Sex-specific mRNA alternative splicing patterns and Dmrt1 isoforms contribute to sex determination and differentiation of oyster.

Alternative splicing (AS) of pre-mRNA is a crucial mechanism that regulates the expression of genes involved in sex determination and differentiation. Despite its importance, AS has been rarely characterized in molluscs. In this study, PacBio Iso-Seq was employed to obtain full-length transcriptome and unveil AS patterns of gonads in the Pacific oyster Crassostrea gigas. A total of 24,783 AS events were identified across 6259 genes, with many enriched in phosphorylation-related processes. Splicing factors were found to drive a high frequency of AS events in gonads. Significant sex-based differences in isoform abundance and the incidence of AS events were observed. Comparative analysis of mature female and male gonads revealed a subset of overlapping differential alternative splicing genes and differentially expressed genes enriched in processes related to microtubule function and cell motility. In addition, the expression levels of sex-biased genes were found correlated with their isoform number in both female and male gonads. A novel isoform of Dmrt1 was identified with male specific expression in mature gonads. This study provides the first comprehensive understanding of full-length transcriptome and AS patterns in molluscan gonads, shedding light on the post-transcriptional regulatory mechanisms underlying sex determination and differentiation in molluscs and potentially across other animals.