【作者】 杨媛媛；

【导师】 王根林；

【作者基本信息】 南京农业大学 ， 动物遗传育种与繁殖， 2006， 硕士

【摘要】 本实验对近年来发现的一种阳离子通道Catsper(cation channel of sperm)的基本特性进行了探讨。通过相对半定量RT-PCR和组织免疫化学的方法分析了该蛋白在不同组织中的表达情况及其表达量与个体发育之间的关系，通过免疫荧光方法对该蛋白在精子上的定位进行了试验，以阐明Catsper蛋白的基本表达特性，为研究超活化的机理及调节机制提供基础。本实验分为两部分： 一、Catsper蛋白在各组织及精子上的表达情况 通过RT-PCR和组织免疫化学对Catsper蛋白在各组织中和精子上的表达进行了探讨，并对睾丸组织和附睾组织中该蛋白的表达量进行了相对定量分析。结果表明，Catsper蛋白特异性的表达于睾丸和附睾组织中，而且在附睾组织中的丰度要大于睾丸组织中的。通过精子免疫荧光化学法对精子上该蛋白的定位进行了试验分析，发现该蛋白主要是定位于精子尾部的主段。本试验为进一步研究Catsper对精子超活化的调节提供了依据。 二、Catsper蛋白表达量与个体发育之间的关系 通过相对定量RT-PCR对Catsper蛋白随个体发育其表达量的变化情况进行了分析。Catsper蛋白的表达量是受到发育所调控的，与个体性成熟之间有密切的关系。发现小鼠catsper蛋白从3周龄开始出现表达，表达量较低，随后表达量随周龄的增长而增加，前几周表达量增加显著(P＜0.05)，6周龄以后表达量增加幅度就变的不显著，本试验为进一步深入研究Catsper蛋白的功能提供了基础。更多还原

【Abstract】 The basic characteristics of Catsper, a cation channel of sperm which was discovered recently, were explored in this thesis. The expression of Catsper in eight tissues and the relation between its expression and individual development were analyzed by RT-PCR and immunohistochemistry. Its expression in sperm was also determined by immunofluorescen-ce. All of the above was to provide evidence for further research of the mechanism of the regulation of sperm hyperactivation. This thesis includes two sections as follows.In the first section , the expression of Catsper in tissue and sperm were examined respectively. The results indicated that Catsper was exclusively expressed in testis and epididymis, and the abundance of Catsper in epididymis was much larger than that in testis. Furthermore, Catsper was mainly located in the principal piece of spermatozoa tail.In the second section, the expression changes of Catsper in different-aged mice were analyzed by RT-PCR. Catsper gene expression was developmently regulated with a direct correlation between Catsper expression and mouse sexual maturation. The results indicated that the expression of Catsper began from 3 week of age, and the expression amount was much low. Then the expression amount increased significantly till 6 week of age, from which the expression increased gradually into a stationary phase.更多还原