Fig. 3.

Functional activation of FLAG-DOPr in KI mice. SNC80 (10 mg/kg; s.c.)-induced locomotor activity was assessed in WT, (A) KI (WT, n = 5; KI, n = 6; *P = 0.0102, **P = 0.0032, and ^φP = 0.0170; one-way ANOVA followed by Tukey’s post hoc test), and (B) KO mice (WT, n = 5–7; KO, n = 6; **P = 0.0038, ***P = 0.0004, and ^φφP = 0.0056; one-way ANOVA followed by Tukey’s post hoc test). DLT II (1 µg; i.t.)-induced antihyperalgesic effects was evaluated in WT, KO, and (C) KI mice using the Hargreaves test and the CFA model of inflammation (WT, n = 11 and ***P = 0.0005; KI, n = 7 and ***P = 0.0004; two-way ANOVA followed by Tukey’s post hoc test compared to KO [n = 11]) or (D) KO mice intrathecally injected with the recombinant adenoassociated virus AAV2/9-CBA-Cre (WT, n = 11 and ****P < 0.0001; KO + AAV2/9-CBA-Cre, n = 8 and *P = 0.0330; two-way ANOVA followed by Tukey’s post hoc test compared to KO [n = 8]). (E and F) The integrity of the G-protein coupling of DOPr (and FLAG-DOPr) was evaluated using the ³⁵S-GTPγS binding assay on coronal sections of olfactory bulb (20 µm) from WT, KI, and KO mice. ³⁵S-GTPγS binding density is shown in color scale (E). Quantification of binding signal was carried out on olfactory bulb sections (WT, n = 7 and ***P = 0.0004; KO, n = 4 and P = 0.1457; KI, n = 4 and *P = 0.0182; one-sample Wilcoxon test) from three independent experiments. Data are expressed as fold increase in GTPγS binding for nontreated (basal) and DLT II-treated sections (activated). Nonspecific binding was assessed in the presence of 10 µM GTPγS. n.s., not significant.