Alcohol preference in zebra fish: a novel paradigm

酗酒是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。然而，允许快速筛查斑马鱼大脑与酒精相关的功能变化的高通量行为测试(或允许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。该提案的目标是开发这样的行为范例。将设计一种新颖的Y形水槽系统，在该系统中，斑马鱼将在缓慢的水流中游泳，并在水箱的左或右目标臂之间进行选择。该范式是多才多艺的，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。测试中行为的量化将通过使用红外光电池探测器将行为数据馈送到计算机来自动化，计算机将监测和记录行为反应以及控制刺激(例如，酒精和条件性刺激)的传递。自动化将使我们能够增加并行运行的设备数量，从而使该系统适用于高通量筛选。最终，我们将使用该范例来筛选和鉴定由随机突变(乙基亚硝脲或ENU诱导点突变)产生的突变斑马鱼，这是一种正向遗传方法，将导致识别与急性和慢性酒精相关行为相关的基因。斑马鱼特别适合作为我们目标的模式生物，因为它体积小，多产(每次产卵200条)，易于控制其环境，易于传递酒精等水溶性物质(鱼类通过鳃和皮肤吸收)，以及可用的大量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有很高的同源性，我们希望拟议的研究将有助于揭示人类急性和慢性酒精影响的机制，最终导致开发有效的治疗方法。

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.