Alcohol preference in zebra fish: a novel paradigm

酒精滥用是最具破坏性和最昂贵的疾病之一。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作用机制。然而，能够快速筛选与酒精相关的斑马鱼大脑功能改变的高通量行为测试（或能够快速检测与酒精相关机制相关的基因突变引起的神经生物学变化的测试）几乎完全不可用。该提案的目标是发展这样的行为范式。将设计一种新颖的y型水箱系统，斑马鱼将在缓慢的水流中游泳，并在水箱的左臂或右臂之间选择。这个范例是通用的，将使我们能够测试运动功能，酒精偏好/回避，以及酒精奖励的位置偏好。测试中的行为量化将通过使用红外光电池探测器自动实现，将行为数据输入计算机，计算机将监控和记录行为反应，并控制刺激的传递（例如酒精和条件刺激）。自动化将允许我们增加并行运行的设备数量，从而使系统对高通量筛选有用。最后，我们将使用该范式筛选和鉴定随机突变（乙基-亚硝基-尿素或ENU诱导的点突变）产生的斑马鱼突变体，这是一种前向遗传方法，将导致鉴定涉及急性和慢性酒精相关行为的基因。斑马鱼特别适合作为我们研究目的的模式生物，因为它体积小，多产的性质（每次产卵200多条鱼），易于控制其环境，易于输送水溶性物质，如酒精（鱼通过鳃和皮肤吸收），以及大量的遗传信息和遗传工具。由于在斑马鱼中发现的几个基因与人类具有高度的同源性，我们希望所提出的研究将有助于揭示人类急性和慢性酒精效应的机制，最终导致有效治疗的发展。

DESCRIPTION Alcohol abuse is one of the most devastating and costly diseases. 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. Zebra fish (Danio rerio) may be a useful model system with which the mechanisms of acute and chronic effects of alcohol can be investigated. However, high throughput behavioral tests allowing fast screening of alcohol related functional alterations of the brain of zebra fish (or tests allowing quick detection of neurobiological changes induced by mutations of genes associated with alcohol related mechanisms) are almost completely unavailable. The goal of the proposal is to develop such behavioral paradigms. A novel Y-tank system will be designed in which zebra fish will swim against a slow water current and choose between the left or right goal arms of the tank. The paradigm is versatile and will enable us to test motor function, alcohol preference/avoidance, and alcohol rewarded place preference. Quantification of behavior hi the test will be automated with the use of infrared photocell detectors feeding the behavioral data to computers that will monitor and record the behavioral responses as well as control the delivery of stimuli (e.g., alcohol and conditioned stimuli). Automation will allow us to increase the number of apparati running in parallel thus making the system useful for high throughput screening. Ultimately, we will use the paradigm to screen and identify mutant zebra fish generated by random mutagenesis (ethyl-nitroso-urea or ENU induced point mutations), a forward genetic approach that will lead to the identification of genes involved in acute and chronic alcohol related behaviors. Zebra fish is particularly appropriate as a model organism for our purpose because of its small size, prolific nature (200+ fish per spawning), ease of controlling of its environment, ease of delivery of water soluble substances such as alcohol (fish absorb it through their gills and skin), and the huge amount of genetic information and genetic tools available. As several genes discovered in zebra fish show high homology to human, we hope the proposed studies will facilitate the unraveling of the mechanisms of acute and chronic alcohol effects in human ultimately resulting in the development of efficient treatments.