Metabolite profiling of whole murine embryos reveals metabolic perturbations associated with maternal valproate-induced neural tube closure defects

Birth Defects Res. 2017 Jan 30;109(2):106-119. doi: 10.1002/bdra.23583.


Background: Valproic acid (VPA) is prescribed therapeutically for multiple conditions, including epilepsy. When taken during pregnancy, VPA is teratogenic, increasing the risk of several birth and developmental defects including neural tube defects (NTDs). The mechanism by which VPA causes NTDs remains controversial and how VPA interacts with folic acid (FA), a vitamin commonly recommended for the prevention of NTDs, remains uncertain. We sought to address both questions by applying untargeted metabolite profiling analysis to neural tube closure (NTC) stage mouse embryos.

Methods: Pregnant SWV dams on either a 2 ppm or 10 ppm FA supplemented diet were injected with a single dose of VPA on gestational day E8.5. On day E9.5, the mouse embryos were collected and evaluated for NTC status. Liquid chromatography coupled to mass spectrometry metabolomics analysis was performed to compare metabolite profiles of NTD-affected VPA-exposed whole mouse embryos with profiles from embryos that underwent normal NTC from control dams.

Results: NTDs were observed in all embryos from VPA-treated dams and penetrance was not diminished by dietary FA supplementation. The most profound metabolic perturbations were found in the 10ppm FA VPA-exposed mouse embryos, compared with the other three treatment groups. Affected metabolites included amino acids, nucleobases and related phosphorylated nucleotides, lipids, and carnitines.

Conclusion: Maternal VPA treatment markedly perturbed purine and pyrimidine metabolism in E9.5 embryos. In combination with a high FA diet, VPA treatment resulted in gross metabolic changes, likely caused by a multiplicity of mechanisms, including an apparent disruption of mitochondrial beta-oxidation. Birth Defects Research 109:106-119, 2017. © 2016 Wiley Periodicals, Inc.

Keywords: folic acid; metabolism; metabolomics; neural tube defects; one-carbon metabolism; teratogen; valproic acid.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Amino Acids / metabolism
  • Animals
  • Carnitine / metabolism
  • Dietary Supplements*
  • Disease Models, Animal
  • Embryo, Mammalian
  • Female
  • Folic Acid / administration & dosage*
  • Lipids / analysis
  • Male
  • Metabolome
  • Mice
  • Neural Tube / abnormalities
  • Neural Tube / drug effects
  • Neural Tube / metabolism
  • Neural Tube Defects / chemically induced
  • Neural Tube Defects / metabolism*
  • Neural Tube Defects / pathology
  • Neurulation / drug effects*
  • Penetrance
  • Pregnancy
  • Purines / metabolism
  • Pyrimidines / metabolism
  • Teratogens / toxicity*
  • Valproic Acid / toxicity*


  • Amino Acids
  • Lipids
  • Purines
  • Pyrimidines
  • Teratogens
  • Valproic Acid
  • Folic Acid
  • Carnitine