Figure 4. Sleep Deprivation Alters Astroglial Ca²⁺ Signals.

(A) Maximum projection images of GCaMP6f⁺ astrocytes at ZT6 (left) & ZT12 (right) for baseline (top; BL) and 6 h sleep deprivation (bottom; SD) conditions from two-photon microscopy. Scale bar = 10 μm. Color bar shows arbitrary fluorescence units (AFU). (B) Mean ΔF/F values from all vigilance states for somata (S) and processes (P) during BL ZT6, SD ZT6, and SD ZT12. ^a, difference from BL ZT6 processes; ^b, difference from SD ZT6 processes (Friedman). (C & E) Mean ΔF/F values during (C) wake and (E) NREMS at ZT6 under BL and SD conditions (Wilcoxon signed rank). (D & F) Distributions of differences in ΔF/F values between processes and their soma at ZT6 under BL and SD conditions for (D) wake and (F) NREMS (Wilcoxon signed rank). Black squares in boxplots denote means (wake: BL = 2.20%, SD = 4.35%; NREMS: BL = 1.21%, SD = 1.24%). Values are means ± s.e.m. for B, C, and E. Group means are from n = 60 somata and n = 6074 process ROIs from 4 mice. *, different from BL. p < 0.05. See also Figure S3.