Female Long-Evans rats were housed with an experienced male overnight and gestational day (GD) 1 was defined by the presence of sperm in the vaginal smear on the following morning. No handling other than routine cage changing occurred prior to breeding. Pregnant female rats were singly-housed and assigned to one of three treatments: ethanol-treated (ET), intubated control (IC), and non-treated control (NC). Prenatal treatments occurred from GD 1 through GD 22. ET dams were intubated using a stainless steel gavage tube with 4.5 g/kg of ethanol in 20 ml/kg of distilled water on a daily basis between 3:00 and 5:00 PM; food and water was available ad libitum. IC dams were pair-fed to an ET dam of similar weight and were intubated daily with maltose-dextrin made isocaloric to the ethanol solution in 20 ml/kg of distilled water. Inclusion of the IC group is to control for both nutritional and stress effects of the intubation procedure (Kelly et al., 2008). ET and IC dams were weighed daily, while NC dams were not handled in any manner. The day of birth was GD 23 and was considered PD 1 and no treatments occurred on this day.
