Rat brain tissue was harvested in a time course during and after binge exposure: 4D (directly after the last dose of alcohol on the fourth day), 4D+2 (2d after the last dose of alcohol), 4D+7, and 4D+28. To examine cell proliferation, rats were administered Bromo-deoxy-Uridine (BrdU; 300mg/kg, i.p; Fluka, Buchs, Switzerland) and sacrificed two hours later (4D injected 1 h after the last dose of alcohol). To investigate cell survival, BrdU (300mg/kg, i.p.) was administered at 4D and the rats were then sacrificed 28 days later (4D+28 days). The 300 mg/kg dose of BrdU was chosen as that which maximally labels dividing cells in PND 35 rats (Cameron and McKay, 2001). Identical to previous reports (Knapp and Crews, 1999; Nixon and Crews, 2004), rats were sacrificed by anesthetic overdose (Nembutal®; MWI Veterinary Supply, Nampa, Idaho) and transcardially perfused with 0.1M phosphate buffered saline (PBS, pH 7.4) followed by 4% paraformaldehyde (pH 7.4 in 0.1M phosphate buffer). Brains were postfixed for 24 h in 4% paraformaldehyde then rinsed and stored in PBS until sectioning. Brains were sectioned coronally, at a thickness
