R; Sack, K

    1991-11-22

    A 15-year-old girl of Turkish descent had for one year complained of severe recurrent fever-associated deep back pains. Since she was three years of age she had suffered from repeated attacks of fever and severe abdominal pain which ceased spontaneously in 1-3 days. On physical examination the sacrum and iliosacral joints were very painful to percussion, and she limped. Radiography revealed symmetric destructive sacroiliitis. Despite the unusual location of the arthritis, the triad of fever, abdominal pain and arthritis, as well as her belonging to an ethnic "at risk" group, pointed to the diagnosis of familial mediterranean fever (FML) or recurrent hereditary polyserositis. This diagnosis was confirmed by a positive metaraminol provocation test in that infusion of metaraminol reproduced the typical pains. Collagen diseases, rheumatic disease, acute porphyria and chronic infectious processes were excluded. The sacroiliitis quickly responded to long-term administration of colchicine, 0.5 mg twice daily. The patient also has Hageman factor deficiency whose significance remains unclear.

  7. Seasonality of Arctic Mediterranean Exchanges

    NASA Astrophysics Data System (ADS)

    Rieper, Christoph; Quadfasel, Detlef

    2015-04-01

    The Arctic Mediterranean communicates through a number of passages with the Atlantic and the Pacific Oceans. Most of the volume exchange happens at the Greenland-Scotland-Ridge: warm and saline Atlantic Water flows in at the surface, cold, dense Overflow Water flows back at the bottom and fresh and cold Polar Water flows out along the East Greenland coast. All surface inflows show a seasonal signal whereas only the outflow through the Faroe Bank Channel exhibits significant seasonality. Here we present a quantification of the seasonal cycle of the exchanges across the Greenland-Scotland ridge based on volume estimates of the in- and outflows within the last 20 years (ADCP and altimetry). Our approach is comparatistic: we compare different properties of the seasonal cycle like the strength or the phase between the different in- and outflows. On the seasonal time scale the in- and outflows across the Greenland-Scotland-Ridge are not balanced. The net flux thus has to be balanced by the other passages on the Canadian Archipelago, Bering Strait as well as runoff from land.

  8. Familial Mediterranean fever in siblings.

    PubMed

    Özçakar, Z Birsin; Erdogan, Beyza Doganay; Elhan, Atilla H; Yalçinkaya, Fatoş

    2012-11-01

    Genetic and environmental factors have been implicated in disease severity and development of amyloidosis in familial Mediterranean fever (FMF). We investigated similarities in clinical characteristics, disease severity, and treatment response within siblings with FMF. The study group consisted of 2 or more siblings who were followed in our center with the diagnosis of FMF. Siblings were evaluated for demographic data, clinical and laboratory disease features, genetic analysis of MEFV mutations, and disease severity score. The intraclass correlation coefficient (ICC), which can be interpreted as the expected correlation between 2 siblings, was used to reflect within-family similarity. The study included 67 pediatric patients from 31 different families. When we investigated the similarity of siblings after adjusting for genetic effects, we found very low ICC with p > 0.05 in the majority of clinical features, disease severity, and colchicine dosages. However, age at disease onset, age at onset of therapy, attack-free acute-phase reactant levels, and presence of amyloidosis were found to be similar within siblings (relatively high ICC with p < 0.05). Siblings with FMF had different clinical findings and disease severity. They had similar amyloidogenic potential, proven by both similar presence of amyloid and increased levels of acute-phase reactants between attacks. Our findings strongly support that genetic factors may be more dominant in the development of amyloidosis.

  9. Myxozoan infections in Mediterranean mariculture.

    PubMed

    Palenzuela, O

    2006-06-01

    Fish mariculture has dramatically expanded in recent years in Mediterranean countries. In this scenario, several pathological problems have logically arisen and parasitological etiologies are increasingly being reported, either as primary or secondary pathogens. Myxozoa is the most diverse and economically important group of fish parasites, and several species are known to cause or contribute to losses in mariculture. Species of the genus Enteromyxum currently constitute the most serious parasitological threat. Some unusual biological characters, such as wide host spectrum and direct fish-to-fish transmission, together with high virulence for some host species, combine a dangerous cocktail which is emerging in recent years. Closed-system (recirculation) and heated-water locations are especially sensitive to chronic infections by these parasites, which can cause serious mortality and even discourage culture of some fish species at certain locations (i.e, Diplodus puntazzo). The presentation presents an overview of recent advances in research of marine myxozoans, focusing mainly in the most pathogenic, Enteromyxum spp. The incidence of these and other emerging infections, and the design of potential strategies for control will be introduced.

  10. Evidence by a voltage clamp study of an electrically mediated block to polyspermy in the egg of the ascidian Phallusia mammillata.

    PubMed

    Goudeau, H; Depresle, Y; Rosa, A; Goudeau, M

    1994-12-01

    Eggs of the ascidian Phallusia mammillata were voltage clamped (from -100 to +60 mV) and inseminated with a low or heavy sperm concentration. From inseminations with low sperm concentration (1 x 10(6) sp/ml), we found that fertilization currents occurred between -100 and +40 mV: they were always inward and displayed an analogous pattern whatever the clamped voltage. We established that the percentages of inseminated eggs that produced a fertilization current varied as a function of the clamped voltage. These percentages were not statistically different from 100% at clamped voltages between -100 and -30 mV, they decreased to 68 and 56% at clamped Vm of -10 and 0 mV, respectively, but were not statistically different from 0% at clamped Vm between +10 and +40 mV. We never obtained any egg electrical response at a clamped voltage of +50 mV. Almost all eggs (96%) which responded electrically were penetrated by one or several spermatozoa. These eggs were resuming meiosis (81 to 50%) at values of clamped Vm between -100 and 0 mV, respectively. At clamped Vm between +10 and +50 mV, the percenta