Transcriptomic analysis revealed gene expression profiles during the sex differentiation of Chinese tongue sole (Cynoglossus semilaevis)

Sex differentiation in aquatic fish is important both for theoretical study and practical production, as growth dimorphism frequently appears in different sexes, especially in marine fish. The deciphered genome, identification of the male-determining gene dmrt1 and established genotypic sex screening method make Chinese tongue sole (Cynoglossus semilaevis) an ideal model to study sex differentiation in fish. In this study, comparative gonadal transcriptomic analyses were conducted for genetic females and males at 48, 68, and 108 days post hatching (dph), representing pre-, during- and post-gonadal differentiation stages, although the gonad is not completely differentiated and isolable in 48 and 68 dph individuals, while it is in 108 dph individuals. Altogether, 28 libraries were constructed, and a mean of 46.64 M clean reads was obtained. Differentially expressed gene (DEG) analysis revealed that 179 genes had similar expression patterns in males and females in all three stages. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis suggested that the enriched pathways included ubiquitin-mediated proteolysis, lysosomes, and RNA degradation. Moreover, weighted gene coexpression network analyses (WGCNA) identified 14 modules, one of which was closely correlated with female differentiation, exhibiting female-biased expression in all three stages (48, 68, 108 dph). An illustrated core gene interaction network of this module identified 50 genes, most of which are on W chromosomes. Six genes, including two ubiquitin conjugating enzymes, were selected for further investigation, and their female-biased expression was confirmed in even earlier stages, at 10 and 30 dph. These data facilitate our understanding of sex differentiation in fish and provide a genomic rationale for screening candidate genes (preferentially W-linked genes) that could be involved in the female differentiation process.