Effect of intraperiaqueductal grey (PAG) and rostroventral medulla (RVM) microinjection of HU210 (CB1/2 receptor agonist, 4 μg/animal) on withdrawal reflexes to mechanical von Frey hair stimulation in P10, P21, and P40 rats. (A) Intra-PAG HU210 significantly decreased spinal reflex excitability compared with vehicle responses in all ages tested (P < 0.0001; 2-way analysis of variance [ANOVA]). This effect was more pronounced in P10 animals compared with P40s. (B) Intra-PAG HU210 significantly increased mechanical thresholds in P21 and P40 rats (P < 0.0001; 2-way ANOVA). This effect is more pronounced in P21 and P40 animals compared with P10. (C) Intra-RVM HU210 decreased spinal reflex excitabilities compared with vehicle responses in all ages tested (P < 0.0001, 2-way ANOVA). This effect was strongest in P21 animals. (D) Intra-RVM HU210 significantly increased mechanical thresholds compared with vehicle responses in all ages tested (P < 0.0001, 2-way ANOVA). This effect was strongest for P21 animals. Four to 8 animals per drug per age group. Data shown here represent the mean ± SEM. **** = P < 0.0001, between drug comparisons, 2-way ANOVA with Bonferroni multiple comparisons. # and #### = P < 0.05 and P < 0.0001, respectively, between age comparison, 2-way ANOVA with Bonferroni multiple comparisons.