(a) (top) Typical electrophysiological recording showing the increase in firing rate of VTA DA cell induced by 500 mg/kg i.v. alcohol injection in WT mice. (bottom) Example responses of a VTA DA neuron to consecutive doses of alcohol. Arrowheads indicate the time of the injection. (b) (left) Mean ± SEM DA cell firing frequency modification after injection of saline and the indicated ethanol dose (saline, n = 28; 125 mg/kg, n = 13; 250 mg/kg, n = 14; 500 mg/kg, n = 14; 750 mg/kg, n = 7). Arrowheads indicate the time of the injection. (right) Dose-response curve of ethanol-elicited responses for the same groups of neurons. ***p < 0.001, **p < 0.01, Wilcoxon paired test from baseline. Horizontal lines indicate significant dose effect (one-way ANOVA). (c) (top) Example responses of a VTA DA neuron to consecutive injections of alcohol, nicotine and combined alcohol + nicotine. Arrowheads indicate the time of the injections. (bottom) Barplot of the maximum of firing frequency variation from baseline (mean ± SEM) after alcohol (black), nicotine (white), or combined alcohol + nicotine injections (gray). ***p < 0.001, Wilcoxon paired test from baseline is indicated within each vertical bar. Difference between groups is indicated above the horizontal lines. ***p < 0.001, Wilcoxon paired test between groups. (d) (left) Examples of electrophysiological recordings of the basal activity of a DA neuron of a nic− (black) and a nic+ (gray) mouse. Insets: Enlarged views of two or more action potentials. (right) Barplot of the mean frequency and %SWB for nic− (black, n = 92) and nic+ mice (gray, n = 65). *p < 0.05, Wilcoxon test. (e) Dose-response curve of ethanol-elicited DA cell responses for nic− (black) and nic+ (gray) mice. Mean ± SEM of variation from baseline in firing frequency. Horizontal lines indicate significant dose effect and vertical lines indicate treatment effect (***p < 0.001, *p < 0.5, two-way ANOVA. Nic−: saline: n = 36; 125 mg/kg: n = 19; 250 mg/kg: n = 22; 500 mg/kg: n = 17; 750 mg/kg: n = 9; nic+: saline: n = 19; 125 mg/kg: n = 3; 250 mg/kg: n = 9; 500 mg/kg: n = 9; 750 mg/kg: n = 12).