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广泛的疾病，构成了一个重大的公共卫生问题。 在美国，酗酒的流行率从1992年的3.03%上升到2002年的4.65%。 酒精使用占全球所有死亡人数的1.5%，是美国第三大可预防的死亡原因。此外，在美国，与酗酒有关的经济成本正在上升，目前估计约为1 840亿美元。这些研究结果清楚地表明，必须制定旨在减少人力和财力成本的治疗战略。关于确定酒精滥用和依赖的神经生物学相关因素的研究提供了一个框架以及开发药物疗法的具体目标，可以帮助实现这一目标。一个潜在的领域，以集中这种目标是血清素系统，调节酒精的强化作用。尽管对乙醇和血清素系统之间的相互作用进行了多年的研究，但确切的作用机制仍不清楚。5-HT 1A受体是控制细胞外5-羟色胺浓度的主要细胞成分之一，也用于调节兴奋性和抑制性神经传递。在这个应用中，我们提出了一个全面的检查乙醇对5-HT 1A受体在猴子模型的慢性乙醇自我管理的影响。我们将在不同水平的分析中评估受体：1）使用体外受体放射自显影的受体密度，2）使用激光捕获显微切割和定量PCR的基因转录，3）使用免疫沉淀方案，然后进行多维液相色谱和MALDI ToF/ToF质谱的信号转导和运输。使用猴子自我管理模式，非常接近人类饮酒行为的模型，这些方法将提供新的发现乙醇对这个受体系统的影响，可能有助于解释一些文献中报道的差异，并可以确定潜在的药理学目标。重要的是，拟议的研究还旨在为我提供必要的培训和工具，以便在未来全面研究类似的系统。 公共卫生相关性：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。显然需要制定旨在减少人力和财力成本的治疗战略。这里提出的关于酒精滥用和依赖的神经生物学相关性的研究提供了一个框架，从中开发出有助于实现这一目标的靶向药物疗法。

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.