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Biosynthesis of avian perivitelline membrane ZPC protein and regulation of sperm receptor activity

禽卵周膜ZPC蛋白的生物合成及精子受体活性的调节

基本信息

批准号：
11660280
负责人：
MORI Makoto
金额：
$ 2.24万
依托单位：
Shizuoka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1999
资助国家：
日本
起止时间：
1999 至 2000
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11660280/
关键词：
Perivitelline membrane ZPC Ovary Granulosa Cell Testosterone Quail 精子卵胞排卵卵子オビダクチン

项目摘要

Avian perivitelline membrane, an investment homologous to the zona pellucida of mammalian oocytes, is composed of at least two glycoproteins. In the present study, we have shown hat one of the components, a glycoprotein homologous to mammalian ZPC, is produced and secreted by the granulosa cells of developing follicles of the quail ovary. from the following experiments.1. we evaluated the expression and regulation of ZPC in quail granulosa cells in vivo and in vitro. Western blot analysis of the SDS-solubilized granulosa layer using antiquail ZPC antiserum showed that the amount of ZPC increased in parallel with follicular development. Northern blot analysis of total RNA using complemental DNA of quail ZPC showed that the increase in messenger RNA expression was also correlated with follicular development.2. To investigate the regulation of ZPC production, the granulosa cells were cultured in a medium containing steroid hormones such as progesterone, estradiol-17 β or testosterone.3. By measuring ZPC protein and messenger RNA with Western blot and Northern blot analyses, respectively, we found that addition of testosterone stimulated not only ZPC protein production but also ZPC messenger RNA expression in the granulosa cells.4. These results suggested that testosterone stimulates ZPC protein production at the gene transcription level.

禽卵周膜是一种与哺乳动物卵母细胞透明膜同源的包埋物，由至少两种糖蛋白组成。在本研究中，我们已经表明帽子的一个组成部分，糖蛋白同源哺乳动物ZPC，产生和分泌的颗粒细胞的卵泡发育的鹌鹑卵巢。从下面的实验中.1.我们在体内和体外评估了ZPC在鹌鹑颗粒细胞中的表达和调节。用antiquail ZPC抗血清对SDS溶解的颗粒层进行Western印迹分析，结果表明ZPC的量与卵泡发育平行增加。利用鹌鹑ZPC的互补DNA对总RNA进行北方印迹分析，结果表明mRNA表达的增加也与卵泡发育相关.为了研究ZPC的产生，将颗粒细胞培养在含有类固醇激素如孕酮、雌二醇-17 β或睾酮的培养基中.通过Western blot和北方印迹分析检测颗粒细胞中ZPC蛋白和信使RNA的表达，我们发现睾酮不仅刺激颗粒细胞中ZPC蛋白的产生，而且还刺激颗粒细胞中ZPC信使RNA的表达.这些结果表明，睾酮在基因转录水平上刺激ZPC蛋白的产生。