REGULATION OF GENE EXPRESSION IN STRIATAL NEURONS

纹状体神经元基因表达的调节

• 批准号: 6625445
• 负责人: QIANG WANG
• 金额: $ 17.99万
• 依托单位: UNIVERSITY OF MISSOURI KANSAS CITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-12-01 至 2004-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6625445
• 关键词: AP1 protein; NMDA receptors; antisense nucleic acid; cAMP response element binding protein; calcium binding protein; calcium ion; calmodulin dependent protein kinase; corpus striatum; dopamine; dopamine antagonists; dynorphins; enkephalins; gene induction /repression; gene targeting; genetically modified animals; glutamate receptor; glutamates; in situ hybridization; inhibitor /antagonist; laboratory mouse; neural plasticity; neurogenetics; neuropharmacology

DESCRIPTION(Adapted from applicant's abstract): Receptor-mediated gene expression (stimulus-transcription coupling) in matured CNS neurons is thought to be an important component in processing adaptive changes in neuronal physiology (neuroplasticity) related to a variety of normal and abnormal neural activities. As a key part of basal ganglia, striatum is among the major central sites for the investigation of such gene expression. Our recent studies both in vivo and in vitro show that metabotropic glutamate receptors (mGiuRs), which are densely expressed in striatal projection neurons, are involved in the regulation of opioid gene expression in striatal neurons (preprodynorphin in striatonigral, and preproenkephalin in striatopallidal, neurons). The regulation is facilitatory in nature and may be mediated through selective activation of group I mGluRs, which are positively coupied to phosphoinositide hydrolysis (PI), rather than group II or III mGluRs, which are negatively coupled to adenylate cyclase. Based on these encouraging hndings, a series of experiments was proposed in this project to explore and characterize the regulation of striatal opioid peptide gene expression by the group I mGluRs in vivo and to dissect intracellular signaling pathways responsible for the group I-sensitive stimulus-transcription coupling in vitro. Our working hypothesis is that activation of the investigator-coupled group I mGluRs upregulates opioid gene expression in striatal neurons and that a investigator-sensitive cascade serves as the signaling pathway bridging surface group I mGluR stimulation to nuclear gene expression. With multidisciplinary approaches, this hypothesis will be tested in three aims which are to (1) confirm and characterize pharmacological profiles of the facilitatory regulation of opioid gene expression in the striatum with the group I selective agonists/antagonists, and investigate possible pre- and postsynaptic mechanisms underlying the group I regulation of striatal opioid expression in a well-characterized in vivo rat model, (2) differentiate the relative importance of the two group I subtypes, mGluR1 and mGluR5, in this event in vivo using the subtype-selective agonists/antagonists, antisense oligos and mutant mice (mGluR1/5 knockouts), and aboutL identify the intraceliular effectors (Ca2+, CaMK, CREB, CBP and AP-1) bridging group I mGluR stimulation on the membrane to opioid gene expression in the nucleus in vitro in primary striatal neuronal cultures with pharmacological (inhibitors/activators) and molecular (antisense oligos, Ca2+ image, enzymatic assay and DNA binding activity) approaches. We will rely on quantitative in situ hybridization to analyze mRNA expression in vivo and invitro. Accomplishment of this project will improve our current understanding of transcriptional regulation of gene expression in matured CNS neurons. Since inducible gene expression is conceived to participate in the development of psychoplasticity, data from this project will provide valuable insight into celiular/molecular mechanisms for various mental illnesses (motor, psychiatric and cognitive impairments).

描述(改编自申请人的摘要)： 成熟期受体介导的基因表达(刺激-转录偶联) 中枢神经系统神经元被认为是处理适应性的重要组成部分 与多种正常人相关的神经生理学(神经可塑性)的变化 以及异常的神经活动。纹状体是基底神经节的重要组成部分。 在研究这种基因表达的主要中心位置中。 我们最近在体内和体外的研究表明，代谢性谷氨酸 纹状体投射神经元中密集表达的受体(MGiuRs)， 参与纹状体神经元阿片类基因表达的调节 (纹状体中的前强啡肽原和纹状体中的前脑啡肽原， 神经元)。这项规定在性质上是便利的，可以通过 选择性激活I组mGluR，与其正向偶联 磷脂酰肌醇水解(PI)，而不是II或III组mGluR，后者是 与腺苷环化酶负偶联。基于这些令人鼓舞的消息，一个 在这个项目中提出了一系列的实验来探索和表征 I型mGluRs对纹状体阿片肽基因表达的调节 并剖析细胞内信号通路，这些信号通路负责 I组敏感的体外刺激-转录偶联。我们的工作 假设是研究人员偶联的I组mGluRs的激活 上调纹状体神经元阿片类基因的表达 研究者敏感的级联作为信号通路的桥接面 I组mGluR刺激核基因表达。与多学科合作 方法，这一假设将在三个目标进行检验，这三个目标是：(1) 确认和描述促进剂的药理学特征 选择性给药对纹状体阿片基因表达的调节 激动剂/拮抗剂，并研究可能的突触前和突触后机制 纹状体阿片样蛋白表达的I组调控 在体大鼠模型的特点，(2)区分相对重要性 两个I组亚型，mGluR1和mGluR5，在体内使用 亚型选择性激动剂/拮抗剂、反义寡核苷酸和突变小鼠 (mGluR1/5基因敲除)和大约L识别细胞内效应物(钙离子， CaMK、CREB、CBP和AP-1)桥接膜上mGluR刺激 阿片类基因在体外培养的纹状体神经元中的表达 含有药理学(抑制物/激活剂)和分子(反义)的培养 寡核苷酸、钙离子图像、酶分析和DNA结合活性)方法。我们 将依靠定量原位杂交来分析细胞中的mRNA表达 活体和体外。这个项目的完成将改善我们目前的状况 对成熟中枢神经系统基因表达转录调控的认识 神经元。由于可诱导的基因表达被认为参与了 心理可塑性的发展，这个项目的数据将提供有价值的 深入了解各种精神疾病的细胞/分子机制(运动、 精神和认知障碍)。