Diethylstilbestrol affects the expression of GPER in the gubernaculum testis

Abstract

Recent evidence suggested a positive correlation between environmental estrogens (EEs) and high incidence of abnormalities in male urogenital system. EEs are known to cause the abnormalities of testes development and testicular descent. Diethylstilbestrol (DES) is a nonsteroidal synthetic estrogen that disrupts the morphology and proliferation of gubernacular cells, and its nongenomic effects on gubernaculum testis cells may be mediated by G protein-coupled estrogen receptor (GPER). In this study, we detected the expression of GPER in mouse gubernacular testis and investigated the effects of DES on the expression of GPER in gubernaculum testis cells. RT-PCR analysis revealed that GPER mRNA was expressed in the gubernaculum. GPER protein was detected in the parenchymal cells of the gubernaculum early in development. Furthermore, we demonstrate that GPER inhibitor G15 relieved DES-induced inhibition of GPER expression in gubernaculum testis cell, but ER inhibitor ICI 182780 had the converse effects on DES-induced inhibition of GPER expression in these cells. These data suggest that the effects of DES on mouse gubernaculum testis cells are mediated at least partially by the regulation of GPER expression.

Keywords: DES; EEs; GPER; gubernaculums testis.

Figure 1

Expression of GPER in mouse gubernaculums testis. A. Immunohistochemical staining of GPER in gubernaculums testis. a. GD19, b. PD0, c. PD3, d. PD7 (× 400). B. Immunoelectron microscopy showing colloidal gold particles positively stained by GPER antibody. C. RT-PCR analysis of GPER and GAPDH expression in gubernaculums testis. D. Western blot analysis of GPER protein expression in mouse gubernaculums testis at different stages of testis descent. E. Densitometry analysis of the blots shown in D (n=3). GAPDH served as loading control. *P<0.05, **P<0.01 versus PD0 control.

Figure 2

Western blot analysis of the expression of GPER in mouse gubernaculum testis cells. A. Cells were treated with DES or additionally pretreated with ICI 182780 or G15 as indicated, and GPER expression was detected by Western blot analysis. B. Densitometry analysis of the blots shown in A (n=3). *P<0.05, **P<0.01 versus the corresponding control in the left, ▽▽P<0.01, ▽P<0.05 versus DES in the same time point group.