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Gonadal transcriptomes associated with sex phenotypes provide potential male and female candidate genes of sex determination or early differentiation in Crassostrea gigas, a sequential hermaphrodite mollusc
Background
In the animal kingdom, mollusca is an important phylum of the Lophotrochozoa. However, few studies have investigated the molecular cascade of sex determination/early gonadal differentiation within this phylum. The oyster Crassostrea gigas is a sequential irregular hermaphrodite mollusc of economic, physiological and phylogenetic importance. Although some studies identified genes of its sex-determining/−differentiating pathway, this particular topic remains to be further deepened, in particular with regard to the expression patterns. Indeed, these patterns need to cover the entire period of sex lability and have to be associated to future sex phenotypes, usually impossible to establish in this sequential hermaphrodite. This is why we performed a gonadal RNA-Seq analysis of diploid male and female oysters that have not changed sex for 4 years, sampled during the entire time-window of sex determination/early sex differentiation (stages 0 and 3 of the gametogenetic cycle). This individual long-term monitoring gave us the opportunity to explain the molecular expression patterns in the light of the most statistically likely future sex of each oyster.
Results
The differential gene expression analysis of gonadal transcriptomes revealed that 9723 genes were differentially expressed between gametogenetic stages, and 141 between sexes (98 and 43 genes highly expressed in females and males, respectively). Eighty-four genes were both stage- and sex-specific, 57 of them being highly expressed at the time of sex determination/early sex differentiation. These 4 novel genes including Trophoblast glycoprotein-like, Protein PML-like, Protein singed-like and PREDICTED: paramyosin, while being supported by RT-qPCR, displayed sexually dimorphic gene expression patterns.
Conclusions
This gonadal transcriptome analysis, the first one associated with sex phenotypes in C. gigas, revealed 57 genes highly expressed in stage 0 or 3 of gametogenesis and which could be linked to the future sex of the individuals. While further study will be needed to suggest a role for these factors, some could certainly be original potential actors involved in sex determination/early sex differentiation, like paramyosin and could be used to predict the future sex of oysters.
Keyword(s)
RNA-Seq, Gonad transcriptome, Early sex differentiation, Sex determination, Irregular hermaphrodite, Oyster
Full Text
File | Pages | Size | Access | |
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Publisher's official version | 16 | 2 Mo | ||
Additional file 1 Eighty-four genes identified in the C. gigas transcriptome by RNA-Seq Differential gene Expression Analysis (DEA): Name, NCBI gene ID, fold change (male vs female), p-value, FDR ... | - | 19 Ko | ||
Additional file 2 Statistical bilateral Wilcoxon tests performed for RT-qPCR experiments at each gametogenetic stage between sexes. Significance at p < 0.05. | 1 | 391 Ko | ||
Additional file 3. Primers used for RT-qPCR: list, sequences, number of bases, blasted name of the gene amplified, R2 and % of PCR efficiency. | - | 11 Ko |