Fig. 1
Spontaneous Ca2+ events during 5-min periods following long-term chemogenetic manipulation of astrocytes. a Timeline overview of the experimental procedure. b Two-photon confocal image of CA1 astrocytes expressing DREADDs, fused to an mCherry protein for visualization (red) and the fluorescent Ca2+ indicator OGB-1 AM (green). Scale bar of 50 µm is marked in white. c Representative traces of individual astrocytes before and 5 min after CNO application. Graph (d) shows the average amount of Ca2+ events per 5-min imaging periods recorded from CA1 astrocytes before, during and after CNO (10 µM) application. RM Two-Way ANOVA shows a significant difference between groups. Bonferroni’s post-hoc analysis shows significant differences in the Gq-DREADD transduced mice 5 to 35 min after CNO application compared to control transduced slices (control vector vs. Gq-DREADD; 5 min after CNO p = 0.0006; 15 min after CNO, p = 0.0377; 35 min after CNO, p = 0.0410; Gq-DREADD, n = 5; control vector, n = 4), while no significant differences were observed for the Gi-DREADD during nor after CNO application (Bonferroni’s post-hoc analysis, control vector vs. Gi-DREADD; 5 min after CNO p = 0.8973; 15 min after CNO, p = 0.9766; 35 min after CNO, p > 0.9999; 20 min after wash-out, p = 0.5380; 30 min after wash-out, p > 0.9999; n = 4). *p < 0.05; ***p < 0.001. Data are represented as the mean ± SEM, Gq-DREADD, n = 5; Gi-DREADD, n = 4; and control vector, n = 4