PROTEIN(S) INVOLVED IN NEUROTRANSMISSION

参与神经传递的蛋白质

• 批准号: 2244503
• 负责人: Jean Chen Shih
• 金额: $ 29.45万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-03-01 至 1996-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2244503
• 关键词: PROTEIN; INVOLVED; NEUROTRANSMISSION

5-hydroxytryptamine (5-HT, serotonin) and its receptors have been implicated in the etiology and therapeutic treatment of human mental depression. A basic knowledge of 5-HT receptors will help to better understand the molecular mechanism of 5-HT mediated neural function and will provide insights into the molecular basis of mental depression. The objectives of this project are to investigate the structure and function relationships, the genomic organization and the regulation of human 5-HT-2 receptor(s). These objectives will be accomplished by using the rat brain 5-HT-2 receptor cDNA cloned recently. The specific aims of the proposed study are outlined below. 1. To clone 5-HT-2 receptor and related cDNAs from human brain. Various 5-HT-2 receptor subtypes and related cDNA clones will be isolated and sequenced. The pharmacological profiles and the second messenger systems linked to the expressed receptors (phosphoinositol (PI) turnover, CA++ release, or adenylate cyclase) will be characterized. 2. To map the chromosomal location of the 5-HT-2 receptor gene. This aim will be accomplished by somatic cell mapping and in situ hybridization of the chromosome. 3. To isolate human genomic 5-HT-2 receptor clones. 4. To construct the restriction map and to sequence human 5-HT-2 receptor genomic clones. 5. To identify and to analyze promoter regions of this gene. This aim will be accomplished by RNase protection, primer extension, and chloramphenicol acetyltransferase (CAT) reporter gene assay. The cis- elements, trans-acting factors and the functions of these elements will be systematically analyzed. 6a. To investigate if 5-HT-2 receptor gene is regulated by protein kinase C (PKC) and/or protein kinase A (PKA). The effects of TPA (12-0- tetradecanoyl-phorbol-14 acetate, activator of PKC) on the 5-HT-2 receptor gene expression will be studied by using CAT report gene assay and footprinting analysis. The ability of 1-(5-isoquinolylsulfonyl)-2- methylpiperazine (H7) to block TPA activation will be studied. Similar studies will be performed using activators of PKA such as forskolin and 8- bromodibutyryl cAMP. 6b. To investigate if antagonist induced 5-HT-2 receptor down regulation is at the transcriptional level.

5-羟色胺(5-羟色胺，5-羟色胺)及其受体已被 与人类精神疾病的病因和治疗有关 抑郁症。5-羟色胺受体的基础知识将有助于更好地 了解5-羟色胺介导的神经功能和分子机制 将提供对精神抑郁的分子基础的洞察。这个 该项目的目标是研究其结构和功能。 人类5-羟色胺-2的相互关系、基因组结构及其调控 受体(S)。这些目标将通过使用大鼠的大脑来实现 新近克隆的5-羟色胺-2受体基因。建议的具体目标 研究概述如下。 1.从人脑组织中克隆5-羟色胺-2受体及其相关基因。五花八门 将分离5-羟色胺-2受体亚型和相关的cDNA克隆 已排序。药理研究概况与第二信使系统 与表达的受体(磷脂酰肌醇(PI)周转率，CA++)相连 释放，或腺苷环化酶)将被表征。 2.5-羟色胺-2受体基因的染色体定位。这一目标 将通过体细胞定位和原位杂交来完成 染色体。 3.分离人基因组5-HT-2受体克隆。 4.人5-羟色胺受体限制性内切酶图谱的构建及测序 基因组克隆。 5.鉴定和分析该基因的启动子区域。这一目标 将通过RNase保护、引物扩展和 氯霉素乙酰转移酶(CAT)报告基因分析。顺位- 要素、反式作用因素和这些要素的功能将是 系统地分析了。 6A。5-羟色胺-2受体基因是否受蛋白激酶调控 C(PKC)和/或蛋白激酶A(PKA)。TPA的影响(12-0- PKC激活剂)对5-羟色胺-2受体的激活作用 基因表达将通过CAT报告基因分析和 足迹分析。研究了1-(5-异喹啉磺酰)-2-羟基-4-甲基-4-甲基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-4-羟基-2- 将研究甲基哌嗪(H7)阻断TPA激活的作用。类似 研究将使用PKA的激活剂，如Forskolin和8- 溴化丁基营。 6B。研究拮抗剂是否诱导5-羟色胺-2受体下调 是在转录水平上。