Fig. 1

Metabolomic changes during sleep–wake states. a Real-time sleep state analysis, DG dissection, and metabolomic analysis using mass spectrometry. b Timing of tissue sampling (ZT = 0 indicates start of light cycle). c Average duration between detection of target sleepwake state and tissue sampling. d PLS-DA plot showing significant separation of the Wake group from the REM and NREM groups. Each point reflects one mouse, and ellipses represent 95% CIs. REM, n = 10 mice; NREM, n = 10 mice; Wake, n = 11 mice. e Top 5% of metabolites with the highest VIP scores. f Enrichment analysis of the top 5% of metabolites with the highest VIP scores listed in order of their statistical significance (i.e., smallest p-value at top). g-i Volcano plots of p-values of each metabolite from permutation analysis: REM vs. Wake (G), NREM vs. Wake (H), and REM vs. NREM (I). Red dashed line, p = 0.05; In, increase; De, decrease. j Venn diagram indicating numbers of significantly changed metabolites for each comparison (e.g., REM/Wake means significantly changed, either positively or negatively, in REM vs. Wake groups). k Fold change of each metabolite (*|fold change|> 1.5 and p < 0.05, **|fold change|> 1.5 and p < 0.01), sorted from the highest fold change for REM vs. Wake. Error bars, 95% CIs. *p < 0.05, **p < 0.01