GONADOTROPIN RECEPTOR REGULATION IN VIVO AND IN VITRO

该项目的长期目标是提供基本的 对卵巢细胞促性腺激素受体调节的认识 它最终将适用于生育率问题 监管。这个应用程序主要涉及 确定一种细菌的化学和生物学特性 从血小板源中部分纯化的因子 生长因子(PDGF)制剂。这种材料有助于 增强FSH刺激的促黄体生成素受体诱导分化 颗粒细胞的诱导。它似乎是一种独特的 我们所处的人类转化生长因子β 暂定为转化生长因子-β样蛋白 材料(转化生长因子-B-微球)。为了扩大这些发现，这项建议是 涉及三个具体目的：1)阐明该化学物质 A)电泳法纯化转化生长因子-β-微球蛋白的性质 均一性，b)测定氨基酸序列 通过气相微测序技术获得材料，以及c)通过 用于纯化的转化生长因子-β-LM抗体的制备 2)研究转化生长因子-β的生物学特性。 通过a)更详细地分析转化生长因子-β-脂蛋白是如何增强 卵泡刺激素刺激功能性黄体生成素/人绒毛膜促性腺激素受体的诱导 表征其促生长或抑制生长的特性 颗粒细胞或卵丘细胞的原代培养与卵巢癌 C)利用转化生长因子-B-M和抗-转化生长因子-B-M 免疫沉淀法测定卵巢组织中 细胞自身产生转化生长因子-β-M；3)分离和鉴定 FSH和EGF调控的猪MR 27,000组分 颗粒细胞，并确定其主要生理作用。

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 primarily with determining the chemical and biological characteristics of a factor which has been partially purified from platelet-derived growth factor (PDGF) preparations. This material facilitates differentiation by enhancing FSH-stimulated LH receptor induction in granulosa cells. It appears to be a unique species of human Transforming Growth Factor Beta which we are provisionally designating Transforming Growth Factor-Beta-Like Material (TGF-B-LM). To extend these findings, this proposal is concerned with three specific aims: 1) to elucidate the chemical properties of TGF-B-LM by a) purifying it to electrophoretic homogeneity, b) determining the amino acid sequence of the material by gas-phase microsequenator technology, and c) by generating antibodies to TGF-B-LM for application to purification procedures; 2) to investigate the biological properties of TGF-B- LM by a) analyzing in greater detail how TGF-B-LM potentiates FSH-stimulated induction of functional LH/hCG receptor, b) characterizing its growth-promoting or inhibiting properties upon primary cultures of granulosa or cumulus cells and ovarian cancer cell lines and c) utilizing TGF-B-LM and anti-TGF-B-LM immunoprecipitation approaches to determine whether ovarian cells themselves produce TGF-B-LM; 3) to isolate and identify an FSH- and EGF-modulated Mr 27,000 component from porcine granulosa cells and determine its principal physiological role.