摘要
目的:探讨C末端驱动蛋白1(KIFC1)在精子顶体形成过程中的时空表达特征及其功能。方法:利用流式分选及免疫荧光技术,分析KIFC1在精子顶体形成过程中的表达和定位。运用体外GC2-spd细胞系筛选出具有高干扰效率的KIFC1的小干扰RNA(RNAi1),将混有0.5%台盼蓝的KIFC1 RNAi1,显微注射至3周龄Balb/c小鼠的睾丸生精小管内,同一只对侧睾丸注射阴性Con-RNAi;在注射后3周后,取附睾尾部精子进行形态学分析。结果:精子变形早期,KIFC1蛋白主要表达于精子细胞的细胞质中;变形中期,KIFC1蛋白表达于顶体泡及顶体;变形晚期,KIFC1蛋白主要表达于残余体内;成熟期,KIFC1蛋白在精子表达消失。RNAi1组的精子头部畸形率明显高于对照[(32.12±0.25)%vs(7.06±1.25)%,P<0.01]。结论:KIFC1在精子顶体形成过程中可能发挥重要作用,主要影响精子顶体的形成。
Objective: To investigate the spatio-temporal expression and function of the end-directed KIFC1(kinesin family member C1) protein during acrosomogenesis. Methods: The expression and location of KIFC1 were analyzed by flow separation and immunofluorescence, and the small interference RNA(RNAi1) of KIFC1 with high-interference efficiency was screened using in vitro GC2-spd cell lines. The KIFC1 RNAi1 mixed with 0.5% trypan blue solution was microinjected into the testicular seminiferous tubules and negative Con-RNAi1 into the contralateral testis of a 3-week-old Balb/c mouse, followed by morphological analysis of the sperm collected from the testis tail at 3 weeks after injection. Results: The expression of KIFC1 was observed mainly in the cytoplasm of the sperm cells in the early stage of sperm deformation, in the acrosomal vesicle and acrosome in the middle stage, and in the residual body in the late stage, but vanished during sperm maturation. The sperm head deformity rate was significantly higher in the RNAi1 than in the negative control group([32.12 ± 0.25]% vs [7.06 ± 1.25]%, P < 0.01). Conclusion: The KIFC1 protein may play an important role in the formation of spermatozoa, mainly affecting acrosomogenesis.
作者
智二磊
李朋
姚晨成
汪小波
洪燕
李铮
ZHI Er-lei;LI Peng;YAO Chen-cheng;WANG Xiao-bo;HONG Yan;LI Zheng(Department of Andrology,Shanghai General Hospital,Shanghai Jiao Tong University,Shanghai 200080,China)
出处
《中华男科学杂志》
CAS
CSCD
北大核心
2019年第12期1059-1065,共7页
National Journal of Andrology
基金
国家自然科学基金(81701524)~~
关键词
C末端驱动蛋白1
顶体形成
小干扰RNA
睾丸显微注射
kinesin family member C1(KIFC1)
acrosomogenesis
small interference RNA
testicular micro-injection
