Pfeiffenberger Cory, Lear Bridget C, Keegan Kevin P, Allada Ravi
Cold Spring Harb Protoc. 2010 Nov 1;2010(11):pdb.prot5520. doi: 10.1101/pdb.prot5520.
Adult behavioral assays have been used with great success in Drosophila melanogaster to identify circadian rhythm genes. In particular, the locomotor activity assay can identify altered behavior patterns over the course of several days in small populations, or even individual flies. Sleep is a highly conserved behavior that is required for optimal performance and, in many cases, life of an organism. Drosophila demonstrate a behavioral state that shows traits consistent with sleep: periods of relative behavioral immobility that coincide with an increased arousal threshold after ~5 min of inactivity, regulated by circadian and homeostatic mechanisms. However, because flies do not produce brain waves recordable by electroencephalography, sleep researchers use behavior-based paradigms to infer when a fly is asleep, as opposed to awake but immobile. Data on Drosophila activity can be collected using an automated monitoring system to provide insight into sleep duration, consolidation, and latency, as well as sleep deprivation and rebound. This protocol details the use of Counting Macro, an Excel-based program, to process data created with the Drosophila Activity Monitoring (DAM) System from TriKinetics for sleep analyses. Specifically, it details the steps necessary to convert the raw data created by the DAM System into sleep duration and consolidation data, broken down into the light (L), dark (D), light:dark cycling (LD), and constant darkness (DD) phases of a behavior experiment.
成年行为分析在黑腹果蝇中已被成功用于识别昼夜节律基因。特别是,运动活动分析可以在几天内从小群体甚至单个果蝇中识别出行为模式的改变。睡眠是一种高度保守的行为,对于生物体的最佳表现乃至在许多情况下的生存都是必需的。果蝇表现出一种行为状态,其特征与睡眠一致:在静止约5分钟后,相对行为静止期与唤醒阈值增加相吻合,这受昼夜节律和稳态机制调节。然而,由于果蝇不会产生可通过脑电图记录的脑电波,睡眠研究人员使用基于行为的范式来推断果蝇何时入睡,而不是清醒但静止不动的时候。可以使用自动监测系统收集果蝇活动的数据,以深入了解睡眠时间、巩固情况和潜伏期,以及睡眠剥夺和反弹情况。本方案详细介绍了使用基于Excel的程序Counting Macro来处理由TriKinetics的果蝇活动监测(DAM)系统生成的数据以进行睡眠分析。具体来说,它详细说明了将DAM系统创建的原始数据转换为睡眠时间和巩固数据所需的步骤,这些数据分为行为实验的光照(L)、黑暗(D)、光照:黑暗循环(LD)和持续黑暗(DD)阶段。