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. 2021 Jul 13;27(11):1313–1326. doi: 10.1111/cns.13704

FIGURE 7.

FIGURE 7

The effects of chemogenetic inhibition of GPER‐positive neurons in the RVM on the pain threshold and the activity of RVM neurons in pain persisting mice. (A) The protocol of the experiments. CNO (3 mg·kg‐ 1 per day, i.p.) or saline (i.p.) injection is performed from 10 to 14 days after incision surgery. (B) The schematic showing the injection of pAAV‐hSyn‐DIO‐hM4D(Gi)‐mCherry into the RVM of Gper‐Cre mice. (C) Successful expression of hM4D(Gi)‐mCherry in the RVM after viral infection. Scale bar, 100 μm. (D) and (E) The PWT and PWL of the daily baseline before CNO or saline administration in three groups mice: Pain recovery mice, Pain persisting + Saline mice and Pain persisting + CNO mice (n = 6 in each group, *p < 0.05, ***p < 0.001, ****p < 0.0001, between Pain persisting + CNO mice and Pain persisting + Saline mice; ##p < 0.01, ###p < 0.001, ####p < 0.0001, between Pain recovery mice and Pain persisting + Saline mice, two‐way repeated measures ANOVA followed by the Bonferroni post hoc test). (F) Representative immunofluorescence images showing the distribution of c‐Fos+ neurons in the RVM of the three groups mice on 15 days after incision surgery. Green: c‐Fos+ neurons. All scale bars = 100 μm. (G) Statistical analysis of the number of c‐Fos+ neurons in the RVM of three groups mice (n = 3 in each group, *p < 0.05, **p < 0.01, one‐way ANOVA)