摘要
目的探讨配子生成素结合蛋白2(GGNBP2)和配子生成素(GGN)在小鼠睾丸生殖过程中的细胞定位以及作用,初步阐明GGNBP2缺失引起睾丸生精功能障碍的机制。方法采用免疫沉淀方法检测GGNBP2与GGN相互作用;免疫荧光、免疫组化检测GGNBP2与GGN细胞定位;PCR检测野生型Ggnbp2基因敲除型小鼠睾丸生殖细胞GGN mRNA表达;Western blot检测其GGN蛋白表达。结果 GGNBP2与GGN相互作用,两种蛋白共同定位于精母细胞和精子细胞(Golgi phase,Cap phase)中的细胞核和胞浆,精子细胞(Acrosome phase,Maturation phases)中的顶体和尾部,GGNBP2缺失引起GGNmRNA水平和蛋白水平减低,在精母细胞中GGN定位改变。结论 Ggnbp2基因敲除引起小鼠生精功能缺陷可能通过降低GGN表达,改变GGN细胞定位,DNA双链损伤修复机制受损。
Objective To explore the localization and function of GGNBP2and GGN in mouse germ cells during spermatogenesis,and preliminarily clarify the mechanism by which GGNBP2loss results in spermatogenesis arrest. Methods Immunoprecipitation was performed to detect the GGNBP2and GGN interaction.Immunofluorescence and immunohistochemistry were applied to localize GGNBP2and GGN in spermatocytes and spermatids.PCR and western blot were used to detect GGN mRNA protein expression in wild type and Ggnbp2KO mice germ cells.Results There existedan interaction between GGNBP2and GGN.These two proteins were co-localized at nucleus and cytoplasm of spermatocytes and spermatids (Golgi,cap phase),and at acrosome and tail of the spermatids (acrosome and maturation phase).Knockout of GGNBP2downregulated the expression of GGN and changed its cell location.Conclusion Spermatogenesis dysfimction caused by Ggnbp2knockout might be attributed to GGN downregulation,GGN translocation,and compromised DNA double-stranded break repair.
作者
郭凯敏
梁作文
田润辉
刘凌云
Guo Kaimin;Liang Zuowen;Tian Runhui;Liu Lingyun(Department ofAndrology,the First Hospital of Jilin University,Jiling 130021,Changchun,China;Department of Psychology,the First Hospital of Jilin University)
出处
《中国男科学杂志》
CAS
CSCD
2018年第4期3-8,共6页
Chinese Journal of Andrology
