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

Calli Bennett, Jacalyn Green, Mae Ciancio, Joanna Goral, Lenore Pitstick, Matt Pytynia, Alice Meyer, View ORCID ProfileNeha Kwatra, View ORCID ProfileNafisa M. Jadavji
doi: https://doi.org/10.1101/2020.12.13.422535
Calli Bennett
1Biomedical Sciences Program, College of Graduate Studies, Midwestern University, Glendale, US
2College of Osteopathic Medicine, Midwestern University, Glendale, US
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Jacalyn Green
3Biochemistry and Molecular Genetics, College of Graduate Studies, Midwestern University, Downers Grove, US
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Mae Ciancio
4Biomedical Sciences Program, College of Graduate Studies, Midwestern University, Downers Grove, US
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Joanna Goral
5Anatomy, College of Graduate Studies, Midwestern University, Downers Grove, US
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Lenore Pitstick
3Biochemistry and Molecular Genetics, College of Graduate Studies, Midwestern University, Downers Grove, US
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Matt Pytynia
4Biomedical Sciences Program, College of Graduate Studies, Midwestern University, Downers Grove, US
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Alice Meyer
5Anatomy, College of Graduate Studies, Midwestern University, Downers Grove, US
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Neha Kwatra
1Biomedical Sciences Program, College of Graduate Studies, Midwestern University, Glendale, US
6College of Dental Medicine, Midwestern University, Glendale, US
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  • ORCID record for Neha Kwatra
Nafisa M. Jadavji
1Biomedical Sciences Program, College of Graduate Studies, Midwestern University, Glendale, US
7College of Veterinary Medicine, Midwestern University, Glendale, US
8Department of Neuroscience, Carleton University, Ottawa, Canada
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  • ORCID record for Nafisa M. Jadavji
  • For correspondence: njadav@midwestern.edu
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Abstract

Background Recent evidence suggests that sex plays a role when there are deficiencies in one-carbon metabolism, however, the impact on brain tissue remains unknown.

Objective The aim of the study was to examine the impact of sex differences and dietary folic acid deficiency on brain tissue in adult mice.

Methods Male and female C57Bl/6J mice were placed on a folic acid deficient (FD) or control diet (CD) at six weeks of age. Mice were maintained on these diets for six months, after which animals were euthanized and brain tissue and serum were collected for analysis. Serum folate levels were measured. In brain tissue, hippocampal volume and morphology including cortical area 1 and 3 (CA1; CA3), and dentate gyrus thickness were measured. Apoptosis within the hippocampus was assessed using active caspase-3 immunofluorescence staining. Additionally, cortical acetylcholine metabolism was measured in brain tissue using immunofluorescence staining of acetylcholinesterase (AChE) and choline acetyltransferase (ChAT).

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 CA3 region. Interestingly, there was a sex difference in the levels of apoptosis within the CA1 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 through increased apoptosis and changes acetylcholine metabolism within the cortex. The data from our study indicate a sex difference in apoptosis and differences in hippocampal morphology and choline metabolism as a result of dietary folic acid deficiency.

Competing Interest Statement

The authors have declared no competing interest.

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The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC 4.0 International license.
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Posted December 13, 2020.
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Dietary folic acid deficiency impacts hippocampal morphology and cortical acetylcholine metabolism in adult male and female mice
Calli Bennett, Jacalyn Green, Mae Ciancio, Joanna Goral, Lenore Pitstick, Matt Pytynia, Alice Meyer, Neha Kwatra, Nafisa M. Jadavji
bioRxiv 2020.12.13.422535; doi: https://doi.org/10.1101/2020.12.13.422535
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Dietary folic acid deficiency impacts hippocampal morphology and cortical acetylcholine metabolism in adult male and female mice
Calli Bennett, Jacalyn Green, Mae Ciancio, Joanna Goral, Lenore Pitstick, Matt Pytynia, Alice Meyer, Neha Kwatra, Nafisa M. Jadavji
bioRxiv 2020.12.13.422535; doi: https://doi.org/10.1101/2020.12.13.422535

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