Fig. 1: Microglia regulate sleep through P2Y12–Gi signaling.
From: Microglia regulate sleep through calcium-dependent modulation of norepinephrine transmission
a, Schematic for the chemogenetic experiment. b, Confocal images from the prefrontal cortex showing hM4Di expression in Iba1+ microglia (Extended Data Fig. 1). Scale bar, 50 µm. c, Quantification of efficiency and specificity (mean ± s.e.m.; n = 3 mice). ‘Overlap’: HM4Di–HA+ and Iba1+. d, Example chemogenetic experiments. Shown are the EEG spectrogram (normalized by the maximum of each session; Freq., frequency), EMG amplitude (Ampl.) and brain states (color-coded). e, Summary of the percentages of time in each brain state following CNO and saline injection (mean ± s.e.m.; n = 8 mice: 3 female and 5 male, recorded between ZT6 and ZT11). *P < 0.05, **P < 0.01 (two-way ANOVA with Bonferroni correction; NREM: Ptreatment < 0.0001, Ptime < 0.0001, *P0.5h = 0.044, **P1h = 0.0087, **P1.5h = 0.0097; wake: Ptreatment < 0.0001, Ptime < 0.0001, *P1h = 0.020, *P1.5h = 0.022, *P2.5h = 0.024; REM: Ptreatment = 0.13, Ptime < 0.0001). f,g, Mean episode duration (f) and episode number per hour (g) for each brain state within 3 h of CNO or saline injection. Each circle indicates data from one mouse (mean ± s.e.m.; n = 8 mice). **P < 0.01, ***P < 0.001 (paired two-tailed t-test; in f, ***P = 0.0001; in g, **PNREM = 0.0091, **Pwake = 0.0070, **PREM = 0.0056). h, Schematic for P2Y12 antagonist infusion. i,j, Confocal images (i) and quantification (j) of P2Y12 expression in Iba1+ microglia in the prefrontal cortex (n = 4 mice). Scale bar, 50 μm. ‘Overlap’: Iba1+ and P2Y12+. k, Percentage of time in each brain state following PSB0739 (PSB) and ACSF infusion (mean ± s.e.m.; n = 6 mice: 1 female and 5 male). *P < 0.05, **P < 0.01 (two-way ANOVA with Bonferroni correction; NREM: Ptreatment = 0.0005, Ptime < 0.0001, **P2h = 0.0053, **P4.5h = 0.0021; wake: Ptreatment = 0.0011, Ptime < 0.0001, *P2h = 0.014, **P4.5h = 0.0037; REM: Ptreatment = 0.23, Ptime < 0.0001). l,m, Mean episode duration (l) and episode number per hour (m) for each brain state within 5 h of PSB0739 or ACSF infusion. Each circle indicates data from one mouse (mean ± s.e.m.; n = 6 mice). *P < 0.05 (two-tailed Wilcoxon signed rank test, *P = 0.031).
