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. 2023 Feb 7;9:16. doi: 10.1038/s41421-022-00498-9

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© The Author(s) 2023

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. 8 — a Schematic diagram depicting fiber photometry recording of extracellular adenosine level and population Ca²⁺ signal of astrocytes in LC/SLD region of the brainstem during the sleep–wake cycle. b Top to bottom, EEG power spectrogram, EMG (scale, 1 mV), GCaMP fluorescence (scale, 1 z-score), and GRAB_Ado fluorescence (scale, 1 z-score). c Fiber photometry recording of extracellular adenosine level in LC/SLD in CD73-KO mice during the sleep–wake cycle. d GRAB_Ado signal in WT and CD73-KO mice in different brain states. Each line represents data from one recording. n = 12 sessions from 5 mice in the WT group; n = 20 sessions from 6 mice in the CD73-KO group. Wake: P = 0.52; NREM: P = 0.47; REM: P = 0.22; Wilcoxon rank-sum test. e–l Same as Fig. 3g–n, respectively, except that experiments were performed in LC/SLD. hM₃Dq group, n = 7 mice; tdTomato group, n = 8 mice. The statistical method used was two-way repeated measures ANOVA, followed by Tukey’s post hoc multiple comparison test. In f, hM₃Dq-CNO vs tdTomato-CNO: Wake, P = 0.47; NREM, P = 0.97; REM, **P = 0.0024. hM₃Dq-Saline vs hM₃Dq-CNO: Wake, P = 0.34; NREM, P = 0.98; REM, ***P < 0.001. tdTomato-Saline vs tdTomato-CNO: Wake, P = 0.99; NREM, P = 0.97; REM, P = 0.94. In g, hM₃Dq-CNO vs tdTomato-CNO: Wake, P = 0.35; NREM, P = 0.42; REM, *P = 0.045. hM₃Dq-Saline vs hM₃Dq-CNO: Wake, *P = 0.030; NREM, *P = 0.034; REM, **P = 0.0011. tdTomato-Saline vs tdTomato-CNO: Wake, P = 0.27; NREM, P = 0.25; REM, P = 0.70. In h, hM₃Dq-CNO vs tdTomato-CNO: Wake, P = 0.12; NREM, P = 0.41; REM, *P = 0.045. hM₃Dq-Saline vs hM₃Dq-CNO: Wake, *P = 0.021; NREM, *P = 0.021; REM, P = 0.28. tdTomato-Saline vs tdTomato-CNO: Wake, P = 0.88; NREM, P = 0.19; REM, P = 0.95. In j, hM₃Dq-CNO vs tdTomato-CNO: delta, **P = 0.0021; theta, P = 0.088; sigma, ***P < 0.001; beta, ***P < 0.001. hM₃Dq-Saline vs hM₃Dq-CNO: delta, **P = 0.0085; theta, P = 0.46; sigma, ***P < 0.001; beta, ***P < 0.001. tdTomato-Saline vs tdTomato-CNO: delta, P = 0.81; theta, P = 0.97; sigma, P = 0.53; beta, P = 0.58. In l, hM₃Dq-CNO vs tdTomato-CNO: delta, P = 0.38; theta, **P = 0.0013; sigma, P = 0.98; beta, P = 0.89. hM₃Dq-Saline vs hM₃Dq-CNO: delta, P = 0.36; theta, ***P < 0.001; sigma, P = 0.97; beta, P = 0.73. tdTomato-Saline vs tdTomato-CNO: delta, P = 0.67; theta, P = 0.84; sigma, P = 0.93; beta, P = 1.0. m–t Same as e–l, respectively, except that experiments were performed in CD73-KO mice. hM₃Dq group, n = 9 mice; tdTomato group, n = 7 mice. The statistical method used was two-way repeated measures ANOVA, followed by Tukey’s post hoc multiple comparison test. In n, hM₃Dq-CNO vs tdTomato-CNO: Wake, **P = 0.0020; NREM, *P = 0.023; REM, ***P < 0.001. hM₃Dq-Saline vs hM₃Dq-CNO: Wake, P = 0.064; NREM, P = 0.80; REM, ***P < 0.001. tdTomato-Saline vs tdTomato-CNO: Wake, P = 0.40; NREM, P = 0.40; REM, P = 0.98. In o, hM₃Dq-CNO vs tdTomato-CNO: Wake, P = 0.32; NREM, P = 0.34; REM, **P = 0.0077. hM₃Dq-Saline vs hM₃Dq-CNO: Wake, P = 0.051; NREM, *P = 0.050; REM, ***P < 0.001. tdTomato-Saline vs tdTomato-CNO: Wake, P = 0.68; NREM, P = 0.67; REM, P = 0.93. In p, hM₃Dq-CNO vs tdTomato-CNO: Wake, P = 0.098; NREM, P = 0.80; REM, P = 0.23. hM₃Dq-Saline vs hM₃Dq-CNO: Wake, P = 0.11; NREM, P = 0.12; REM, P = 0.53. tdTomato-Saline vs tdTomato-CNO: Wake, P = 1.0; NREM, P = 0.13; REM, P = 0.77. In r, hM₃Dq-CNO vs tdTomato-CNO: delta, *P = 0.015; theta, P = 0.95; sigma, ***P < 0.001; beta, **P = 0.0015. hM₃Dq-Saline vs hM₃Dq-CNO: delta, **P = 0.0042; theta, P = 0.81; sigma, ***P < 0.001; beta, ***P < 0.001. tdTomato-Saline vs tdTomato-CNO: delta, P = 0.91; theta, P = 0.98; sigma, P = 0.90; beta, P = 0.96. In t, hM₃Dq-CNO vs tdTomato-CNO: delta, P = 1.0; theta, P = 0.18; sigma, P = 1.0; beta, P = 0.85 hM₃Dq-Saline vs hM₃Dq-CNO: delta, P = 0.72; theta, P = 0.22; sigma, P = 0.95; beta, P = 1.0. tdTomato-Saline vs tdTomato-CNO: delta, P = 1.0; theta, P = 0.99; sigma, P = 0.99; beta, P = 0.99.