178 - Impact of Choline Insufficiency on Ethanol Metabolism in a Prenatal Alcohol Exposure Mouse Model

Publication Number: 178.5490

Bosco Bakwatanisa, Case Western Reserve University School of Medicine, Cleveland, OH, United States; Junwoo Suh, Case Western Reserve University School of Medicine, Cleveland, OH, United States; Stephanie Ford, UH Rainbow Babies and Children's Hospital, Case Western Reserve University School of Medicine, Cleveland, OH, United States; Michiko Watanabe, Case Western Reserve University, Kent, OH, United States; Michael Jenkins, Case Western Reserve University School of Medicine, Cleveland, OH, United States; Andrew M. Rollins, Case Western Reserve University, Cleveland, OH, United States

Background: Prenatal alcohol exposure (PAE) is associated with developmental abnormalities, including congenital heart defects. Studies suggest that choline supplementation, an essential nutrient involved in embryogenesis, mitigates some of PAE’s detrimental effects. During pregnancy, however, the demand for choline escalates and many women of reproductive age do not meet recommended choline intake. We have previously shown that a choline deficient diet (CDD) mouse model with PAE during gastrulation induces congenital heart disease in embryos.

Objective: To characterize a choline-deficient mouse model and assess how this state influences blood alcohol concentrations (BAC).

Design/Methods: (i). Choline Assay: C57Bl/6J female mice were given a CDD for 4-5 weeks. Serum choline concentrations were measured at each time point using the Sigma Aldrich MAK508 choline assay. (ii). Ethanol Assay: Mice on standard diet received 2 gavage doses of 0.01 ml/g 30% ethanol, administered 2 hrs apart, to simulate one binge drinking episode. BAC was measured at 2, 3, and 4 hours after dosing using the Sigma Aldrich MAK481 ethanol assay kit to assess ethanol pharmacokinetics. (iii) Ethanol Assay on Choline-Insufficient Mice: The ethanol assay will be used to assess ethanol metabolism of mice on a choline-deficient diet compared to that of mice on a standard diet. Analysis of variance (ANOVA) will be used to compare between group serum choline levels and BAC over time.

Results: Samples from 8 mice were analyzed in triplicate for each group. Serum choline concentrations decreased by 31% between weeks 4 and 5 (Figure 1A). BAC in standard chow-fed mice peaked at 2 hrs after the ethanol doses, at a concentration of 42.29 mg/dL, with a gradual decline but above zero even at 4 hrs (Figure 1B). Ethanol concentrations on CDD are in process as of this submission.

Conclusion(s): Our preliminary results indicated that dams had a continued reduction in serum choline levels after 4-5 weeks on a choline deficient diet. This may have implications for fetal development in the context of PAE. The ethanol assay shows that the alcohol ingested is consistent with previous ethanol exposure mouse models and can be related to human exposure. This study underscores the importance of choline in prenatal health, and future studies will determine if choline insufficiency exacerbates the metabolic burden of alcohol exposure. These findings provide a foundation for exploring dietary interventions in pregnant women at risk of alcohol consumption, with the goal of reducing the incidence of congenital heart defects and other malformations.

Figure 1: Serum Choline and Blood Alcohol Concentrations in Mice
Panel A shows serum choline concentrations (ng/µL) in mice after 4 and 5 weeks on a choline-deficient diet (CDD). Concentrations in Week 5 are generally lower than in Week 4, indicating a decrease over time. Panel B displays blood alcohol concentrations (BAC, mL/g) in mice measured 2, 3, and 4 hours after gavage feeding with two doses (0.01 mL/g) administered 2 hours apart. The decrease in BAC over time reflects alcohol metabolism. Yellow diamonds indicate the mean concentration in each group.