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. 2023 Apr 27;14:2435. doi: 10.1038/s41467-023-38180-7

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 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. 1 — a Schematic of changes in neuronal activity of the LHb-RMTg-VTA pathway in chronic and acute stress responses. DA, dopamine; GABA, gamma-aminobutyric acid; GLU, glutamate. b, c Expression of AADC/VGLUT2 (b) and AADC/TH/TPH2 (c) in the LHb by FISH. d Pie chart of the percentage of VGLUT2-expressing AADC-positive cells in the LHb by FISH (n = 3 independent mice). e, Experimental schematic of acute tail suspension (TS)-induced stress, followed by FISH analysis. f Expression of AADC/c-Fos/VGLUT2 in the LHb of naïve and TS model mice by FISH. g–i Percentages of c-Fos-expressing AADC-positive cells (n = 8 independent Naïve mice and n = 11 independent TS mice) (g), c-Fos-expressing VGLUT2-positive cells (n = 8 independent Naïve mice and n = 11 independent TS mice) (h) and c-Fos-expressing cells in total cells in the LHb by FISH (n = 8 independent Naïve mice and n = 11 independent TS mice) (i). j Expression of c-Fos/TH in the VTA (top) and c-Fos/GAD1 in the RMTg (bottom) in naïve and TS model mice by FISH. k Percentages of c-Fos-expressing TH-positive cells in the VTA (left; n = 8 independent Naïve mice and n = 11 independent TS mice) and c-Fos-expressing GAD1-positive cells in the RMTg (right; n = 8 independent Naïve mice and n = 11 independent TS mice) of naïve and TS model mice. l Schematic of acute stress-mediated alterations in neuronal activity in the LHb-RMTg-VTA pathway. Magnified images of the regions of interest (yellow squares) are shown next to the main images in (b), (c) and (f). Unless otherwise stated, statistical comparisons were performed using a two-tailed unpaired t test. **p < 0.01 and ***p < 0.001. Data are presented as the mean values ± s.e.m. Source data are provided as a Source Data file.