The Role of a Specific Epigenetic Mechanism in Cocaine-induced Drug-seeking Behav

描述（由申请人提供）：本研究计划的重点是研究组蛋白去乙酰化酶3 （HDAC3）在体内作为情境药物相关记忆形成的潜在负调节因子的作用。先前的研究表明，HDAC3在体外分别通过与NCoR1和14-3-3蛋白相互作用，与hdac4和hdac5形成多蛋白复合物。我们假设HDAC3与体内海马和伏隔核（NAc）中的hdac4和/或5结合，转移到细胞核并使组蛋白去乙酰化，抑制与可卡因诱导的条件位置偏好（CPP）获得相关的长期情境记忆形成所必需的转录。了解与上下文药物相关的记忆形成有关的HDAC亚型将有助于理解长期记忆形成和药物寻找行为的分子机制，并可能最终导致靶向染色质修饰酶的选择性药物治疗来治疗药物成瘾。具体目标探讨Hdac3缺失对其他hdac表达及组蛋白乙酰化的影响。假说：HDAC3在体内与HDAC4、5形成复合体，促进核定位和组蛋白去乙酰化。具体目标2。探讨HDAC3在可卡因诱导CPP获得中的作用。假设：HDAC3功能的丧失将显著促进可卡因诱导CPP的获得。具体目标3。研究HDAC3是否作为可卡因诱导CPP获得的关键负调节因子。假设：HDAC3和/或NCoR1的过度表达将阻止可卡因诱导的CPP的获得。研究培训计划设计：1)对申请人进行行为药理学技术培训，使其能够有效地研究情境药物相关记忆形成的表观遗传机制；2)培养申请人成为一名独立研究者，使其将来能够熟练地在学术界担任实验室科学家、导师和首席研究者；3)强调负责任的研究行为，包括在实验室实验中使用动物的人道待遇和伦理数据分析，研究生和本科生的指导，与其他博士后研究员和主要研究人员建立合作关系，熟练设计和执行实验。这些技能将使申请人在他作为学术研究教授的职业生涯的下一阶段进行自己的独立调查。

DESCRIPTION (provided by applicant): The focus of this research proposal is to examine the role of histone deacetylase 3 (HDAC3) in vivo as a potential negative regulator of context-drug associated memory formation. Previous studies have shown that HDAC3 forms multi-protein complexes with HDACs 4 and 5 in vitro, via interactions with NCoR1 and 14-3-3 protein, respectively. We hypothesize that HDAC3 associates with HDACs 4 and/or 5 in vivo, in the hippocampus and nucleus accumbens (NAc), to translocate to the nucleus and deacetylates histones, repressing transcription necessary for long-term contextual memory formation associated with the acquisition of cocaine-induced conditioned place preference (CPP). Understanding the HDAC isoforms involved in context-drug associated memory formation will contribute to an understanding of the molecular mechanisms of long-term memory formation and drug-seeking behaviors and may eventually lead to selective pharmacotherapies which target chromatin modifying enzymes to treat drug addiction. Specific Aim 1. To examine the effects of Hdac3 deletion on the expression of other HDACs and histone acetylation. Hypothesis: HDAC3 forms a complex with HDAC4 and 5 in vivo to facilitate nuclear localization and histone deacetylation. Specific Aim 2. To examine the role of HDAC3 in acquisition of cocaine-induced CPP. Hypothesis: Loss of HDAC3 function will significantly facilitate the acquisition of cocaine-induced CPP. Specific Aim 3. To examine whether HDAC3 serves as a critical negative regulator of cocaine- induced CPP acquisition. Hypothesis: Over-expression of HDAC3 and/or NCoR1 will prevent acquisition of cocaine-induced CPP. The research training program will be designed: 1) for the applicant to be trained in behavioral pharmacology techniques, allowing him to effectively study epigenetic mechanisms of context-drug associated memory formation; 2) to train the applicant to be an independent investigator so he may be proficient in the future as a laboratory scientist, mentor and lead investigator in academia; and 3) to emphasize responsible conduct of research, including the humane treatment of animals used in laboratory experiments and ethical data analysis, mentoring of graduate and undergraduate students, building collaborations with other post- doctoral fellows and principal investigators and proficient design and execution of experiments. Those skills will allow the applicant to carry on independent investigations of his own during the next phase of his career as an academic research professor. PUBLIC HEALTH RELEVANCE: The philosophy of drug addiction has changed drastically from early misconceptions that addicts are "simply unable to control themselves." It has become clear that the long-term intake of abusive substances fundamentally alters the neurochemistry of an addict such that he/she becomes dependent on the drug for either physical or mental well-being. Thus a complete understanding of the persistent neurological changes underlying drug dependency, such as epigenetic mechanisms of drug-induced alterations in gene expression, may reveal novel and specific pharmacotherapy targets for addiction treatments.