Fig. 4

Protective effects of RS504393 on postnatal ethanol-induced loss of β-endorphin neurons in hypothalamus of rats. Postnatal rats were pretreated with RS504393 (1 mg/kg; 1 h before the first feeding) daily and fed from PND 2–6 with ethanol-containing milk formula (AF), pair-fed isocaloric milk formula (PF), or left undisturbed in the litter with mother (AD). β-Endorphin neurons were processed for immunofluorescence staining and colored green. Representative images of β-endorphin positive neurons in the hypothalamus of different treatment groups are shown in A, B for neonatal rats (PND6) and adult rats (PND60) in D, E. Histograms C, F represent mean ± SEM values of β-endorphin neuronal number (relative to AD using the ImageJ analysis) and are analyzed using two-way ANOVA with Tukey’s multiple comparison post hoc test. Significant differences between the groups are marked with *, p < 0.05; **, p < 0.01