Fig. 2
ACC CRISPR/Cas9 mediated Drd1 KO modulate pain related behaviors. a Schematic diagram of experimental design. b Representative brain slice sections obtained from Cas9-EGFP mice injected with AAV-hSyn-Cre + AAV-Drd1-sgRNA-mCherry into the ACC (dashed box 1). The striatum (dashed box 2) is shown for comparison. Scale bars = 500 μM for full brain section; 100 μM for ACC and striatum sections. c No difference between control and ACC_D1-KO mice in the open field for distance traveled over the 30 min observation period (two-way ANOVA, main effect of genotype: F1,22 = 0.01, p = 0.91; main effect of time: F3,67 = 77.94, p < 0.0001; genotype x time interaction: F1,109 = 1.85, p = 0.11). d ACC_D1-KO mice spend significantly more time in the center of the open field compared with control mice (t-test, t21 = 2.48, p = 0.02). e No difference between control and ACC_D1-KO mice in total time spent (s) in the open arms of the elevated plus maze (t-test, t22 = 0.07, p = 0.95). f No difference between control and ACC_D1-KO mice in the time spent in the light zone as measured using the dark/light emergence test (t-test, t19 = 0.15, p = 0.88). g ACC_D1-KO and control mice spend more time in the new arm of the Y-maze (left graph) compared with previously visited arms (arms A and B) (two-way ANOVA, main effect of arm: F2,57 = 10.74, p < 0.001; main effect of genotype: F1,57 = 0.00, p = 0.99; arm x genotype interaction: F2,57 = 0.01, p = 0.99). There was no difference between the genotypes for percentage of visits to the new arm (t19 = 0.07, p = 0.94) or total distance travelled in the Y-maze (right graph, t22 = 0.15, p = 0.88). h ACC_D1-KO mice have lower mechanical thresholds before, but not following CFA injection when compared with control mice (two-way ANOVA, main effect of genotype: F1,68 = 0.48, p = 0.49; main effect of day: F1,68 = 50.89, p < 0.001; genotype x day: F1,68 = 10.08, p = 0.02). All behaviors were conducted with control (n = 12) and ACC_D1-KO (n = 12) mice. Data in all graphs represent the mean and standard error of the mean (SEM). *p < 0.05, **p < 0.01, ****p < 0.0001