?1 ADRENERGIC RECEPTOR ACTIVATION & EXPRESSION

?1 肾上腺素能受体激活

• 批准号: 6592300
• 负责人: CURTIS A MACHIDA
• 金额: $ 11.11万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6592300
• 关键词: Mammalia; animal tissue; biotechnology; cardiovascular system; nervous system; nucleic acids

The ?-adrenergic receptors (?-ARs) are important modulators in the sympathetic control of various metabolic processes in the central (CNS) and peripheral nervous system. These receptors are localized at multiple sites throughout the nervous system, and serve as important regulators of CNS-mediated behavior and several neural functions, including mood, memory, neuroendocrine control, and stimulation of autonomic function. My laboratory has cloned, sequenced, and expressed the rat and rhesus macaque ?1-adrenergic receptor genes. We have defined the ?1-AR transcriptional start sites, promoter elements, and polyadenylation sites used for expression in the C6 glioma cell line, and have identified potential enhancer and repressor regions that influence ?1-AR expression. We have demonstrated that the inducible cAMP early repressor (ICER) is induced in C6 cells by ?1-AR agonists, and that ICER interacts with the ?1-AR promoter to down-regulate ?1-AR mRNA transcription, w ith resul tant diminution of cell surface receptor. We have also shown that the rat ?1-AR mRNA contains two functional polyadenylation sites, and that transcript selection occurs in C6 cells undergoing isoproterenol-induced ?1-AR mRNA down-regulation. This process may be regulated by post-transcriptional control mechanisms, potentially with the use of RNA stability proteins interacting with an AU-rich ?1-AR 3' untranslated region. Current research is centered on identifying genomic elements and trans-activators of ?1-AR transcription, with the use of ?1-AR luciferase-reporter recombinants, transfection assays, DNA mobility shift and DNase footprinting assays. The overall focus of this project is to precisely identify functional regulatory elements and factors involved in the regulation of the ?1-adrenergic receptor gene, and to analyze the transcriptional activation of the ?1-adrenergic receptor subtype during receptor down-regulation and hormonal stimulation. Research will initially utiliz e rodent tissues, prior to the use of nonhuman primate tissues, in the identification of ?1-AR transactivators. FUNDING NIH HL42358, and American Heart Association Established Investigator Award PUBLICATIONS Li Z, Vaidya RVA, Alvaro JD, Iredale PA, Hsu R, Hoffman G, Fitzgerald L, Curran PK, Machida CA, Fishman PH, Duman RS. Protein kinase C-mediated down-regulation of ?1-AR gene expression in rat C6 glioma cells. Mol Pharmacol 54:14-21, 1998. Yang YF, Kirigiti P, Li B, Deshmane S, Machida CA. Differential selection of ?1-adrenergic receptor transcripts during agonist-induced down-regulation. Soc Neurosci Abstr 24:600, 1998 (abstract 237.20).

？ - 肾上腺素能受体（？-ARS）是重要的调节剂 中央各种代谢过程的交感神经控制 （CNS）和周围神经系统。 这些受体位于 整个神经系统中的多个站点，并作为重要 中枢神经系统介导的行为和几种神经功能的调节剂， 包括情绪，记忆，神经内分泌控制和刺激 自主功能。 我的实验室已经克隆，测序了，并且 表示大鼠和恒河猴1-肾上腺素能受体基因。 我们 已经定义了1-AR的转录起始站点，启动子元素， 和用于在C6神经胶质瘤细胞中表达的聚腺苷酸化位点 线，并确定了潜在的增强剂和阻遏物区域 这种影响是1-ar的表达。 我们已经证明了 1-AR诱导诱导型营地早期阻遏物（ICER）在C6细胞中诱导 激动剂，那个ICER与？1-AR启动子互动 下调？1-ar mRNA转录，降低了 细胞表面受体。 我们还表明了大鼠1-ar mRNA 包含两个功能性聚腺苷酸位点，该转录本 选择发生在接受异丙肾上腺素诱导的C6细胞中？1-ar mRNA下调。 这个过程可能受 转录后控制机制，有可能使用 RNA稳定性蛋白与富含AU的1-AR 3'相互作用 未翻译区域。 当前的研究集中于确定 1-AR转录的基因组元素和反式激活剂， 使用？1-AR荧光素酶 - 重组剂重组，转染测定法， DNA迁移率转移和DNase足迹测定。 整体重点 该项目的精确确定功能调节 调节的元素和因素是1-肾上腺素能 受体基因，并分析 受体下调期间1-肾上腺素能受体亚型和 激素刺激。 研究最初将利用啮齿动物 在使用非人类灵长类组织之前的组织 识别？1-AR反式激活剂。 资助NIH HL42358，以及 美国心脏协会成立研究者奖出版物 Li Z，Vaidya RVA，Alvaro JD，Iredale PA，Hsu R，Hoffman G，Fitzgerald L，Curran PK，Machida CA，Fishman PH，Duman RS。 蛋白激酶 C介导的1-AR基因表达的下调大鼠C6胶质瘤 细胞。 Mol Pharmacol 54：14-21，1998。Yang YF，Kirigiti P，Li B， DESHMANE S，MACHIDA CA。 1-肾上腺素能的差异选择 激动剂引起的下调期间的受体转录本。 Soc Neurosci Abstr 24：600，1998（摘要237.20）。