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Identification and biosynthesis of sperm receptor in quail oocyte

鹌鹑卵母细胞精子受体的鉴定及生物合成

基本信息

批准号：
09660300
负责人：
MORI Makoto
金额：
$ 0.51万
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：
1997
资助国家：
日本
起止时间：
1997 至 1998
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09660300/
关键词：
Vitelline membrane Sperm Receptor Fertilization Quail Fowl Follicle Cell division

项目摘要

The avian oocyte is bounded by the thin translucent membrane called vitelline membrane. The vitelline membrane consists of three layers, the inner and outer layers and the continuous membrane. Only the inner layer of vitelline membrane surrounds the oocyte at the time of ovulation. We have found that the granulosa cells of the preovulatory follicles contain 33 kDa glycoprotein (ZPC) one of the major components of the inner layer in quail egg. Though the physiological importance of ZPC for fertilization was pointed out as the candidate of "sperm receptor" in quail, the details in the regulation of ZPC production in granulosa cells during ovulation cycle remains obscure. To investigate the regulatory mechanism of ZPC production we have studied the expression of ZPC during the ovulation cycle by Western blot and Northern blot analyses. Moreover, we cultured the granulosa cells with or without various hormones and growth factors to study the endocrine and paracrine control of ZPC production. From the Western blot and Northern blot analyses in in-vivo study, the expression of ZPC in the granulosa cells of growing follicles increased during follicular maturation with the highest activity in the largest follicle. To investigate the effects of various hormones and growth factors on the ZPC production, the granulosa cells obtained from the three largest follicles were cultured for 72 hr with FSH, LH, EGF, IGF- I, IGF- II, TGF- alpha, TGF- beta, progesterone, estradiol, or testosterone. The most prominent effects on stimulation of ZPC production was observed in the granulosa cells isolated from the third largest follicle cultured by the addition of FSH.Other hormones or growth factors caused no significant increase. These data illustrate that quail granulosa cells produce ZPC in response to FSH.

鸟类卵母细胞被称为卵黄膜的薄的半透明膜所包围。卵黄膜由三层组成，即内层、外层和连续膜。排卵时卵母细胞周围只有内层卵黄膜。我们发现鹌鹑排卵前卵泡颗粒细胞中含有33 kDa糖蛋白（ZPC），它是鹌鹑卵内层的主要成分之一。尽管ZPC作为鹌鹑“精子受体”的候选者在受精过程中具有重要的生理学意义，但其在排卵周期中对颗粒细胞ZPC产生的调控机制尚不清楚。为了研究ZPC产生的调节机制，我们通过Western印迹和北方印迹分析研究了ZPC在排卵周期中的表达。此外，我们培养的颗粒细胞有或没有各种激素和生长因子，研究ZPC的生产的内分泌和旁分泌控制。体内研究中的Western blot和北方印迹分析表明，ZPC在生长卵泡颗粒细胞中的表达随着卵泡成熟而增加，且在最大卵泡中的表达活性最高。为了研究各种激素和生长因子对ZPC产生的影响，将从三个最大卵泡获得的颗粒细胞与FSH、LH、EGF、IGF-1、IGF- II、TGF-α、TGF-β、孕酮、雌二醇或睾酮一起培养72小时.在从第三大卵泡中分离的颗粒细胞中观察到最显著的刺激ZPC产生的效果，通过添加FSH培养。这些数据表明，鹌鹑颗粒细胞产生ZPC响应FSH。