Gene Expression and -Drug-Induced Sensitivity to Alcohol

基因表达和药物诱导的酒精敏感性

• 批准号: 6623623
• 负责人: RICHARD A RADCLIFFE
• 金额: $ 30.5万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6623623
• 关键词: RNase protection assay; alcohols; behavioral genetics; behavioral habituation /sensitization; chemosensitizing agent; dopamine; dopamine receptor; functional /structural genomics; gene environment interaction; gene expression; genetic strain; haloperidol; laboratory rat; male; methamphetamine; microarray technology; neurogenetics; polymerase chain reaction

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.

描述（由申请人提供）：对酒精的行为敏感性可以是 修饰是环境和遗传因素的函数。这 这种作用的基础是神经元适应反应的能力 环境扰动，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 过程中的过程可能是酒精中毒病因的重要特征。证据 表明作用于多巴胺（DA）神经元的药物能够改变 对啮齿动物中酒精的行为反应。因此，一次后24小时 DA拮抗剂氟哌啶醇或间接DA激动剂的处理 甲基苯丙胺分别对酒精或多或少敏感 单剂量酒精后的一天，“快速”的耐受性发生。这些 急性药物反应将被利用以研究全球基因 在DA途径中发生的表达变化，导致酒精改变 灵敏度。具体目标1将充分表征对 急性治疗后24小时的酒精（失去矫正反射） 氟哌啶醇，酒精或甲基苯丙胺在复制的高和低位 酒精敏感的大鼠菌株。基因表达分析将在 特定目标2。在给药后，将在8小时时处死大鼠 每种药物的单一最佳剂量以及纹状体和腹中脑将是 解剖。 RNA将从这些结构中提取，基因表达将 可以通过使用Affymetrix Genechip系统来确定。这将提供 共有16个实验条件由不同的药物/基因型组成 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。基因 对于改变的酒精反应，将通过聚类和 比较表达曲线的判别分析程序 以及16条条件中的行为反应。具体的目标3将 通过使用核糖核酸酶来验证重要的基因表达变化 保护测定和/或实时定量PCR。结果 拟议的研究将提供对神经适应过程的见解 有助于酗酒，还将提供更多信息 进一步研究与酒精相关的神经适应。