Regulation of Gene Expression in Striatal Neurons

纹状体神经元基因表达的调控

• 批准号: 7528126
• 负责人: QIANG WANG
• 金额: $ 28.23万
• 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 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7528126
• 关键词: 1-Phosphatidylinositol 3-Kinase; Agonist; Antisense Oligonucleotides; Basal Ganglia; Cell Nucleus; Cognition Disorders; Corpus striatum structure; Couples; Coupling; Cyclic AMP-Responsive DNA-Binding Protein; DNA; Data; Development; Dorsal; Electrophoretic Mobility Shift Assay; Event; Functional disorder; Gene Expression; Gene Expression Regulation; Genetic Transcription; Immunohistochemistry; In Vitro; Investigation; Knock-out; Label; Mediating; Membrane; Mental disorders; Metabotropic Glutamate Receptors; Methods; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases; Modeling; Molecular; Motor; Mutant Strains Mice; Nerve Degeneration; Neuronal Plasticity; Neurons; Nucleus Accumbens; Opioid; Opioid Peptide; Pathway interactions; Pharmacotherapy; Phosphorylation; Physiological; Physiology; Play; Process; Protein Binding; Protein Kinase; Protein Kinase C; Protein Tyrosine Kinase; RNA Interference; Rattus; Receptor Signaling; Regulation; Relative (related person); Role; Series; Signal Pathway; Signal Transduction; Signaling Protein; Site; Small Interfering RNA; Stimulus; Structure; Surface; Surveys; Testing; Ventral Striatum; Western Blotting; calmodulin-dependent protein kinase II; in vivo; inhibitor/antagonist; innovative technologies; interdisciplinary approach; mRNA Expression; novel; postsynaptic; pre-prodynorphin; preproenkephalin; prodynorphin; proenkephalin; promoter; protein kinase C kinase; putamen; receptor; relating to nervous system; research study; transcription factor; tyrosine receptor

DESCRIPTION (provided by applicant): Receptor-mediated gene expression in matured CNS neurons remains a hot topic in elucidating molecular mechanisms for the development of neuroplasticity. As a key structure of the basal ganglia, the striatum is among the major central sites for investigating such gene expression aimed to unravel striatal mechanisms underlying a variety of mental illnesses (psychiatric and cognitive disorders). Metabotropic glutamate receptors (mGluRs) are densely expressed in the striatal projection neurons. From studies conducted in the last period, we have established that Ca2+ -Iinked group I mGluRs play a profound role in regulating opioid peptide gene expression (prodynorphin and proenkephalin) in the rat striatum. In the effort of identifying intracellular signaling pathways transducing group I mGluR signals to the gene expression, we recently found that activation of group I receptors activates mitogen-activated protein kinases (MAPKs). As an information superhighway between the surface membrane and the nucleus, the MAPK pathway therefore likely couples group I mGluRs to gene expression. In this continuation proposal, a series of experiments was proposed to evaluate an overarching hypothesis that group I mGluRs activate MAPK cascades to facilitate opioid gene expression in striatal neurons. Using multidisciplinary approaches, this hypothesis will be tested both in vivo and in vitro n striata neurons in four general experiments: (1) define the role of group I receptors in the regulation of MAPKs by characterizing group I I mGluR-regulated phosphorylation of a major MAPK subclass, extracellular signal-regulated kinase 1 and 2 (ERKI/2), in a I well-characterized in vivo rat model, (2) differentiate the relative importance of the two group I subtypes, mGluR1 and I mGluR5, in this event using the subtype-selective agonists/antagonists, antisense oligos and mutant mice (mGluR1 or 5l knockouts) in vivo, (3) identify signaling pathways transducing group I mGluR signals to MAPK/ERK by evaluating roles of Ca 2+ signals and protein kinases (CaMKII, PI3-kinase, PKC, and tyrosine kinases) in our primary striatal neuronal culture model with selective inhibitors, antisense oligos and silencer siRNAs, and (4) define physiological roles of group I mGluRactivated MAPK/ERK cascades in the regulation of striatal gene expression via evaluating the importance of the two transcription factor targets of MAPK/ERK, Elk-1 and CREB, for group I/ERK-sensitive prodynorphin and proenkephalin gene expression in vivo and in vitro. Accomplishment of this project will provide a new interpretation of receptor and signal transduction mechanisms underlying inducible gene expression in matured CNS neurons. Since inducible gene expression is conceived to be an important component of the development of neuroplasticity, data from this project can ultimately contribute to the development of novel pharmacotherapies, by targeting group ImGluRs and MAPK cascades, for the treatment of various mental illnesses resulted from striatal dysfunctions.

描述（由申请人提供）： 成熟的中枢神经系统神经元中受体介导的基因表达仍然是阐明神经可塑性发展的分子机制的热门话题。作为基底神经节的关键结构，纹状体是研究这种基因表达的主要中心部位之一，该基因表达旨在揭示各种精神疾病（精神病和认知障碍）的纹状体机制。代谢性谷氨酸受体（mglurs）在纹状体投影神经元中密集表达。从上一时期进行的研究中，我们确定了Ca2+ i -I -I -I -I -I -I -I -Glurs在调节大鼠纹状体中的阿片类肽基因表达（Prodynorphin和Proenkephalin）中起着重要作用。为了鉴定细胞内信号通路通道I MGLUR信号转换为基因表达，我们最近发现，I组受体的激活激活了有丝分裂原激活的蛋白激酶（MAPKS）。作为表面膜和核之间的信息高速公路，MAPK途径可能会夫妻组与基因表达。在此持续提案中，提出了一系列实验，以评估一个总体假设，即MGlurs激活MAPK级联反应以促进纹状体神经元中阿片类药物的表达。 Using multidisciplinary approaches, this hypothesis will be tested both in vivo and in vitro n striata neurons in four general experiments: (1) define the role of group I receptors in the regulation of MAPKs by characterizing group I I mGluR-regulated phosphorylation of a major MAPK subclass, extracellular signal-regulated kinase 1 and 2 (ERKI/2), in a I well-characterized in vivo rat model, (2) differentiate the relative importance of the two group I subtypes, mGluR1 and I mGluR5, in this event using the subtype-selective agonists/antagonists, antisense oligos and mutant mice (mGluR1 or 5l knockouts) in vivo, (3) identify signaling pathways transducing group I mGluR signals to MAPK/ERK by evaluating roles Ca 2+信号和蛋白激酶（CAMKII，PI3-激酶，PKC和酪氨酸激酶）在我们的主要纹状体神经元培养模型中，具有选择性抑制剂，反义寡素和沉默的siRNA和（4）通过IMGLURURATIVENTINIC ERARTATIC MAPARTIAT INDERATION CASCADES CASC CASC的生理效果（4），均具有较高的型号。 I/ERK敏感性的Prodynorphin和Proenkephalin基因表达的MAPK/ERK，ELK-1和CREB的两个转录因子靶标的在体内和体外。该项目的完成将提供对成熟中枢神经系统神经元中诱导基因表达的受体和信号转导机制的新解释。由于诱导基因表达被认为是神经塑性发展的重要组成部分，因此该项目的数据最终可以通过靶向组IMGLURS和MAPK CASCADES来促进新型药物疗法的发展，以治疗侧面功能障碍引起的各种精神疾病。