CONTROL OF RELAXIN SECRETION FROM SINGLE LUTEAL CELLS

卵巢分泌类固醇激素和一系列蛋白质。 荷尔蒙对控制疾病非常重要 生殖功能。然而，目前人们对此知之甚少。 调节卵巢释放的细胞和分子途径 蛋白质荷尔蒙。在这些拟议的研究中，更完整的 了解一种这样的荷尔蒙，蛋白质荷尔蒙松弛素， 将尝试通过表征分泌物和 抑制分泌功能的细胞内信号通路， 并对结构-功能关系进行了分析。 这些目标将通过使用反向溶血斑块来实现 化验。这项检测是基于抗体导向、补体- 与激素分泌相邻的红细胞的介导裂解，因此 使激素释放的微观可视化成为可能 培养中的单个黄体细胞。该计划的主要目标 建议的研究是：(A)评估以下各项的相对重要性： 和差异反应，刺激性(前列腺素)和 用于松弛的抑制性(促黄体生成素，自身抑制)促分泌剂 释放；(B)描述影响、相互作用和 秒的相对重要性和差异化响应 信使通路，如环核苷酸、钙 动员、蛋白激酶C活化与花生四烯酸 代谢物，并确定黄体细胞是否具有兴奋性 细胞；及(C)测定黄体的分泌特性 含有松弛素但不释放松弛素的细胞(“沉默”) 细胞“)。为了实现这些目标，分泌特征 单一激素-分泌物的数量将被联合监测 (在适当的情况下)采用免疫细胞化学技术。它是 预计将获得有关以下方面的有意义的信息 细胞和分子的同一性、重要性和相互作用 抑制卵巢激素释放的机制。此外，新的 对不释放的分泌细胞之谜的洞察将是 收获了。这种知识最终不仅可以用来识别 卵巢疾病的原因(和正在开发的治疗方法)， 也是为了找出调节人类卵巢的新策略 功能。

The ovary secretes both steroid hormones and a spectrum of protein hormones that are of great importance to the control of reproductive function. However, little is presently known of the cellular and molecular pathways that regulate release of ovarian protein hormones. In these proposed studies, a more complete understanding of one such hormone, the protein hormone relaxin, will be attempted by characterizing the secretagogues and intracellular signalling pathways that subserve secretory function, together with an analysis of structure-function relationships. These objectives will be met by use of a reverse hemolytic plaque assay. This assay is based on antibody-directed, complement- mediated lysis of erythrocytes adjacent to hormone secretors, thus enabling the microscopic visualization of hormone release from individual luteal cells in culture. The primary objectives of the proposed studies are: (a) to evaluate the relative importance of, and differential response to, stimulatory (prostaglandins) and inhibitory (LH, auto-suppression) secretagogues for relaxin release; (b) to characterize the influence, interactions and relative importance of, and differential response to, second messenger pathways such as cyclic nucleotides, calcium mobilization, protein kinase C activation and arachidonic acid metabolites, and to determine if luteal cells function as excitable cells; and (c) to determine the secretory characteristics of luteal cells that contain relaxin but do not release relaxin ("silent" cells"). To accomplish these goals, the secretory characteristics of single hormone-secretors will be monitored, in combination (where appropriate) with immunocytochemical techniques. It is anticipated that meaningful information will be derived concerning the identity, importance and interactions of cellular and molecular mechanisms that subserve ovarian hormone release. Moreover, new insights into the enigma of non-releasing secretory cells will be gained. This knowledge can eventually be used not only to identify the causes of (and developing treatments for) ovarian pathologies, but also to identify new strategies for regulating human ovarian function.