Methyl donor-deficient diet during development can affect fear and anxiety in adulthood in C57BL/6J mice

PLoS One. 2014 Aug 21;9(8):e105750. doi: 10.1371/journal.pone.0105750. eCollection 2014.


DNA methylation is one of the essential factors in the control of gene expression. Folic acid, methionine and choline (methyl donors)--all nutrients related to one-carbon metabolism--are known as important mediators of DNA methylation. A previous study has shown that long-term administration of a diet lacking in methyl donors caused global DNA hypermethylation in the brain (Pogribny et al., 2008). However, no study has investigated the effects of a diet lacking in methyl donors during the developmental period on emotional behaviors such as fear and anxiety-like behavior in association with gene expressions in the brain. In addition, it has not been elucidated whether a diet supplemented with methyl donors later in life can reverse these changes. Therefore, we examined the effects of methyl donor deficiency during the developmental period on fear memory acquisition/extinction and anxiety-like behavior, and the relevant gene expressions in the hippocampus in juvenile (6-wk) and adult (12-wk) mice. We found that juvenile mice fed a methyl-donor-deficient diet had impaired fear memory acquisition along with decreases in the gene expressions of Dnmt3a and Dnmt3b. In addition, reduced anxiety-like behavior with decreased gene expressions of Grin2b and Gabar2 was observed in both the methyl-donor-deficient group and the body-weight-matched food-restriction group. After being fed a diet supplemented with methyl donors ad libitum, adult mice reversed the alteration of gene expression of Dnmt3a, Dnmt3b, Grin2b and Gabar2, but anxiety-like behavior became elevated. In addition, impaired fear-memory formation was observed in the adult mice fed the methyl-donor-deficient diet during the developmental period. Our study suggested that developmental alterations in the one-carbon metabolic pathway in the brain could have effects on emotional behavior and memory formation that last into adulthood.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Anxiety* / chemically induced
  • Anxiety* / metabolism
  • Behavior, Animal*
  • DNA (Cytosine-5-)-Methyltransferases / biosynthesis
  • DNA Methylation*
  • DNA Methyltransferase 3A
  • DNA Methyltransferase 3B
  • Diet / adverse effects*
  • Fear*
  • Female
  • Gene Expression Regulation
  • Hippocampus / metabolism*
  • Hippocampus / pathology
  • Male
  • Mice
  • Receptors, N-Methyl-D-Aspartate / biosynthesis


  • Dnmt3a protein, mouse
  • NR2B NMDA receptor
  • Receptors, N-Methyl-D-Aspartate
  • DNA (Cytosine-5-)-Methyltransferases
  • DNA Methyltransferase 3A

Grants and funding

This work was supported by Japan Society for the Promotion of Science KAKENHI Grant Number 24791196. DM received this funding. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.