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Pyrosequencing of Mytilus galloprovincialis cDNAs: tissue-specific expression patterns.

ABSTRACT: BACKGROUND: Mytilus species are important in marine ecology and in environmental quality assessment, yet their molecular biology is poorly understood. Molecular aspects of their reproduction, hybridisation between species, mitochondrial inheritance, skewed sex ratios of offspring and adaptation to climatic and pollution factors are priority areas. METHODOLOGY/PRINCIPAL FINDINGS: To start to address this situation, expressed genetic transcripts from M. galloprovincialis were pyrosequenced. Transcripts were isolated from the digestive gland, foot, gill and mantle of both male and female mussels. In total, 175,547 sequences were obtained and for foot and mantle, 90% of the sequences could be assembled into contiguous fragments but this reduced to 75% for the digestive gland and gill. Transcripts relating to protein metabolism and respiration dominated including ribosomal proteins, cytochrome oxidases and NADH dehydrogenase subunits. Tissue specific variation was identified in transcripts associated with mitochondrial energy metabolism, with the digestive gland and gill having the greatest transcript abundance. Using fragment recruitment it was also possible to identify sites of potential small RNAs involved in mitochondrial transcriptional regulation. Sex ratios based on Vitelline Envelop Receptor for Lysin and Vitelline Coat Lysin transcript abundances, indicated that an equal sex distribution was maintained. Taxonomic profiling of the M. galloprovincialis tissues highlighted an abundant microbial flora associated with the digestive gland. Profiling of the tissues for genes involved in intermediary metabolism demonstrated that the gill and digestive gland were more similar to each other than to the other two tissues, and specifically the foot transcriptome was most dissimilar. CONCLUSIONS: Pyrosequencing has provided extensive genomic information for M. galloprovincialis and generated novel observations on expression of different tissues, mitochondria and associated microorganisms. It will also facilitate the much needed production of an oligonucleotide microarray for the organism.

SUBMITTER: Craft JA

PROVIDER: S-EPMC2810337 | biostudies-literature | 2010

REPOSITORIES: biostudies-literature

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Pyrosequencing of Mytilus galloprovincialis cDNAs: tissue-specific expression patterns.

Craft John A JA Gilbert Jack A JA Temperton Ben B Dempsey Kate E KE Ashelford Kevin K Tiwari Bela B Hutchinson Tom H TH Chipman J Kevin JK

PloS one 20100125 1

<h4>Background</h4>Mytilus species are important in marine ecology and in environmental quality assessment, yet their molecular biology is poorly understood. Molecular aspects of their reproduction, hybridisation between species, mitochondrial inheritance, skewed sex ratios of offspring and adaptation to climatic and pollution factors are priority areas.<h4>Methodology/principal findings</h4>To start to address this situation, expressed genetic transcripts from M. galloprovincialis were pyrosequ ...[more]

PMID: 20111607

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Project description:Direct comparison of the transcriptional patterns between male and female in the digestive gland of a natural population of the marine mussel Mytilus galloprovincialis sampled in the Bizerta Lagoon, Tunisia, across November 2007 -February March 2008 (four stages, winter peak). Background: Seasonal environmental changes may affect the physiology of Mytilus galloprovincialis (Lam.), an intertidal filter-feeder bivalve occurring commonly in Mediterranean and Atlantic coastal areas. We investigated seasonal variations in relative transcript abundance of the digestive gland and the Digestive gland (gonads) of males and females. To identify gene expression trends, we used a medium-density cDNA microarray (1.7 K probes) in dual-color competitive hybridization analyses. Results: Hierarchical clustering of digestive gland microarray data showed two main branches, distinguishing profiles associated with the “hot” months (May–August) from the other months. Genes involved in chitin metabolism, associated with mussel nutrition and digestion, showed higher expression during summer. Moreover, we found different gene expression patterns in the digestive glands of males and females during the four stages of mussel gonadal development. Microarray data from gonadal transcripts also displayed clear patterns during the different developmental phases with peak relative mRNA abundance at the ripe phase (stage III) for both sexes. Conclusion: These data showed a clear temporal pattern in gene expression profiles of mussels sampled over an annual cycle. Physiological response to thermal variation, food availability, and reproductive status across months may contribute to variation in gene expression. Test/reference design (female/male). Direct comparison of RNA extracts obtained from the Digestive gland tissue of female and male animals. Two (male, female) x four conditions (gonad developmental stage 1, stage 2, stage 3, stage 4). Dual color competitive hybridizations with label swap. Single individuals. Four biological replicates. One replicate per array.

Dataset Information

Pyrosequencing of Mytilus galloprovincialis cDNAs: tissue-specific expression patterns.

Publications

Pyrosequencing of Mytilus galloprovincialis cDNAs: tissue-specific expression patterns.

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