bZIP Repression of Adrenergic Receptor RNA in Neurons

bZIP 抑制神经元中肾上腺素能受体 RNA

• 批准号: 7027080
• 负责人: CURTIS A MACHIDA
• 金额: $ 14.96万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-03-05 至 2008-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7027080
• 关键词: DNA binding protein; RNA interference; RNase protection assay; androgen receptor; ecdysone; gel mobility shift assay; gene expression; gene induction /repression; laboratory rat; messenger RNA; neurons; polymerase chain reaction; site directed mutagenesis; small interfering RNA; transcription factor; western blottings; yeast two hybrid system

DESCRIPTION (provided by applicant): Adrenergic receptors (ARs) serve as important regulators of central nervous system- (CNS-) mediated behavior and several neural functions, including mood, memory, neuroendocrine control, and stimulation of autonomic function. Regulation of neurotransmitter receptor efficacy and control of neuronal plasticity, as evidenced in receptor desensitization and chronic down-regulation mechanisms, are recognized as part of the adaptive process in the brain that occurs during depression and antidepressant therapy. Beta-AR down-regulation occurs during chronic treatment with antidepressants, suggesting that the dysregulation of the beta`-adrenergic receptor (beta1-AR) subtype, the predominant (beta-AR subtype in the brain, may be associated with depression. Furthermore, the central administration of (beta-AR agonist isoproterenol in the brain induces antidepressant-like effects in the rat, and appears to specifically involve the (beta1-AR subtype. My laboratory has examined the molecular mechanisms underlying beta1AR mRNA down-regulation following agonist induction, and have identified potential transcriptional and post-transcriptional control mechanisms. We have recently identified a transcriptional represser region in the beta1-AR gene, encompassing positions -396 to -367 relative to the translational start site, and have identified the potential represser molecule as a novel bZIP-like transcription factor. This novel transcription factor contains a basic DNA-binding domain, including a leucine zipper motif (bZIP-like), and a phosphatase motif similar to those contained within the RNA polymerase II C-terminal domain (CTD) phosphatases. The specific aims of this application are to validate the bZIP-like transcription factor as a represser of beta1-AR expression in neonatal rat cortical neurons and to test the hypothesis that this factor is a determinant in the agonist-mediated down-regulation of beta1-AR mRNAs. The primary experimental objectives of this proposal are: 1) to verify interaction of the bZIP-like transcription factor in the repression of the beta1-AR promoter in neonatal rat cortical neurons, 2) to mutagenize the phosphatase motif and basic DNA-binding domain in the transcription factor for potential development of a dominant negative mutant, 3) to identify other potential partners in the bZIP-like transactivator complex formed during beta1-AR transcriptional repression, and 4) to determine the role of the bZIP-like transcription factor in molecular mechanisms underlying agonist-mediated beta1-AR mRNA down-regulation in neonatal rat cortical neurons using both ecdysone-inducible expression and RNA interference systems. This information may provide important insights in the regulation of beta-ARs in the brain and the potential role of these neurotransmitter receptors in depression. This R21 grant application is submitted under PA-03-107 ("NIH Exploratory/Developmental Grant [R21] Program").

描述（由申请人提供）：肾上腺素能受体（ARS）是中枢神经系统（CNS-）介导的行为和几种神经功能的重要调节剂，包括情绪，记忆，神经内分泌控制和刺激自主功能。正如受体脱敏和慢性下调机制所证明的神经递质受体功效和神经元可塑性的控制，被认为是在抑郁症和抗抑郁药治疗过程中发生的大脑自适应过程的一部分。 β-AR下调发生在抗抑郁药长期治疗期间，表明β“肾上腺素能受体（beta1-ar）亚型的失调失调，主要是（大脑中的β-ar-ar subtype）（可能与抑郁症相关，可能与抑郁症相关。 and appears to specifically involve the (beta1-AR subtype. My laboratory has examined the molecular mechanisms underlying beta1AR mRNA down-regulation following agonist induction, and have identified potential transcriptional and post-transcriptional control mechanisms. We have recently identified a transcriptional represser region in the beta1-AR gene, encompassing positions -396 to -367 relative to the translational start site,并确定了潜在的抑制剂分子为新型的BZIP样转录因子。该应用的具体目的是验证新生大鼠皮质神经元中BBZIP样转录因子作为BetA1-AR表达的阻遏物，并检验该因子在激动剂介导的BetA1-AR mRNA中的决定因素是决定因素。该提案的主要实验目标是：1）在新生大鼠皮质神经元中抑制BZIP样转录因子的相互作用，2）在磷酸酶基序和基本的DNA结合域中的基本DNA结合域在潜在的负面突变中的转移中的转移，以识别磷酸酶基序和基本的DNA结合域在3中的转移，3） beta1-ar转录抑制和4），以确定BZIP样转录因子在使用Ecdysone-诱导性表达和RNA Interperience Systemp的新生大鼠皮质神经元中的激动剂介导的BetA1-AR mRNA下调的分子机制中的作用。这些信息可能会在调节大脑中的β-ARS调节以及这些神经递质受体在抑郁症中的潜在作用提供重要见解。该R21赠款申请根据PA-03-107（“ NIH探索/发展赠款[R21]计划”提交）。