Role of TARPs in Addiction-Related Plasticity

TARP 在成瘾相关可塑性中的作用

• 批准号: 7407952
• 负责人: Carrie Ferrario
• 金额: $ 4.68万
• 依托单位: ROSALIND FRANKLIN UNIV OF MEDICINE & SCI
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-11-25 至 2010-11-24
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7407952
• 关键词: AMPA Receptors; American; Amphetamines; Animal Model; Behavior; Biochemical; Brain; Brain region; Co-Immunoprecipitations; Cocaine; Coculture Techniques; Cues; Data; Development; Glutamates; Health; Knowledge; Learning; Link; Neuronal Plasticity; Nucleus Accumbens; Patients; Phosphorylation; Phosphotransferases; Play; Prefrontal Cortex; Protein Isoforms; Rattus; Role; Saline; Self Administration; Self-Administered; Surface; Synapses; Synaptic Receptors; System; Targeted Research; Techniques; Testing; Tissues; Western Blotting; Withdrawal; addiction; alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; amino 3 hydroxy 5 methylisoxazole 4 propionate; base; design; drug craving; genetic regulatory protein; immunocytochemistry; novel; psychostimulant; research study; therapy development; trafficking

DESCRIPTION (provided by applicant): Understanding the cellular mechanisms by which addictive substances modify the brain is a key target of research that is designed to provide a basis for development of treatment strategies for addicted patients. Our hypothesis is that a critical aspect of addiction is abberant glutamatergic neuronal plasticity involving mechanisms common to learning, such as LTP and LTD. The experiments proposed in this application will test this hypothesis by examining the role of transmembrane AMPA receptor regulatory proteins (TARPs) in both AMPA receptor trafficking and addiction-related plasticity in the nucleus accumbens (NAc), a brain region that plays a critical role in the development and persistence of addiction. I propose to investigate 1) the interactions between TARP isoforms and AMPA receptor subunits at the cellular level, 2) the role of TARP phosphorylation in AMPA receptor trafficking in the NAc, 3) the relationship between increases in AMPA receptor surface expression associated with enhanced drug-craving and TARP phosphorylation and surface expression in an animal model of addiction. TARPs have not previously been examine in the NAc. First, I will use immunocytochemical and quantitative co-immunoprecipitation techniques to determine the distribution of TARP isoforms and their association with AMPA receptor subunits in the NAc. Second, I will determine the effect of AMPA receptor synaptic incorporation on the synaptic incorporation of TARPs using immunocytochemistry and on TARP phosphorylation using Western blotting and isoelecrtric focusing in a novel NAc-prefrontal cortex co-culture system. Finally, I will examine the relationship between enhanced cue elicited drug-craving after withdrawal from cocaine self-administration and TARP phosphorylation and surface expression. To this end, NAc tissue from rats previously allowed to self-administer cocaine or saline will be collected after withdrawal and several biochemical techniques will be used to determine TARP phosphorylation, kinase activity, and surface expression of AMPA receptors and TARPs. Taken together, data from these experiments will provide essential and novel knowledge about the mechanisms underlying AMPA receptor trafficking in the NAc and will contribute to our understanding of glutamatergic plasticity associated with addiction. As many as 3 million Americans will use cocaine within their lifetimes, and addiction to cocaine (as well as other psychostimulants such as amphetamines) continues to be a health concern at the national level. The experiments proposed in this application will provide knowledge about the cellular mechanisms of AMPA receptor trafficking, and will also provide a link between alterations in the glutamatergic system and behaviors indicative of addiction, such as enhanced drug craving.

描述（由申请人提供）：了解成瘾物质改变大脑的细胞机制是研究的一个关键目标，旨在为成瘾患者的治疗策略提供基础。我们的假设是，成瘾的一个关键方面是异常的神经元可塑性，涉及学习的共同机制，如LTP和LTD。本申请中提出的实验将通过检查跨膜AMPA受体调节蛋白（TARPs）在AMPA受体运输和成瘾相关可塑性中的作用来验证这一假设。这一大脑区域在成瘾的发展和持续中起着关键作用。我建议调查1）之间的相互作用TARP亚型和AMPA受体亚基在细胞水平上，2）的作用，TARP磷酸化的AMPA受体在NAc运输，3）之间的关系，在AMPA受体表面表达增加与增强药物渴求和TARP磷酸化和表面表达成瘾的动物模型。TARP以前没有在NAC中进行过检查。首先，我将使用免疫细胞化学和定量免疫共沉淀技术，以确定分布的TARP亚型和他们的协会与AMPA受体亚单位在NAC。第二，我将确定AMPA受体突触掺入的影响，在一个新的NAC-前额叶皮层共培养系统中，使用免疫细胞化学和TARP磷酸化，使用Western印迹和等电聚焦的TARP的突触掺入。最后，我将研究可卡因自我管理和TARP磷酸化和表面表达的增强线索引起的药物渴求之间的关系。为此，将在戒断后收集来自先前允许自我施用可卡因或盐水的大鼠的NAc组织，并将使用几种生化技术来确定TARP磷酸化、激酶活性以及AMPA受体和TARP的表面表达。两者合计，从这些实验的数据将提供必要的和新的知识的机制AMPA受体贩运的NAc，并将有助于我们了解与成瘾相关的神经元可塑性。多达300万美国人将在他们的一生中使用可卡因，可卡因成瘾（以及其他精神兴奋剂，如安非他明）仍然是国家一级的健康问题。本申请中提出的实验将提供关于AMPA受体运输的细胞机制的知识，并且还将提供在药物能系统的改变与指示成瘾的行为（例如增强的药物渴望）之间的联系。