Fig. 2: Cellular resolution calcium imaging of mPFC neurons during the two choice operant assay.

a Top row: Schematic of the viral strategy used to label mPFC neurons with GCaMP6f. Bottom row: Imaging setup of GRIN lens placement in the mPFC, including representative images showing GRIN lens placement (left), GCaMP6f expression (middle, GCaMP6f in green; DAPI in blue) in the mPFC at (from left to right) 4x, 20x, and 60x magnification. Right panel shows nuclear exclusion of GCaMP6f. Scale bars: 500 µm, 250 µm and 25 µm. A pie chart showing the number of neurons recorded from each male (b, n = 459 neurons, 9 mice) and female mouse (d, n = 570 neurons, 6 mice). Reconstruction of GRIN lens placement in the nine male (c) and six female (e) mice using WholeBrain software⁸⁴. Each colored line corresponds to the same colored slice in the pie chart and shows the position of the lens in the Allen Mouse Brain Common Coordinate Framework. Coordinates are relative to bregma. f A schematic of the two choice operant assay events to which mPFC neuronal activity was aligned, from left to right, trial start, social choice, sucrose choice, social reward, and sucrose reward. Top row: Heatmaps of average normalized fluorescence of all mPFC neurons that are significantly modulated (excited or inhibited) by each task event in male (g) and female (h) mice. Neurons are sorted by the time of maximum fluorescence across each task event. Bottom row: Average normalized fluorescence traces of the neurons from the corresponding heatmap that are significantly modulated by each task event. Shaded error regions indicate ± SEM. Proportions of total recorded neurons that are modulated by the various task events in male (i) and female (j) mice (male: n = 459 neurons, 9 mice; female: n = 570 neurons, 6 mice). Mouse schematic (f) adapted from Open Clipart by lemmling.