MOLECULAR DISTRIBUTION OF HUMAN ADRENERGIC RECEPTORS

人肾上腺素能受体的分子分布

• 批准号: 3368222
• 负责人: Debra Anne Schwinn
• 金额: $ 29.72万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1993
• 资助国家: 美国
• 起止时间: 1993-09-01 至 1998-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3368222
• 关键词: adrenergic receptor; autoradiography; blood vessels; brain; cardiovascular disorder; cardiovascular function; cell bank /registry; congestive heart failure; diabetes mellitus; heart; heart transplantation; human tissue; hypertension; in situ hybridization; messenger RNA; nucleic acid sequence; polymerase chain reaction; postmortem; radiotracer; spinal cord

The long term objective of this project is to relate adrenergic receptor (AR) subtype molecular distribution to hormone/drug interaction and evoked human physiologic responses in health and disease. Within this context, the overall focus of this grant is characterization of the molecular distribution of human AR subtypes. ARs play key roles in the regulation of physiologic processes such as myocardial blood flow, smooth muscle contraction, liver metabolism, and control of systemic arterial blood pressure, and have been shown to be altered in number and function in various disease states. Recently the principal investigator has discovered striking species heterogeneity in the distribution of AR subtypes, making characterization and distribution of AR subtypes in human tissue essential. Therefore we propose characterizing the distribution of all nine cloned AR subtypes in human tissue in order to test the hypothesis that AR subtype specific changes occur during diseases of the cardiovascular system. Selective agonists and antagonists have not yet been developed for each AR subtype, hence direct techniques such as autoradiography and ligand binding cannot be used for this project. The principal investigator is unique in having human sequence for all nine AR subtype genes and access to human tissues, enabling in situ hybridization techniques to be employed in localizing the distribution of mRNA encoding AR subtypes to specific cell layers in human tissue sections. Therefore the first major aim is evaluation of the distribution of AR subtype mRNA in normal tissue from the cardiovascular system, peripheral tissues, and central nervous system will be used for an extensive survey of human AR localization; preliminary studies using tissue sections from human renal artery, hippocampus, [prostate, and spinal cord] demonstrate the viability of RNA in tissue obtained at rapid autopsy. The second major aim of this proposal is evaluation of changes in distribution of AR subtypes in human cardiovascular disease. Selected human tissue from numerous patients with and without coincident cardiovascular diseases is available to the principal investigator for discarded surgical specimens. Surgical specimens will be used to quantitate AR subtype mRNA changes (compared with normal tissue) in patients with selected coincident diseases (see specific aims for a concise list of selected diseases and tissues proposed for this study). Many drugs used in modulating cardiovascular responses are agonists and antagonists of ARs; therefore defining the location of various AR subtypes in human tissues enhances the development of more selective pharmacological agents designed for specific physiological responses. In addition, quantitation of changes in mRNA encoding distinct AR subtypes in specific tissues for a given disease (combined with future direct studies of receptor protein once selective ligands and antibodies are available), is critical for ultimately understanding mechanisms of human diseases involving the adrenergic nervous system.

该项目的长期目标是将肾上腺素受体 (AR) 激素/药物相互作用的亚型分子分布和 引起人类健康和疾病的生理反应。 在此范围内 背景下，这笔赠款的总体重点是表征 人类 AR 亚型的分子分布。 AR 在其中发挥着关键作用 调节心肌血流、平滑肌等生理过程 肌肉收缩、肝脏代谢和全身动脉的控制 血压，并已被证明数量和功能发生改变 在各种疾病状态下。 最近，首席研究员 发现 AR 分布存在显着的物种异质性 亚型，对 AR 亚型进行表征和分布 人体组织必不可少。 因此我们建议表征 所有九种克隆 AR 亚型在人体组织中的分布，以便 检验 AR 亚型特定变化发生期间的假设 心血管系统疾病。 选择性激动剂和 尚未针对每种 AR 亚型开发出拮抗剂，因此直接 放射自显影和配体结合等技术不能用于 这个项目。 首席研究员的独特之处在于拥有人类 所有九个 AR 亚型基因的序列以及进入人体组织的途径， 使原位杂交技术能够用于定位 编码 AR 亚型的 mRNA 分布到特定细胞层 人体组织切片。 因此，首要目标是评估 AR亚型mRNA在正常组织中的分布 心血管系统、周围组织、中枢神经系统 将用于人类 AR 定位的广泛调查； 使用人类肾动脉的组织切片进行初步研究， 海马、[前列腺和脊髓] 证明了 RNA 的活力 在快速尸检获得的组织中。 本次活动的第二个主要目标 提案是评估人类 AR 亚型分布的变化 心血管疾病。 从众多患者中精选的人体组织 有或没有同时患有心血管疾病的人均可获得 废弃手术标本的主要研究者。 外科 样本将用于定量 AR 亚型 mRNA 变化（比较 与正常组织）在患有选定的重合疾病的患者中（参见 选定疾病和组织的简明清单的具体目标 为这项研究提出的）。 许多药物用于调节心血管 反应是 AR 的激动剂和拮抗剂；因此定义 各种 AR 亚型在人体组织中的位置促进了发育 专为特定目的而设计的更具选择性的药物制剂 生理反应。 此外，mRNA变化的定量 针对特定疾病在特定组织中编码不同的 AR 亚型 （结合未来对受体蛋白的直接研究一旦选择性 配体和抗体可用），对于最终至关重要 了解涉及肾上腺素能的人类疾病的机制 神经系统。