Liang Hongwei, Meng Yan, Cao Lihuan, Li Xiang, Zou Guiwei
Key Laboratory of Aquatic Genomics, Ministry of Agriculture, Yangtze River Fisheries Research Institute, Chinese Academy of Fisheries Science, Wuhan, Hubei 430223, China; and Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan, Hubei 430223, China; and Corresponding authors. Emails:
Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan, Hubei 430223, China.
Reprod Fertil Dev. 2019 Aug;31(9):1425-1433. doi: 10.1071/RD19045.
Little is known about sex determination and differentiation in the Chinese soft-shelled turtle Pelodiscus sinensis. R-Spondin 1 (RSPO1), a candidate sex-determining gene, is an important regulator of ovarian differentiation in animals. Exogenous drugs can affect sex differentiation. In this study we cloned the RSPO1 gene from P. sinensis (psRSPO1) and analysed its expression profile. The psRSPO1 gene exhibited sequence identity with RSPO1 genes from other species. RSPO1 protein-based phylogenetic analysis showed that psRSPO1 in P. sinensis is closely related to RSPO1 proteins from other turtles. psRSPO1 showed abundant expression in adult brain and gonads, with higher levels in females than males. We also evaluated the effects of three finaconcentration of 2.5, 5.0 and 10mgmL-1 exogenous oestradiol (E2) and aromatase inhibitor (letrozole) on the expression of psRSPO1, external embryo morphology, growth status of embryos and the sex ratio when the drugs were injected to eggs during incubation. The expression of psRSPO1 was upregulated and downregulated by exogenous oestradiol and letrozole respectively, despite inconsistent expression trends at different embryo development times. External embryo morphology, growth status and sex ratio were affected by both exogenous oestradiol and the aromatase inhibitor. Feminisation was induced by oestradiol, but inhibited by letrozole. These results will contribute to studies of the potential molecular mechanisms underlying sex differentiation and sex control in the Chinese soft-shelled turtle.
中华鳖性别决定和分化的相关情况鲜为人知。R- 斯普林蛋白 1（RSPO1）作为一个候选性别决定基因，是动物卵巢分化的重要调节因子。外源性药物会影响性别分化。在本研究中，我们从中华鳖中克隆了RSPO1基因（psRSPO1）并分析了其表达谱。psRSPO1基因与其他物种的RSPO1基因具有序列同一性。基于RSPO1蛋白的系统发育分析表明，中华鳖中的psRSPO1与其他龟类的RSPO1蛋白密切相关。psRSPO1在成年中华鳖的脑和性腺中表达丰富，雌性中的表达水平高于雄性。我们还评估了三种浓度（2.5、5.0 和 10mg/mL）的外源性雌二醇（E2）和芳香化酶抑制剂（来曲唑）在孵化期间注射到卵中时对psRSPO1表达、胚胎外部形态、胚胎生长状况和性别比例的影响。尽管在不同胚胎发育时期表达趋势不一致，但外源性雌二醇和来曲唑分别上调和下调了psRSPO1的表达。外源性雌二醇和芳香化酶抑制剂均影响胚胎外部形态、生长状况和性别比例。雌二醇诱导雌性化，但来曲唑抑制雌性化。这些结果将有助于对中华鳖性别分化和性别控制潜在分子机制的研究。
