Zebrafish model: molecular basis of acute EtOH tolerance

描述(由申请人提供)：急性功能性耐受(AFT)是一种众所周知的现象，被认为影响酒精中毒风险风险的个体差异，但AFT的潜在分子底物仍然是一个谜。在这一应用中，我们建议检验斑马鱼(Danio Rerio)将对乙醇造成的行为干扰表现出AFT的假设，主要目标是在AFT的分子基础研究中表征和发展斑马鱼作为模式生物。斑马鱼为遗传研究提供了许多优势，包括世代时间短、后代数量多、成本和空间需求低、关于斑马鱼遗传学的信息量越来越多，以及可以轻松完成遗传操作。斑马鱼在AFT研究中的另一个重要优势是，由于乙醇是通过与保持罐溶液的平衡来给药的，因此可以很容易地保持血液和大脑中稳定的乙醇浓度；本质上，这是一个极其简单的乙醇夹。已经在金鱼身上证明了AFT对某些行为的作用，但还没有在斑马鱼身上进行过研究。该项目的具体目标是为获得AFT筛选几种既定的斑马鱼行为，通过测试几个近交系斑马鱼品系来确定AFT是否存在自然发生的遗传变异，并开发用于AFT的自动化、高通量突变筛选方法。未来的研究将在大量诱变斑马鱼上进行高通量筛选，目标是绘制斑马鱼AFT的分子基础图。

DESCRIPTION (provided by applicant): Acute functional tolerance (AFT) to ethanol is a well known phenomenon that is thought to influence individual differences in risk liability for alcoholism, but the underlying molecular substrates of AFT remain a mystery. In this application, we are proposing to test the hypothesis that the zebrafish (Danio rerio) will exhibit AFT to the behavioral disruptions produced by ethanol, with the main goal of the proposal to characterize and develop the zebrafish as a model organism in the study of the molecular basis of AFT. Zebrafish provide numerous advantages for genetic studies including short generation time, large numbers of offspring, low cost and space demands, a growing body of information on the genetics of zebrafish, and the ease with which genetic manipulations can be accomplished. Another important advantage of zebrafish in AFT studies is that a steady-state blood and brain ethanol concentration can easily be maintained since the ethanol is administered via equilibration with the holding tank solution; in essence, an extremely simple ethanol clamp. AFT for certain behaviors has been demonstrated in goldfish, but it has not been examined in zebrafish. The Specific Aims of this project are to screen several established zebrafish behaviors for the acquisition of AFT, determine if there is naturally occurring genetic variation for AFT by testing several inbred zebrafish strains, and to develop methods for automated, high-throughput mutagenesis screens for AFT. Future studies will implement the high-throughput screens on large numbers of mutagenized zebrafish with the goal of mapping the molecular basis of AFT in zebrafish.