GONADOTROPIN RECEPTOR REGULATION IN VIVO AND IN VITRO

该项目的长期目标是提供一个基本的了解 卵巢细胞促性腺激素受体的调节， 适用于生育调节问题。 本申请是 关注继续长期项目的三个一般性问题 目标，并总结/包含本提案的具体目标：a） 某些生长因子在关键卵巢细胞分化中的作用 过程如FSH依赖性LH受体诱导，B）是否 促性腺激素和/或性类固醇可能反过来调节受体， 人群生长因子，和c）使用无血清培养 研究完整细胞中受体调节的条件。 具体目标 该项目包括以下实验目标：1） 测试血小板衍生生长因子（PDGF）的均质制剂， FSH刺激的LH受体诱导和随后的LH 猪和大鼠颗粒细胞受体依赖性细胞功能 单层，2）检查非PDGF成分的可能性， 血小板提取物可能促进FSH刺激的LH受体诱导 和/或选择的后续LH受体依赖性细胞功能，3） 表征颗粒细胞PDGF受体结合并研究PDGF受体 调节生殖激素在颗粒细胞单层，4）， 表征颗粒细胞表皮生长因子（EGF）受体结合 并研究EGF受体调节生殖激素在体内和 体外，和5）扩大我们的研究，无血清培养环境和 培养的颗粒细胞的性能，通过a）研究 低密度脂蛋白（LDL）和高密度脂蛋白（HDL） 生长因子/促性腺激素刺激的类固醇生成，和B） 表征颗粒细胞亚群培养中的反应 优先富集或缺乏特定的膜受体。

The long-term objective of the project is to provide a basic understanding of ovarian cellular gonadotropin receptor regulation which will eventually be applicable to the problem of fertility regulation. This application is concerned with three general issues which continue the long-term project objective and summarize/encompass the specific aims of this proposal: a) the role of certain growth factors in key ovarian cellular differentiation processes such as FSH-dependent LH receptor induction, b) whether gonadotropins and/or sex steroids might, in turn, modulate receptor populations for growth factors, and c) the use of serum-free culture conditions to study receptor regulation in intact cells. The specific aims of the project consist of the following experimental objectives: 1) to test homogenous prepatations of platelete-derived growth factor (PDGF) upon FSH-stimulated LH receptor induction and upon subsequent LH receptor-dependent cell functions in porcine and rat granulosa cell monolayers, 2) to examine the possibility that a non-PDGF component of platelet extracts might facilitate FSH-stimulated LH receptor induction and/or selected subsequent LH receptor dependent cell functions, 3) to characterize granulosa cell PDGF receptor binding and study PDGF receptor modulation by reproductive hormones in granulosa cell monolayers, 4) to characterize granulosa cell epidermal growth factor (EGF) receptor binding and study EGF receptor modulation by reproductive hormones in vivo and in vitro, and 5) to extend our study of the serum-free culture milieu and the performance of cultured granulosa cells by a) investigating the effects of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) upon growth factor/gonadotropin-stimulated steroidogenesis, and b) characterizing the responses in culture of granulosa cell sub-populations preferentially enriched or lacking in a particular membrane receptor.