Fig. 3: Effect of microglia Gi activation on sleep depends on increased intracellular Ca2+.
From: Microglia regulate sleep through calcium-dependent modulation of norepinephrine transmission
a, Schematic for two-photon Ca2+ imaging with p130PH or p130PHR134L expression. b, Confocal images of p130PH–mCherry expression in Iba1+P2Y12+ microglia in the prefrontal cortex. Scale bar, 50 μm. c, z-scored Ca2+ activity averaged across all microglia imaged from mice expressing p130PH (CNO, n = 44; saline, n = 31; from 5 mice: 3 female and 2 male) or p130PHR134L (CNO, n = 32; saline, n = 29; from 5 mice: 2 female and 3 male). Dashed line, time of injection. d–f, Population summary of the CNO-induced change in mean Ca2+ level (d) and the amplitude (e) and frequency (f) of Ca2+ events in soma and processes (difference between before and after injection) in mice expressing p130PH or p130PHR134L. Each circle indicates data from one cell. p130PHCNO: n = 44 somas, n = 789 processes; p130PHsaline: n = 31 somas, n = 457 processes; p130PHR134LCNO: n = 32 somas, n = 595 processes; p130PHR134Lsaline: n = 29 somas, n = 564 processes. **P < 0.01, ****P < 0.0001 (one-way ANOVA with Holm–Šídák’s test; d, P < 0.0001, ****P < 0.0001; e, Psoma = 0.65, Pprocesses < 0.0001, ****P < 0.0001; f, Psoma = 0.50, Pprocesses < 0.0001, ***P = 0.0007, ****P < 0.0001, **P = 0.0024). g,h, Effect of microglia Gi activation on sleep in mice expressing p130PHR134L (g; n = 8 mice: 4 female and 4 male) or p130PH (h; n = 8 mice: 3 female and 5 male). *P < 0.05, **P < 0.01 (two-way ANOVA with Bonferroni correction; p130PHR134L: NREM: Ptreatment < 0.0001, Ptime < 0.0001, *P0.5h = 0.038, **P1h = 0.0036; wake: Ptreatment = 0.0002, Ptime < 0.0001, *P0.5h = 0.044, **P1h = 0.0051, *P1.5h = 0.019; REM: Ptreatment = 0.036, Ptime < 0.0001; p130PH: NREM: Ptreatment = 0.94, Ptime < 0.0001; wake: Ptreatment = 0.95, Ptime < 0.0001; REM: Ptreatment = 0.97, Ptime < 0.0001). i, Changes in each brain state induced by chemogenetic activation (difference between CNO and saline injections, averaged across 3 h after injection) in mice expressing p130PHR134L or p130PH. Each circle indicates data from one mouse (p130PHR134L, n = 8 mice; p130PH, n = 8 mice). **P < 0.01 (unpaired two-tailed t-test, **PNREM = 0.0055, **Pwake = 0.0082). Data are presented as the mean ± s.e.m.
