HORMONE ACTION IN THE OVARY: ROLE OF CALCIUM IONS

卵巢中的激素作用：钙离子的作用

• 批准号: 3313965
• 负责人: JOHANNES D VELDHUIS
• 金额: $ 14.95万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-04-01 至 1995-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3313965
• 关键词: angiotensin II; calcium flux; cell differentiation; cyclic AMP; diacylglycerols; exocytosis; fluorescence microscopy; follicle stimulating hormone; granulosa cell; hormone regulation /control mechanism; intracellular transport; protein kinase C; second messengers; single cell analysis; steroid hormone biosynthesis; swine; tissue /cell culture; video recording system

Our commitment to studying the role of calcium effector pathways in ovarian differentiation will be continued by focusing on the regulatory actions of the calcium-phospholipid-diacylglycerol (PKC) second-messenger system in a well defined and established in vitro model of swine granulosa cells that retain responsivity to a variety of hormonal agonists. We will particularly investigate the manner in which FSH and angiotensin II (presumptively acting respectively through distinct cyclic AMP and PKC intracellular signaling pathways) alter intracellular calcium ion concentrations in single porcine granulosa cells (Time-resolved fluorescence video-microscopy); how hormonal agonists trigger increased secretory activity by single ovarian cells (reverse hemolytic plaque assay of relaxin secretion by pig luteal cells); employ a microperifusion system to test the temporal concordance between intracellular calcium-ion concentration changes and secretory events; probe the causal association between increased intracellular calcium-ion concentrations and resultant secretion using experimentally induced calcium transients in permeabilized cells; and begin to dissect the important interactions between the calcium effector signaling pathway and the cyclic AMP second messenger pathway on sterol-metabolizing enzymes. These emphases reflects work carried out under my parallel RCDA, in which I have has occasion to learn appropriate single-cell and reproductive biology techniques that can now be applied to this R01 renewal. The significance of the calcium second-messenger system in the ovary will therefore be explored at the single cell level (Specific Aim I), in relation to calcium's causal relevance to hormone secretion (Specific Aim II), and in relation to the interactions between the PKC and classical cyclic AMP effector pathways (interactions to be studied at the level of steroidogenesis). Overall, we anticipate that these approaches will yield significant and interesting new information regarding ovarian cellular physiology, which will enhance our physiological understanding or reproductive biology and fertility regulation in higher mammals.

我们致力于研究钙效应途径在卵巢中的作用 将继续关注监管行动的差异化 钙-磷脂-二酰基甘油（PKC）第二信使系统 明确和建立的猪颗粒细胞体外模型 保持对多种激素激动剂的反应性。 我们将 特别研究 FSH 和血管紧张素 II 的作用方式 （推测分别通过不同的环 AMP 和 PKC 起作用 细胞内信号通路）改变细胞内钙离子 单个猪颗粒细胞中的浓度（时间分辨 荧光视频显微镜）；荷尔蒙激动剂如何触发增加 单个卵巢细胞的分泌活性（反向溶血斑测定 猪黄体细胞分泌的松弛素）；采用微灌注系统 测试细胞内钙离子之间的时间一致性 浓度变化和分泌事件；探究因果关系 细胞内钙离子浓度增加和由此产生的 使用实验诱导的钙瞬变在透化细胞中进行分泌 细胞；并开始剖析钙之间的重要相互作用 效应信号通路和环AMP第二信使通路 甾醇代谢酶。 这些重点反映了已开展的工作 在我的平行 RCDA 下，我有机会学习适当的 单细胞和生殖生物学技术现在可应用于 这次R01更新。 钙第二信使系统的意义 因此，将在单细胞水平上探索卵巢中的（具体 目标 I)，关于钙与激素分泌的因果关系 （具体目标 II），以及关于 PKC 和 经典的环 AMP 效应通路（相互作用待研究） 类固醇生成水平）。 总的来说，我们预计这些方法 将产生有关卵巢的重要且有趣的新信息 细胞生理学，这将增强我们的生理理解或 高等哺乳动物的生殖生物学和生育力调节。