Roles of estrogen receptors during sexual reversal in Pelodiscus sinensis

Abstract

Abstract The Chinese soft-shelled turtle, Pelodiscus sinensis, exhibits distinct sexual dimorphism, with the males growing faster and larger than the females. During breeding, all-male offspring can be obtained using 17β-estradiol (E2). However, the molecular mechanisms underlying E2-induced sexual reversal have not yet been elucidated. Previous studies have investigated the molecular sequence and expression characteristics of estrogen receptors (ERs). In this study, primary liver cells and embryos of P. sinensis were treated with ER agonists or inhibitors. Cell incubation experiments revealed that nuclear ERs (nERs) were the main pathway for the transmission of estrogen signals that was EB upregulated the expression of Rspo1, whereas AS downregulated. The expression of Dmrt1 was enhanced after AS + G-1 treatment, indicating that the regulation of male genes may not be through a single estrogen receptor, but a combination of ERs. As to embryos, only the ERα agonist (EB) remarkably promoted the expression levels of Rspo1, Wnt4, and β-catenin, whereas the ERα inhibitor (AS) had a suppressive effect. Additionally, Dmrt1, Amh, and Sox9 expression levels were downregulated after ERβ inhibitor (PHTPP) treatment. GPER agonist G-1 has a significant promotion effect on Rspo1, Wnt4, and β-catenin, while the inhibitor G-15 has no effect on male-related genes. Overall, these results suggest that ERs play different roles during sexual reversal in P. sinensis and ERa may be the main carrier of estrogen-induced sexual reversal in P. sinensis. Further studies need to be performed to analyze the mechanism of ER action.