RESPONSES OF SUBPOPULATIONS OF MUSCARINIC RECEPTORS

毒蕈碱受体亚群的反应

• 批准号: 3115769
• 负责人: JOHN ELLIS
• 金额: $ 12.56万
• 依托单位: UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
• 依托单位国家: 美国
• 财政年份: 1985
• 资助国家: 美国
• 起止时间: 1985-02-01 至 1991-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3115769
• 关键词: Alzheimer's disease; alkylation; anticholinergic agent; autoradiography; brain cell; cell population study; histochemistry /cytochemistry; immunochemistry; laboratory mouse; laboratory rat; muscarinic receptor; neural information processing; neurochemistry; neurons; phosphorylation; radioassay; receptor binding; stimulus /response; tissue /cell culture

Reduced cholinergic function in the central nervous system contributes to memory disturbances, including those associated with senile dementia of the Alzheimer type. Previous studies by ourselves and others have demonstrated conclusively that distinct subtypes of muscarinic receptors are present in varying proportions in different brain regions. Classical receptor theory dictates that the most useful receptor classification schemes be based on the affinities of antagonists and several selective antagonists are now available (including pirenzepine, AF-DX 116, and quaternary antagonists). Past characterizations of muscarinic responses may have been confounded by tissue-related differences, as the different responses were studied in different tissues or preparations. This proposal intends to correlated sophisticated assays of binding and response in a model in vitro system, so as to relate the responses to given subpopulations. We will evaluate multiple muscarinic responses and receptor binding in parallel, in identical preparations (CNS neurons in primary culture); thereby, the problems of between-tissue differences will be eliminated and the affinities of a given antagonists in the different response assays and binding assays can be compared directly. The ability to generate neuronal cultures from regions of the brain that possess very different distributions of subpopulations will test the uniformity of the relationship between response and receptor subtype. The results of classical pharmacological methods, such as Schild analysis of competitive antagonists, will be supplemented by an independent procedure: selective protection of given subpopulation(s) of receptors from irreversible blockage by alkylating antagonists. Autoradiographic studies will determine whether subpopulations of muscarinic receptors are localized to specific types of (immunocytochemically identified) neurons. Studies of phosphorylation of muscarinic receptors are aimed at determining whether different subpopulations are differentially phosphorylated, desensitized, and down-regulated. The interactions of different kinases in regulating the sensitivity of the receptor(s) will also be investigated. It is expected that these studies will lead to a better understanding of the functions and regulation of different subpopulations of muscarinic receptors.

中枢神经系统胆碱能功能降低 导致记忆障碍，包括相关的记忆障碍 患有阿尔茨海默型老年痴呆症。 之前的研究 我们自己和其他人已经最终证明了这种独特的 毒蕈碱受体的亚型存在于不同的 不同大脑区域的比例。 经典受体理论 表明最有用的受体分类方案是 基于拮抗剂的亲和力和一些选择性 拮抗剂现已上市（包括哌仑西平、AF-DX 116、 和四元拮抗剂）。 毒蕈碱的过去特征 反应可能被组织相关的因素所混淆 差异，因为在不同的环境中研究了不同的反应 组织或制剂。 该提案旨在关联 体外模型中结合和反应的复杂测定 系统，以便将响应与给定的亚群联系起来。 我们 将评估多种毒蕈碱反应和受体结合 平行地，在相同的准备中（初级中枢神经系统神经元） 文化）;因此，组织间差异的问题将 被消除以及给定拮抗剂的亲和力 可以比较不同的反应测定和结合测定 直接地。 从区域生成神经元培养物的能力 具有非常不同的分布的大脑 亚群体将检验关系的一致性 反应和受体亚型之间。 经典的结果 药理学方法，例如 Schild 竞争分析 拮抗剂，将由独立程序补充： 选择性保护特定受体亚群免受 烷化拮抗剂的不可逆阻断。 放射自显影 研究将确定毒蕈碱亚群是否 受体定位于特定类型 （免疫细胞化学鉴定的）神经元。 研究 毒蕈碱受体的磷酸化旨在确定 不同亚群是否存在差异 磷酸化、脱敏和下调。 这 不同激酶在调节敏感性中的相互作用 还将研究受体。 预计这些 研究将有助于更好地理解功能和 毒蕈碱受体不同亚群的调节。