REGULATION OF GENE EXPRESSION IN STRIATAL NEURONS

纹状体神经元基因表达的调节

• 批准号: 6680917
• 负责人: QIANG WANG
• 金额: $ 18.13万
• 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/6680917
• 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（根据申请人的摘要改编）： 受体介导的基因表达（刺激转录耦合）成熟 CNS神经元被认为是处理自适应的重要组成部分 与多种正常的神经元生理学（神经塑性）的变化 和异常的神经活动。作为基底神经节的关键部分，纹状体是 在研究这种基因表达的主要中央部位。 我们最近在体内和体外研究表明代谢型谷氨酸 受体（mgiurs），在纹状体投影神经元中密集表达， 参与纹状体神经元中阿片类药物基因表达的调节 （纹状体中的前肾上球和纹状体质合的前舌蛋白， 神经元）。该法规本质上是促进的，可以通过 选择性激活I组mglurs，它们是积极的惠值 磷酸肌醇水解（PI），而不是II组或III组，是 负耦合与腺苷酸环化酶。基于这些令人鼓舞的Hndings 在该项目中提出了一系列实验来探索和表征 I组对纹状体阿片类肽基因表达的调节 体内并剖析负责的细胞内信号传导途径 I组对体外的刺激转录耦合。我们的工作 假设是研究者耦合组I mglurs的激活 上调纹状体神经元中的阿片类药物基因表达， 研究者对敏感的级联反应是桥接表面的信号通路 I组刺激核基因表达。具有多学科 方法是，该假设将以三个目标进行检验，这是（1） 确认和表征促进性的药理特征 使用I选择性的纹状体调节阿片类药物基因表达 激动剂/拮抗剂，并研究可能的突触前和突触后机制 I组在A中调节纹状体阿片类药物表达 体内大鼠模型的特征良好，（2）区分相对重要性 在这两个I组亚型mglur1和mglur5中 亚型选择性激动剂/拮抗剂，反义寡聚和突变小鼠 （MGLUR1/5敲除），并且大约识别肾上腺内效应子（Ca2+， CAMK，CREB，CBP和AP-1）在膜上桥接i Mglur刺激 原发性纹状体神经元中的细胞核中阿片类药物基因表达 具有药理（抑制剂/激活剂）和分子（反义）的培养物 寡聚，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 Vivo和Invitro。完成该项目将改善我们的当前 了解成熟CN中基因表达的转录调节 神经元。由于诱导基因表达被认为参与 心理塑性的发展，该项目的数据将提供宝贵的 深入了解各种精神疾病的自旋/分子机制（运动， 精神病和认知障碍）。