Fig. 1

Characterization of the exposure paradigm. a Time line for alcohol exposures and experiments. Pups, along with dams, were exposed to alcohol for 4 h/day via vapor inhalation from P3 to P5 (gray bar). Samples collected from P3–P6 were taken either during periods of withdrawal or peak blood alcohol concentrations (BAC) as indicated and were used for PCR (pink text) and/or immunohistochemistry (IHC, blue text). Each collected IHC time point included staining for IBA-1/microglia, GFAP/astrocytes, and neuronal loss. IHC was also performed for neuronal apoptosis but only on P4 and only in the cerebellar vermis region. Additional time points were included beyond the exposure paradigm for PCR, IHC, and behavior (purple text). b Pups were weighed daily and the average weight per pup was recorded (n = 3–12 litters). c Nursing ability was quantified as the percentage of pups in each litter with milk visible in their stomachs each morning prior to exposure (n = 5–6 litters). d Similar litter sizes were assigned to each treatment group on P3 (n = 17 litters). e Pup blood alcohol concentrations (n = 4 pups from 4 litters) were each measured at the beginning (0 h), middle (2 h), and end (4 h) of the exposure, as well as at several time points after. The green line represents the 4-h exposure period. Peak maternal BAC was assessed at 4 h (n = 3 dams exposed after pups had been weaned). (***p < 0.001)