Chicago Center for Reproductive Research

描述（由申请人提供）：本申请寻求芝加哥生殖研究中心（U 54）的资金。该中心将由四个项目和三个核心组成，并将对生殖轴中的类固醇激素、促性腺激素和胰岛素信号通路进行多学科研究。项目1，临床研究项目，将确定一种变异的类固醇生成酶17 β-羟基类固醇脱氢酶5型（17 β-HSD 5），负责卵巢睾酮生产的主要酶，在多囊卵巢综合征（PCOS）中的作用。卵巢睾酮分泌对体内促黄体生成激素（LH）和胰岛素水平的调节反应与17 β-HSD 5基因型有关，并可能为特定遗传性状导致的PCOS表型提供第一个直接证据。项目2的目的是利用一系列雌激素受体（ER）突变体和基因敲除（KO）小鼠，确定细胞系和转基因小鼠中雌激素调节中枢生殖轴的机制。在促性腺激素释放激素（GnRH）神经元或促性腺激素中携带ER-α和/或ER-β的条件性敲除的小鼠将确定雌激素负反馈控制的机制。项目3，密切补充项目2，将测试的假设，孕酮受体（PR）介导的孕酮对GnRH神经元的抑制作用，并确定参与这种调节机制的PR的解剖位置。计划进行特定实验以确定GnRH神经元是否表达PR并介导GnRH脉动性的抑制。项目4旨在确定胰岛素调节在促性腺激素特异性基因的发育和表达中的重要性。目的将使用胰岛素受体和cAMP反应元件（CREB）结合蛋白（CBP）的条件S，以及探索胰岛素信号传导在CBP募集到转录复合物中的作用。 分子技术核心将为研究者提供及时获取DNA序列和常规和/或专业分子生物学程序的性能，用于生产转基因小鼠的试剂以及对他们构建的小鼠品系进行组织学分析。 配体测定核心将为涉及人类受试者和动物模型的研究提供激素测定。拟议中的芝加哥生殖研究中心还将利用芝加哥大学和西北大学的现有核心。美国国立卫生研究院资助的一般临床研究中心（GCRC）在芝加哥大学，其中博士罗伯特L。罗森菲尔德是副项目主任，也将是中心参与者的重要资源。预计在该中心进行的研究将导致增加对生殖信号通路的理解。

DESCRIPTION (provided by applicant): This application seeks funding for a Chicago Center for Reproductive Research (U54). The Center will consist of four projects and three cores and will carry out multidisciplinary investigations of steroid hormone, gonadotropin, and insulin signaling pathways in the reproductive axis. Project 1, the clinical research project, will determine the role of a variant steroidogenic enzyme 17beta-hydroxysteroid dehydrogenase type 5 (17beta-HSD5), the major enzyme responsible for ovarian testosterone production, in the polycystic ovary syndrome (PCOS). Ovarian testosterone secretion in response to in vivo manipulation of luteinizing hormone (LH) and insulin levels will be related to 17beta-HSD5 genotype and may provide the first direct evidence for a PCOS phenotype resulting from a specific genetic trait. Project 2 will aim to determine the mechanism of estrogen regulation in the central reproductive axis in cell lines and transgenic mice using a series of estrogen receptor (ER) mutants and knockout (KO) mice. Mice bearing a conditional knock-out of ER-alpha and/or ER-beta in either the gonadotropin releasing hormone (GnRH) neuron or gonadotrope will determine the mechanism of negative feedback control by estrogen. Project 3, closely complementing Project 2, will test the hypothesis that progesterone receptors (PRs) mediate the inhibitory effects of progesterone on the GnRH neuron, and determine the anatomic locations of PRs involved in this regulatory mechanism. Specific experiments are planned to determine whether GnRH neurons express PRs and mediate inhibition of GnRH pulsatility. Project 4 aims to determine the importance of insulin regulation in the development and expression of gonadotrope-specific genes. The aims will use conditional s of the insulin receptor and the cAMP response element (CREB) binding protein (CBP) as well as explore the role of insulin signaling in CBP recruitment to the transcription complex. The Molecular Technology Core, will provide investigators with timely acquisition of DNA sequence and performance of routine and/or specialized molecular biology procedures, the reagents for production of transgenic and mice and the histological analysis of the mouse lines they construct. The Ligand Assay Core, will provide hormone assays for studies involving human subjects and animal models. The proposed Chicago Center for Reproductive Research will also utilize existing cores at the University of Chicago and Northwestern University. The NIH-funded General Clinical Research Center (GCRC) at the University of Chicago, where Dr. Robert L. Rosenfield is Associate Program Director, will also be an important resource for Center participants. It is anticipated that the research performed under this Center will lead to an increased understanding of signaling pathways in reproduction.