Alcohol and zebra fish: Behavioral screens for mutants

描述（由申请人提供）：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式生物。斑马鱼（Danio rerio）可能是一个有用的模型系统，可以研究酒精的急性和慢性效应的机制。在这里，新的行为测试范式将开发的量化酒精对新奇的诱导，竞争，社会和捕食者引起的行为的影响。这些测试有望挖掘大脑可能被酒精改变的各种功能。这些测试可能使我们能够检测一系列化学诱变剂诱导的遗传变化，这是一种向前的遗传方法。斑马鱼特别适合作为我们研究的模式生物。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大，易于控制环境因素）。第二，酒精的传递很简单（鱼通过鳃和皮肤吸收这种物质）。第三，大量的遗传信息和复杂的遗传工具可用于产生突变和识别新基因。第四，所发现的基因与人类具有很高的同源性。然而，尽管有这些优点，表型筛选方法，特别是斑马鱼的行为往往缺乏。重要的是，我们认为，一个成功的正向遗传学研究的基础在于适当的表型测试范式的发展。此外，也许测试大脑功能变化的最好和最简单的方法之一是行为分析。因此，该提议的主要目标是首先开发检测酒精诱导的变化的新的行为范例，其次，在技术上和程序上完善这些范例以用于突变体的高通量筛选。本提案的另一个目标是使用新开发的行为筛选产生和鉴定第一个突变体，然后建立稳定的突变斑马鱼品系。最终，也就是说，在该提案的任期结束后，拟议的研究将导致在斑马鱼中识别与酒精相关行为相关的新基因，希望这将有助于解开人类急性和慢性酒精影响的机制。

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.