中华鳖SOX9基因在胚胎期性腺中的表达模式及在性逆转中的表达变化  

The expression pattern of SOX9 gene during embryonic development and its expression changes in sex reversal in Pelodiscus sinensis

作　　者：王莉[1,2] 孙伟[1] 储经乾 刘颖[1] 史思瑞 葛楚天[1] 钱国英[1,2] 

机构地区：[1]浙江万里学院生物与环境学院,浙江宁波315100 [2]上海海洋大学水产与生命学院,上海201306

出　　处：《水产学报》2014年第9期1286-1293,共8页Journal of Fisheries of China

基　　金：国家自然科学基金(31101884);浙江省自然科学基金(LY14C190008);浙江省公益性技术应用研究项目(2013C32054);宁波市科技创新团队(2012B82016);宁波市自然科学基金(2012A610142)

摘　　要：SOX9基因在脊椎动物性别决定和性腺分化中扮演重要的调控作用。从mRNA和蛋白水平分析中华鳖SOX9基因在不同组织中的差异性表达、在胚胎性腺和成年睾丸中的细胞定位以及在性逆转中的表达变化,研究SOX9基因在中华鳖性别分化中的调控作用。Real-time PCR结果显示,SOX9基因在中华鳖雄性性腺中特异性表达。免疫荧光染色分析显示,SOX9蛋白在雄性18期胚胎性腺中开始表达,随着性腺的发育,SOX9蛋白定位于性腺Sertoli前体细胞细胞核中;而在雌性胚胎性腺并未见其表达。此外,在雌激素诱导的雄性向雌性性逆转胚胎中,SOX9基因显著下调,而在芳香化酶抑制剂诱导的雌性向雄性性逆转胚胎中,SOX9基因表达则显著上升。研究表明,SOX9基因为中华鳖雄性特异性基因,参与雄性性腺的发育过程,可能在中华鳖早期性别分化过程中起调控作用。The molecular mechanism of sex determination in Pelodiscus sinensis has been a long-standing mystery. In many vertebrate species,SOX9 gene is required for male sexual differentiation. In this study,the mRNA and protein expression of SOX9 gene and its cellular localization in gonads of different developmental stages of both sexes were examined to investigate the involvement of SOX9 in sexual differentiation in P.sinensis. Real-time PCR results showed that,among different tissues,SOX9 gene highly expressed in testis tissue. Immunofluorescence analysis revealed that SOX9 protein started to express at embryonic stage 18,and localized in the nuclei of pre-sertoli cells in male embryonic gonads. However,no SOX9 protein was detected in female gonads during embryonic development. Moreover,SOX9 protein expression significantly decreased in estrogen-induced male to female sex reversal. Whereas SOX9 protein was triggered to highly express in aromatase inhibitor-induced female to male sex reversal. In conclusion,this study indicates that SOX9 gene is a male-specific gene and participates in the male gonadal development,which is likely to play a key role during primary male sex determination in P. sinensis.

关 键 词：中华鳖 SOX9基因 性别分化 性逆转 

分 类 号：Q785[生物学—分子生物学] S966.5[农业科学—水产养殖]