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

禽卵周膜是一种卵母细胞外基质，与哺乳动物的透明膜或鱼类的绒毛膜同源，由ZPC和ZP 1组成。以前的研究表明，其中一个成分，一种与哺乳动物ZPC同源的糖蛋白，在鹌鹑卵巢发育卵泡的颗粒细胞中被睾酮刺激产生，然而，对鸟类卵周膜的另一个成分，ZP 1的分子生物学知之甚少，特别是缺乏有关基因表达的信息。我们克隆了日本鹌鹑的ZP 1基因，并检测了其基因表达。利用RT-PCR和快速扩增技术从成年雌性鹌鹑肝脏中克隆了一个编码ZP 1基因的cDNA。它编码一个934个氨基酸的蛋白质，与鸡ZP 1具有最大的同源性（87%的同一性）。其N端128 ~ 571个氨基酸是独特的，与其它哺乳动物的同源性不到40%。RT-PCR扩增880个碱基，表明肝脏中的ZP 1 mRNA仅限于成熟的产蛋雌性。ZP 1 mRNA的表达刺激体内治疗与己烯雌酚在未成熟的女性以及男性。这些结果表明雄激素和雌激素协同调节鹌鹑卵周膜蛋白的形成。此外，使用ZP 1转录诱导在男性或未成熟的女性作为环境雌激素的生物标记进行了讨论。

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.