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. 2021 Jul 30;12:4646. doi: 10.1038/s41467-021-24915-x

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — a Subcortical brain regions were collected during the day when mice were asleep (Sleep_day, n = 6 mice) and at night when mice were awake (Wake_night, n = 6 mice) for metabolomic analysis. b A heatmap of lipid species showing significant Sleep_day/Wake_night expression differences. The p value for each lipid was obtained by compare the difference between Sleep_day and Wake_night. LCFAs, long chain saturated fatty acids, p = 0.019; LC-MUFAs long chain monounsaturated fatty acids, p = 0.006; LC-PUFAs long chain polyunsaturated fatty acids, p = 0.016; DA dicarboxylic fatty acids, p = 0.01; LCACs long chain saturated acyl carnitines, p = 0.005; MUACs monounsaturated acyl carnitines, p = 0.004; PUACs polyunsaturated acyl carnitines, p = 0.022; Carnitine, p = 0.036; ACs acylcholines, p = 0.048; HFAs dihydroxy fatty acids, p = 0.008; EC endocannabinoids, p = 0.04; PLP phospholipids, p = 0.018; PC phosphatidylcholines, p = 0.01; PE phosphatidylethanolamine, p = 0.019; PS phosphatidylserine, p = 0.024; PG phosphatidylglycerol, p = 0.019; LP lysophospholipid, p = 0.026; PL plasmalogen, p = 0.034; LyPL lysoplasmalogen, p = 0.044; MG monoacylglycerol, p = 0.021; DAG diacylglycerol, p = 0.031; CERs ceramides, p = 0.04; HexCERs hexosylceramides, p = 0.018; dhSMs dihydrosphingomyelins, p = 0.043; SMs sphingomyelins, p = 0.021; SPHs sphingosines, p = 0.003; Sterol, p = 0.042. c A pathway analysis indicated that lipid-related pathways were most affected by Sleep_day/Wake_night. d A volcano plot of measured sphingolipids; differentially represented metabolites with >1.3-fold change (Sleep_day vs Wake_night) and p values <0.10 are indicated in color; n.s. nonsignificant. e Sleep deprivation promoted subcortical ceramide accumulation. Subcortical region was sampled at ZT12 (timepoint of light off). p = 0.035 for Ctl vs SD12. Ctl control mice, n = 8; SD6, mice with 6 h of sleep deprivation, n = 8; SD12, mice with 12 h of sleep deprivation, n = 8. f Tight correlation between brain ceramide level and wakefulness. Wakefulness was defined with video recording for 1 h before sampling (ZT12). Pearson’s correlation, p value was calculated using two-sided hypothesis test. Eight mice. *p < 0.05; two-sided unpaired t-test for b, d, and e. Data are reported as mean ± SEM. See also Supplementary Fig. 4.