摘要
目的:研究RhoA在男性生殖系统中的表达和功能。方法:采用RT-PCR来检测RhoA的mRNA表达,Western blot和免疫组化用来检测RhoA蛋白在人睾丸,附睾及成熟精子中的表达。免疫荧光用来观察这个蛋白在人精子上的定位。豌豆凝集素(PSA)染色用来确定精子顶体反应率。结果:只能在附睾中检测到RhoA的mRNA,而RhoA蛋白不仅在附睾上皮中表达,在人精子上也有表达。它位于人精子的顶体和尾部,并且随着顶体反应转移到精子的赤道部及顶体后区。同时特异的RhoA抗体能够显著地抑制精子发生顶体反应(P<0.01)。结论:结果表明RhoA可能是在附睾成熟过程中带到人精子质膜表面,从而使精子获得发生顶体反应的能力。
Objective：To investigate the expression and possible function of RhoA in human male reproduction. Methods： RT-PCR was used to detect RhoA mRNA, Western blot and Immunohistochemistry were used to detect the RhoA protein expression in human testis, epididymis and mature spermatozoa. The localization of this protein on human spermatozoa was determined by indirect im- munofluorescent staining using anti-RhoA antibody. The percentage of acrosome reaction was determined by PSA staining. Results： RhoA mRNA was only detected in the epididymis,but its protein could be expressed in mature sperm as well as in the epididymal epithelium. RhoA was localized on human sperm entire acrosome and the flagellum,and it was translocated to the equatorial regions and post-acrosomal region after acrosome reacted. Meanwhile,the percentage of acrosome reacted sperm was decreased in comparison to pre-immune control after treatment with specific anti-RhoA antibody （P 〈 0.01）. Concluslon：The results suggest that RhoA might be absorbed or incorporated into the human sperm plasma membrane during epididymal transition and could help human sperm to de- velop the ability to undergo acrosome reaction.
出处
《南京医科大学学报（自然科学版）》
CAS
CSCD
北大核心
2008年第7期827-830,835,共5页
Journal of Nanjing Medical University(Natural Sciences)
基金
国家973项目(2006CB5004002,2006CB944002)
关键词
附睾分泌蛋白
顶体反应
精子成熟
受精
epididymal secretory protein
acrosome reaction
sperm maturation
fertilization
