Dietary folic acid deficiency impacts hippocampal morphology and cortical acetylcholine metabolism in adult male and female mice

Nutr Neurosci. 2022 Oct;25(10):2057-2065. doi: 10.1080/1028415X.2021.1932242. Epub 2021 May 27.


Objective: One-carbon (1C) metabolism is a metabolic network that integrates nutritional signals with biosynthesis, redox homeostasis, and epigenetics. There are sex differences in hepatic 1C metabolism, however, it is unclear whether sex differences in 1C impact the brain. The aim of this study was to investigate if sex modulates the effects of dietary folic acid deficiency, the main component of 1C, in brain tissue using a mouse model.

Methods: Male and female C57Bl/6J mice were placed on a folic acid deficient (FD) or control diet (CD) at six weeks until six months of aged. After which brain tissue and serum were collected for analysis. In brain tissue, hippocampal volume, morphology, and apoptosis as well as cortical acetylcholine metabolism were measured.

Results: Male and female FD mice had reduced serum levels of folate. Both males and females maintained on a FD showed a decrease in the thickness of the hippocampal CA1-CA3 region. Interestingly, there was a sex difference in the levels of active caspase-3 within the CA3 region of the hippocampus. In cortical tissue, there were increased levels of neuronal ChAT and reduced levels of AChE in FD females and male mice.

Conclusions: The results indicated that FD impacts hippocampal morphology and cortical neuronal acetylcholine metabolism. The data from our study indicate that there was only one sex difference and that was in hippocampal apoptosis. Our study provides little evidence that sex modulates the effects of dietary folate deficiency on hippocampal morphology and cortical neuronal acetylcholine metabolism.

Keywords: One-carbon metabolism; acetylcholine; apoptosis; choline; cortex; folic acid; hippocampus; sex differences.

MeSH terms

  • Acetylcholine / metabolism
  • Carbon
  • Caspase 3 / metabolism
  • Diet
  • Female
  • Folic Acid
  • Folic Acid Deficiency*
  • Hippocampus / metabolism
  • Humans
  • Male


  • Carbon
  • Folic Acid
  • Caspase 3
  • Acetylcholine