Gene Expression and -Drug-Induced Sensitivity to Alcohol

描述（由申请人提供）：对酒精的行为敏感性可能是 作为环境和遗传因素的函数进行修饰。的 这种效应的潜在基础是神经元适应反应的能力 包括但不限于酒精暴露， 这种反应的具体性质取决于基因组成。这些 过程可能是酒精中毒病因学的一个重要特征。证据 表明作用于多巴胺（DA）神经元的药物能够改变多巴胺神经元的功能。 啮齿动物对酒精的行为反应因此，在单次注射后24小时， 用DA拮抗剂氟哌啶醇或间接DA激动剂治疗 甲基苯丙胺，大鼠变得对酒精或多或少敏感， 并且在单剂量酒精后一天“快速”产生耐受性。这些 急性药物反应将被用来研究全球基因 DA通路中发生的表达变化导致酒精改变 灵敏度具体目标1将充分描述行为反应， 急性治疗后24小时酒精（翻正反射丧失） 氟哌啶醇、酒精或甲基苯丙胺在重复近交系中的高和低 酒精敏感大鼠品系。基因表达分析将在 具体目标2。将在施用以下化合物后8小时处死大鼠： 每种药物的单一最佳剂量以及纹状体和腹侧中脑将被 解剖。RNA将从这些结构中提取，基因表达将 可以通过使用Affyssin基因芯片系统来确定。这将提供 总共16个实验条件，包括不同的药物/基因型 组合。据推测，这些条件中的每一个都将显示 不同但重叠的基因表达谱。那些基因 重要的改变酒精反应将确定聚类和 表达谱比较中的判别分析程序 以及16种情况下的行为反应。第3章将 使用核糖核酸酶验证重要的基因表达变化 保护测定和/或实时定量PCR。的结果 拟议的研究将提供深入了解神经适应过程， 有助于酒精滥用，并将提供更多的信息， 进一步研究酒精相关的神经适应。

DESCRIPTION (provided by applicant): Behavioral sensitivity to alcohol can be modified as a function of both environmental and genetic factors. The underlying basis of this effect is the ability of neurons to adapt in response to environmental perturbations including, but not limited to alcohol exposure, and the specific nature of this response is dependent on genetic makeup. These processes may be an important feature in the etiology of alcoholism. Evidence indicates that drugs acting on dopamine (DA) neurons are able to alter the behavioral response to alcohol in rodents. Thus, 24 hrs after a single treatment with the DA antagonist haloperidol or the indirect DA agonist methamphetamine, rats become more or less sensitive to alcohol, respectively, and "rapid" tolerance develops one day after a single dose of alcohol. These acute drug responses will be exploited to investigate the global gene expression changes occurring in DA pathways that contribute to altered alcohol sensitivity. Specific Aim 1 will fully characterize the behavioral response to alcohol (loss of righting reflex) 24 hrs after acute treatment with haloperidol, alcohol, or methamphetamine in replicate inbred High and Low Alcohol Sensitive rat strains. Gene expression analyses will be performed in Specific Aim 2. Rats will be sacrificed at 8 hrs following administration of a single optimal dose of each drug and the striatum and ventral midbrain will be dissected. RNA will be extracted from these structures and gene expression will be determined with the use of the Affymetrix GeneChip system. This will provide a total of 16 experimental conditions comprised of different drug/genotype combinations. It is postulated that each of these conditions will show different, but overlapping gene expression profiles. The genes that are important in the altered alcohol response will be identified by clustering and discriminative analysis procedures in a comparison of the expression profiles and the behavioral responses among the 16 conditions. Specific Aim 3 will validate important gene expression changes with the use of ribonuclease protection assays and/or real-time quantitative PCR. The results of the proposed studies will offer insight into the neuroadaptive processes that contribute to alcohol abuse and will also provide more information from which to further investigate alcohol related neuroadaptation.