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. 2025 Nov;155(11):3781-3790.
doi: 10.1016/j.tjnut.2025.07.013. Epub 2025 Jul 24.

Contrasting Effects of Phosphatidylcholine and Betaine Supplementation on Embryonic Development in a Mouse Model of the MTHFD1 R653Q Variant

Karen E Christensen  1 , Marie-Lou Faquette  1 , Aaron T Gebert  2 , Vafa Keser  1 , Teodoro Bottiglieri  2 , Rima Rozen  3
Affiliations

Contrasting Effects of Phosphatidylcholine and Betaine Supplementation on Embryonic Development in a Mouse Model of the MTHFD1 R653Q Variant

Karen E Christensen et al. J Nutr. 2025 Nov.

Abstract

Background: Folate and choline are interconnected nutrients important for embryonic development. Genetic variants in folate enzymes, such as MTHFD1 R653Q (c.1958 G>A, rs2236225), may increase dietary choline requirements.

Objectives: To evaluate the impact of phosphatidylcholine or betaine supplementation on embryonic development and maternal choline metabolism in the Mthfd1S+/- mouse model for the MTHFD1 653QQ genotype.

Methods: Female Mthfd1S+/+ (wild type [WT]) and Mthfd1S+/- (heterozygous [HET]) mice were fed amino acid-defined control (Ctrl), phosphatidylcholine-supplemented (PtdCho), or betaine-supplemented (BTN) diets for 4-6 wk before mating and during pregnancy (18-33 mice/group). Ctrl contained 2.5 g choline bitartrate/kg diet (based on AIN-93G). Supplemented diets contained additional phosphatidylcholine or betaine to obtain the molar equivalent of 3 times the free choline of Ctrl. Embryos were examined for anatomical developmental delays and morphological defects at embryonic day 10.5. One-carbon metabolites (homocysteine, choline, betaine, methionine, cystathionine, S-adenosylmethionine, S-adenosylhomocysteine) were measured in maternal plasma and liver by liquid chromatography-mass spectrometry (LC-MS/MS). The MTHFR protein in maternal liver was assessed by Western blot. Data were analyzed by binary logistic regression or 2-way analysis of variance.

Results: PtdCho increased developmental delays, compared with Ctrl, in litters of WT (170%) and HET (125%) mothers (Pdiet < 0.05). BTN decreased delays 53% compared with Ctrl in HET mother's litters (Pdiet (HET) < 0.05). Defects decreased 71% in BTN-fed HET mother's litters compared to WT (P < 0.05). BTN increased total pregnancy losses 360% (Pdiet < 0.01); PtdCho had no effect. Both diets decreased maternal plasma homocysteine (30-50%, Pdiet < 0.05), increased liver BTN (220-290%, Pdiet < 0.0001), and decreased S-adenosylmethionine/S-adenosylhomocysteine (WT: 20-30%, HET: 48-61%, Pdiet < 0.05). MTHFR protein expression decreased 25% in PtdCho maternal liver (Pdiet < 0.01).

Conclusions: Phosphatidylcholine and betaine supplementation had contrasting effects on reproductive outcomes. PtdCho increased developmental delays. BTN reduced delays and defects in the Mthfd1S+/- mouse model, but may impair the establishment of pregnancy. The timing, form, and dosage of choline supplementation may be important and require further investigation.

Keywords: MTHFD1; betaine; choline; developmental defects; developmental delay; embryo; neural tube defects; phosphatidylcholine.

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Conflict of interest statement

Conflict of interest The authors report no conflicts of interest.

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