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）： 成熟细胞中受体介导的基因表达（刺激-转录耦合） 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 与多种正常神经元相关的神经生理学变化（神经可塑性） 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 是研究此类基因表达的主要中心位点之一。 我们最近的体内和体外研究表明，代谢型谷氨酸 受体（mGiuRs），在纹状体投射神经元中密集表达， 参与纹状体神经元阿片类基因表达的调节 （纹状体黑质中的前强啡肽和纹状体苍白球中的前脑啡肽原， 神经元）。该监管本质上是便利性的，可以通过 选择性激活 I 组 mGluR，其正耦合到 磷酸肌醇水解 (PI)，而不是 II 组或 III 组 mGluR，它们是 与腺苷酸环化酶负耦合。基于这些令人鼓舞的发现， 该项目提出了一系列实验来探索和表征 I 组 mGluR 对纹状体阿片肽基因表达的调节 体内并剖析负责的细胞内信号通路 体外 I 组敏感刺激转录耦合。我们的工作 假设是研究者偶联的 I 组 mGluR 的激活 上调纹状体神经元中的阿片类基因表达 研究人员敏感级联作为信号通路桥接表面 I 组 mGluR 刺激核基因表达。具有多学科 方法，该假设将在三个目标上进行检验，即（1） 确认并表征促进剂的药理学特征 纹状体中阿片类药物基因表达的 I 组选择性调控 激动剂/拮抗剂，并研究可能的突触前和突触后机制 纹状体阿片类药物表达的 I 组调节的基础 充分表征的体内大鼠模型，(2) 区分相对重要性 在本次体内实验中，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 敲除），以及大约 L 识别细胞内效应器（Ca2+、 CaMK、CREB、CBP 和 AP-1) 桥接 I 组 mGluR 刺激膜 原代纹状体神经元细胞核中阿片类基因的体外表达 具有药理学（抑制剂/激活剂）和分子（反义 寡核苷酸、Ca2+ 图像、酶测定和 DNA 结合活性）方法。我们 将依靠定量原位杂交来分析 mRNA 表达 体内和体外。该项目的完成将改善我们目前的 了解成熟中枢神经系统基因表达的转录调控 神经元。由于诱导型基因表达被认为参与 心理可塑性的发展，该项目的数据将提供有价值的 深入了解各种精神疾病（运动、 精神和认知障碍）。