Dataset Information

Comparative and quantitative proteomics reveals the adaptive strategies of oyster larvae to ocean acidification

ABSTRACT: Acidification of seawater due to anthropogenic CO2, called as ocean acidification (OA), makes most coastal environments unfavorable for oysters. This is a serious socio-economical issue for China which is accounting for >70% of the world’s edible oyster supply. Understanding of OA effects at proteome level could lead to a better aquaculture management. Here, we present an iTRAQ based protein profiling analysis for the detection and quantification of proteome change under OA in one of the early life stages of a commercially important oyster species, Crassostrea hongkongensis. The completion of the genome sequence for oysters enabled us to confidently quantify over 1500 proteins in the larval oyster. Over 10% of proteome was altered in response to OA process at pH 7.6. Analysis of differentially expressed proteins and their associated pathways indicated that up-regulation of calcification, metabolic processes, cytoskeletal functions, oxidative stress and cell signaling processes might have been used by larvae to acclimate under OA. Although the expressions of cytoskeletal and signal transduction proteins were down-regulated in response to OA, there was no marked alternation in developmental processes such as metamorphic success. This study suggests that the estuarine edible oyster possess an adequate short-term adaptability or acclimation mechanism at proteome level to tolerate near-future OA scenario.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Crassostrea Gigas (pacific Oyster) (crassostrea Angulata)

SUBMITTER: Dineshram Ramadoss

LAB HEAD: V Thiagarajan

PROVIDER: PXD002138 | Pride | 2015-10-29

REPOSITORIES: Pride

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Comparative and quantitative proteomics reveal the adaptive strategies of oyster larvae to ocean acidification.

Dineshram R R Quan Q Q Sharma Rakesh R Chandramouli Kondethimmanahalli K Yalamanchili Hari Krishna HK Chu Ivan I Thiyagarajan Vengatesen V

Proteomics 20151201 23-24

Decreasing pH due to anthropogenic CO2 inputs, called ocean acidification (OA), can make coastal environments unfavorable for oysters. This is a serious socioeconomical issue for China which supplies >70% of the world's edible oysters. Here, we present an iTRAQ-based protein profiling approach for the detection and quantification of proteome changes under OA in the early life stage of a commercially important oyster, Crassostrea hongkongensis. Availability of complete genome sequence for the pac ...[more]

PMID: 26507238

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Project description:Background Triploidy can occur in all species but is often lethal in birds and mammals. In amphibian, invertebrates and numerous species of fishes, triploid animals are viable and undistinguishable from diploid individuals. Gametogenesis is often affected and most animals are sterile for at least one sex, and gametes for the other sex are often unfertile. Although the majority of triploid oysters are sterile (beta individuals, 3nb), a low but persistent proportion of male and female animals produce gametes (alpha individuals, 3na). Thus, oysters constitute a unique model to study the effect of triploidy on germ cells development of both male and females. In this study, we used microarray to study the consequences of polyploidy on triploid oyster germ cells mitosis and meiosis. Results We compared the transcriptome of gonads of 3na and 3nb oyster gonads over the course of gametogenesis to the transcriptome of diploid (2n) oyster gonads. This study allowed us to reveal an increase in DNA repair and apoptosis through the NF-kappaB pathway, and a decrease in actin remodeling and chromatin remodeling in all 3n oysters. The comparison of 3na and 3nb individuals with 2n revealed that a pachytene checkpoints may be responsible for the success in gametogenesis of 3na individuals and for the observed delay in gametogenesis. However, the sterility of 3nb individuals can be explained by a disruption of sex determinism mechanisms. Indeed 3nb females express male-specific genes including enkurin and an Elav-like gene, and 3nb males express female-specific genes including Forkhead box L2 and beta-catenin. Conclusions Our results bring back to the front of the research field the questions of genetic sex determinism, mitosis/meiosis control, pachytene checkpoint, and cell type specific DNA damage pathways. Furthermore, this study identifies numerous new candidate genes which function should now be studied in details in oysters and in other triploid animals in order to elucidate the complex mechanisms involved in the regulation of triploid cells division. Triploid spats were obtained by crossing tetraploid males and diploid females in the ifremer experimental hatchery (La tremblade, Charente Maritime, France). We performed microarray analysis on a total of 35 individual triploid gonads that can be grouped as follow: 3n stage 0 (4 individuals), 3n alpha Stage 1 (8 individuals), 3n beta Stage 1 (8 individuals), 3n alpha Stage 3 male (4 individuals), 3n beta Stage 3 male (3 individuals), 3n alpha Stage 3 female (4 individuals), and 3n beta stage 3 female (4 individuals).

Project description:Background The Pacific oyster Crassostrea gigas (Mollusca, Lophotrochozoa) is an alternative and irregular protandrous hermaphrodite. Little is known about the genetic and phenotypic bases of sex determinism 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 mollusks gametogenesis should provide a better understanding of the key factors involved in sex differentiation and the regulation of oyster reproduction. Results Expression of the 31,918 ESTs was studied in gonads of oysters cultured along the French Atlantic coasts over a yearly reproductive cycle. Principal component analysis and hierarchical clustering first showed a significant divergence in gene expression patterns of males and females beginning when gonial mitosis started. Early expressed male-specific genes included bindin and female-specific genes included foxL2, a pancreatic lipase related protein, cd63 and vitellogenin. ANOVA analysis of the data further revealed 2482 genes differentially expressed during the course of males and/or females gametogenesis. The expression of 434 genes could be localized in either germ cells or somatic cells of the gonad by comparing the transcriptome of gonads to the transcriptomes of striped oocytes and somatic tissues. Analysis of the annotated genes revealed conserved molecular mechanisms between mollusks and mammals. Genes involved in chromatin condensation, DNA replication and repair, mitosis and meiosis regulation, transcription, translation and apoptosis were expressed in both male and female gonads. Additional lists of genes more specifically involved in either spermatogenesis (meiotic phase, in spermatozoids and mature sperm formation and in flagella structure and movement) or oogenesis (female sex differentiation, transcriptional regulation, cell cycle regulation and oocytes maturation) were also generated. Conclusions Our study provides novel insight into spermatogenesis and oogenesis in an alternative hermaphrodite bivalve, the Pacific oyster C. gigas. We identified genes opening new perspectives for functional studies of the signaling pathways implicated in gonad differentiation and development. Individual gonad oysters were sampled in 3 sites: site 1 = Locmariaquer (Brittany, France), site 2 = Baie des Veys (Normandy, France), and site 3 = Argenton (Brittany, France). 8 undifferentiated stage 0 gonads from site 1 were processed. 4 individuals from site 1 were processed for each gonad developmental stage and sex: stage 1 male, stage 1 female, stage 2 male, stage 2 female, stage 3 male, and stage 3 female. Pools of stage 3 females were prepared with individuals from site 1 and site 2 for biological validation of the results. Striped gonads from stage 3 females were prepared for prediction of gene expression localization.

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