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. 2022 Mar 22;27(6):2901–2913. doi: 10.1038/s41380-022-01513-x

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© The Author(s) 2022

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/..

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Fig. 1 — a Illustration of unpredictable chronic stress protocol and phenotype assessment, with one stressor treatment randomly chosen per day, lasting for 4 weeks. b Residence time in each site of open field (blue, less time; red, more time). c Behavioral analysis of control (n = 10) and stress group mice (n = 11) in open field test showed significant decrease in both time spent in central area (unpaired Student’s t-test, P < 0.0001) and entries into central area in stress group (unpaired Student’s t-test, P = 0.0009), but no obvious change in traveling distance between groups (unpaired Student’s t-test, P = 0.4896). d Residence time in each site of elevated plus-maze (blue, less time; red, more time) of control and stress groups. e Control (n = 10) and stress groups (n = 11) in elevated plus-maze showed significant decrease in time spent in open arm (unpaired Student’s t-test, P < 0.0001) and entries into open arm in stress group (unpaired Student’s t-test, P < 0.0001), but no obvious change in traveling distance between groups (unpaired Student’s t-test, P = 0.9215). f Body weight monitored after chronic stress period showed no obvious change compared with control group (n = 9 in each group, unpaired Student’s t-test, P = 0.7125), and 24-h food intake after overnight fasting decreased in stress group (n = 9, unpaired Student’s t-test, P = 0.0014) compared with control group (n = 9). g Average respiration exchange ratio (RER) decreased in stress group compared with control group (unpaired student’s t-test, P = 0.0003, n = 9 mice in each group); RER curve shifted after chronic stress (two-way ANOVA, P = 0.0002, F (1, 16) = 21.95, with Bonferroni correction). h Significant decreases in 24-h energy expenditure (EE) curve and average EE were observed in stress group (two-way ANOVA, P = 0.0057, F (1, 16) = 20.2, with Bonferroni correction; unpaired Student’s t-test; P = 0.0060). i Increased c-fos expression in dmVMH under chronic stress (unpaired Student’s t-test, P < 0.001, n = 5 mice in each group). Scale bar, 100 µm. j Schematic of GCaMP6s injection and expression in VMH SF-1 neuron. Scale bar, 100 µm. k, l Representative in vivo calcium fluorescence traces and heat maps demonstrate enhanced spontaneous calcium signals (dF/F) of SF-1 neurons in mice of stress group compared with control (6 mice in each group, unpaired Student’s t-test, P < 0.001). Data are means ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001. CLAMS comprehensive laboratory animal monitoring system.