锌指蛋白185 在小鼠睾丸支持细胞中的表达与定位

目的:检测锌指蛋白185在小鼠睾丸支持细胞中的表达,探讨其与精子发生的关系。方法:提取睾丸和支持细胞总RNA,经半定量RT-PCR法检测ZNF185的转录水平;提取睾丸和支持细胞的蛋白质,利用Western blot分析ZNF185的表达量;制备支持细胞爬片,采用免疫荧光技术检测ZNF185的定位。结果:(1)半定量RT-PCR结果显示:在睾丸支持细胞中扩增出ZNF185基因条带。(2)Western blot结果显示:ZNF185在支持细胞中的表达量显著低于在睾丸组织中的表达量。(3)免疫荧光结果显示:ZNF185主要定位于支持细胞胞质中。结论:ZNF185可能参与支持细胞结构与功能的调控,进而影响精子发生过程。

The Expression and Localization of Zinc Finger Protein 185 in the SertoliCells of the Mouse Testis

Objective:To detect the expression of Zinc finger protein 185 in the sertoli cells of the mouse testis and to investigatethe relationships between ZNF185 and spermatogenesis.Methods:The total RNA was extracted from the testis and sertoli cells cells, thetranscription level of ZNF185 was detected by using semi-quantitative RT-PCR assay; proteins were extracted from the testis and sertolicells, the expression level of ZNF185 was analyzed by Western blot; the coverglass-cultured cells were prepared, the positioning ofZNF185 was detected by immunofluorescence technique.Results:(1) The semi-quantitative RT-PCR showed that ZNF185 gene wasamplified in the sertoli cells of the testis. (2) The result of Western blot showed that the expression of ZNF185 in the sertoli cells wassignificantly lower than that in the testis. (3) Immunofluorescence showed that ZNF185 was mainly located in the cytoplasmof the sertolicells.Conclusion:ZNF185 may affect the process of spermatogenesis by regulating the structure and function of the sertoli cells.