PURIFICATION OF LOW M FSH BINDING INHIBITOR

卵巢卵泡液中低分子量因子的存在 抑制FSH与受体的结合，提示局部调节FSH受体的表达。 促性腺激素作用 这种调制预计将发挥 在控制卵泡发生和选择卵泡中的重要作用 卵泡排卵。 该项目的具体目标是： 从牛卵泡液中纯化该因子，和B）测定其 化学特性、作用机制和药理潜力。 将牛卵泡液超滤以获得脱盐部分 含有分子量在500和5000之间的组分。 将从该级分纯化推定的调节肽。 伴随凝胶过滤、离子交换和反相（RP-HPLC） 液相色谱法 将通过RP-HPLC薄层色谱法测定纯度 色谱法和质谱法。 化学特性（组成和 序列）的纯化将通过手动Edman 降解和质谱分析。 将通过以下方式检查作用机制： 研究：1）纯化的抑制剂对FSH与 受体，2）放射性标记的抑制剂与性腺（Sertoli和 颗粒细胞）组织和3）抑制剂影响其他 培养的性腺（支持和颗粒）细胞中对FSH的反应（即， 激动剂对拮抗剂活性）。 最后，药理学潜力将 通过检查纯化的抑制剂的功效来研究， 阻断FSH在体内的作用。 该项目的长期目标是 鉴定天然存在的FSH抑制剂，并提供 有关其性质、作为药物的药理潜力的信息 避孕药和有用性的基础研究FSH的作用。 这些研究将有助于了解人类卵巢功能， 功能障碍，因为FSH作用的局部调节剂是一种潜在的药物， 调节卵泡生成和人类生育力的靶点。

The presence of a low molecular weight factor in ovarian follicular fluid which inhibits the binding of FSH to receptor suggests local modulation of gonadotropin action. Such modulation would be expected to play a significant role in the control of folliculogenesis and selection of follicles for ovulation. The specific aims of this project are: a) to purify this factor from bovine follicular fluid and b) determine its chemical identity, mechanisms of action and pharmacological potential. Bovine follicular fluid will be ultrafiltered to obtain a desalted fraction containing components of molecular weights between 500 and 5,000. Purification of the putative regulatory peptide from this fraction will be accompanied by gel filtration, ion-exchange and reverse-phase (RP-HPLC) liquid chromatography. Purity will be determined by RP-HPLC, thin-layer chromatography and mass spectroscopy. Chemical identity (composition and sequence) of the purified factor will be accomplished by manual Edman degradation and mass spectroscopy. Mechanism of action will be examined by studying: 1) effects of purified inhibitor on the binding of FSH to receptor, 2) binding of radiolabeled inhibitor to gonadal (sertoli and granulosa cells) tissue and 3) ability of inhibitor to affect other responses to FSH in cultured gonadal (sertoli and granulosa) cells (i.e., agonist vs antagonist activity). Finally, the pharmacologic potential will be investigated by examining the efficacy of the purified inhibitor in blocking FSH actions in vivo. The long term objectives of this project are to identify a naturally occurring inhibitor of FSH and to provide information regarding its nature, pharmacologic potential as a contraceptive agent and usefulness in basic research on the action of FSH. These studies will be relevant to understanding human ovarian function and dysfunction since a local modulator of FSH action is a potential agent or target for the regulation of folliculogenesis and human fertility.