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. 2022 Jul 22;13:4229. doi: 10.1038/s41467-022-31917-w

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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 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. 5 — a Chemogenetic inhibiting apMPOA-recipient neurons in ARC/PVH individually in wild type mice by injecting AAV1-Cre-GFP in apMPOA and DREAD-Gi in the ARC and PVH. Transsynaptic neurons in the PVH (b) and ARC (e) expressed hM4Di-mCherry. Image representative of n = 8 mice. Scale bar, 100 µm. c Chemogenetic inhibiting postsynaptic PVH neurons significantly increased food intake after CNO injection under all thermal conditions compared to PBS. Interaction effect: F_2,13(temperature × treatment) = 6.56, *p = 0.014; main effect: F_2,13(temperature) = 4.95, *p = 0.026; F_1,14(treatment) = 56.82, ***p < 0.001. Pairwise comparisons showed that chemogenetic inhibiting apMPOA-recipient PVH neurons increased food intake in all thermal conditions (***p < 0.001, **p < 0.01, *p < 0.05). Within-group tests showed significant simple effect of temperature in control group (F(2,13) = 5.08, *p = 0.023), but not in the taCasp3 group (F(2,13) = 2.42, p = 0.13). Two-way repeated-measure ANOVA and post hoc LSD multiple comparison, n = 8 animals. f Chemogenetic inhibiting postsynaptic ARC neurons significantly decreased food intake. Interaction effect: F_2,11(temperature × treatment) = 0.46, p = 0.64; main effect: F_2,11(temperature) = 8.81, **p = 0.006; F_1,12(treatment) = 11.78, **p = 0.005. Pairwise comparisons showed that chemogenetic inhibiting apMPOA-recipient ARC neurons decreased food intake in all thermal conditions (all **p < 0.01). Within-group tests showed significant simple effect of temperature in the control group (F(2,11) = 4.82, *p = 0.031), and the taCasp3 group (F(2,11) = 5.13, *p = 0.027). Two-way repeated-measure ANOVA and post hoc LSD multiple comparison, n = 8 animals. d, g The controlled groups that expressed mCherry did not show food intake changes between CNO and PBS injections. PVH-mCherry (d): interaction effect: F_2,9(temperature × treatment) = 0.47, p = 0.64; main effect: F_2,9(temperature) = 35.98, ***p < 0.001: F_1,12(treatment) = 1.55, p = 0.24. ARC-mCherry (g): interaction effect: F_2,9(temperature × treatment) = 1.5, p = 0.28; main effect: F_2,9(temperature) = 18.15, ***p = 0.001; F_1,10(treatment) = 1.97, p = 0.19. Two-way repeated-measure ANOVA and post hoc LSD multiple comparison, n = 6 animals for each group. All error bars show SEM.