热应激对猪睾丸Hsp60和Hsp90表达与定位的影响

Effects of Heat Stress on the Expression and Localization of Hsp60 and Hsp90 in Boar Testes

建立了环境热应激和睾丸局部热处理猪动物模型,用qRT-PCR和Western blot法检测了热休克蛋白家族成员Hsp60和Hsp90 mRNA和蛋白表达水平的变化,用免疫组织化学方法检测Hsp60和Hsp90在猪睾丸的组织细胞学定位。qRT-PCR和Western blot结果显示,与对照组相比,环境热应激组和局部睾丸热应激组Hsp60和Hsp90 mRNA和蛋白的表达量均显著升高。免疫组织化学结果显示,对照组Hsp60主要表达在精母细胞和支持细胞的细胞质,环境热应激组和局部睾丸热应激组Hsp60免疫阳性从精母细胞的细胞质转移至精原细胞的细胞核。热处理前后Hsp90定位于精原细胞和长形精子细胞。环境高温热应激和局部睾丸热刺激导致Hsp60和Hsp90表达升高,提示Hsp60和Hsp90在热应激猪睾丸精子发生过程中发挥重要作用。

In this study,environmental heat stress and local testicular heating models were established in boars.The expression of Hsp60 and Hsp90 mRNA and protein was detected by qRT-PCR and Western blot.The localization of Hsp60 and Hsp90 in boar testes was conducted by immunohistochemistry method.qRT-PCR and Western blot results showed that the expression of Hsp60 and Hsp90 was significantly increased in environmental hyperthermia group and local testicular heating group compared with the control group.Immunohistochemistry results showed that Hsp60 was primarily expressed in the cytoplasm of spermatocytes and Sertoli cells in control group.In hyperthermia group and local testicular heating group,Hsp60 immunopositive staining was translocated from the cytoplasm of spermatocyte to the nucleus of spermatogonia.Hsp90 was localized in spermatogonia and elongated spermatid.Environmental heat stress and local testicular heating lead to increased expression of Hsp60 and Hsp90,which indicated that Hsp60 and Hsp90 play an important role in spermatogenesis of boar testes under heat stress conditions.