GONADOTROPIN RECEPTOR REGULATION IN VIVO AND IN VITRO

该项目的长期目标是提供基本的 了解卵巢细胞促性腺激素受体调节 这最终将适用于生育问题 调控 本申请主要涉及 确定化学和生物特性的一个 已从血小板衍生物中部分纯化的因子 生长因子（PDGF）制剂。 这种材料有助于 通过增强FSH刺激的LH受体分化 颗粒细胞中的诱导。 它似乎是一种独特的 人类转化生长因子β 暂时命名为转化生长因子β样 材料（TGF-B-LM）。 为了推广这些发现，本建议 涉及三个具体目标：1）阐明化学 通过a）将其纯化至电泳， B）确定所述多肽的氨基酸序列， 材料的气相微测序仪技术，和c）通过 产生用于纯化的TGF-B-LM抗体 方法：2）研究TGF-β的生物学特性， a）更详细地分析TGF-β-LM如何增强 FSH刺激的功能性LH/hCG受体的诱导，B） 表征其生长促进或抑制特性， 颗粒细胞或卵丘细胞和卵巢癌的原代培养物 c）利用TGF-β-LM和抗TGF-β-LM 免疫沉淀方法来确定卵巢是否 细胞本身产生TGF-β-LM; 3）分离和鉴定一种 来自猪的FSH和EGF调节的Mr 27，000组分 颗粒细胞，并决定其主要生理作用。

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.