Persistent changes in brain expression after withdrawal

长期接触酒精会导致身体依赖的发展。在去除乙醇后，会出现可能危及生命的戒断综合症。戒断综合症的各个方面也受到性别的影响。利用微阵列分析，我们建议通过识别长期酒精暴露和戒断过程中调节戒断综合症严重程度的mRNAs，来识别在调节戒断综合症严重程度方面具有重要意义的特定基因。我们将采用独特的戒断严重程度模型，即易戒断癫痫和耐戒断品系(WSP和WSR)，它们代表了近交系的八个等位基因，这些近交系是在几代人中选择性地培育出戒断严重程度高或低的近交系。需要检验的假设是，戒断引起的大脑基因表达变化是调节神经适应和身体依赖的关键因素。慢性酒精暴露后调控的基因可能会增加戒断风险，或者对戒断起到保护作用。我们还假设，基因图谱将显示出性别特有的调控。由于自愿饮酒和戒断严重程度之间的遗传关系相反，对照WSP和WSR动物之间的比较可能会发现调节酒精消费的基因。雄性和雌性WSP或WSR小鼠将暴露在醉人水平的乙醇中并退出。前额叶皮质将从存在或不存在严重戒断的小鼠身上获得；基因表达的时间模式将使用微阵列分析在更长的时间过程中识别。具体目标1将通过表征WSP小鼠mRNA表达的差异，鉴定可能介导神经适应过程的mRNA转录本，这些过程是导致男性和女性身体依赖和戒断综合症发展风险的基础。特定目标2将通过表征WSR小鼠的mRNA表达差异，识别可能介导神经适应过程的mRNA转录本，这些转录本可以防止两性身体依赖和戒断综合症的发展。具体目标3将通过表征对照男性和女性WSP和WSR之间的mRNA表达差异，确定可能作为调节自愿饮酒的候选的mRNA转录本。这项研究的长期目标是了解慢性酒精暴露的有害反应(即导致戒断发作的身体依赖)如何在基因水平上受到调控的机制。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 Male and female WSP or WSR mice will be exposed to intoxicating levels of ethanol and withdrawn.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.