Effects of ethanol on 5-HT1A receptor density, gene expression and function

描述(由申请人提供)：酒精滥用和依赖是一种普遍存在的疾病，构成了一个重大的公共卫生问题。美国酗酒的流行率从1992年的3.03%上升到2002年的4.65%。饮酒占全球死亡人数的1.5%，是美国第三大可预防的死亡原因。此外，在美国，与酗酒相关的经济成本正在上升，目前估计约为1840亿美元。这些发现清楚地表明，有必要制定旨在减少人力和财政成本的治疗战略。关于确定酒精滥用和依赖的神经生物学相关性的研究为开发有助于实现这一目标的药物疗法提供了框架和具体目标。调节酒精强化作用的5-羟色胺系统是关注这一靶点的一个潜在领域。尽管对乙醇和5-羟色胺系统之间的相互作用进行了多年的研究，但其确切的作用机制仍不清楚。5-HT1a受体是控制细胞外5-羟色胺浓度的主要细胞成分之一，也起着调节兴奋性和抑制性神经传递的作用。在这项应用中，我们提出了一个全面的检测乙醇对5-HT1a受体的影响的慢性酒精自我给药猴子模型。我们将在不同的分析水平上评估受体：1)受体密度使用体外受体放射自显影，2)基因转录使用激光捕获显微切割和定量聚合酶链式反应，3)信号转导和运输使用免疫沉淀法，随后使用多维液相色谱和MALDI ToF/ToF质谱仪。使用非常接近模拟人类饮酒行为的猴子自我给药范式，这些方法将为乙醇对这一受体系统的影响提供新的发现，可能有助于解释文献中报告的一些差异，并可以确定潜在的药物治疗靶点。重要的是，拟议的研究还旨在为我提供必要的培训和工具，以便在未来全面审查类似的系统。 与公共卫生相关：酗酒和依赖是一种普遍存在的疾病，构成了一个重大的公共卫生问题。显然需要旨在减少人力和财政成本的治疗战略。这里提出的关于确定酒精滥用和依赖的神经生物学相关性的研究为开发有助于实现这一目标的靶向药物疗法提供了一个框架。

DESCRIPTION (provided by applicant): Alcohol abuse and dependence are widespread disorders and constitute a significant public health concern. The prevalence of alcohol abuse has increased in the United States from 3.03% in 1992 to 4.65% in 2002. Alcohol use is responsible for 1.5% of all deaths globally and is the third leading preventable cause of death in the United States. In addition, the economic costs associated with alcohol abuse are on the rise in the United States with current estimates at approximately $184 billion. These findings make it clear that a need for treatment strategies aimed at reducing human and financial costs is crucial. Research on the identification of the neurobiological correlates of alcohol abuse and dependence provides a framework as well as specific targets for which to develop pharmacotherapies that can help to achieve this goal. One potential area to focus this targeting is the serotonin system which modulates the reinforcing effects of alcohol. Despite years of research on the interaction between ethanol and the serotonin system, the exact mechanisms of action remain unclear. The 5-HT1A receptor is one of the main cellular constituents controlling extracellular serotonin concentration and also serves to modulate excitatory and inhibitory neurotransmission. In this application, we propose a comprehensive examination of ethanol's effects on the 5-HT1A receptor in a monkey model of chronic ethanol self-administration. We will evaluate the receptor at various levels of analysis: 1) receptor density using in vitro receptor autoradiography, 2) gene transcription using laser capture microdissection and quantitative PCR and 3) Signal transduction and trafficking using an immunoprecipitation protocol followed by multidimensional liquid chromatography and MALDI ToF/ToF mass spectrometry. Using a monkey selfadministration paradigm that very closely models human drinking behavior, these approaches will provide novel findings on ethanol's effects on this receptor system, may help to explain some of the discrepancies reported in the literature and can identify potential pharmacotherapeutic targets. Importantly, the proposed research is also designed to provide me with the training and tools necessary to comprehensively examine similar systems in the future. PUBLIC HEALTH RELEVANCE: Alcohol abuse and dependence are widespread disorders and constitute a significant public health concern. A need for treatment strategies aimed at reducing human and financial costs is evident. The research proposed here on the identification of the neurobiological correlates of alcohol abuse and dependence provides a framework from which to develop targeted pharmacotherapies that can help to achieve this goal.