Extended Data Fig. 8: Regulation of GCG neurons by oral signals of ingestion.

a, Left, PRLH (Prlh^CreRosa^Tom, red) and GCG (Gcg^GFP, green) neurons are intermingled but non-overlapping in the cNTS. Scale bar = 100 µm. Right. Quantification of overlap for Prlh^CreRosa^Tom (red) and Gcg^GFP (green) cells in the cNTS (n = 3 mice). b, Left, fiber placement and GcaMP6s expression in GCG neurons. Right, example traces of calcium dynamics from GCG neurons during self-paced Ensure, Intralipid, or glucose consumption. c, Example traces of calcium dynamics from GCG neurons during self-paced sucralose, saline, or water consumption, or dry licking at an empty sipper. d, Left, PSTH of GCG neuron responses during just the first lick bout of the trial, aligned to the first lick of the trial, for the indicated solutions. Right, mean z-scores (0–10 s) during the first lick bout. e, Left, PSTH aligned to the first lick of the bout (averaged across all lick bouts) during Ensure consumption for GCG and PRLH neurons. Right, mean z-scores (0–10 s) during Ensure consumption for GCG and PRLH neurons. f, Left, PSTH aligned to the last lick of the bout (averaged across all lick bouts) during Ensure consumption for GCG and PRLH neurons. Right, mean decrease in z-score (0–15 s after last lick of each bout) during Ensure consumption for GCG and PRLH neurons. g, Mean Pearson Correlation Coefficient (PCC) for the relationship between the instantaneous lick rate each second during Ensure consumption and the z-scored change in activity across the entire 30 min trial for GCG and PRLH neurons. h, Comparison of the mean z-scores (0–30 min after lickometer access) for PRLH neurons (from Fig. 1) and GCG neurons (from Fig. 5). i, Left, PSTH of GCG neuron activity aligned to the first lick (averaged across all lick bouts). Right, mean response (0–10 s after first lick) averaged across all licking bouts. j, Comparison of the mean z-scores (0–10 s after first lick in bout) for PRLH neurons (from Fig. 2) and GCG neurons (from Fig. 5). k, Mean PCC for the relationship between the cumulative licks performed in the preceding time intervals and the z-scored change in activity across the entire 30 min trial of self-paced Ensure consumption. l, Mean z-score per lick (mean z-score over 30 min trial/total number of licks) for each tastant. m, Mean z-score per lick (mean z-score 0–10 s of each bout divided by the number of licks in the same time frame) stratified by bout size for all tastants from GCG neurons. Each data point is an averaged value from a single animal. n, Left, mean z-score per lick by bout size for glucose consumption in WT and Trpm5^−/− mice (GCG neurons). Middle, mean z-score per lick by bout size for sucralose consumption in WT and Trpm5^−/− mice. Right, mean z-score per lick by bout size for Intralipid consumption in WT and Trpm5^−/− mice. o, Left, PSTH of GCG neuron responses during feeding tube insertion into the esophagus (0–30 s). Right, mean z-scores (0–30 s) during feeding tube insertion. p, Left, example trace of calcium dynamics from GCG neurons during self-paced chow consumption. Right, example trace of calcium dynamics from GCG neurons during self-paced HFD consumption. q, Top, PSTH of PRLH neuron responses during self-paced chow or HFD consumption, or presentation of a non-food object (black), aligned to moment of food presentation. Bottom, cumulative fraction of total bites during the trial. r, Left, mean z-scores (0–10 min) after food/object access. Right, mean z-scores (0–30 min) after food/object access. NS, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. Data are mean ± sem. Statistics are shown in Supplementary Table 1.