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. Author manuscript; available in PMC: 2023 Dec 19.
Published in final edited form as: Cell Rep. 2023 Oct 3;42(10):113188. doi: 10.1016/j.celrep.2023.113188

Figure 6. MC3R is required for the orexigenic action of ghrelin on AgRP neurons.

Figure 6.

(A) 1-h food intake of AgRP-Cre and AgRP-specific MC3R KO male mice given saline or ghrelin injection (1.6 mg/kg, i.p.) (n = 7–8 mice for all groups).

(B and C) Representative images of cFos immunostaining, and quantifications of cFos-positive cell number in the ARC of AgRP-Cre and AgRP-specific MC3R KO male mice in response to saline or ghrelin injection. Scale bar, 100 μm (n = 4–5 mice for each group).

(D) Schematic showing the time course of ghrelin injection and food presence in fiber photometry experiment

(E‒H) Traces and quantifications of averaged dF/F0 (%) GCaMP6s signal in AgRP neurons in WT and MC3R KO male mice in response to ghrelin and food cue (n = 4–5 mice for all groups).

(I) qPCR analysis of hypothalamic GHSR mRNA levels in fed versus 24-h-fasted WT and MC3R KO male mice (n = 3–5 mice for all groups).

(J‒L) RNAscope analysis of Agrp and Ghsr mRNA expression, and quantifications of the number of Ghsr transcripts in each AgRP neuron and the percentage of AgRP neurons coexpressing Ghsr in the ARC of fed versus 24-h-fasted AgRP-Cre and AgRP-specific MC3R KO male mice. Scale bar, 100 μm (n = 3–4 mice for all groups).

Data are plotted as mean, and all error bars represent the SEM. ns, non-significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 in unpaired Student’s t test and two-way ANOVA with Sidak’s post hoc test.