Alcohol and zebra fish: Behavioral screens for mutants

描述(申请人提供)：酗酒是社会必须面对的最具破坏性和代价最高的疾病之一。其发病机制尚不清楚，缺乏有效的治疗方法。研究这种复杂疾病的一种方法是使用模型生物。斑马鱼(Danio Rerio)可能是一个有用的模型系统，可以用来研究酒精的急性和慢性影响的机制。在这里，将开发新的行为测试范式，以量化酒精对新奇诱导的、争斗的、社交的和捕食者诱导的行为的影响。这些测试希望利用酒精可能改变的大脑的不同功能。这些测试可能使我们能够检测到一系列化学诱变剂诱导的基因变化，这是一种向前遗传的方法。斑马鱼特别适合作为我们的目标模式生物。首先，它可以在实验室中大量饲养、繁殖和测试(体积小，每次产卵的后代数量多，易于控制环境因素)。其次，饮酒很简单(鱼通过鳃和皮肤吸收这种物质)。第三，大量的遗传信息和先进的遗传工具可用于产生突变和识别新基因。第四，发现的基因与人类有很高的同源性。然而，尽管有这些优点，特别是斑马鱼行为的表型筛选方法往往是缺乏的。重要的是，我们认为成功的正向遗传学研究的基础在于开发适当的表型测试范例。此外，也许测试大脑功能变化的最好、最简单的方法之一是行为分析。因此，这项建议的主要目标是首先开发新的行为范式来检测酒精诱导的变化，其次从技术和程序上改进这些范式，以便高通量筛选突变。这项建议的另一个目标是利用新开发的行为筛选技术产生和鉴定第一批突变斑马鱼，然后建立稳定的突变斑马鱼品系。最终，即在该提案有效期结束后，拟议的研究将导致在斑马鱼中发现与酒精相关行为相关的新基因，这有望有助于揭开人类酒精对急性和慢性影响的机制。

DESCRIPTION (provided by applicant): Alcohol abuse is one of the most devastating and costly diseases society has to face. Its mechanisms are not well understood and effective treatment is lacking. One way to study this complex disease is with the use of model organisms. Zebrafish (Danio rerio) may be a useful model system with which the mechanisms of acute and chronic effects of alcohol can be investigated. Here new behavioral test paradigms will be developed for the quantification of alcohol effects on novelty induced, agonistic, social, and predator elicited behaviors. The tests are hoped to tap into diverse functions of the brain that may be modified by alcohol. The tests may enable us to detect a range of chemical mutagen induced genetic changes, a forward genetic approach. Zebrafish is particularly appropriate as a model organism for our purpose. First, it can be kept, bred, and tested in large numbers in the laboratory (small size, large number of offspring per spawning, ease of controlling environmental factors). Second, alcohol delivery is simple (fish absorb this substance through their gills and skin). Third, a large amount of genetic information and sophisticated genetic tools are available allowing the generation of mutations and identification of novel genes. And fourth, the genes discovered show high homology to human. Despite these advantages, however, phenotypical screening methods especially for behavior of zebrafish are often lacking. Importantly, we argue that the foundation of a successful forward genetic study lies in the development of appropriate phenotypical test paradigms. Furthermore, perhaps one of the best and simplest ways to test functional changes in the brain is behavioral analysis. Thus, the primary goal of this proposal is first to develop novel behavioral paradigms that detect alcohol induced changes, and second, to technically and procedurally refine these paradigms for high throughput screening of mutants. Another goal of this proposal is to generate and identify the first mutants with the use of the newly developed behavioral screens and then to establish stable mutant zebrafish lines. Ultimately, i.e. after the tenure of the proposal, the proposed studies will lead to the identification of novel genes relevant for alcohol related behaviors in zebrafish, which is hoped will facilitate the unraveling of the mechanisms of acute and chronic alcohol effects in human.