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

Fig. 1. Glutamatergic apMPOA neurons respond to altered dietary states.

Fig. 1

a Scheme of feeding test in a temperature-controlled chamber. b Differences of 2 h food intake among the three ambient temperatures. NT neutral temperature, HT high temperature, LT low temperature. One-way ANOVA and post hoc least significant difference (LSD) multiple comparison, *p = 0.019, **p = 0.004, ***p < 0.001, n = 15 animals for each group. c–f C-Fos expressions in the apMPOA in three dietary states: 12 h fasting, 2 h refeeding and no fasting. AVPe, anteroventral periventricular nucleus; VMPO, ventromedial preoptic nucleus; MnPO, median preoptic nucleus; 3V, the 3rd ventricle. Scale bar, 100 µm. One-way ANOVA and post hoc LSD multiple comparison, *p = 0.048, ***p < 0.001, n = 4 animals for each group. g, h Colocalizations of fasting-induced C-Fos (green) with vGluT2+ (red) and GAD2+ neurons (red) in vGluT2::Ai9 (g) and GAD2::Ai9 mice (h). Images were obtained from transgenic mice by crossing Ai9 (Cre-dependent tdTomato reporter) with vGluT2-cre (g), and Ai9 with GAD2-cre (h). Scale bar, 100 µm. i Percentage of C-Fos+ neurons that are vGluT2+ and GAD2+. n = 5 animals for each group. j Percentage of vGluT2+ and GAD2 + neurons that express C-Fos. n = 5 animals for each group. k Scheme of fiber photometry setup, and representative coronal section of apMPOA neurons expressing GCaMP6s. Image representative of n = 5 mice. Scale bar, 100 µm. l, m GcaMP6s (mean, green line; SEM, green shading) and control (mean, purple line; SEM, purple shading) signal changes (ΔF/F) in response to feeding in GAD2-cre (l) and vGluT2-cre mice (m) after 24 h fasting. n = 5 animals for each group. n The mean ΔF/F in a 5 min window after the onset of feeding. T test (two-sided), ***p < 0.001, n = 5 animals for each group. All error bars show SEM.