Molecular characterization and functional analysis of Esr1 and Esr2 in gonads of Chinese soft-shelled turtle (Pelodiscus sinensis)

Estrogens and their receptors play crucial roles in regulating the gonadal development of vertebrates. To clarify the roles of estrogen receptors in the gonadal development of turtles, estrogen receptors (Esr1 and Esr2) in Chinese soft-shelled turtle (Pelodiscus sinensis) were identified and characterized, and their function in gonads was investigated by intraperitoneal injection of agonist propylpyrazoletriol (PPT) and diarylpropionitrile (DPN), and antagonist ICI 182,780 (ICI). Ps-Esr1 encoded a 588 amino acid protein and Ps-Esr2 encoded a 556 amino acid protein. The two receptors contained classic domains, including the DNA-binding domain and ligand-binding domain, and amino acid sequences showed high homology with other turtles. Ps-Esr1 showed the highest expression in the testis, followed by the ovary and liver. However, Ps-Esr2 showed the highest expression in the ovary, followed by the brain and testis. Ps-Esr1 expression showed an up-regulation trend in gonadal differentiation. Histomorphometric analysis showed that the number of follicles increased in female juvenile turtles treated with DPN or PPT. In addition, Tsc2, GnRH, and Fsh beta were up-regulated in ovaries of turtles treated with agonists, while Sycp3 and Picalm were up-regulated in testes of turtles treated with agonists. Treatment with the antagonist decreased the number of sperm in matured turtles. Stra8, Scyp3, Dmc1, Picalm, Evl, Boule, and Cdk1 were up-regulated in testis after antagonist treatment. The results indicated that Esr1 might play an important role in gonadal differentiation, and the two estrogen receptors might be involved in the spermato-genesis of the turtle. These results could provide a reference for further research on the function of the estrogen signal in male vertebrates.