Automated Sleep Deprivation Setup Using a Shaking Platform in Mice

Abstract

The functions of sleep remain largely unclear, and even less is known about its role in development. A general strategy to tackle these questions is to disrupt sleep and measure the outcomes. However, some existing sleep deprivation methods may not be suitable for studying the effects of chronic sleep disruption, due to their lack of effectiveness and/or robustness, substantial stress caused by the deprivation method, or consuming a large quantity of time and manpower. More problems may be encountered when applying these existing protocols to young, developing animals, because of their likely heightened vulnerability to stressors, and difficulties in precisely monitoring sleep at young ages. Here, we report a protocol of automated sleep disruption in mice using a commercially available, shaking platform-based deprivation system. We show that this protocol effectively and robustly deprives both non-rapid-eye-movement (NREM) sleep and rapid-eye-movement (REM) sleep without causing a significant stress response, and does not require human supervision. This protocol uses adolescent mice, but the method also works with adult mice. Graphical abstract Automated sleep deprivation system. The platform of the deprivation chamber was programmed to shake in a given frequency and intensity to keep the animal awake while its brain and muscle activities were continuously monitored by electroencephalography and electromyography.

Keywords: Adolescence; Automated system; Mice; Shaking platform; Sleep deprivation.

Figure 1.. Automated sleep deprivation system and the customized home-cage.

A. Schematics of the automated sleep deprivation system with simultaneous EEG/EMG recording. B. Sleep deprivation chamber. C. Customized home-cage.

Figure 2.. Software interface of Shaker Driver

Video 1.. Setting up Shaker Driver for the SD and Ctrl protocols

Video 2.. Programmed platform shaking during an SD session

Figure 3.. Representative data.

A. Reduction of NREM or REM sleep amount (s) during 4 h Ctrl/SD relative to their respective baseline sleep during the same ZT period. The Ctrl protocol began immediately at dark phase onset and lasted for 4 h (ZT 12–16), and the SD protocol was performed in early light phase onset (ZT 2–6) in adolescent mice (P35–P42). N = 4 mice in Ctrl; 9 mice in SD. Welch’s t-test, ΔNREM, t = 10.63, df = 5.13, P = 0.0001; ΔREM, t = 8.74, df = 10.99, P = 0.000003. B. Five days of SD did not induce significant stress compared to the Ctrl protocol. N = 7 mice in Ctrl; 10 mice in SD. Welch’s t-test, t = 0.3331, df = 14.14, P = 0.74. Data are shown as means ± S.E.M. * P < 0.05; ** P < 0.01; *** P < 0.001; n.s., not significant. SD values were extracted from the data published in Bian et al. (2022).