Timed pregnant Sprague-Dawley dams were obtained from the Center for Behavioral Teratology, San Diego State University Animal Care facilities. Gestational day (GD) 0 was designated as the day when a seminal plug was detected. Dams were housed individually in plastic cages, exposed to a 12:12 hour cycle of light and dark in a temperature- and humidity-controlled room, and received food (LabDiet® 5001, Richmond, IN, containing 2.25 g choline chloride/kg diet) and water ad libitum.
Pregnant dams were randomly assigned to one of three treatment groups: ethanol-exposed (EtOH), yoked pair-fed (PF), or ad libitum control (LC). Ethanol-exposed dams received 6.0 g/kg/day (28.5% v/v) ethanol, PF dams received an isocaloric maltose dextrin solution to control for the calories from alcohol, and LC dams received vehicle (saline), via daily oral gavage from GD 5–20. Daily food intake was measured for the EtOH dams; each PF dam was matched to an EtOH dam of similar weight and food intake was correspondingly yoked. Within each of the 3 treatment groups, dams were randomly assigned to receive either a choline supplementation (choline chloride, Blachem, New Hampton, NY; 250 mg/kg/day) or a vehicle control (saline, Sigma, Milwaukee, WI), added to the daily intubation formula. This administration increases daily choline intake by 2–3 times that of controls.
Animals were monitored until the expected day of delivery GD 22 (PD 0) and the day of birth was recorded. On PD 1, litters were culled to 10 pups (5 males and 5 females when possible). Data on litter characteristics and birth weights have been previously reported (Thomas et al., 2009 (link)). Blood alcohol levels over a 24-hour period were obtained from a separate group of pregnant rats on gestational days 5 and 20. Importantly, choline supplementation did not influence blood alcohol concentrations, which peaked at 345 mg/dL (Thomas et al., 2009 (link)), indicating that choline does not alter the amount of fetal alcohol exposure.
Pregnant dams were randomly assigned to one of three treatment groups: ethanol-exposed (EtOH), yoked pair-fed (PF), or ad libitum control (LC). Ethanol-exposed dams received 6.0 g/kg/day (28.5% v/v) ethanol, PF dams received an isocaloric maltose dextrin solution to control for the calories from alcohol, and LC dams received vehicle (saline), via daily oral gavage from GD 5–20. Daily food intake was measured for the EtOH dams; each PF dam was matched to an EtOH dam of similar weight and food intake was correspondingly yoked. Within each of the 3 treatment groups, dams were randomly assigned to receive either a choline supplementation (choline chloride, Blachem, New Hampton, NY; 250 mg/kg/day) or a vehicle control (saline, Sigma, Milwaukee, WI), added to the daily intubation formula. This administration increases daily choline intake by 2–3 times that of controls.
Animals were monitored until the expected day of delivery GD 22 (PD 0) and the day of birth was recorded. On PD 1, litters were culled to 10 pups (5 males and 5 females when possible). Data on litter characteristics and birth weights have been previously reported (Thomas et al., 2009 (link)). Blood alcohol levels over a 24-hour period were obtained from a separate group of pregnant rats on gestational days 5 and 20. Importantly, choline supplementation did not influence blood alcohol concentrations, which peaked at 345 mg/dL (Thomas et al., 2009 (link)), indicating that choline does not alter the amount of fetal alcohol exposure.
