Extended Data Fig. 5: Ca2+ signaling evoked by PV interneuron in mPFC slices.
From: GABAergic signaling to astrocytes in the prefrontal cortex sustains goal-directed behaviors
(a) Schematic drawing of an mPFC slice indicating the targeted area with AAV2.5-GFAP-cyto-GCaMP6f (green shaded area) in PV-tdTomato mice, and the depolarization protocol used to stimulate PV interneurons (120 ms @ 5 Hz, 30 s; firing rate per depolarization pulse ~ 50 Hz), mimicking theta-gamma cycles; PV cells (red) and surrounded by astrocytes (green). (b) Confocal images of NeuN labeling (blue), cyto-GCaMP6f (green) and PV-tdTomato with S100 labeling (red and white, respectively) from a, showing the selective expression of GCaMP6f in astrocytes. Scale bar, 20 μm. (c) Representative fluorescence images of cyto-GCaMP6f labeling astrocytes before (pre stim) and after (post stim) PV depolarization. Scale bar, 15 μm. (d) Representative Ca2+ traces showing an enhanced Ca2+ signaling from surrounding astrocytes by PV stimulation (control), and after perfusion of CGP55845 (5 μm) (PV stimulation denoted by red line). (e) Summary graph showing astrocyte Ca2+ event frequency and amplitude before (light color bars) and after the PV stimulation (dark color bars) in control, after bath application of CGP55845, and Ip3r2-/- mice. *** P <0.001. One-Way ANOVA two-sided (see Source Data file). Data shown are mean±SEM. Box plots indicate median and whiskers indicate the 25th and 75th percentiles, respectively.
