Fig. 5: Rasd2 overexpression in DRD2^PrL-NAcc neurons decreases depression-like behavior.

a Schematic of experimental design and timeline. b Representative image for the identification of Drd2-cre genotypes. c Schematic diagram of microinjection of viral vectors into mouse PrL and NAcc sites (top, left); Representative images of EGFP (top, middle) and fDIO-mCherry (top, right) injection sites in the NAcc and PrL, respectively; Representative images of mCherry- (middle, left) and EGFP- (middle, middle) positive neurons in the PrL; Representative image of mCherry and EGFP co-labeled in the PrL (middle, right), arrows indicate co-expressed neurons.; Representative images of mCherry- (bottom, left) and EGFP- (bottom, middle) positive neurons in the NAcc; Representative image of mCherry and EGFP co-labeled in the NAcc (bottom, right). d Representative images showing co-localization of RASD2 and DRD2 in NAcc 3 weeks after virus injection (left, scale bar = 20 μm) and the number of DRD2-positive neurons that co-express RASD2 in NAcc (Right). N = 4 mice per group. Two-tailed unpaired t-test: t₍₂₂₎ = 2.101, p = 0.0474. Representative tracks (e) and heatmaps (f) displaying mouse movement in the open field test. Total distance moved (g), line crosses (h), rearing (i), and center entries (j) in the open field test. N = 10 mice per group. Two-tailed unpaired t-test: distance, t₍₁₈₎ = 0.9870, p = 0.3367; line crosses, t₍₁₈₎ = 0.0605, p = 0.9524; rearing, t₍₁₈₎ = 0.3031, p = 0.7653; center entries, t₍₁₈₎ = 0.8985, p = 0.3808. Duration of immobility (k), climbing (l), and swimming (m), and the number of fecal boli (defecation) (n) in the forced swim test. N = 10 mice per group. Two-tailed unpaired t-test: immobility, t₍₁₈₎ = 3.658, p = 0.0018; climbing, t₍₁₈₎ = 2.384, p = 0.0284; swimming, t₍₁₈₎ = 2.115, p = 0.0486; defecation, t₍₁₈₎ = 0.5153, p = 0.6128. o Graph represents sucrose preference (%) before (Day 0) and after (Day 23) treatment. N = 10 mice for each group. Two-way ANOVA (factor 1: time; factor 2: treatment): F_{time×treatment (1, 36)} = 3.817, p = 0.0586; F_{time (1, 36)} = 1.321, p = 0.2581; F_{treatment (1,36)} = 3.817, p = 0.0586. Tukey’s HSD post hoc test: after treatment, p_{(WT vs. Drd2-cre)} = 0.0426. p Duration of immobility in the tail suspension test. N = 10 mice per group. Two-tailed unpaired t-test: t₍₁₈₎ = 3.300, p = 0.0040. The data are expressed as mean ± s.e.m. *p < 0.05. 5-d UMS 5-day unpredictable mild stress, OFT open field test, FST forced swim test, TST tail suspension test, IF immunofluorescence, WT wild type, Cg1 cingulate cortex 1, PrL prelimbic cortex, IL infralimbic cortex, NAcc NAc core, DRD2 dopamine D2 receptor.