Fig. 6: Miniscope Ca²⁺ imaging reveals two populations of LS^Nts neurons that are activated and inhibited during feeding.

A Schematic showing Ca²⁺ imaging of individual LS^Nts neurons with a head-mounted miniature microscope. B Ca²⁺ dynamics of 10 representative LS^Nts cells during feeding. Scale bar: 1 z score. C Ca²⁺ responses of 290 LS^Nts cells aligned to the first lick during free feeding of Ensure. Approximately 31% of cells were activated, and 32% of cells were inhibited. D The time to the peak of activated cells (n = 88) was shorter than the time to the trough of inhibited cells (n = 86). Mann–Whitney U test. ***P < 0.001. E The response half-width of activated cells (n = 88) was shorter than that of inhibited cells (n = 86). Mann–Whitney U test. ***P < 0.001. F. Ca²⁺ responses of 130 LS^Nts cells aligned to the first lick under the food-deficient condition. Approximately 40% of cells were activated, and 11% of cells were inhibited. G. Percentages of activated, inhibited and nonresponsive cells under free feeding (left panel) and food omission (right panel) conditions. H Area under the curve of Ca²⁺ activity for activated (left panel) and inhibited (right panel) cells during the free feeding of Ensure and food omission trials. Mann–Whitney U test. Ns, no significant difference. I. Correlation between the response half-width and bout duration for inhibited (left panel) and activated neurons (middle panel). Right panel: The correlation coefficient between the response and behavior of inhibited cells was larger than that of activated cells. Mann–Whitney U test. ***P < 0.001. J Ca²⁺ response of activated (right panel) and inhibited (left panel) cells when aligned to the last lick during free feeding of Ensure. K. Ca²⁺ response of LS^Nts neurons to the sucrose solution and Ensure (n = 143 neurons from 4 mice) grouped by k-means clustering. Vertical dashed line: first lick. L Ca²⁺ response of LS^Nts neurons to Ensure and regular food (n = 124 neurons from 4 mice) grouped by k-means clustering. Vertical dashed line: first lick. M Ca²⁺ response of LS^Nts neurons to Ensure and water (n = 161 neurons from 4 mice) grouped by k-means clustering. Vertical dashed line: first lick. N Percentages of activated, inhibited and nonresponsive cells identified during the free feeding of Ensure, sucrose solution, regular food and water.