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测量单个黑腹果蝇酒精镇静时间的高通量方法。

High-Throughput Method for Measuring Alcohol Sedation Time of Individual Drosophila melanogaster.

作者信息

Sass Tatum N, MacPherson Rebecca A, Mackay Trudy F C, Anholt Robert R H

机构信息

Department of Genetics and Biochemistry and Center for Human Genetics, Clemson University.

Department of Genetics and Biochemistry and Center for Human Genetics, Clemson University;

出版信息

J Vis Exp. 2020 Apr 20(158). doi: 10.3791/61108.

Abstract

Drosophila melanogaster provides an excellent model to study the genetic underpinnings of alcohol sensitivity. In contrast to studies in human populations, the Drosophila model allows strict control over genetic background, and virtually unlimited numbers of individuals of the same genotype can be reared rapidly under well-controlled environmental conditions without regulatory restrictions and at relatively low cost. Flies exposed to ethanol undergo physiological and behavioral changes that resemble human alcohol intoxication, including loss of postural control, sedation, and development of tolerance. Here, we describe a simple, low-cost, high-throughput assay for assessing alcohol sedation sensitivity in large numbers of single flies. The assay is based on video recording of single flies introduced without anesthesia in 24-well cell culture plates in a set-up that enables synchronous initiation of alcohol exposure. The system enables a single person to collect individual ethanol sedation data on as many as 2,000 flies within an 8 h work period. The assay can, in principle, be extended to assess the effects of exposure to any volatile substance and applied to measure effects of acute toxicity of volatiles on other insects, including other fly species.

摘要

黑腹果蝇为研究酒精敏感性的遗传基础提供了一个绝佳的模型。与人类群体研究不同,果蝇模型能够严格控制遗传背景,并且在可控的环境条件下,几乎可以无限制地快速饲养大量相同基因型的个体,不受监管限制且成本相对较低。暴露于乙醇的果蝇会发生类似于人类酒精中毒的生理和行为变化,包括姿势控制丧失、镇静以及耐受性的发展。在这里,我们描述了一种简单、低成本、高通量的方法,用于评估大量单只果蝇的酒精镇静敏感性。该方法基于对未麻醉的单只果蝇在24孔细胞培养板中的视频记录,该设置能够同步开始酒精暴露。该系统使一个人能够在8小时的工作时间内收集多达2000只果蝇的个体乙醇镇静数据。原则上,该方法可以扩展到评估暴露于任何挥发性物质的影响,并应用于测量挥发性物质对其他昆虫(包括其他果蝇物种)的急性毒性影响。

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