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. 2022 Dec 16;48(10):1509–1519. doi: 10.1038/s41386-022-01520-0

Fig. 3. Chemogenetic inhibition or ablation of DRGABA+ neurons increases mechanical nociceptive sensitivity.

Fig. 3

A Schematic representation of DR injection of hM3Dq, hM4Di, and mCherry viruses in GAD2-ires-Cre mice and representative coronal sections of virus infection in the DR from bregma −4.24 to −4.84 mm. Scale bar, 250 μm. B Withdrawal threshold of the hind paw in response to Von Frey mechanical stimulation after chemogenetic regulation of DRGABA+ neurons following CNO administration. Interaction F(2, 36) = 3.453, P = 0.0425; CNO Factor F(1, 36) = 11.59, P = 0.0016; Group Factor F(2, 36) = 4.279, P = 0.0215. n = 7 for mCherry, hM3Dq and hM4Di. C Hind paw withdrawal latency in response to radiant heat after chemogenetic regulation of DRGABA+ neurons. D Representative images of Vgat-positive neurons (red) in the DR 3 weeks after control or taCaspase3 virus injection. Scale bar 100 μm. E Decreased mechanical threshold over time after ablation of DRGABA+ neurons by taCaspase3. Interaction F(1, 10) = 0.05683, P = 0.8164; Time Factor F(1, 10) = 11.34, P = 0.0072; Group Factor F(1, 10) = 23.58, P = 0.0007. n = 5 for control, n = 7 for taCaspase3. F Effect of taCaspase3 virus injection for 3 weeks on thermal withdrawal latency. P = 0.2611, t(10) = 1.191. n = 5 for Control, n = 7 for taCaspase3. G Representative movement traces in the OFT test showing the locations of the mCherry, hM3Dq, and hM4Di groups after CNO administration. H Quantitative results of center area entries (left) and the time spent in the center area (right) during the OFT test after the activation or inhibition of DRGABA+ neurons. Center area entries: F = 2.490, P = 0.1127; the time spent in the center area: F = 1.590, P = 0.2327. n = 7 for mCherry and hM3Dq, n = 6 for hM4Di. I Representative movement traces in the EPM test showing the locations of the mCherry, hM3Dq, and hM4Di groups after CNO administration. J Quantitative results of open arm entries (left) and the time spent in the open arms (right) of the mCherry, hM3Dq, and hM4Di groups after chemogenetic regulation of DRGABA+ neurons following CNO administration. Open arm entries: F = 0.6361, P = 0.5415; the time spent in the open arms: F = 0.05862, P = 0.9433. n = 7 for mCherry and hM3Dq, n = 6 for hM4Di. 2 W: 2 weeks; 3 W: 3 weeks. Data are presented as the Mean ± S.E.M. *P < 0.05, **P < 0.01.