Gene Expression and -Drug-Induced Sensitivity to Alcohol

描述(申请人提供)：行为对酒精的敏感度 作为环境和遗传因素的函数而改变的。这个 这种效应的潜在基础是神经元适应反应的能力。 对环境的干扰，包括但不限于酒精暴露， 而这种反应的具体性质取决于基因构成。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 啮齿动物对酒精的行为反应。因此，在一次单一的 用多巴胺拮抗剂氟哌啶醇或间接多巴胺激动剂治疗 甲基苯丙胺，大鼠对酒精或多或少变得敏感， 单次饮酒一天后，就会出现“快速”耐受性。这些 将利用急性药物反应来研究全球基因 导致酒精改变的DA通路中的表达变化 敏感度。具体目标1将充分描述对以下行为的反应 酒精(翻正反射丧失)急性治疗后24小时 氟哌啶醇、酒精或甲基苯丙胺在高、低复制近交系中的应用 对酒精敏感的老鼠品系。基因表达分析将在 具体目的2.给药后8小时处死大鼠 每种药物的单一最佳剂量将是纹状体和腹侧中脑 被解剖了。RNA将从这些结构中提取出来，基因表达将 使用Affymetrix基因芯片系统进行检测。这将提供 共有16个实验条件，由不同的药物/基因组成 组合。据推测，这些条件中的每一种都将表明 不同但重叠的基因表达谱。这些基因是 在酒精反应改变中的重要作用将通过聚集和 表达谱比较中的判别分析程序 以及16种条件下的行为反应。具体目标3将 使用核糖核酸酶验证重要基因表达的变化 保护性检测和/或实时定量聚合酶链式反应。评选结果 拟议的研究将提供对神经适应过程的洞察， 促进酗酒，并将提供更多信息，从中 以进一步研究与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.