Published: December 14th, 2020
Sleep deprivation is a powerful tool to investigate sleep function and regulation. We describe a protocol to sleep deprive Drosophila using the Sleep Nullifying Apparatus, and to determine the extent of rebound sleep induced by deprivation.
Sleep homeostasis, the increase in sleep observed following sleep loss, is one of the defining criteria used to identify sleep throughout the animal kingdom. As a consequence, sleep deprivation and sleep restriction are powerful tools that are commonly used to provide insight into sleep function. Nonetheless, sleep deprivation experiments are inherently problematic in that the deprivation stimulus itself may be the cause of observed changes in physiology and behavior. Accordingly, successful sleep deprivation techniques should keep animals awake and, ideally, result in a robust sleep rebound without also inducing a large number of unintended consequences. Here, we describe a sleep deprivation technique for Drosophila melanogaster. The Sleep Nullifying Apparatus (SNAP) administers a stimulus every 10s to induce negative geotaxis. Although the stimulus is predictable, the SNAP effectively prevents >95% of nighttime sleep even in flies with high sleep drive. Importantly, the subsequent homeostatic response is very similar to that achieved using hand-deprivation. The timing and spacing of the stimuli can be modified to minimize sleep loss and thus examine non-specific effects of the stimulus on physiology and behavior. The SNAP can also be used for sleep restriction and to assess arousal thresholds. The SNAP is a powerful sleep disruption technique that can be used to better understand sleep function.
Sleep is near universal in animals, yet its function remains unclear. Sleep homeostasis, the compensatory increase in sleep following sleep deprivation, is a defining property of sleep, that has been used to characterize sleep states in a number of animals1,2,3,4,5.
Sleep in the fly has many similarities with human sleep, including a robust homeostatic response to sleep loss4,5. Numerous studies of sleep in the fly have u....
1. Experimental preparation
Canton S (Cs) was used as a wild-type strain. Flies were maintained on a 12 h light: 12 h dark schedule, and sleep deprived for 12 hours overnight. Inspection of the sleep profiles of Cs flies on the baseline day (bs), sleep deprivation day (sd), and two recovery days (rec1 and rec2) (Figure 2A) suggests that flies were effectively sleep deprived in the SNAP, and recovered sleep during the day consistent with observed reports in the literature
Sleep in Drosophila was independently characterized in 2000, by two groups4,5. In these pioneering studies, flies were deprived of sleep by gentle handling (i.e., hand deprivation) and shown to exhibit a robust homeostatic response to overnight sleep deprivation. Importantly, with any sleep deprivation experiment it is crucial to control for potential confounding effects of the method used to keep the animal awake. Hand deprivation studies set the benchm.......
|Locomotor activity tubes
|Fisher Tissue Prep Wax
|Wax used for sealing tubes
|We cut 5mm diameter Pyrex glass tubes into 65mm long tubes to record sleep. Pre-cut tubes can also be purchased.
|Nutri Fly Bloomington Formulation fly food
|Labs might use their own fly food recipe. It is important that sleep be recorded on the same food that flies were reared in.
|Rotary glass cutting tool
|Dremel Multi Pro
|Used to cut 65mm long glass tubes
|DAM System and DAMFileScan software
|Software used to acquire data from DAM monitors and save the acquired data in an appropriate format
|Data acquisition computer
|Idea Centre AIO3
|A equivalent computer from any manufacturer can substitute
|Drosophila Activity Monitors
|These monitors are used to record flies' locomotor activity
|Not essential, but an easy way to monitor environmental conditions in the chamber where sleep is recorded
|A convenient way to control the timing of when the SNAP is turned on and off
|Power Supply Interface Unit for DAM
|Required for data acquisition computers to record Trikinetics locomotor acitvity data
|DAM monitors accept RJ11 jacks
|Used to connect upto 5 DAM monitors
|Telephone cable wire
|Phone cables to acquire data from DAM monitors
|Power supply for the SNAP
|Sleep Nullifying Apparatus
|Washington University School of Medicine machine shop
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