DETERMINANTS OF DOPAMINE RECEPTOR FUNCTION

多巴胺受体功能的决定因素

• 批准号: 6343730
• 负责人: CLARE M BERGSON
• 金额: $ 10.22万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-03-15 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6343730
• 关键词: animal genetic material tag; brain mapping; cell membrane; cell type; chimeric proteins; dendrites; dopamine receptor; genetic library; glycosylation; human genetic material tag; intermolecular interaction; northern blottings; nucleic acid sequence; protein structure function; receptor binding; receptor coupling; receptor expression; tissue /cell culture; yeast two hybrid system

Regulation of mesocortical and striatonigral pathways by drugs aimed at inhibiting or enhancing dopaminergic neurotransmission is a commonly used strategy for treating many psychiatric and neurodegenerative diseases. These drugs act through dopamine receptors to elicit both short and long term changes in ion channel activity, protein kinase activities, and gene expression. The recent discovery of multiple G-protein-linked dopamine receptor subtypes exhibiting either classical "D1"-like or "D2-like" ligand binding and signal transduction properties raises several questions regarding the specific functions served by each receptor. Currently there are at least two D1-like and at least three D2-like DA receptors known. My previous immunohistochemical analyses of several D1- and D2-like subtypes showed that each receptor protein has a unique cellular and subcellular distribution within mesocortical, mesolimbic, and nigrastriatal pathways. These results support the notion that each subtype serves a unique function. However, subtype-specific signal transduction differences have not yet been identified in vivo. Without this functional information, it is difficult to understand the physiologic requirements for multiple D1-like and D2-like receptor subtypes. The goal of my research is to identify molecular and cellular processes that differentially regulate DA receptor function and localization in vivo using the D1-like receptors as a paradigm. One experimental approach involves use of the yeast two-hybrid system to identify proteins that potentially specify subtype-specific coupling to signal transduction systems or subcellular localization. My strategy is to use unique sequences within the closely related hD1 and hD5 subtypes as bait for detecting interacting proteins expressed in human brain. In complementary approaches, I will use subtype-specific monoclonal antibodies and affinity chromatography to isolate interacting proteins. Subtype-specific antibodies will also be used to confirm receptor interaction with proteins identified in the two-hybrid screen through immunoprecipitation experiments. This research will lead to a better understanding of the molecular organization of the DA receptor system, and identify new molecular targets for treating psychiatric and movement disorders.

通过针对的药物调节中皮质和纹状体途径 抑制或增强多巴胺能神经传递是一种常用的 治疗许多精神病和神经退行性疾病的策略。 这些药物通过多巴胺受体起作用，以引起短而长的 离子通道活性，蛋白激酶活性和基因的项变化 表达。最近发现多种G蛋白连接的多巴胺 受体亚型，表现出经典的“ D1”或“ D2样” 配体结合和信号转导特性提高了几个 有关每个受体提供的特定功能的问题。 目前至少有两个D1状和至少三个D2样DA 受体已知。我以前对几个D1-的免疫组织化学分析 和D2样的亚型表明，每个受体蛋白具有独特的 中皮层中的细胞和亚细胞分布 和黑质途径。这些结果支持每个人的观念 亚型具有独特的功能。但是，亚型特异性信号 转导差异尚未在体内鉴定。没有 这些功能信息，很难理解 多种D1状和D2样受体的生理要求 亚型。我的研究的目的是鉴定分子和细胞 差异调节DA受体功能的过程和 使用D1样受体作为范式在体内定位。一 实验方法涉及使用酵母两杂交系统 确定可能指定亚型特异性耦合的蛋白质 信号转导系统或亚细胞定位。我的策略是 在密切相关的HD1和HD5亚型中使用独特的序列 作为检测人脑中表达的相互作用蛋白的诱饵。在 互补方法，我将使用亚型特异性单克隆 分离相互作用蛋白的抗体和亲和色谱。 亚型特异性抗体也将用于确认受体 通过在两性杂交屏幕中鉴定的蛋白质相互作用 免疫沉淀实验。这项研究将导致更好 了解DA受体系统的分子组织， 并确定用于治疗精神病和运动的新分子靶标 疾病。