Zebrafish model: molecular basis of acute EtOH tolerance

斑马鱼模型：急性乙醇耐受的分子基础

• 批准号: 6879229
• 负责人: RICHARD A RADCLIFFE
• 金额: $ 7.7万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6879229
• 关键词: alcoholism /alcohol abuse; alternatives to animals in research; behavior test; behavioral /social science research tag; behavioral genetics; biomedical automation; digital imaging; drug administration rate /duration; drug tolerance; ethanol; ethology; genetic screening; high throughput technology; method development; model design /development; pharmacogenetics; substance abuse related behavior; zebrafish

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.

描述（由申请人提供）：对乙醇的急性功能耐受性（AFT）是一种众所周知的现象，被认为会影响酒精中毒风险倾向的个体差异，但AFT的潜在分子底物仍然是一个谜。在这个应用程序中，我们建议测试的假设，即斑马鱼（Danio rerio）将表现出AFT的乙醇产生的行为中断，该建议的主要目标是表征和开发的斑马鱼作为模式生物在AFT的分子基础的研究。斑马鱼为遗传学研究提供了许多优势，包括世代时间短，后代数量大，成本低，空间需求小，斑马鱼遗传学信息量不断增加，以及易于完成遗传操作。斑马鱼在AFT研究中的另一个重要优势是可以容易地维持稳态血液和脑乙醇浓度，因为乙醇是通过与保持罐溶液平衡来施用的;本质上，是一种非常简单的乙醇钳。某些行为的AFT已在金鱼中得到证实，但尚未在斑马鱼中进行检查。本项目的具体目标是筛选几种已建立的斑马鱼行为，以获得AFT，通过测试几种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，高通量诱变筛选的方法。未来的研究将在大量的诱变斑马鱼上进行高通量筛选，以定位斑马鱼AFT的分子基础。