qRT-PCR analysis of expression ofCcl2,NOS-2,TNF-α,Tnfrsf1aandCD74during the first 48 hours of alcohol withdrawal. Alcohol-treated rats were fed the Lieber-DeCarli liquid alcohol diet for 35 days prior to forced withdrawal. qRT-PCR was performed on CeA and DVC samples. All transcripts were found to have a significant treatment effect by two-factor ANOVA, and to change significantly as a result of alcohol treatment and withdrawal. (A) CeA mean ∆∆CT values for the transcripts Ccl2, NOS-2, TNF-α and Tnfrsf1a and CD74 in control (C; N = 10), chronically alcohol exposed (E; N = 5), and 4 hours (4 h W; N = 5) and 48 hours (48 h W, N = 3) withdrawn rats. (B) DVC mean ∆∆ CT values (C N = 11; E = 5, 4 h W n = 7, 48 h N = 5). A ∆∆CT = 1 is a doubling of control mRNA levels. Error bars represent the +/−SEM. *Significant treatment effect identified via two-way ANOVA (P ≤ 0.05); **Significant treatment effect identified via two-way ANOVA (P ≤ 0.01); ***Significant treatment effect identified via two-way ANOVA (P ≤ 0.005); † Significant post hoc Tukey’s test versus control, (P ≤ 0.01.) ANOVA, two-way analysis of variance; Ccl2, chemokine (C-C motif) ligand 12; CeA, central nucleus of the amygdala; DVC, dorsal vagal complex; NOS-2, inducible nitric oxide synthase; qRT-PCR, quantitative reverse transcription polymerase chain reaction.
Freeman et al. Journal of Neuroinflammation 2012 9:97 doi:10.1186/1742-2094-9-97