Persistent changes in brain expression after withdrawal

慢性乙醇暴露会导致身体依赖的发展。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，我们建议通过鉴定受慢性酒精暴露和长时间戒断反应调节的mrna，来确定在男女戒断综合征严重程度调节中起重要作用的特定基因。我们将采用独特的戒断严重程度模型，戒断发作倾向系和戒断发作抗性系（WSP和WSR），它们代表了来自自交系的八向杂交的等位基因，这些等位基因选择性地经过几代繁殖以达到高或低戒断严重程度。需要验证的假设是，戒断导致的大脑基因表达改变是介导神经适应和身体依赖的关键因素。基因调控后，慢性乙醇暴露可能增加戒断风险，或发挥保护对戒断。我们还假设基因谱将显示性别特异性调节。由于自愿饮酒与戒断严重程度之间的遗传反比关系，比较对照组WSP和WSR动物可能会发现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。在存在或不存在严重戒断的情况下，将从小鼠身上采集前额皮质；在基因表达的时间模式将确定在一个延长的时间过程中使用微阵列分析。特异性目的1将通过表征WSP小鼠mRNA表达的差异，确定可能介导神经适应过程的mRNA转录物，这些神经适应过程是雄性和雌性身体依赖和戒断综合征发展风险的基础。特异性目的2将通过表征WSR小鼠mRNA表达的差异，鉴定可能介导神经适应过程的mRNA转录物，这些神经适应过程可以防止身体依赖和戒断综合征在两性中的发展。特异性目标3将通过表征对照组男性和女性WSP和WSR之间mRNA表达的差异，确定介导自愿饮酒的mRNA转录物。本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）如何在遗传水平上受到调节的机制。这一知识可能有助于制定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 Male and female WSP or WSR mice will be exposed to intoxicating levels of ethanol and withdrawn.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.