?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).

什么？-肾上腺素能受体（？AR）是免疫调节剂中的重要调节剂。 交感神经对中枢各种代谢过程的控制 (CNS)和外周神经系统。 这些受体位于 在整个神经系统的多个部位， CNS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 自主神经功能 我的实验室已经克隆，测序， 老鼠和恒河猴说，1-肾上腺素能受体基因 我们 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，并确定了潜在的增强子和阻遏物区域 这种影响力？1-AR表达。 我们已经证明， 诱导型cAMP早期阻遏物（ICER）诱导C6细胞？1-AR 激动剂，并与ICER相互作用？1-AR启动子， 下调？1-AR mRNA的转录，从而减少 细胞表面受体 我们还证明了老鼠？1-AR mRNA 含有两个功能性多聚腺苷酸化位点， 选择发生在C6细胞经历异丙肾上腺素诱导？1-AR mRNA下调。 这一过程可以通过以下方式进行调节： 转录后控制机制，可能与使用 RNA稳定性蛋白质相互作用的AU丰富的？1-AR 3' 非翻译区 目前的研究主要集中在识别 基因组元件和反式激活因子？1-AR转录，带有 使用？1-AR转移酶-报告基因重组体，转染测定， DNA迁移率变化和DNA酶足迹分析。 总体重点 该项目的目的是准确识别功能监管 要素和因素参与的监管？1-肾上腺素能 受体基因，并分析转录激活的 ? 1-肾上腺素能受体亚型在受体下调和 荷尔蒙刺激 研究最初将利用啮齿动物 在使用非人灵长类动物组织之前， 身份证？1-AR反式激活因子。 资助NIH HL 42358，以及 美国心脏协会设立的研究者奖出版物 Li Z，Vaidya RVA，Alvaro JD，Iredale PA，Hsu R，霍夫曼G，菲茨杰拉德 L，Curran PK，町田CA，Fishman PH，Duman RS. 蛋白激酶 C介导的下调？1-AR基因在大鼠C6胶质瘤中的表达 细胞 Mol Pharmacol 54：14-21，1998. Yang YF，Kirigiti P，Li B， Deshmane S，町田CA. 差异选择？1-肾上腺素能 受体转录物在激动剂诱导的下调。 SoC Neurosci Abstr 24：600，1998（摘要237.20）。