Figure 1.

Cocaine exposure downregulates miR-125b expression. SH-SY5Y cells were cultured overnight and then differentiated for 3–5 d. A, Dopaminergic phenotype of these cells was monitored via neurite length measurement by phase-contrast microscopy. B, Western blot analysis of DAT expression in differentiated SH-SY5Y cells. C, DAT and TH expression in differentiated SH-SY5Y cells were detected by confocal microscope. Data are representative of three independent experiments. D, Differentiated SH-SY5Y cells were treated with varying concentrations of cocaine. Cells were harvested and miRNA expression was analyzed by qPCR. miR-125b expression was normalized to the 5S rRNA levels. Relative expression of miR-125b in cocaine-treated cells was compared to the untreated control cells and are plotted as fold change. E, Mice were injected (i.p.) with a single dose of cocaine (20 mg/kg weight of mice) or saline and animals were sacrificed. NAc tissues were used for miRNA isolations and were subjected to qPCR. miR-125b expression was normalized to the 5s rRNA levels and relative expression of miR-125b in the NAc of cocaine-treated mice (n = 6) are expressed as fold changes to the NAc of saline-treated mice (n = 6). *p < 0.05 for the comparison of cocaine-treated samples versus untreated controls.