Extended Data Fig. 4: Effect of systemic σ receptor ligands on motor behaviour.
From: Structures of the σ2 receptor enable docking for bioactive ligand discovery
a, Response of mice to a von Frey filament after spared nerve injury (SNI). All five ligands are compared to their respective vehicles (PD-144418 10 mg/kg (n = 5) and 30 mg/kg (n = 5) vs. kolliphor (n = 5), one-way ANOVA, F(2, 12) = 7.49, p = 0.008; Z4446724338 10 mg/kg (n = 10) and 20 mg/kg (n = 5) vs cyclodextrin (n = 10), one-way ANOVA, F(2, 22) = 25.12, p < 0.001; Z4857158944 10 mg/kg (n = 5) and 20 mg/kg (n = 5) vs cyclodextrin (n = 10), one-way ANOVA, F(2, 17) = 5.10, p = 0.02; Z1665845742 10 mg/kg (n = 10) and 20 mg/kg (n = 5) and PB28 30 mg/kg (n = 10) vs saline (n = 10), one-way ANOVA, F(3, 31) = 6.18, p = 0.002; asterisks define individual group differences to respective vehicle control using Dunnett’s multiple comparisons Post-hoc test; ns = not significant, * p < 0.05, ** p < 0.01, *** p < 0.001). Data shown are mean ± s.e.m. Data for higher doses and vehicles is replotted from Fig. 4. b, No sedation or motor impairment on the rotarod was observed after drug treatments compared to vehicle at 1 h (Z1665845742 10 mg/kg (n = 5) and Z4857158944 20 mg/kg (n = 5) vs saline (n = 5), one-way ANOVA, F(2, 12) = 1.04, p = 0.38; Z4446724338 10 mg/kg (n = 5) vs kolliphor (n = 5), unpaired two-tailed Student’s t-test, t(8) = 0.47, p = 0.65) or 24 h post-injection (Z1665845742 10 mg/kg (n = 5) and Z4857158944 20 mg/kg (n = 5) vs saline (n = 5), one-way ANOVA, F(2, 12) = 0.45, p = 0.65; Z4446724338 10 mg/kg (n = 5) vs kolliphor (n = 5), unpaired two-tailed Student’s t-test, t(8) = 0.72, p = 0.49); ns = not significant. Data shown are means ± s.e.m. c, Response of SNI mice to a von Frey filament after repeated injections of Z4446724338 10 mg/kg (n = 5). Mechanical thresholds were assessed 1 h and 24 h after four separate injections. Data shown are means ± s.e.m. normalized to each mouse’s SNI baseline. d, Response of SNI mice to a von Frey filament after repeated injections of Z4857158944 10 mg/kg (n = 5). Mechanical thresholds were assessed 1 h and 24 h after four separate injections. Data shown are means ± s.e.m. normalized to each mouse’s SNI baseline. e. Quantification of the expression levels of Sigmar1 (σ1) and Tmem97 (σ2) in wildtype (WT) and SNI mice detected by in situ hybridization (n = 3 mice per group). Representative images can be found in panel f. Data shown are mean ± s.e.m.; unpaired two-tailed Student’s t-test— Sigmar1: t(4) = 0.5, p = 0.64; Tmem97: t(4) = 1.0, p = 0.37; ns = not significant. AU = arbitrary units. f, in situ hybridization of mouse dorsal root ganglion (DRG) sections for Sigmar1 (σ1) and Tmem97 (σ2) genes illustrates expression in myelinated (Nefh-positive; blue) and unmyelinated (Acpp-positive; red) subsets of sensory neurons and no change after SNI
