MOLECULAR BIOLOGY OF HORMONES THAT REGULATE REPRODUCTION

调节生殖的激素的分子生物学

• 批准号: 3073506
• 负责人: KELLY E MAYO
• 金额: $ 6.35万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-02-01 至 1996-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3073506
• 关键词: animal tissue; brain; clone cells; developmental genetics; estrus; gene expression; genetic regulatory element; gonadotropin releasing factor; granulosa cell; hormone regulation /control mechanism; in situ hybridization; inhibin; laboratory rat; neuroendocrine system; peptide hormone; placenta; reproductive hormone; transfection

The goal of this project is to examine the expression and regulation of genes encoding brain and gonadal hormones that are involved in controlling mammalian reproduction. Four polypeptide hormones that modulate the synthesis and secretion of the pituitary gonadotropins have been described. These are the hypothalamic peptide gonadotropin-releasing hormone (GnRH), the related gonadal hormones inhibin and activin, and the recently described ovarian hormone follistatin. Inhibin, activin, and follistatin regulate the secretion of follicle-stimulating hormone (FSH) and GnRH regulates the secretion of FSH and lutienizing hormone (LH) from the pituitary. Because FSH and LH play a crucial role in modulating the gametogenic and steroidogenic functions of the gonads, these regulatory hormones are likely to be important for homeostasis in the reproductive system. We have isolated cDNA clones from the rat for all of these peptide hormones: GnRH, follistatin, and the alpha- and beta-chains of inhibin/activin. We have used these cDNAs to examine expression of the corresponding mRNAs in reproductive tissues using in situ hybridization histochemistry, and have begun to explore the regulation of these genes during the rat estrous cycle and during pregnancy. We now propose to extend these studies in two directions. Firstly, we plan to explore the molecular mechanisms by which these genes are regulated. We will complete in vivo experiments to examine changes in GnRH, inhibin, and activin mRNA levels during the estrous cycle and in response to specific hormones. We will develop in vitro assays, including ovarian organ cultures and primary rat granulosa cell cultures, to directly examine the effects of steroid and polypeptide hormones on inhibin gene expression. The granulosa cells will also be utilized for transfection studies in which inhibin and activin gene regulatory elements will be characterized. Secondly, we plan to examine the biological significance of expression of these genes in several novel tissues. The inhibin/activin beta-chain mRNA has been found in diverse tissues including the brain, pituitary, hematopoietic system, and the placenta. This is particularly intriguing in light of its similarity to the transforming growth factor-beta family of growth and differentiation regulators. We will investigate when in development the inhibin/activin beta-chain gene is expressed in these tissues, and determine whether the gene is regulated in a manner similar to that observed in the gonads. We will also examine extra-gonadal expression of the rat follistatin gene, pursuing our recent observation that this gene is highly expressed in decidual tissue early in pregnancy. To aid in these studies, we will generate immunological reagents specific for the activin and follistatin proteins. We expect these experiments to enhance our understanding of the roles these hormones play in regulating normal reproduction, and to provide a framework for examining how alterations in their activities might lead to reproductive disorders or dysfunction.

该项目的目标是检查表达和调节 编码参与控制大脑和性腺激素的基因 哺乳动物的繁殖。 调节四种多肽激素 已经描述了垂体促性腺激素的合成和分泌。 这些是下丘脑肽促性腺激素释放激素（GnRH）， 相关的性腺激素抑制素和激活素，以及最近 描述了卵巢激素卵泡抑素。 抑制素、激活素和卵泡抑素 调节促卵泡激素 (FSH) 和 GnRH 的分泌 调节 FSH 和黄体生成素 (LH) 的分泌 垂体。 因为 FSH 和 LH 在调节 性腺的配子生成和类固醇生成功能，这些调节 激素可能对生殖系统的稳态很重要 系统。 我们从大鼠中分离出所有这些肽的 cDNA 克隆 激素：GnRH、卵泡抑素以及α链和β链 抑制素/激活素。 我们使用这些 cDNA 来检查 使用原位杂交在生殖组织中检测相应的 mRNA 组织化学，并开始探索这些基因的调控 在大鼠发情周期和妊娠期间。 我们现在建议 将这些研究扩展到两个方向。 首先，我们计划探索 调节这些基因的分子机制。 我们将完成 体内实验检查 GnRH、抑制素和激活素 mRNA 的变化 发情周期期间的水平以及对特定激素的反应。 我们 将开发体外测定，包括卵巢器官培养和原代培养 大鼠颗粒细胞培养物，直接检查类固醇和 多肽激素对抑制素基因表达的影响。 颗粒细胞将 也可用于转染研究，其中抑制素和激活素基因 监管要素将得到表征。 其次，我们计划考察 这些基因在几种新颖的药物中表达的生物学意义 组织。 抑制素/激活素 β 链 mRNA 已在多种体内发现 组织包括脑、垂体、造血系统和 胎盘。 鉴于其与以下内容的相似性，这一点特别有趣： 生长和分化的转化生长因子-β家族 监管机构。 我们将调查抑制素/激活素的开发时间 β链基因在这些组织中表达，并确定是否 基因的调节方式与在性腺中观察到的类似。 我们 还将检查大鼠卵泡抑素基因的性腺外表达， 根据我们最近的观察，该基因在 妊娠早期的蜕膜组织。 为了帮助这些研究，我们将 生成针对激活素和卵泡抑素的特异性免疫试剂 蛋白质。 我们希望这些实验能够加深我们对 这些激素在调节正常生殖中发挥作用，并提供 一个框架，用于检查其活动的改变如何可能导致 生殖障碍或功能障碍。