ABSTRACT
DNA methylation is an important way to regulate gene expression in eukaryotes. In order to reveal the role of DNA methylation in the regulation of germ cell-specific piwi gene expression during spermatogenesis of Japanese flounder (Paralichthys olivaceus), the expression profiles of piwil1 (piwi-like 1) and piwil2 (piwi-like 2) genes in the gonads of female, male, and sex-reversed pseudo-male P. olivaceus were analyzed, and the dynamic of DNA methylation was investigated. As a result, piwil1 and piwil2 genes were highly expressed in the testis of both male and pseudo-male P. olivaceus, with significant variation among male individuals. The DNA methylation levels in the promoter regions of both piwil1 and piwil2 were negatively correlated with their expression levels, which may contribute to the transcriptional regulation of piwi genes during spermatogenesis. There was also sperm quality variation among male P. olivaceus, and the sperm curvilinear velocity was positively correlated with the expression of both piwil1 and piwil2 genes. These results indicated that the DNA methylation in piwil1 and piwil2 promoter regions may affect the initiation of piwi gene transcription, thereby regulating gene expression and further affecting the spermatogenesis process and gamete quality in P. olivaceus.
Subject(s)
Argonaute Proteins , DNA Methylation , Flounder , Spermatogenesis , Spermatozoa , Animals , Male , Argonaute Proteins/genetics , Argonaute Proteins/metabolism , Flounder/genetics , Flounder/metabolism , Spermatozoa/metabolism , Spermatogenesis/genetics , Female , Promoter Regions, Genetic , Testis/metabolism , Gene Expression Regulation , Fish Proteins/genetics , Fish Proteins/metabolismABSTRACT
Steroidogenesis controls the conversion of cholesterol into steroid hormones through the complex cascade reaction of various enzymes, which play essential roles in sexual differentiation and gonadal development in vertebrates, including teleosts. Japanese flounder (Paralichthys olivaceus) and Chinese tongue sole (Cynoglossus semilaevis) are important marine cultured fishes in China and have remarkable sexual dimorphism with bigger females and sex reversal scenarios from female to neo-male. Several steroidogenic genes have been analyzed individually in the two species, but there is a lack of information on the coordinated interaction of steroidogenic gene regulation. Therefore, in this study, through genomic and transcriptomic analysis, 39 and 42 steroidogenic genes were systematically characterized in P. olivaceus and C. semilaevis genomes, respectively. Phylogenetic and synteny analysis suggested a teleost specific genome duplication origin for cyp19a1a/cyp19a1b, hsd17b12a/hsd17b12b, ara/arb and esr2a/esr2b but not for star/star2 and cyp17a1/cyp17a2. Comparative transcriptome analysis revealed conserved expression patterns for steroidogenic genes in P. olivaceus and C. smilaevis gonads; star/star2, cyp11a/cyp11c, cyp17a1/cyp17a2, cyp21a, hsd3b1, hsd11b and hsd20b were strongly expressed in testis, while cyp19a1a and hsd17b genes were highly expressed in ovaries. Only a few genes were differentially expressed between male and neo-male testis of both P. olivaceus and C. semilaevis, and even fewer genes were differentially regulated in the brains of both species. Network analysis indicated that cyp11c, cyp17a1 and hsd3b1 actively interacted with other steroidogenic genes in P. olivaceus and C. semilaevis, and may play a more sophisticated role in the steroid hormone biosynthesis cascade. The coordinated interaction of steroidogenic genes provided comprehensive insights into steroidogenic pathway regulation with a global biological impact, as well as sexual development in teleost species.
