MOLECULAR DISTRIBUTION OF HUMAN ADRENERGIC RECEPTORS

人肾上腺素受体的分子分布

• 批准号: 2225203
• 负责人: Debra Anne Schwinn
• 金额: $ 22.33万
• 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/2225203
• 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.

本项目的长期目标是将肾上腺素能受体 (AR)亚型分子分布与激素/药物相互作用， 诱发健康和疾病时的人体生理反应。 在这 在这种情况下，这项赠款的总体重点是表征 人AR亚型的分子分布。 AR在以下方面发挥着关键作用： 调节生理过程，如心肌血流量，平滑 肌肉收缩、肝脏代谢和全身动脉的控制 血压，并已被证明在数量和功能上发生变化 在不同的疾病状态。 最近，首席研究员 发现了AR分布中惊人的物种异质性 亚型，使AR亚型的特征和分布， 人体组织必需品 因此，我们建议将 所有九种克隆的AR亚型在人体组织中的分布， 检验AR亚型特异性变化发生在 心血管系统疾病。 选择性激动剂和 尚未针对每种AR亚型开发拮抗剂，因此直接 诸如放射自显影和配体结合的技术不能用于 这个项目 首席研究员是独一无二的， 所有九个AR亚型基因的序列和人类组织的访问， 使得原位杂交技术能够用于定位 编码AR亚型的mRNA在特定细胞层中的分布， 人体组织切片 因此，第一个主要目标是评估 AR亚型mRNA在正常组织中的分布 心血管系统、外周组织和中枢神经系统 将用于人类AR定位的广泛调查; 使用来自人肾动脉的组织切片的初步研究， 海马、[前列腺和脊髓]证明了RNA的活力 在快速尸检中获得的组织中。 第二个主要目的是 建议是评估人类AR亚型分布的变化 心血管疾病 从众多病人身上挑选的人体组织 有或没有并发心血管疾病， 废弃手术标本的首席研究员。 手术 标本将用于定量AR亚型mRNA的变化（比较 与正常组织）的患者（见 选定疾病和组织简明清单的具体目标 为本次研究提供）。 许多用于调节心血管疾病的药物 反应是AR的激动剂和拮抗剂;因此， 不同AR亚型在人体组织中的定位增强了 更有选择性的药物， 生理反应。 此外，定量mRNA的变化， 在特定组织中编码特定疾病的不同AR亚型 （结合未来对受体蛋白的直接研究， 配体和抗体是可用的），对于最终 了解人类疾病的机制，涉及肾上腺素能 神经系统