Figure 1. Nutrient intake is necessary and sufficient for sustained AgRP neuron inhibition.

(A) Schematic of experiment in (B-C). Fasted mice were presented with caged chocolate and then available chocolate during photometry recording from AgRP neurons.

(B) Calcium signal from AgRP neurons in fasted mice presented first with caged chocolate (gray) and then available chocolate (red) (n=7 mice).

(C) Quantification of ΔF/F from (B). Times shown are 5-min windows immediately after chocolate presentation (5 min), immediately prior to chocolate removal (20 min), and 10 min following chocolate removal (30 min). * P = 0.02 compared to caged chocolate (Holm-Sidak's multiple comparisons test adjusted p-value)

(D) Schematic of the experimental set-up for AgRP photometry recording during intragastric nutrient infusion for 24 min.

(E) Calcium signal from AgRP neurons in fasted mice during intragastric infusion with water (black) or Ensure liquid diet (red). Gray denotes infusion (n = 7 mice for water; n = 5 mice for Ensure).

(F) Quantification of ΔF/F from (E). Times shown are 5-min windows from the early part of infusion (5 min), the end of infusion (20 min), and 10 min following the end of infusion (30 min). *P < 0.05, **P = <0.01, ***P < 10-3 compared to water infusion at the indicated time point (Holm-Sidak's multiple comparisons test, adjusted p-value).

(G) Quantification of ΔF/F at the end of infusion following the first and last intragastric exposures to water (black) and Ensure (red). Infusions were separated by approximately 7 weeks.

(B and E) Traces represent mean ± SEM

(C,F,G) ■ denotes individual mice. Bars represent mean ± SEM

See also Figure S1.