Extended Data Fig. 2: supSDH astrocytic Ca2+ responses to several stimuli using in vivo imaging.
a, Schematic illustration of in vivo imaging of the superficial SDH in mice using two-photon microscopy. The chamber was attached at the L3−4 vertebral level. b, Immunohistochemical identification of GCaMP6m-expressing cells using cell-type markers (SOX9 and GFAP, NeuN, and IBA1). Note that GCaMP6m-expressing cells were immunolabeled by SOX9 and GFAP but not by NeuN or IBA1, indicating that GCaMP6m is expressed selectively in astrocytes. The data are representative of three independent experiments. c, Peak amplitude of astrocytic [Ca2+]i in the left SDH was high during the first 15 min (but not 55–60 min) after intraplantar injection of capsaicin (n = 59 ROIs, 3 mice). A von Frey filament (vF; 0.6 or 2.0 g) was applied to the left hindpaw before (pre) and 60 min after the injection. These light mechanical stimuli did not elicit astrocytic [Ca2+]i increases. Kruskal-Wallis test with Dunn’s multiple comparisons test. d, Representative Ca2+ traces in individual supSDH astrocytes in mice injected with capsaicin into the ipsilateral (left) and contralateral side (right). Synchronous and persistent Ca2+ responses were observed in the ipsilateral but not in the contralateral side. Data show the mean ± s.e.m. Scale bar, 20 µm. See source data for statistical parameters.
