Figure 4
Role of Rhes in the modulation of cocaine induced-cAMP/PKA pathway activation in DStr and NAc and in the induction of the immediate-early gene expression. (A) Quantitative reverse transcription-PCR to evaluate Rhes mRNA expression in DStr (n = 6) and NAc (n = 6) from wild-type mouse samples. (B,C) Analysis of 32 kDa (B) and 47 kDa (C) Rhes protein abundance in DStr (n = 5) and NAc (n = 4) from wild-type mouse lysates. Unpaired t test. **p < 0.01, compared to DStr group. (D–F) Evaluation of pGluA1protein levels, following 30 mg/kg cocaine treatment in WT and KO mice at different time points post-cocaine injection in DStr (10 min: vehicle, n = 6 WT/3 KO; cocaine, n = 6 WT/4 KO; 30 min: vehicle, n = 5/genotype; cocaine, n = 5/genotype; 60 min: vehicle, n = 4/genotype; cocaine, n = 5/genotype) (D–F) and NAc (10 min: vehicle, n = 6 WT/3 KO; cocaine, n = 6/genotype; 30 min: vehicle, n = 5/genotype; cocaine, n = 5/genotype; 60 min: vehicle, n = 4/genotype; cocaine, n = 5/genotype) (D'–F'). *p < 0.05, **p < 0.01, ***p < 0.0001, compared to vehicle-treated group within genotype, Uncorrected Fisher’s LSD test. Top panels show representative blots. All representative blots shown in the figures arise from cut-out and pasted bands for reassembling the image. Of note, the representative bands come from the same films for each graph. G,I Representative images of radioactive in situ hybridization (ISH) on mouse prefrontal cortex (G) and caudate putamen (I) using specific antisense riboprobes for the immediate-early genes Arc, Zif 268, and Homer 1a. Densitometric analysis of autoradiograms reported in the histogram show the quantification of Arc, Zif 268, and Homer 1a expression levels in the prefrontal cortex (H) and caudate putamen (J) of adult wild-type and KO mice treated with cocaine or vehicle. Scale bar 500 μm. *p < 0.05, **p < 0.01, compared to vehicle-treated group within genotype, Uncorrected Fisher’s LSD test. Values are expressed as mean ± SEM of relative optical density (ROD). Genotypes and treatments are as indicated.
