Persistent changes in brain expression after withdrawal

慢性酒精暴露导致身体依赖的发展。去除乙醇后，会出现戒断综合征，可能危及生命。戒断综合征的各个方面也受到性别的影响。使用微阵列分析，我们建议确定特定的基因的重要性，在调节的严重程度，在两种性别的戒断综合征，通过确定的mRNA调节长期酒精暴露和退出一个较长的时间过程。我们将采用独特的戒断严重性模型，即戒断癫痫倾向和耐药品系（WSP和WSR），它们代表了来自近交系小鼠品系的八向杂交的等位基因，这些品系选择性地经过几代繁殖，以获得高或低的戒断严重性。有待检验的假设是，由戒断引起的大脑基因表达的改变是介导神经适应和身体依赖的关键因素。慢性乙醇暴露后调节的基因可能会增加戒断风险，或对戒断产生保护作用。我们还假设基因图谱将显示性别特异性调控。由于自愿饮酒和戒断严重程度之间的逆遗传关系，对照WSP和WSR动物之间的比较可以确定调节酒精消耗的基因。雄性和雌性WSP或WSR小鼠将暴露于中毒水平的乙醇并退出。将在存在或不存在严重戒断的情况下从小鼠收获前额叶皮质;将使用微阵列分析在延长的时间过程中鉴定基因表达的时间模式。具体目标1将通过表征WSP小鼠中mRNA表达的差异，鉴定可能介导神经适应过程的mRNA转录物，该过程是雄性和雌性动物发生身体依赖和戒断综合征风险的基础。具体目标2将通过表征WSR小鼠中mRNA表达的差异来鉴定可能介导神经适应过程的mRNA转录物，所述神经适应过程保护两性免于身体依赖和戒断综合征的发展。具体目标3将通过表征对照组雄性和雌性WSP和WSR之间mRNA表达的差异来鉴定作为介导自愿饮酒的假定候选者的mRNA转录物。这项研究的长期目标是了解慢性乙醇暴露的有害反应（即导致戒断发作的身体依赖）在遗传水平上的调节机制。这些知识可能有助于开发治疗酒精依赖的新策略。

Chronic ethanol exposure results in the development of physical dependence. After ethanol removal, a withdrawal syndrome is exhibited that can be life threatening. Aspects of the withdrawal syndrome are also influenced by gender. Using microarray analysis, we propose to identify specific genes of importance in modulating the severity of the withdrawal syndrome in both genders by identifying mRNAs regulated by chronic alcohol exposure and withdrawal over an extended time course. We will employ unique models of withdrawal severity, the Withdrawal Seizure- Prone and -Resistant lines (WSP and WSR), that represent alleles from an eight-way cross of inbred mouse lines selectively bred over generations for high or low withdrawal severity. The hypothesis to be tested is that altered brain gene expression that results from withdrawal is a critical factor mediating neuroadaptation and physical dependence. Genes regulated following chronic ethanol exposure might promote withdrawal risk, or exert protection against withdrawal. We also hypothesize that gene profiles will show sex-specific regulation. Due to the inverse genetic relationship between voluntary ethanol drinking and withdrawal severity, comparison between control WSP and WSR animals may identify genes modulating alcohol consumption. Male and female WSP or WSR mice will be exposed to intoxicating levels of ethanol and withdrawn. Prefrontal cortex will be harvested from mice in the presence or absence of severe withdrawal; temporal patterns in gene expression will be identified over an extended time course using microarray analysis. Specific Aim 1 will identify mRNA transcripts that may mediate neuroadaptative processes that underlie risk for the development of physical dependence and aspects of the withdrawal syndrome in both males and females by characterizing differences in mRNA expression in WSP mice. Specific Aim 2 will identify mRNA transcripts that may mediate neuroadaptative processes that protect against the development of physical dependence and aspects of the withdrawal syndrome in both sexes by characterizing differences in mRNA expression in WSR mice. Specific Aim 3 will identify mRNA transcripts that are putative candidates for mediating voluntary alcohol consumption by characterizing differences in mRNA expression between control male and female WSP and WSR. The long-term goal of this research is to understand mechanisms underlying how deleterious responses to chronic ethanol exposure (i.e. physical dependence leading to withdrawal seizures) are regulated at the genetic level. This knowledge may help in the development of new strategies for the treatment of alcohol dependence.