Skip to main content
. Author manuscript; available in PMC: 2023 Sep 1.
Published in final edited form as: Nat Neurosci. 2023 Jan 23;26(3):458–469. doi: 10.1038/s41593-022-01250-y

Fig. 2 |. Silencing VIP INs during NREM sleep prevents S1 hyperactivation and nociceptive allodynia.

Fig. 2 |

a, Representative two-photon images of VIP INs expressing PSAM4-GlyR-EGFP and PNs expressing GCaMP6s (7 mice). b, Timeline for chemogenetic inhibition of VIP INs after SNI, at ZT2 (rest phase) or ZT14 (active phase), in vivo Ca2+ imaging, and behavioral testing. c, PN Ca2+ activity in SNI mice with VIP INs inhibited at ZT2 (n = 152 cells from three mice) or ZT14 (n = 182 cells from four mice). Wake, F(1, 996) = 5.31, P = 0.021; NREM, F(1, 996) = 25.12, P < 0.0001. d, Nociceptive thresholds under various conditions (n = 7, 5, 5, 4 mice for von Frey, n = 5, 5, 5, 4 mice for other tests). Daily VIP inhibition at ZT2, but not ZT14, reduces punctate (ZT2 vs. SNI, P = 0.0006, 0.0003, < 0.0001 on day 14, 21, and 28, respectively), dynamic (P = 0.0025, 0.0004, < 0.0001), cold (P = 0.0008, 0.0003, 0.0069) and hot (P = 0.0027, 0.0044, 0.011) allodynia after SNI. IR, infrared radiant. e, Experimental design for brain state-dependent inhibition of VIP INs expressing eNpHR. f, Nociceptive thresholds under various conditions (n = 6, 6, 5, 5, 5, 4 mice). Daily VIP inhibition in NREM, but not wake or REM, reduces punctate (NREMZT2–10 vs. SNI, P = 0.018, 0.015, 0.038; NREMZT10–2 vs. SNI, P < 0.0001), dynamic (NREMZT2–10 vs. SNI, P < 0.0001, < 0.0001, = 0.0002; NREMZT10–2 vs. SNI, P < 0.0001), cold (NREMZT2–10 vs. SNI, P = 0.0012, 0.0003, < 0.0001; NREMZT10–2 vs. SNI, P = 0.0023, 0.034, 0.010), and hot (NREMZT2–10 vs. SNI, P = 0.0006, < 0.0001, 0.0004; NREMZT10–2 vs. SNI, P = 0.0026, 0.0019, 0.0052) allodynia after SNI. Mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001; NS, not significant; by two-sided two-way ANOVA followed by Bonferroni’s test (c, d, f). Detailed statistics in Supplementary Table 1.