Fig. 4.

Chemogenetic inhibition of AgRP^ARC neurons reduces PVN neuronal activity without affecting plasma corticosterone levels in fasted animals: Number of mCherry positive neurons (A), FosB (left y scale and color bars) and double mCherry/FosB (right y scale and black bars) expression in the ARC (B), representative photomicrographs (×10 and insert ×20) of double immunostaining mCherry/FosB in the ARC (C), body weight change (D), plasma leptin (E), glucose (F) and corticosterone (G) levels, PVN FosB expressing neurons (H) and representative photomicrographs (×10 and insert ×40) of FosB in the PVN (I) of adult (8–12 weeks) male Agrp-IRES-Cre mice (n = 14–18) that received intra ARC injection of AAV-DIO-mCherry or AAV-DIO-hM4D(Gi)-mCherry submitted to 36 h of fasting and treated with 5 i.p. injections CNO (1 mg/kg) or saline. The i.p. injections were performed each 8 h and the animals were euthanized at 10am as described in the Additional file 1: Fig. S1. Data are expressed as mean ± SEM and were analyzed by two-way ANOVA followed by post hoc Tukey’s multiple comparison (D and G) or unpaired t test or Mann–Whitney (A, B, E, F and H). *p < 0.05 vs saline; ^#p < 0.05 vs Agrp-IRES-Cre::AAV-DIO-mCherry