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）：了解成瘾性物质修饰大脑的细胞机制是研究的关键目标，旨在为成瘾患者开发治疗策略的基础。我们的假设是成瘾的一个关键方面是涉及LTP和LTD等学习的机制的宽大谷氨酸能神经元可塑性。本应用程序中提出的实验将通过检查AMPA受体运输和成瘾相关的可塑性在八核（NAC）（NAC）中的跨膜AMPA受体调节蛋白（TARPS）的作用来检验这一假设，这是一个在大脑区域（NAC），这是一个在成立和持久性成瘾的发展中起着至关重要的作用。我建议研究1）在细胞水平上TARP同工型与AMPA受体亚基之间的相互作用，2）TARP磷酸化在NAC中的AMPA受体运输中的作用，3）3）与增强的药物抗磷酸化和TARP磷酸化和tarp磷酸化表达相关的AMPA受体表面表达的增加与动物模型的增强表达相关。篷布以前尚未在NAC中检查。首先，我将使用免疫细胞化学和定量共免疫沉淀技术来确定TARP同工型的分布及其与NAC中AMPA受体亚基的关联。其次，我将确定使用免疫细胞化学的AMPA受体突触掺入对粉尘的突触掺入的影响，并使用Western印迹和等质异常聚焦在新型的NAC-Prefrontal Cortrontal Cortex共培养系统中使用Western印迹和等质激素。最后，我将研究从可卡因自我给药和tarp磷酸化和表面表达的可卡因自我给药后提出的提示提示引起的药物捕获之间的关系。为此，戒断后将收集以前允许自动加工可卡因或盐水的NAC组织，并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TARPS的表面表达。综上所述，这些实验的数据将提供有关NAC中AMPA受体运输的机制的基本知识，并将有助于我们对与成瘾相关的谷氨酸能塑性的理解。多达300万美国人将在其一生中使用可卡因，而对可卡因（以及其他心理刺激剂（如苯丙胺））的成瘾仍然是国家一级的健康问题。本应用程序中提出的实验将提供有关AMPA受体运输的细胞机制的知识，还将提供谷氨酸能系统的改变与表明成瘾的行为之间的联系，例如增强的药物渴望。