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：
13660284
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：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13660284/
关键词：
quail oocyte vitelline membrane liver ZP1 estrogen gene PCR

项目摘要

Avian perivitelline membrane, an oocyte extracellular matrix homologous to the zona pellucida in mammals or chorion in fish, is composed of ZPC and ZP1. Previous studies have indicated that one of the components, a glycoprotein homologous to mammalian ZPC, is produced in the granulosa cells of the developing follicles of quail ovary on stimulation with testosterone.However, little is known about the molecular biology of the other component of the avian perivitelline membrane, ZP1, and information about gene expression is particularly lacking. We have cloned the ZP1 in Japanese quail and examined its gene expression. A cDNA encoding quail ZP1 was isolated from the livers of mature females using reverse transcription-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. It encoded a 934-amino acid protein that showed greatest homology (87% identity) with the chicken ZP1. From 128 to 571 amino acid from its N-terminal is unique, so only less than 40% identity with other mammals. RT-PCR amplification of 880 bases indicated that the ZP1 mRNA in the liver was restricted to mature laying females. The expression of ZP1 mRNA was stimulated by in-vivo treatment with diethylstilbestrol in immature females as well as males. These results suggested that androgens and estrogens coordinately regulate the formation of quail perivitelline membrane proteins. In addition, the use of ZP1 transcriptional induction in males or immature females as a biological marker of environmental estrogens is discussed.

禽类卵周膜是一种卵母细胞细胞外基质，与哺乳动物的透明带或鱼类的绒毛膜同源，由 ZPC 和 ZP1 组成。先前的研究表明，其中一种成分是一种与哺乳动物 ZPC 同源的糖蛋白，在睾酮刺激下，在鹌鹑卵巢发育中的卵泡的颗粒细胞中产生。然而，人们对另一种成分的分子生物学知之甚少。鸟类卵周膜、ZP1 和有关基因表达的信息尤其缺乏。我们在日本鹌鹑中克隆了 ZP1 并检查了其基因表达。使用逆转录聚合酶链式反应 (RT-PCR) 和 cDNA 末端快速扩增，从成熟雌性鹌鹑的肝脏中分离出编码鹌鹑 ZP1 的 cDNA。它编码了一个 934 个氨基酸的蛋白质，与鸡 ZP1 显示出最大的同源性（87% 同一性）。其N端的128至571个氨基酸是独特的，因此与其他哺乳动物的同一性只有不到40%。 880个碱基的RT-PCR扩增表明肝脏中的ZP1 mRNA仅限于成熟产蛋雌性。在未成熟雌性和雄性体内用己烯雌酚进行体内处理可刺激 ZP1 mRNA 的表达。这些结果表明雄激素和雌激素协调调节鹌鹑卵周膜蛋白的形成。此外，还讨论了在雄性或未成熟雌性中使用 ZP1 转录诱导作为环境雌激素的生物标记。