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宫内暴露于氟他胺后 Sprague-Dawley 大鼠生精上皮细胞中连接蛋白 43 的表达。

Connexin 43 expression in Sprague-Dawley rat seminiferous epithelium after in utero exposure to flutamide.

机构信息

Department of Pediatric Urology, Children's Hospital of Chongqing Medical University , Chongqing , China and.

出版信息

Syst Biol Reprod Med. 2014 Oct;60(5):257-62. doi: 10.3109/19396368.2014.921738. Epub 2014 May 27.

Abstract

This study explored the expression of connexin 43 (Cx43) in the testes of prepubertal Sprague-Dawley (SD) rats following in utero flutamide (Flu) exposure. Connexins constitute the major protein type in gap junctions. Connexin 43, the most prominent connexin family member expressed by testes, is localized at the base of seminiferous tubules in humans and rodents, and may be involved in fertility. Flutamide was injected subcutaneously into pregnant SD rats on gestational days 12-21 (25 mg/kg/day). Immunohistochemistry, Western blotting, and real-time PCR was used to investigate the distribution and the expression of Cx43 protein and mRNA in the testis on postnatal day 20 (PD20). Following Flu-exposure, Cx43 was observed between Sertoli cells in the seminiferous tubules. On PD20, no Cx43 protein was expressed by the spermatogonial cell layer of the seminiferous tubules in the controls, but was observed in the Flu-exposed group. Western blotting showed that Cx43 was expressed at significantly lower levels in Flu-exposed testes than controls on PD20 (p < 0.001). On PD20, levels of Cx43 mRNA in undescended Flu-exposed testes were significantly lower than in controls (p < 0.05) and descended Flu-exposed testes (p < 0.01). After Flu-exposure in the rat embryonic period, Cx43 mRNA and protein expression were downregulated, and its distribution in the seminiferous tubules was abnormal.

摘要

本研究探讨了在宫内暴露于氟他胺(Flu)后,未成年 Sprague-Dawley(SD)大鼠睾丸中连接蛋白 43(Cx43)的表达。连接蛋白构成间隙连接的主要蛋白类型。连接蛋白 43 是睾丸中表达最丰富的连接蛋白家族成员,在人类和啮齿动物中位于生精小管的基部,可能与生育能力有关。氟他胺在妊娠第 12-21 天(25mg/kg/天)经皮下注射到怀孕的 SD 大鼠中。免疫组织化学、Western blot 和实时 PCR 用于研究生后第 20 天(PD20)睾丸中 Cx43 蛋白和 mRNA 的分布和表达。在 Flu 暴露后,在生精小管的支持细胞之间观察到 Cx43。在 PD20,对照组的生精小管的精原细胞层没有表达 Cx43 蛋白,但在 Flu 暴露组中观察到。Western blot 显示,在 PD20,Flu 暴露的睾丸中 Cx43 的表达水平明显低于对照组(p<0.001)。在 PD20,未下降的 Flu 暴露睾丸中的 Cx43 mRNA 水平明显低于对照组(p<0.05)和下降的 Flu 暴露睾丸(p<0.01)。在大鼠胚胎期暴露于 Flu 后,Cx43 mRNA 和蛋白表达下调,其在生精小管中的分布异常。

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