Fig 4. Knockdown of alc/AMPKβ in the nervous system disrupts sleep.

(A-B) Activity (A) and sleep (B) profiles over a 24-hour period for control genotypes elav>Dcr-2 (n = 128) and UAS-alc-RNAi/+ (n = 126) versus elav>Dcr-2, alc-RNAi (n = 126). All data obtained from second 24-hour cycle to allow for acclimatization. Activity and sleep are shown in bins of 30 minutes. White and black bars represent ZT time, 12 hours light and 12 hours dark, respectively. (C) Total sleep (min) in flies with pan-neuronal alc knockdown compared to controls. Total sleep is significantly reduced when alc is knocked down in the nervous system. (D) Comparison of activity outside sleep periods between alc knockdown animals and controls shows that pan-neuronal knockdown of alc increases activity during daytime, but not nighttime. (E) Motion-bout length (min) is significantly increased during the day in alc knockdown flies compared to controls. (F) Average sleep-bout length (min) is significantly reduced during both day- and night-time in alc knockdown flies. (G) The number of sleep bouts per day and night increases when alc is knocked down in the nervous system. (H) Duration of the longest sleep bout (min) is significantly shorter in alc knockdown animals than in controls. (I, J) Distribution of length of sleep bouts for control genotypes elav>Dcr-2/+ and UAS-alc-RNAi/+ versus elav>Dcr-2, alc-RNAi during the day (I) and during the night (J). Flies in which alc has been knocked down pan-neuronally only generate a small proportion of sleep bouts of >150 min, whereas the majority of sleep bouts in controls are of >150 min. For controls, lines were crossed to w¹¹¹⁸. Error bars indicate SEM. Kruskal-Wallis test with Dunn’s post-hoc testing was used to determine statistical significance: ***p<0.001, ****p<0.0001, versus the control.