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. 2023 May 22;14:2939. doi: 10.1038/s41467-023-38583-6

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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. 3 — a Four representative video snapshots taken before and during the photostimulation of Glut⁺ CnF neurons for 1 s, which triggers running. The experimental strategy is similar to Fig. 2a. b Raw (DiaEMG) and integrated (Int.) diaphragmatic electromyogram and locomotor speed during photostimulations. c Diaphragmatic frequency and amplitude at rest (CTL), during CnF photostimulation at increasing frequencies, and following light offset (OFF). Amplitudes are normalized to CTL. From left to right: frequency, p = 0,007; p = 0,1046; p = 0,9782; p = 0,7305; p = 0,0392; amplitude, p = 0,0229; p = 0,4756; p = 0,074; p = 0,2066; p = 0,0739. d Enlarged EMG recording showing the “pre-loco” and the “loco” phases. e Diaphragmatic frequency and amplitude in CTL and during the “pre-loco” phase at increasing CnF photostimulation frequencies. From left to right: frequency, p = 0,0156; p = 0,1128; p = 0,9795; p = 0,1639; amplitude, p = 0,3377; p = 0,108; p = 0,3298; p = 0,5486. f Left: similar quantifications during the “loco” phase. From left to right: frequency, p = 0,0067; p = 0,1021; p = 0,9477; p = 0,6789; amplitude, p = 0,0156; p = 0,2892; p = 0,0747; p = 0,6608. Right: diaphragm frequency and amplitude in relation to locomotor speed. Note that 32% of frequency and 41% of amplitude correlate with locomotor speed (R²). CnF photostimulation frequencies are also indicated by the colored circles (blue: 15 Hz, green: 20 Hz, orange: 30 Hz, Red: 40 Hz). g Representative EMG recordings during subthreshold CnF photostimulations that do not initiate running but upregulate breathing frequency and amplitude. h Quantifications of diaphragm frequency and amplitude in CTL and during subthreshold CnF stimulations. Frequency: CTL vs 5 Hz, p = 0,00895; 5 Hz vs 10 Hz, p = 0,0618; CTL vs 10 Hz, p = 0,0109. Amplitude: CTL vs 5 Hz, p = 0,148; 5 Hz vs 10 Hz, p = 0,047; CTL vs 10 Hz, p = 0,0746. In graphs, gray circles are means of 3 trials per animal, colored circles are means ± SD across n mice, * denotes p < 0,05, ** denotes p < 0,01, and ns denotes p < 0,05. All tests are matched-pairs t tests. For all graphs, source data are provided as a Source data file. See also Figs. S1, S4, S5, S6, and S7.