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Metabolomic study of disease progression in scrapie prion infected mice; validation of a novel method for brain metabolite extraction

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Abstract

Introduction

Prion disease is a form of neurodegenerative disease caused by the misfolding and aggregation of cellular prion protein (PrPC). The neurotoxicity of the misfolded form of prion protein, PrPSc still remains understudied. Here we try to investigate this issue using a metabolomics approach.

Objectives

The intention was to identify and quantify the small-in-size and water-soluble metabolites extracted from mice brains infected with the Rocky Mountain Laboratory isolate of mouse-adapted scrapie prions (RML) and track changes in these metabolites during disease evolution.

Methods

A total of 73 mice were inoculated with RML prions or normal brain homogenate control; brains were harvested at 30, 60, 90, 120 and 150 days post-inoculation (dpi). We devised a high-efficiency metabolite extraction method and used nuclear magnetic resonance spectroscopy to identify and quantify 50 metabolites in the brain extracts. Data were analyzed using multivariate approach.

Results

Brain metabolome profiles of RML infected animals displayed continuous changes throughout the course of disease. Among the analyzed metabolites, the most noteworthy changes included increases in myo-inositol and glutamine as well as decreases in 4-aminobutyrate, acetate, aspartate and taurine.

Conclusion

We report a novel metabolite extraction method for lipid-rich tissue. As all the major metabolites are identifiable and quantifiable by magnetic resonance spectroscopy, this study suggests that tracking of neurochemical profiles could be effective in monitoring the progression of neurodegenerative diseases and useful for assessing the efficacy of candidate therapeutics.

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Data availability

The metabolomics data reported in this paper are available at Metabolomics Workbench, study ID1871.

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Funding

This work was supported by CIHR (MOP137094), by the Canada Foundation for Innovation (NIF21633) and by the Alberta Prion Research Institute.

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Authors and Affiliations

  1. Department of Biochemistry, University of Alberta, Edmonton, AB, Canada

    Ze-Lin Fu, Ghazaleh Eskandari-Sedighi, David Westaway & Brian D. Sykes

  2. Centre for Prions and Protein Folding Diseases, University of Alberta, Edmonton, AB, Canada

    Ze-Lin Fu, Ghazaleh Eskandari-Sedighi, Jing Yang & David Westaway

  3. National High Field Nuclear Magnetic Resonance Centre (NANUC), Edmonton, AB, Canada

    Pascal Mercier

Authors
  1. Ze-Lin Fu
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  2. Pascal Mercier
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  3. Ghazaleh Eskandari-Sedighi
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  4. Jing Yang
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  5. David Westaway
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  6. Brian D. Sykes
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Contributions

DW, BDS and ZLF conceived and designed the research. JY and GES collected the samples. GES performed western blot analysis. ZLF conducted the metabolite extraction, NMR experiments and analyzed the data with PC. ZLF wrote the manuscript with DW and BDS.

Corresponding author

Correspondence to Brian D. Sykes.

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Conflict of interest

The authors declare that there is no conflict of interest.

Ethical approval

All the care and use of animals followed the Canadian Council on Animal Care (CCAC) guidelines and the protocols were approved by the animal care use committee for Health Sciences Laboratory Animal Services at the University of Alberta. The protocol used is AUP00000357.

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Fu, ZL., Mercier, P., Eskandari-Sedighi, G. et al. Metabolomic study of disease progression in scrapie prion infected mice; validation of a novel method for brain metabolite extraction. Metabolomics 16, 72 (2020). https://doi.org/10.1007/s11306-020-01690-2

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