REGULATION OF LH RECEPTORS AND PROGESTERONE SECRETION

本研究的目的是为了更好地了解 调节黄体酮分泌的因素 黄体，以开发更好的避孕药和治疗 不孕不育。这项拟议的研究有五个具体目标。1)至 确定MLH和MCG是否有不同的受体介导 在灵长类黄体细胞中的作用，罗丹明标记的激素将 用来测量黄体生成素受体的侧向迁移率 激光漂白回收。如果MCG，而不是MLH，会使 在灵长类黄体细胞中的促黄体激素受体，将被确定是否 MCG还会导致孕酮分泌的更大增加 而不是MLH。这项研究的结果具有非常重要的意义 对不孕不育治疗的启示。2)确定 两种类固醇激素来源的卵泡来源 黄体细胞，亲脂染料将被用来专门标记 排卵前卵泡或卵泡内的卵泡膜或颗粒细胞 出血体。黄体组织将在月经期间采集。 发情周期的黄体早期、中期和晚期以及 大小黄体细胞的起源根据 荧光、亲脂性染料的存在。3)至 阐明生长因子在控制分裂中的作用， 黄体细胞的分化和功能，细胞的数量 胰岛素样生长因子-I、胰岛素样生长因子II、表皮生长因子和成纤维细胞生长因子的受体将在 发情周期的不同阶段。原位杂交法 Cna探针将用于定位生长因子的mrna。 以确定哪些细胞类型合成了这些因子。后续 研究将评估生长因子的作用和控制 它们在黄体细胞纯化制剂中的分泌。4)至 两种类固醇激素合成的调控研究 黄体细胞，初步研究将评估脂蛋白 路径。钙的作用将用“膜片钳”来研究。 评估钙通道数量和活性的程序 荧光染料将被用来监测细胞内钙离子 轮班。钙调素和蛋白质的抑制物和刺激物 激酶C活性将被用来评估这些蛋白的活性 第二个信使。5)最后，黄体细胞的数量，其 受体含量和对各种激素的反应性将是 在两种类固醇生成黄体细胞类型中确定 细胞水平的变化发生在细胞的过渡 动情周期的黄体到黄体 怀孕了。

The goal of this research is to obtain a better understanding of the factors which regulate secretion of progesterone from the corpus luteum in order to develop better contraceptives and treat infertility. The proposed research has five specific aims. 1) To determine if mLH and mCG have different receptor-mediated actions in primate luteal cells, rhodamine-labeled hormones will be used to measure the lateral mobility of the receptor for LH by laser photobleach recovery. If mCG, but not mLH, immobilizes the receptor for LH in primate luteal cells, it will be determined if mCG also causes a greater increase in secretion of progesterone than mLH. The results of this study have very important implications for treatment of infertility. 2) To determine the follicular source of origin of the two types of steroidogenic luteal cells, lipophilic dyes will be used to specifically label thecal or granulosal cells in the preovulatory follicle or the corpus hemorrhagicum. Luteal tissue will be collected during the early, mid and late luteal phases of the estrous cycle and the origins of large and small luteal cells determined based on the presence of the fluorescent, lipophilic dyes. 3) To elucidate the role of growth factors in controlling the division, differentiation and function of luteal cells, the number of receptors for IGF-I, IGF-II, EGF and FGF will be determined at different stages of the estrous cycle. In situ hybridization of cRNA probes will be used to localize mRNA for the growth factors to determine which cell types synthesize these factors. Subsequent studies will evaluate the actions of growth factors and control of their secretion in purified preparations of luteal cells. 4) To study control of steroidogenesis in the two types of steroidogenic luteal cells, initial studies will evaluate the lipoprotein pathway. The role of calcium will be studied using "patch clamp" procedures to evaluate the number and activity of calcium channels and fluorescent dyes will be used to monitor intracellular calcium shifts. Inhibitors and stimulators of calmodulin and protein kinase C activity will be used to evaluate the activity of these second messengers. 5) Finally, numbers of luteal cells, their receptor content and responsiveness to various hormones will be determined in the two steroidogenic luteal cell types to define at the cellular level changes occurring during transition of the corpus luteum of the estrous cycle to the corpus luteum of pregnancy.