Bi Ye, Liu Mingxi, Tu Wenjiao, Wu Yibo, Guo Xuejiang, Zhou Zuomin, Sha Jiahao
State Key Laboratory of Reproductive Medicine, Department of Histology and Embryology, Nanjing Medical University, Nanjing, Jiangsu 210029, China.
J Biomed Res. 2012 Mar;26(2):110-6. doi: 10.1016/S1674-8301(12)60020-7.
This study investigated the expression and distribution of 2810408A11Rik in mouse testis and sperm, and explored its role in spermatogenesis and sperm function. The expression levels of 2810408A11Rik mRNA in multiple tissue samples were analyzed using bioinformatic resources and RT-PCR technique. A specific rabbit polyclonal antibody was prepared by prokaryotic expression of 2810408A11Rik recombinant protein and utilized for animal immunization. Western blotting, immunohistochemistry and immunofluorescence were used to detect the expression and distribution of 2810408A11Rik. The results of the bioinformatic analysis and RT-PCR showed that 2810408A11Rik mRNA was specifically expressed in mouse testis, and 2810408A11Rik protein included a protein phosphatase inhibitor domain. Western blotting assays, immunohistochemistry and immunofluorescence confirmed the expression of 2810408A11Rik protein in mouse testis, especially in post-meiosis round and long spermatids, and that it is localized in the acrosome and the post-nucleus area of sperm. Our findings suggest that 2810408A11Rik may play an important role in spermatogenesis, sperm capacitation and fertilization.
本研究调查了2810408A11Rik在小鼠睾丸和精子中的表达与分布，并探讨了其在精子发生和精子功能中的作用。利用生物信息资源和逆转录-聚合酶链反应（RT-PCR）技术分析了多个组织样本中2810408A11Rik mRNA的表达水平。通过原核表达2810408A11Rik重组蛋白制备了特异性兔多克隆抗体，并用于动物免疫。采用蛋白质免疫印迹法、免疫组织化学和免疫荧光法检测2810408A11Rik的表达与分布。生物信息分析和RT-PCR结果显示，2810408A11Rik mRNA在小鼠睾丸中特异性表达，且2810408A11Rik蛋白包含一个蛋白磷酸酶抑制结构域。蛋白质免疫印迹分析、免疫组织化学和免疫荧光证实了2810408A11Rik蛋白在小鼠睾丸中的表达，尤其是在减数分裂后的圆形和长形精子细胞中，且其定位于精子的顶体和核后区域。我们的研究结果表明，2810408A11Rik可能在精子发生、精子获能和受精过程中发挥重要作用。
