Delgado Nataly M, Sierra Carmen Mariana, Arzola Abraham, Saldes Erick Benjamin, Han Kyung-An, Sabandal Paul Rafael
Department of Biological Sciences, University of Texas at El Paso.
Department of Biological Sciences, University of Texas at El Paso;
J Vis Exp. 2020 May 18(159). doi: 10.3791/61123.
Alcohol use disorder (AUD) remains a serious problem in our society. To develop effective interventions for addiction, it is important to understand the underlying neurobiological mechanisms, for which diverse experimental approaches and model systems are needed. The main ingredient of alcoholic beverages is ethanol, which causes adaptive changes in the central nervous system and behavior upon chronic intake. Behavioral sensitization (i.e., escalated responses) in particular represents a key adaptive change underlying addiction. Most ethanol-induced behavioral sensitization studies in animal models have been conducted on the locomotor activating effect of ethanol. A prominent effect of ethanol is behavioral disinhibition. Behavioral sensitization to the disinhibition effect of ethanol, however, is underrepresented. To address this issue, we developed the Flypub assay that allows measuring the escalated increase in disinhibited courtship activities upon recurring ethanol exposure in Drosophila melanogaster. Here, we report the step-by-step Flypub assay including assembly of ethanol exposure chambers, setup of the assay station, criteria for fly care and collection, ethanol delivery, quantification of disinhibited courtship activities, data processing and statistical analysis. Also provided are how to troubleshoot critical steps, overcome limitations and expand its utility to assess additional ethanol-induced behaviors. The Flypub assay in combination with powerful genetic tools in Drosophila melanogaster will facilitate the task of discovering the mechanism underlying ethanol-induced behavioral sensitization.
酒精使用障碍（AUD）在我们的社会中仍然是一个严重的问题。为了开发有效的成瘾干预措施，了解潜在的神经生物学机制很重要，为此需要多种实验方法和模型系统。酒精饮料的主要成分是乙醇，长期摄入后会引起中枢神经系统和行为的适应性变化。行为敏化（即反应增强）尤其代表成瘾背后的关键适应性变化。动物模型中大多数乙醇诱导的行为敏化研究都是针对乙醇的运动激活作用进行的。乙醇的一个显著作用是行为去抑制。然而，对乙醇去抑制作用的行为敏化研究较少。为了解决这个问题，我们开发了Flypub试验，该试验可以测量果蝇反复接触乙醇后去抑制求偶活动的增强。在此，我们报告了Flypub试验的详细步骤，包括乙醇暴露室的组装、试验站的设置、果蝇饲养和收集的标准、乙醇输送、去抑制求偶活动的量化、数据处理和统计分析。还提供了如何对关键步骤进行故障排除、克服局限性以及扩展其用途以评估其他乙醇诱导行为的方法。Flypub试验与果蝇强大的遗传工具相结合，将有助于发现乙醇诱导行为敏化背后机制的任务。
