Molecular Mechanisms of Cellular Plasticity in a Mouse Model of Excessive Alcohol

描述(由申请人提供)：本次申请的目标是在R.Adron Harris博士的指导下为Igor Ponomarev博士提供职业发展机会。得克萨斯大学奥斯汀分校优越的研究环境、师资和设施将使他能够磨练年轻研究员的技能，并学习脑显微注射和激光捕获显微解剖等细胞神经科学技术。这些技术将使波诺马廖夫博士能够识别和剖析单个神经元群体，进行基因表达分析，并应用干扰RNA和药理学试剂来测试差异调控候选基因的功能。根据该奖项接受的培训将为波诺马廖夫博士在学术科学领域的职业生涯做好更好的准备，并称赞他在行为神经遗传学和功能基因组学方面的专业知识。由于过量饮酒是形成酒精依赖的先决条件，因此确定与高酒精摄入量相关的分子靶点，以了解酒精中毒进展的机制是很重要的，酒精中毒进展机制是介导从受控饮酒向酒精滥用和酒精依赖的转变。这项提议的总体目标是确定遗传因素和酗酒对特定脑区基因表达的影响，这些脑区被认为参与了酒精消费的调节，以及投射到这些脑区的不同群体的多巴胺神经元。我们将使用脑显微注射、激光捕获微缺陷和cDNA微阵列来研究高饮酒倾向的遗传选择小鼠大脑区域和神经元中的基因表达模式。我们假设，与酒精相关的神经回路的不同大脑区域和神经元群体可以通过它们各自的基因表达模式以及它们对遗传选择和饮酒的不同转录反应来区分。这些研究的长期目标是确定单个神经元群体在酒精行为中的作用，并确定特定细胞类型的酒精敏感基因和基因产物作为酒精中毒的潜在治疗靶点。

DESCRIPTION (provided by applicant): The goal of the current application is to provide Dr. Igor Ponomarev with career development opportunities under the mentorship of Dr. R. Adron Harris. The excellent research environment, faculty and facilities at the University of Texas at Austin will allow him to sharpen his skills as a young investigator and to learn cellular neuroscience techniques such as brain microinjections and laser capture microdissection. These techniques will allow Dr. Ponomarev to identify and dissect individual neuronal populations for gene expression analysis and also to apply interference RNA and pharmacological agents to test functionality of differentially regulated candidate genes. Training under this award will better prepare Dr. Ponomarev for a career in academic science and compliment his expertise in behavioral neurogenetics and functional genomics. Because excessive alcohol consumption is a prerequisite for the development of alcohol dependence, it is important to identify molecular targets associated with high alcohol intake in order to understand the mechanisms of alcoholism progression, which mediate the switch from controlled alcohol consumption to alcohol abuse and alcohol dependence. The overall goal of this proposal is to determine the effects of genetic factors and drinking high amounts of alcohol on gene expression in specific brain regions believed to be involved in regulation of alcohol consumption and different populations of dopamine neurons projecting to these brain regions. We will use brain microinjections, laser capture microdisections and cDNA microarrays to study patterns of gene expression in brain regions and neurons of genetically selected mice with predisposition to high alcohol consumption. We hypothesize that different brain regions and neuronal populations of alcohol-related neurocircuitry can be distinguished by their individual patterns of gene expression and by their distinct transcriptional responses to genetic selection and alcohol drinking. The long-term goals of these studies are to determine roles of individual neuronal populations in alcohol actions and to identify cell type - specific alcohol-sensitive genes and gene products as potential therapeutic targets for alcoholism.