Filošević Ana, Al-Samarai Sabina, Andretić Waldowski Rozi
Department of Biotechnology, University of Rijeka, Rijeka, Croatia.
Front Mol Neurosci. 2018 Feb 5;11:25. doi: 10.3389/fnmol.2018.00025. eCollection 2018.
can be used to identify genes with novel functional roles in neuronal plasticity induced by repeated consumption of addictive drugs. Behavioral sensitization is a relatively simple behavioral output of plastic changes that occur in the brain after repeated exposures to drugs of abuse. The development of screening procedures for genes that control behavioral sensitization has stalled due to a lack of high-throughput behavioral tests that can be used in genetically tractable organism, such as . We have developed a new behavioral test, FlyBong, which combines delivery of volatilized cocaine (vCOC) to individually housed flies with objective quantification of their locomotor activity. There are two main advantages of FlyBong: it is high-throughput and it allows for comparisons of locomotor activity of individual flies before and after single or multiple exposures. At the population level, exposure to vCOC leads to transient and concentration-dependent increase in locomotor activity, representing sensitivity to an acute dose. A second exposure leads to further increase in locomotion, representing locomotor sensitization. We validate FlyBong by showing that locomotor sensitization at either the population or individual level is absent in the mutants for circadian genes , , and . The locomotor sensitization that is present in and mutant flies is in large part not cocaine specific, but derived from increased sensitivity to warm air. Circadian genes are not only integral part of the neural mechanism that is required for development of locomotor sensitization, but in addition, they modulate the intensity of locomotor sensitization as a function of the time of day. Motor-activating effects of cocaine are sexually dimorphic and require a functional dopaminergic transporter. FlyBong is a new and improved method for inducing and measuring locomotor sensitization to cocaine in individual . Because of its high-throughput nature, FlyBong can be used in genetic screens or in selection experiments aimed at the unbiased identification of functional genes involved in acute or chronic effects of volatilized psychoactive substances.
可用于鉴定在反复摄入成瘾性药物所诱导的神经元可塑性中具有新功能作用的基因。行为敏化是反复接触滥用药物后大脑中发生的可塑性变化的一种相对简单的行为输出。由于缺乏可用于遗传易处理生物体（如 ）的高通量行为测试，控制行为敏化的基因筛选程序的开发陷入停滞。我们开发了一种新的行为测试方法FlyBong，它将挥发性可卡因（vCOC）递送至单独饲养的果蝇，并对其运动活动进行客观量化。FlyBong有两个主要优点：它是高通量的，并且允许比较单个果蝇在单次或多次接触前后的运动活动。在群体水平上，接触vCOC会导致运动活动短暂且浓度依赖性增加，这代表对急性剂量的敏感性。第二次接触会导致运动进一步增加，这代表运动敏化。我们通过证明在生物钟基因 、 和 的突变体中，群体或个体水平上均不存在运动敏化来验证FlyBong。 和 突变果蝇中存在的运动敏化在很大程度上不是可卡因特异性的，而是源于对暖空气的敏感性增加。生物钟基因不仅是运动敏化发展所需神经机制的组成部分，此外，它们还根据一天中的时间调节运动敏化的强度。可卡因的运动激活作用具有性别差异，并且需要功能性多巴胺能转运体。FlyBong是一种用于诱导和测量个体对可卡因运动敏化的新的改进方法。由于其高通量性质，FlyBong可用于遗传筛选或选择实验，旨在无偏鉴定参与挥发性精神活性物质急性或慢性作用的功能基因。
