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Characterization of a defensin from the oyster Crassostrea gigas - Recombinant production, folding, solution structure, antimicrobial activities, and gene expression ArchiMer
Gueguen, Yannick; Herpin, Amaury; Aumelas, André; Garnier, Julien; Fievet, Julie; Escoubas, Jean-michel; Bulet, Philippe; Gonzalez, Marcelo; Lelong, Christophe; Favrel, Pascal; Bachere, Evelyne.
In invertebrates, defensins were found in arthropods and in the mussels. Here, we report for the first time the identification and characterization of a defensin (Cg-Def) from an oyster. Cg-def mRNA was isolated from Crassostrea gigas mantle using an expressed sequence tag approach. To gain insight into potential roles of Cg-Def in oyster immunity, we produced the recombinant peptide in Escherichia coli, characterized its antimicrobial activities, determined its solution structure by NMR spectroscopy, and quantified its gene expression in vivo following bacterial challenge of oysters. Recombinant Cg-Def was active in vitro against Gram-positive bacteria but showed no or limited activities against Gram-negative bacteria and fungi. The activity of Cg-Def was...
Tipo: Text Palavras-chave: Crassostrea gigas; Genomic library; Microbiology; Defensin; Oyster.
Ano: 2006 URL: http://archimer.ifremer.fr/doc/2006/publication-2127.pdf
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Characterization of a gonad-specific transforming growth factor-β superfamily member differentially expressed during the reproductive cycle of the oyster Crassostrea gigas ArchiMer
Fleury, Elodie; Fabioux, Caroline; Lelong, Christophe; Favrel, Pascal; Huvet, Arnaud.
To study the complex and non identified host causes of summer mortality in C. gigas, divergent selection criteria based upon summer survival have been applied to produce resistant (R) and susceptible (S) oysterfamilies(Samain et al., 2007). A previous analysis by suppression-subtractive hybridization has characterized 46 differentially-expressedcDNAsbetween R and S families(Huvet et al., 2004). The present study reports the screening, using nylon array, of the 46 oyster SSHcDNAsfrom R and S selected families. Among the few differentially expressed genes characterized between R and S samples, one encodes a Transforming-Growth-Factor-β-related (TGFβ) protein. Members of the TGFβsuperfamily are critical growth factors regulating a variety ofimportant...
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Ano: 2007 URL: http://archimer.ifremer.fr/doc/00083/19425/17036.pdf
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Characterization of GnRH-related peptides from the Pacific oyster Crassostrea gigas ArchiMer
Bigot, Laetitia; Zatylny-gaudin, Celine; Rodet, Franck; Bernay, Benoit; Boudry, Pierre; Favrel, Pascal.
Gonadotropin-releasing hormone (GnRH), a key neuropeptide regulating reproduction in vertebrates has now been characterized in a number of non-vertebrate species. Despite the demonstration of its ancestral origin, the structure and the function of this family of peptides remain poorly known in species as distant as lophotrochozoans. In this study, two GnRH-related peptides (Cg-GnRH-a and CgGnRH-G) were characterized by mass spectrometry from extracts of the visceral ganglia of the Pacific oyster Crassostrea gigas. These peptides showed a high degree of sequence identity with GnRHs of other mollusks and annelids and to a lesser extent with those of vertebrates or with AKH and corazonins of insects. Both the mature peptides and the transcript encoding the...
Tipo: Text Palavras-chave: Mollusk; Bivalve; Oyster; Neuroendocrinology; Reproduction; GnRH.
Ano: 2012 URL: http://archimer.ifremer.fr/doc/00079/19003/17477.pdf
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Development of a Pacific oyster (Crassostrea gigas) 31,918-feature microarray: identification of reference genes and tissue-enriched expression patterns ArchiMer
Dheilly, Nolwenn; Lelong, Christophe; Huvet, Arnaud; Favrel, Pascal.
Background: Research using the Pacific oyster Crassostrea gigas as a model organism has experienced rapid growth in recent years due to the development of high-throughput molecular technologies. As many as 56,268 EST sequences have been sequenced to date, representing a genome-wide resource that can be used for transcriptomic investigations. Results: In this paper, we developed a Pacific oyster microarray containing oligonucleotides representing 31,918 transcribed sequences selected from the publicly accessible GigasDatabase. This newly designed microarray was used to study the transcriptome of male and female gonads, mantle, gills, posterior adductor muscle, visceral ganglia, hemocytes, labial palps and digestive gland. Statistical analyses identified...
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Ano: 2011 URL: http://archimer.ifremer.fr/doc/00048/15956/13397.pdf
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Functional Characterization of a short neuropeptide F-related receptor in a Lophotrochozoa, the mollusk Crassostrea gigas ArchiMer
Bigot, Laetitia; Beets, Isabel; Dubos, Marie-pierre; Boudry, Pierre; Schoofs, Liliane; Favrel, Pascal.
Members of the short neuropeptide F (sNPF) family of peptides and their cognate receptors play key roles in a variety of physiological processes in arthropods. In silico screening of GigasDatabase, a specific expressed sequence tag database from the Pacific oyster Crassostrea gigas, resulted in the identification of a receptor (CgsNPFR-like) phylogenetically closely related to sNPF receptors (sNPFRs) of insects. A reverse endocrinology approach was undertaken to identify the peptide ligand(s) of this orphan receptor. Though structurally distinct from insect sNPFs, three RFamide peptides derived from the same precursor, i.e. GSLFRFamide, SSLFRFamide and GALFRFamide, specifically activate the receptor in a dose-dependent manner, with respective EC50 values...
Tipo: Text Palavras-chave: Oyster; Crassostrea gigas; SNPF; Energy metabolism; Reproduction; Starvation.
Ano: 2014 URL: http://archimer.ifremer.fr/doc/00207/31777/30188.pdf
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Gametogenesis in the Pacific Oyster Crassostrea gigas: A Microarrays-Based Analysis Identifies Sex and Stage Specific Genes ArchiMer
Dheilly, Nolwenn; Lelong, Christophe; Huvet, Arnaud; Kellner, Kristell; Dubos, Marie-pierre; Riviere, Guillaume; Boudry, Pierre; Favrel, Pascal.
Background: The Pacific oyster Crassostrea gigas (Mollusca, Lophotrochozoa) is an alternative and irregular protandrous hermaphrodite: most individuals mature first as males and then change sex several times. Little is known about genetic and phenotypic basis of sex differentiation in oysters, and little more about the molecular pathways regulating reproduction. We have recently developed and validated a microarray containing 31,918 oligomers (Dheilly et al., 2011) representing the oyster transcriptome. The application of this microarray to the study of mollusk gametogenesis should provide a better understanding of the key factors involved in sex differentiation and the regulation of oyster reproduction. Methodology/Principal Findings: Gene expression was...
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Ano: 2012 URL: http://archimer.ifremer.fr/doc/00087/19862/17505.pdf
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Generation and analysis of a 29,745 unique Expressed Sequence Tags from the Pacific oyster (Crassostrea gigas) assembled into a publicly accessible database: the GigasDatabase ArchiMer
Fleury, Elodie; Huvet, Arnaud; Lelong, Christophe; De Lorgeril, Julien; Boulo, Viviane; Gueguen, Yannick; Bachere, Evelyne; Tanguy, Arnaud; Moraga, Dario; Fabioux, Caroline; Lindeque, Penelope; Shaw, Jenny; Reinhardt, Richard; Prunet, Patrick; Davey, Grace; Lapegue, Sylvie; Sauvage, Christopher; Corporeau, Charlotte; Moal, Jeanne; Gavory, Frederick; Wincker, Patrick; Moreews, Francois; Klopp, Christophe; Mathieu, Michel; Boudry, Pierre; Favrel, Pascal.
Background: Although bivalves are among the most-studied marine organisms because of their ecological role and economic importance, very little information is available on the genome sequences of oyster species. This report documents three large-scale cDNA sequencing projects for the Pacific oyster Crassostrea gigas initiated to provide a large number of expressed sequence tags that were subsequently compiled in a publicly accessible database. This resource allowed for the identification of a large number of transcripts and provides valuable information for ongoing investigations of tissue-specific and stimulus-dependant gene expression patterns. These data are crucial for constructing comprehensive DNA microarrays, identifying single nucleotide...
Tipo: Text
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-6770.pdf
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GigaTON: an extensive publicly searchable database providing a new reference transcriptome in the pacific oyster Crassostrea gigas ArchiMer
Riviere, Guillaume; Klopp, Christophe; Ibouniyamine, Nabihoudine; Huvet, Arnaud; Boudry, Pierre; Favrel, Pascal.
Background: The Pacific oyster, Crassostrea gigas, is one of the most important aquaculture shellfish resources worldwide. Important efforts have been undertaken towards a better knowledge of its genome and transcriptome, which makes now C. gigas becoming a model organism among lophotrochozoans, the under-described sister clade of ecdysozoans within protostomes. These massive sequencing efforts offer the opportunity to assemble gene expression data and make such resource accessible and exploitable for the scientific community. Therefore, we undertook this assembly into an up-to-date publicly available transcriptome database: the GigaTON (Gigas TranscriptOme pipeliNe) database. Description: We assembled 2204 million sequences obtained from 114 publicly...
Tipo: Text Palavras-chave: Oyster; Transcriptome; Development; Stress; Tissues; NGS.
Ano: 2015 URL: https://archimer.ifremer.fr/doc/00302/41321/40520.pdf
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In vivo RNA interference in oyster -vasa silencing inhibits germ cell development ArchiMer
Fabioux, Caroline; Corporeau, Charlotte; Quillien, Virgile; Favrel, Pascal; Huvet, Arnaud.
This study investigated the potential of RNA interference, which is technically challenging in bivalve mollusc species, to assess gene function in the oyster Crassostrea gigas. We designed dsRNA targeting the oyster vasa-like gene (Oyvlg), specifically expressed in oyster germ cells. In vivo injection of oyvl-dsRNA into the gonad provokes a knockdown phenotype corresponding to germ cell underproliferation and prematurely arrested meiosis througout the organ. The most severe phenotype observed is sterile. This knockdown phenotype is associated with a decrease in Oyvlg mRNA level of between 39% and 87%, and a strong reduction in OYVLG protein, to an undetectable level. Therefore, Oyvlg appears to be essential for germ cell development in Crassostrea gigas,...
Tipo: Text Palavras-chave: Vasa; RNAi; Marine bivalve; Germline; Crassostrea gigas.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-6641.pdf
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Microarray-Based Identification of Gonad Transcripts Differentially Expressed Between Lines of Pacific Oyster Selected to Be Resistant or Susceptible to Summer Mortality ArchiMer
Fleury, Elodie; Moal, Jeanne; Boulo, Viviane; Daniel, Jean-yves; Mazurais, David; Henaut, Alain; Corporeau, Charlotte; Boudry, Pierre; Favrel, Pascal; Huvet, Arnaud.
Summer mortality of the Pacific oyster Crassostrea gigas is the result of a complex interaction between oysters, their environment, and pathogens. Heredity appears to be a major factor determining the sensitivity of oysters to summer mortality, allowing resistant (R) and susceptible (S) lines to be produced. We conducted genome-wide expression profiling of R and S gonads during the 3-month period preceding a summer mortality event, using a cDNA microarray that we designed. ANOVA analysis revealed that 34 genes were differentially expressed between R and S lines on four dates preceding the mortality event. Annotation of some of these genes highlights reproduction and its allocation and antioxidant defenses as the main pathways that operate differentially...
Tipo: Text Palavras-chave: Mollusca; Bivalvia; Gene expression; CDNA microarrays; Reproduction; Oxidative stress; Energy metabolism.
Ano: 2010 URL: http://archimer.ifremer.fr/doc/00005/11590/8258.pdf
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NPY signalling in the gonad of the oyster Crassostrea gigas : involvement in summer mortality? ArchiMer
Corporeau, Charlotte; Fleury, Elodie; Moal, Jeanne; Boudry, Pierre; Favrel, Pascal; Huvet, Arnaud.
In summer, oysters suffer mortality that decimates this year about 60% of the population in France. We previously demonstrated a genetic basis for summer survival thatallowed us to produce resistant (R) and susceptible (S) oyster familiesby divergent selection (Samainet al., 2007). At the physiological level, R families can survive summer mortality because they are reproductively less active than S families (Fleury et al., 2008).In order to identify the molecular basis of the summer survival,a transcriptomicapproach on R and S families was performed using microarray analysis and 34 differentially-expressed genes were identified between R and S in the gonad (Fleury et al., in prep). Among these genes, we found an EST encoding a neuropeptide Y related...
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Ano: 2008 URL: http://archimer.ifremer.fr/doc/00083/19424/17035.pdf
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Structural and functional characterizations of an Activin type II receptor orthologue from the pacific oyster Crassostrea gigas ArchiMer
Le Quere, Herve; Herpin, Amaury; Huvet, Arnaud; Lelong, Christophe; Favrel, Pascal.
Members of the Transforming Growth factor beta (TGF-beta) superfamily of cell signalling polypeptides are known to play important roles in cell proliferation and differentiation during development and in various physiological processes of most animal clades. Recent findings in the mollusc Crassostrea gigas demonstrate the occurrence of a diversity of TGF-beta signalling components including various ligands, three type I receptors but only a single type II receptor. This report describes the characterization of Cg-ActRII, a new type II receptor displaying homology with vertebrate and Drosophila Activin type II receptors. The use of zebrafish embryo as a reporter organism revealed that, in a way similar to its zebrafish counterpart, overexpression of...
Tipo: Text Palavras-chave: Activin receptor; Nervous tissues; Development; Lophotrochozoa; Mollusc.
Ano: 2009 URL: http://archimer.ifremer.fr/doc/2009/publication-6401.pdf
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Transcriptomic Profiling of Gametogenesis in Triploid Pacific Oysters Crassostrea gigas: Towards an Understanding of Partial Sterility Associated with Triploidy ArchiMer
Dheilly, Nolwenn M.; Jouaux, Aude; Boudry, Pierre; Favrel, Pascal; Lelong, Christophe.
Background: Triploidy can occur in many animal species but is often lethal. Among invertebrates, amphibians and fishes, triploids are viable although often sterile or infertile. Most triploids of the Pacific oyster Crassostrea gigas are almost sterile (named "3n beta'') yet a low but significant proportion show an advanced gametogenesis (named "3n alpha''). These oysters thus constitute an interesting model to study the effect of triploidy on germ cell development. We used microarrays to compare the gonad transcriptomes of diploid 2n and the abovementioned triploid 3n beta and 3n alpha male and female oysters throughout gametogenesis. Results: All triploids displayed an upregulation of genes related to DNA repair and apoptosis and a downregulation of genes...
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Ano: 2014 URL: http://archimer.ifremer.fr/doc/00239/34992/33538.pdf
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