Molecular Regulation of Metabotropic Glutamate Receptors in Striatal Neurons

纹状体神经元代谢型谷氨酸受体的分子调控

基本信息

批准号：
7982679
负责人：
QIANG WANG
金额：
$ 37.5万
依托单位：
UNIVERSITY OF MISSOURI KANSAS CITY
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-12-01 至 2015-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7982679
关键词：
Acute Address Affinity Amino Acids Amphetamines Antibodies Area Autoradiography Basal Ganglia Behavior Behavior Control Behavioral Binding Biochemical Brain C-terminal Cells Chronic Cognition Corpus striatum structure Development Disease model Drug Addiction Emotions Feedback Glutamate Receptor Glutamates Homer protein Immunoblotting In Vitro Injection of therapeutic agent Kinetics Life Link Liquid Chromatography Mediating Mental disorders Metabotropic Glutamate Receptors Modeling Molecular Motivation Motor Activity N-Methyl-D-Aspartate Receptors Neuraxis Neurons Peptides Pharmacotherapy Phosphorylation Phosphoserine Phosphothreonine Phosphotransferases Pilot Projects Plastics Post-Translational Regulation Property Protein Kinase Proteomics RNA Splicing Reaction Regulation Research Project Grants Rewards Role Series Serine Signal Transduction Site Small Interfering RNA Substance Addiction Synapses Tail Testing Variant addiction base behavioral sensitization calmodulin-dependent protein kinase II clinically relevant design drug of abuse in vivo innovation insight interdisciplinary approach mutant nano neurobehavioral novel protein protein interaction psychostimulant public health relevance receptor receptor function research study scaffold stem systems research tandem mass spectrometry trafficking transmission process

项目摘要

DESCRIPTION (provided by applicant): Protein phosphorylation is an important mechanism for post-translational regulation of glutamate receptors. Through phosphorylating a specific amino acid in the intracellular domain, protein kinases regulate anchoring, trafficking, and signaling of a given glutamate receptor. Group I metabotropic glutamate receptors (mGluR1/5) are densely expressed in the striatum, a brain area involved in addictive properties of psychostimlants. The long-form mGluR1/5 splice variants (1a, 5a, and 5b) have a large intracellular C-terminal tail, which provides a basis for direct protein-protein interactions and phosphorylation. In our recent studies, we found that Ca2+/calmodulin-dependent protein kinase II (CaMKII) binds directly to the proximal region of mGluR5a C-terminus. This binding converts mGluR5a into a biochemical substrate for phosphorylation likely at a selective serine site. These findings raise innovative questions as to if CaMKII regulates mGluR1/5 via a direct protein-protein interaction and phosphorylation and if this regulation has a high clinical relevance in a disease model. In this continuation proposal, a series of coherent experiments from molecule to behavior was proposed to confirm the direct binding of CaMKII to mGluR1/5 in vitro and to establish that native CaMKII and mGluR1/5 interact with each other in striatal neurons in vivo. We will characterize if and how Ca2+ regulates the interaction between CaMKII and mGluR1/5 in vitro and in vivo. We will then investigate whether Ca2+-regulated CaMKII-mGluR1/5 interactions regulate 1) signaling efficacy of mGluR1/5, 2) trafficking of the receptors, and 3) interactions of mGluR1/5 with key scaffold Homer proteins, in striatal neurons or heterologous cells. Finally, we will carry out neurobehavioral experiments to define the role of CaMKII-mGluR1/5 interactions in the addictive action of the psychostimulant amphetamine. Our results will provide evidence and insights for a new synaptic model of kinase-regulated mGluRs and for its linkage to a mental illness (substance addiction). They will also ultimately contribute to the development of novel pharmacotherapies, by targeting mGluRs and CaMKII, for treating various mental illnesses, including addiction. PUBLIC HEALTH RELEVANCE: This research project is aimed to elucidate molecular mechanisms underlying the regulation of metabotropic glutamate receptors and roles of the receptor in drugs of abuse. The information obtained through this project is valuable for the development of new pharmacotherapies for mental illnesses stemming from dysfunctional glutamatergic transmission in the central nervous system.

描述（由申请人提供）：蛋白质磷酸化是谷氨酸受体翻译后调节的重要机制。通过在细胞内结构域中磷酸化特定的氨基酸，蛋白激酶调节给定的谷氨酸受体的锚定，运输和信号传导。 I组的代谢型谷氨酸受体（MGLUR1/5）在纹状体中密集表达，这是一种涉及精神刺激物的大脑区域。长形的MGLUR1/5剪接变体（1a，5a和5b）具有较大的细胞内C末端尾巴，这为直接蛋白质蛋白质相互作用和磷酸化提供了基础。在最近的研究中，我们发现Ca2+/钙调蛋白依赖性蛋白激酶II（CAMKII）直接与MGLUR5A C末端的近端区域结合。这种结合将MGLUR5A转化为可能在选择性丝氨酸位点的磷酸化的生化底物。这些发现提出了有关CAMKII是否通过直接蛋白质 - 蛋白质相互作用和磷酸化调节MGLUR1/5的创新问题，以及该调节在疾病模型中是否具有很高的临床相关性。在此持续提案中，提出了一系列从分子到行为的连贯实验，以确认Camkii在体外与MGLUR1/5的直接结合，并确定天然的Camkii和Mglur1/5在体内的纹状体神经元中相互相互作用。我们将表征Ca2+如何调节CAMKII与MGLUR1/5在体外和体内之间的相互作用。然后，我们将研究Ca2+调节的CAMKII-MGLUR1/5相互作用是否调节1）MGLUR1/5，2）受体运输的信号传导功效，以及3）MGLUR1/5在纹状体神经元或杂元细胞中MGLUR1/5与关键支架荷马蛋白的相互作用。最后，我们将进行神经行为实验，以定义Camkii-Mglur1/5相互作用在心理刺激苯丙胺的成瘾作用中的作用。我们的结果将为激酶调节的新突触模型及其与精神疾病的联系（药物成瘾）提供证据和见解。他们还将通过靶向MGLUR和CAMKII来治疗各种精神疾病（包括成瘾），最终将通过靶向新的药物疗法的发展。公共卫生相关性：该研究项目旨在阐明对代谢型谷氨酸受体调节的分子机制以及受体在滥用药物中的作用。通过该项目获得的信息对于开发新的药物治疗，用于开发因中枢神经系统功能失调的谷氨酸能传播而导致的精神疾病。