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. 2019 Dec 4;9:18360. doi: 10.1038/s41598-019-54870-z

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© The Author(s) 2019

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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Age-dependent effects of the reversible blockade of the synaptic transmission from the PVN to the DVC. (a–f) left: Representative recordings of the electrical activity of PVN neurons in the brain slices at ages 24 (n = 17) (a), 39 (n = 10) (c), 44 (n = 10) (d), and >60 (n = 13) weeks (f). Middle left and middle right: BW gain (middle left panel), food intake/BW (middle right panel) during intraperitoneal DOX injection at 26 weeks (BW gain, [F_{1, 70} = 0.117, P > 0.05]; food intake, [F_{1, 56} = 0.003, P > 0.05]) (n = 8, 8) (a), 35 weeks (BW gain, [F_{1, 70} = 7.633, P < 0.01]; food intake, [F_{1, 56} = 0.009, P > 0.05]) (n = 7, 9) (b), 39 weeks (BW gain, [F_{1, 75} = 7.779, P < 0.05]; food intake,[F_{1, 60} = 0.226, P > 0.05]) (n = 8, 9) (c), 44 weeks (BW gain, [F_{1, 65} = 41.042, P < 0.01]; food intake, [F_{1, 52} = 0.058, P > 0.05]) (n = 7, 8) (d), 53 weeks (BW gain, [F_{1, 75} = 31.798, P < 0.01]; food intake, [F_{1, 60} = 2.132, P > 0.05]) (n = 8, 9) (e), and 67 weeks old (BW gain, [F_{1, 30} = 25.738, P < 0.01]; food intake, [F_{1, 30} = 1.610, P > 0.05]) (n = 4, 4) (f). *P < 0.05, **P < 0.01. Two-way ANOVA followed by Tukey’s multiple range test. right panel: Food efficacy (average calculated from BW gain/food intake during DOX administration) at 26 (a), 35 (b), 39 (c), 44 (d), 53 (e), and 67 (f) weeks old. Unpaired t-test. (g,h) The membrane potential in the PVN neurons at 17, 24, 39, 44, and >60 weeks old. *P < 0.05, **P < 0.01, one-way ANOVA followed by Tukey’s multiple range test. n = 9–15 (g). The firing frequency in the PVN neurons at 17, 24, 39, 44, and >60 weeks old. **P < 0.01, one-way ANOVA followed by Tukey’s multiple range test. n = 9–15 (h). (i) Rectal temperature before (Day 0) and after (Day 3) the saline (left panel, control) or DOX (right panel) injection. *P < 0.05, paired t-test. n = 3 each.