?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).

β-肾上腺素能受体（β-AR）是重要的调节剂 交感神经控制中枢的各种代谢过程 （中枢神经系统）和周围神经系统。 这些受体位于 遍布神经系统的多个位点，并起着重要作用 中枢神经系统介导的行为和多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、测序并 表达大鼠和恒河猴α1-肾上腺素能受体基因。 我们 已经定义了 ?1-AR 转录起始位点、启动子元件、 和用于在 C6 神经胶质瘤细胞中表达的聚腺苷酸化位点 线，并已确定潜在的增强子和阻遏子区域 影响 ?1-AR 表达。 我们已经证明了 诱导型 cAMP 早期阻遏蛋白 (ICER) 在 C6 细胞中由 ?1-AR 诱导 激动剂，并且 ICER 与 ?1-AR 启动子相互作用 下调 ?1-AR mRNA 转录，从而减少 细胞表面受体。 我们还表明，大鼠 ?1-AR mRNA 包含两个功能性聚腺苷酸化位点，并且该转录本 选择发生在异丙肾上腺素诱导的 ?1-AR 的 C6 细胞中 mRNA 下调。 这个过程可能会受到监管 转录后控制机制，可能与使用 RNA 稳定性蛋白与富含 AU 的 ?1-AR 3' 相互作用 未翻译区域。 当前的研究集中于识别 ?1-AR 转录的基因组元件和反式激活因子， 使用 ?1-AR 荧光素酶报告基因重组体、转染测定、 DNA 迁移率变化和 DNase 足迹分析。 整体重点 该项目的目的是精确识别功能监管 参与调节β1-肾上腺素能的元素和因素 受体基因，并分析其转录激活 受体下调期间的β1-肾上腺素能受体亚型和 荷尔蒙刺激。 研究最初将利用啮齿动物 组织，在使用非人类灵长类动物组织之前， β1-AR反式激活子的鉴定。 资助 NIH HL42358，以及 美国心脏协会设立研究者奖出版物 Li Z、Vaidya RVA、Alvaro JD、Iredale PA、Hsu R、Hoffman G、Fitzgerald L、Curran PK、町田 CA、Fishman PH、杜曼 RS。 蛋白激酶 C介导的大鼠C6神经胶质瘤中α1-AR基因表达的下调 细胞。 Mol Pharmacol 54:14-21, 1998。Yang YF, Kirigiti P, Li B, Deshmane S，町田 CA。 ?1-肾上腺素能的差异选择 激动剂诱导的下调期间受体转录本。 社会组织 Neurosci Abstr 24:600，1998（摘要 237.20）。