Tokuhiro Keizo, Miyagawa Yasushi, Tanaka Hiromitsu
TANAKA Project, Center for Advanced Science and Innovation, Osaka University, 3-1 Yamadaoka, Osaka 565-0871, Japan.
Asian J Androl. 2008 Sep;10(5):711-8. doi: 10.1111/j.1745-7262.2008.00426.x.
To characterize mouse capping protein alpha3 (CPalpha3) during spermatogenesis and sperm maturation.
We produced rat anti-CPalpha3 antiserum and examined the expression of CPalpha3 in various mouse tissues using Western blot analysis and the localization of CPalpha3 in testicular and epididymal sperm using immunohistochemical analyses. We also examined how the localization of CPalpha3 and beta-actin (ACTB) in sperm changed after the acrosomal reaction by performing immunohistochemical analyses using anti-CPalpha3 antiserum and anti-actin antibody.
Western blot analysis using specific antiserum revealed that CPalpha3 was expressed specifically in testes. Interestingly, the molecular weight of CPalpha3 changed during sperm maturation in the epididymis. Furthermore, the subcellular localization of CPalpha3 in sperm changed dynamically from the flagellum to the post-acrosomal region of the head during epididymal maturation. The distribution of ACTB was in the post-acrosomal region of the head and the flagellum. After inducing the acrosomal reaction, the CPalpha3 and ACTB localization was virtually identical to the localization before the acrosomal reaction.
CPalpha3 might play an important role in sperm morphogenesis and/or sperm function.
在精子发生和精子成熟过程中对小鼠封端蛋白α3(CPα3)进行特征描述。
我们制备了大鼠抗CPα3抗血清,并使用蛋白质免疫印迹分析检测CPα3在各种小鼠组织中的表达,以及使用免疫组织化学分析检测CPα3在睾丸和附睾精子中的定位。我们还通过使用抗CPα3抗血清和抗肌动蛋白抗体进行免疫组织化学分析,研究了顶体反应后精子中CPα3和β-肌动蛋白(ACTB)的定位如何变化。
使用特异性抗血清进行的蛋白质免疫印迹分析显示,CPα3在睾丸中特异性表达。有趣的是,CPα3的分子量在附睾精子成熟过程中发生了变化。此外,在附睾成熟过程中,精子中CPα3的亚细胞定位从鞭毛动态变化到头部顶体后区域。ACTB分布在头部顶体后区域和鞭毛中。诱导顶体反应后,CPα3和ACTB的定位与顶体反应前的定位基本相同。
CPα3可能在精子形态发生和/或精子功能中发挥重要作用。