Fig. 5: N2O-induced L5 activity and antidepressant-like effect requires cortical disinhibition. | Nature Communications

Fig. 5: N2O-induced L5 activity and antidepressant-like effect requires cortical disinhibition.

From: Nitrous oxide activates layer 5 prefrontal neurons via SK2 channel inhibition for antidepressant effect

Fig. 5

a Left, Schematic of GABAergic neurons (circles) scattered amongst pyramidal neurons (triangles). While all interneurons can be labeled with GCaMP6 using the mDlx enhancer (outer magenta), subtypes including PV (purple), SST (orange), and VIP (green) can be specifically labeled using Cre transgenic lines depicted on right. Dendritic targeting SST cells and somatic targeting PV are inhibited (circled negative sign) by VIP interneurons. b Representative two-photon images of peak GCaMP6 signals from interneuron subtypes under room air and N2O (50%). Scale bar, 20 μm. Traces in fig. S13. c Summary of genetically defined interneuron calcium responses under room air and N2O. N2O induces a suppression of PV and SST spontaneous activity but activates VIP activity (two-sided Wilcoxon matched-pairs signed rank: mDlx, n = 133 cells from 5 mice, P = 2.9 × 10−13; PV, n = 145 cells from 7 mice, P = 2.9 × 10−22; SST, n = 63 cells from 4 mice, P = 1.5 × 10−13; VIP, n = 197 cells from 10 mice, P = 0.0003). d Interneuron subtypes coexpressing GCaMP6 and DREADD-hM3Gq recorded under wakefulness, post-CNO injection, and N2O (50%). CNO-hM3Gq induced activation of interneuron subtypes blocked N2O induced suppression of PV (n = 84 cells from 3 mice; Kruskal-Wallis (347): P = 3.0 × 10−76 followed by Dunn’s multiple comparisons, P > 0.99) and SST activity (n = 103 cells from 3 mice; Kruskal-Wallis (68): P = 2.0 x 10−15 followed by Dunn’s multiple comparisons, P > 0.99). VIP cells displayed a similar trend (n = 97 cells from 4 mice; Kruskal-Wallis (93): P = 6.8 x 10−21 followed by Dunn’s multiple comparisons, P = 0.30). e Left, representative GCaMP6 traces of individual L5 responses and summary of all cells (right) under room air, post-CNO injection, and N2O. CNO-induced activation of PV (n = 241 cells from 7 mice; Kruskal-Wallis (348): P = 3.0 x 10−76 followed by Dunn’s multiple comparisons, P > 0.99) or SST (n = 110 cells from 3 mice; Kruskal-Wallis (112): P = 3.3 × 10−25 followed by Dunn’s multiple comparisons, P = 0.001) blocked N2O-induced L5 activation whereas VIP promoted N2O-induced L5 activity (n = 139 cells from 3 mice, Kruskal-Wallis (169): P = 1.8 x 10−37 followed by Dunn’s multiple comparisons, P = 1.4 × 10−14). f TST immobility time under the same conditions. PV (n = 13, Kruskal-Wallis (9): P = 0.01 followed by Dunn’s multiple comparisons, P > 0.99) and SST (n = 17, Kruskal-Wallis (27): P = 1.4 × 0−6 followed by Dunn’s multiple comparisons, P > 0.99) activation by hM3Gq blocked N2O-induced decrease in immobility time in CORT mice. VIP activation prior to N2O produced a significant decrease in immobility time (n = 14, Kruskal-Wallis (10): P = 0.006 followed by Dunn’s multiple comparisons, P = 0.03). Error bars show s.e.m.

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