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. 2021 Sep 15;41(37):7727–7741. doi: 10.1523/JNEUROSCI.2592-20.2021

Figure 4.

Figure 4.

Chemogenetic inhibition or activation of RVM GPER+ neurons differentially modulates itch-related behaviors. A, Schematic showing the injection of AAV-DIO-hM4Di-mCherry into the RVM of female Gper-Cre mice. B, Expression of hM4Di-mCherry in the RVM after viral infection. Scale bar, 50 µm. C, Acute mechanical itch test 30 min following saline or CNO injection (1 mg/kg, i.p.) in mice expressing hM4Di receptors in RVM GPER+ neurons; n = 12 mice for each group; two-way ANOVA followed by Bonferroni's post hoc test. F(1,154) = 99.52; 0.013 g, p < 0.0001; 0.04 g, p = 0.0001; 0.08 g, p < 0.0001; 0.1 g, p = 0.0356; 0.3 g, p = 0.0100. D, E, Effect of pharmacogenetic inhibition of GPER+ neurons in the RVM on scratching behavior induced by intradermal injection of chloroquine (150 µg/20 µL; D) and histamine (500 µg/20 µL; E); n = 10–12 mice for each group; unpaired Student's t test. p = 0.0028, t = 3.382, df = 21 (D). p = 0.0021, t = 3.523, df = 20 (E). F, Time-dependent changes in scratching bouts of DCP-treated Gper-Cre mice after viral infection, CNO were injected intraperitoneally on days 5, 7, and 10 (gray frame), and the control group were injected with saline intraperitoneally; n = 5 mice for each group. Note CNO-treated group had higher scratching bouts than the saline group on the days of treatment (day 5, 7, and 10), although statistical significance was not reached (a: p = 0.1022, t = 0.1845 df = 8; b: p = 0.5456, t = 0.6310, df = 8; c: p = 0.2258, t = 1.312, df = 8). G, Schematic showing the injection of AAV-DIO-hM3Dq-eGFP into the RVM of female Gper-Cre mice. H, Expression of hM3Dq-eGFP in the RVM after viral infection. Scale bar, 50 µm. I, Acute mechanical itch test following saline or CNO injection in mice expressing hM3Dq receptors in RVM GPER+ neurons; n = 9 mice for each group; two-way ANOVA followed by Bonferroni post hoc test. F(1,112) = 7.877; 0.013 g, 0.04 g, 0.08 g, 0.1 g, 0.2 g and 0.8 g, p > 0.9999; 0.3 g, p = 0.4622. J, K, Pharmacogenetic activation of RVM GPER+ neurons decreased scratching behavior induced by chloroquine (200 µg/20 µL; J) and histamine (600 µg/20 µL; K); n = 6–9 mice for each group; CNO (1 mg/kg, i.p.); unpaired Student's t test. p = 0.0012, t = 4.062, df = 14 (J). p = 0.0005, t = 4.916, df = 11 (K). L, Pharmacogenetic activation of RVM GPER+ neurons attenuated spontaneous scratching behavior on day 15 of contact dermatitis mouse models; n = 4–5 mice for each group, unpaired Student's t test. p = 0.0339, t = 2.630, df = 7. NS, Not significant.