Fig. 4: Wide-field optical imaging of intracellular ATP dynamics in cortical neurons.

a Representative ATP signals, with EEG and EMG signals, in the whole cortical area across the sleep-wake states imaged in one example mouse. An imaging window was placed over the cortex. Arrows indicate corresponding time points of imaging at the bottom. A: anterior, P: posterior, R: right, L: left. Scale bar = 1 cm. b Mean ATP signals in the observed cortical area during the wake, non-REM sleep (NREMS), and REM sleep (REMS). *p < 0.05, one-way ANOVA with the Bonferroni post-hoc test. Data are presented as mean ± SEM (n = 5 mice). c Circles depict the six ROIs in which ATP signals were analyzed. RA: right-anterior, RM: right-middle, RP: right-posterior, LA: left-anterior, LM: left-middle, LP: left-posterior. Scale bar = 1 cm. d Mean ATP signals during the wake, NREMS, and REMS states in six ROIs from one example mouse. Colors correspond to ROI circles in (c). *p < 0.05, two-way ANOVA with the Bonferroni post-hoc test (n = 6 ROIs). e ATP signals in six ROIs from one animal for the transitions of NREMS-to-REMS and REMS-to-wake. Transitions occurred at 0 s. Data are from 4-s intervals characterized by state transitions. f Correlation map between six ROIs across the sleep-wake states. The data were averaged from four mice. g Correlation maps between six ROIs in the wake, NREMS, and REMS states. Note that higher correlations were observed between hemisphere-symmetric ROIs (RA-LA, RM-LM, and RP-LP) and neighboring ROIs (e.g., RA-RM or LM-LP). Data are averaged from four mice. h The comparison of correlation coefficients among states. Data are presented as mean ± SEM (n = 4 mice). See also Supplementary Movie 1-3 and Supplementary Fig. 6.