THE ROLE OF BIOACTIVE FSH IN REPRODUCTION

生物活性 FSH 在生殖中的作用

• 批准号: 2198809
• 负责人: AARON JW HSUEH
• 金额: $ 14.73万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-04-01 至 1997-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2198809
• 关键词: autoantibody; bioassay; biological signal transduction; follicle stimulating hormone; glycosylation; hormone binding protein; hormone inhibitor; hormone receptor; human genetic material tag; human tissue; laboratory rat; ligands; luminescence; luteinizing hormone; method development; ovary disorder; polymerase chain reaction; precocious puberty; protein structure function; receptor binding; receptor expression; recombinant proteins; reproduction; site directed mutagenesis; spontaneous abortion; transfection

Recent cloning of cDNAs for LH, FSH and TSH receptors indicated these proteins belong to a sub-family of G-protein-coupled receptors with seven transmembrane regions and a large extracellular domain. We have recently isolated cDNAs for both human LH and FSH receptors and expressed the functional proteins capable of signal transduction in transfected cells. Binding studies indicated that the human receptors, unlike their rodent counterparts, have unique species-specific ligand recognition properties, underlying the importance of using the human receptors to study human physiology and pathophysiology. Because truncated rat LH receptor with only the extracellular region retains full ligand binding, we propose to study the ligand binding and signal transduction of human LH and FSH receptors with altered N-linked carbohydrates and peptide backbone in the extracellular region. Using a recently adapted RNA virus expression system, we will express large amounts of the extracellular region of recombinant human LH and FSH receptors with full ligand binding ability for testing as receptor antagonists in vitro and in vivo. Using the human LH receptor and a reporter gene driven by a gonadotropinresponsive promoter, we have developed a luminescence bioassay for human LH and CG. We will use this bioassay to screen for defective hCG molecules during spontaneous abortion. Because auto-antibodies to TSH receptors have been shown to be involved in different thyroid diseases, we propose to use recombinant human gonadotropin receptors to screen for inhibitory and stimulatory receptor autoantibodies in sera of patients with premature ovarian failure and testotoxicosis, respectively. The present proposal should yield truncated human receptors capable of selectively antagonizing the actions of human LH and FSH in patients,, as well as further our understanding of diseases with defects of gonadotropins and their receptors.

LH，FSH和TSH受体的CDNA最近克隆表明了这些 蛋白质属于G蛋白偶联受体的亚家族，七个 跨膜区域和一个较大的细胞外域。 我们最近有 人LH和FSH受体的分离cDNA，并表达 能够在转染细胞中信号转导的功能蛋白。 结合研究表明，与啮齿动物不同 对应物具有独特的物种特异性配体识别特性， 使用人体受体研究人的重要性 生理学和病理生理学。 因为截短的大鼠LH受体与 只有细胞外区域保留完整的配体结合，我们建议 研究人LH和FSH的配体结合和信号转导 N-连接碳水化合物和肽主链改变的受体在 细胞外区域。 使用最近适应的RNA病毒表达 系统，我们将表达大量的细胞外区域 重组人LH和具有完整配体结合能力的FSH受体 用于在体外和体内作为受体拮抗剂的测试。 使用 人类LH受体和由促性腺激素反应驱动的记者基因 启动子，我们为人LH和CG开发了一种发光生物测定。 我们将使用此生物测定法筛选出有缺陷的HCG分子 自发流产。 因为对TSH受体的自动抗体已经 我们建议使用不同的甲状腺疾病，我们建议使用 重组人促性腺激素受体以筛选抑制性和 早产患者血清中的刺激受体自身抗体 卵巢衰竭和测试毒性。 目前的提议 应该产生能够选择性的人类受体 对抗人LH和FSH对患者的作用，以及 进一步，我们对促性腺激素缺陷和 他们的受体。