Fig. 1: Hippocampal astrocytes respond to anxiogenic environment by intracellular calcium elevation.

a Schematic illustration showing hGFAP-GCaMP6s generation by breeding hGFAP-CreERT2 and floxed-stop-GCaMP6s transgenic mice. b Representative confocal image showing GCaMP6s (green) colocalization in astrocytes (GFAP⁺, purple). Scale bar, 50 μm. Experiment was repeated five-times independently. c Design of VR environment; a 2D image (upper) and 3D image (bottom) as seen by the mouse. d Schematic representation of two-photon imaging (left) and the hippocampal window (right) indicating the imaging area. Scale bar, 1 mm. e Example region of interest (ROI) of calcium signal recording. Yellow circle: recorded cells. Scale bar, 50 μm. Experiment was repeated five-times independently. f Representative Ca²⁺ traces during exploration in VR condition. Pink, blue and white bar indicates center, corner and closed corridor in VR. Red triangles indicate Ca²⁺ peaks. g Number of Ca²⁺ peak per second when mouse explored the VR center, corner, and closed area (n = 87 cells). One-way ANOVA (p = 0.000) followed by post-hoc analysis LSD (p = 0.000 (center vs corner), p = 0.000 (center vs closed), p = 0.492 (corner vs closed)). h Heat map trace of normalized astrocytic GCaMP6 signals when mice enter the center (left) or the corner (right) from the closed corridor (n = 129 events). i Categories and percentages of hippocampal astrocytes according to activity patterns when mice enter the center (upper) or the corner (lower) from the closed area. Black arrow: Direction of movement. The bar graphs depict the mean ± SEM. ***p < 0.001. See Supplementary Data for detailed statistics. Source data are provided as a Source Data file.