[动力蛋白轻链1在小鼠睾丸和精子中的定位及其在精子发生中的作用]

[Location of Dctn1 in the mouse testis and sperm and its role in spermiogenesis].

作者信息

Zheng Bo, Jiang Min, Li Su-Ying, Wu Yi-Bo, Zhu Hui, Zhou Zuo-Min, Sha Jia-Hao

机构信息

Laboratory of Reproductive Medicine, Nanjing Medical University, Nanjing, Jiangsu 210029, China.

出版信息

Zhonghua Nan Ke Xue. 2011 Sep;17(9):799-804.

PMID:21961240

Abstract

OBJECTIVE

To investigate the role of dynactin 1 (Dctn1) in the process of mouse spermiogenesis.

METHODS

Western blot and indirect immunofluorescence were used to analyze the expression and location of Dctn1 in the mouse testis and spermatozoa. The highest efficiency of small interference RNA (siRNA) was verified by GC2-spd cell line in vitro and in vivo studies, respectively. Dctn1 siRNA mixed with the indicator (0.4% trypan blue) was injected into the seminiferous tubules of 3-week-old ICR mice through rete testis microinjection, and negative control siRNA injected into the control testes. The normal group included 3-week-old ICR mice that did not receive any treatment. Spermatozoa were collected from the cauda epididymis 3 weeks after siRNA injection for morphological analysis.

RESULTS

Dctn1 was mainly localized in the tail of spermatozoa. After interference, the sperm tail abnormality in the Dctn1 siRNA group was (23.57 +/- 0.55)%, significantly higher than (12.35 +/- 2.29)% in the control (P < 0.01, n = 3), and it was (3.37 +/- 0.69)% in the normal group.

CONCLUSION

Dctn1 plays an important role in mouse spermiogenesis, and mainly affects the formation of the tail of spermatozoa.

摘要

目的

研究动力蛋白激活蛋白1（Dctn1）在小鼠精子发生过程中的作用。

方法

采用蛋白质免疫印迹法和间接免疫荧光法分析Dctn1在小鼠睾丸和精子中的表达及定位。分别通过体外GC2-spd细胞系和体内实验验证小干扰RNA（siRNA）的最高效率。将Dctn1 siRNA与指示剂（0.4%台盼蓝）混合，经睾丸网显微注射注入3周龄ICR小鼠的生精小管，对照组睾丸注射阴性对照siRNA。正常组包括未接受任何处理的3周龄ICR小鼠。siRNA注射3周后从附睾尾收集精子进行形态学分析。