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Oligodendrocytes pp 37–63Cite as

Myelin: Methods for Purification and Proteome Analysis

Part of the Methods in Molecular Biology book series (MIMB,volume 1936)

Abstract

Molecular characterization of myelin is a prerequisite for understanding the normal structure of the axon/myelin-unit in the healthy nervous system and abnormalities in myelin-related disorders. However, reliable molecular profiles necessitate very pure myelin membranes, in particular when considering the power of highly sensitive “omics”-data acquisition methods. Here, we recapitulate the history and recent applications of myelin purification. We then provide our laboratory protocols for the biochemical isolation of a highly pure myelin-enriched fraction from mouse brains and for its proteomic analysis. We also supply methodological modifications when investigating posttranslational modifications, RNA, or myelin from peripheral nerves. Notably, technical advancements in solubilizing myelin are beneficial for gel-based and gel-free myelin proteome analyses. We conclude this article by exemplifying the exceptional power of label-free proteomics in the mass-spectrometric quantification of myelin proteins.

Key words

  • Myelin
  • Oligodendrocyte
  • White matter
  • Density gradient ultracentrifugation
  • Mass spectrometry
  • Proteome/proteomics
  • Proteoform
  • Lipidome/lipidomics
  • Transcriptome/transcriptomics
  • Cyclic nucleotide phosphodiesterase (CNP)
  • Proteolipid protein (PLP)
  • Myelin basic protein (MBP)
  • Cholesterol
  • Demyelination
  • Nerve conduction

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  • DOI: 10.1007/978-1-4939-9072-6_3
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Acknowledgment

We thank K.-A. Nave for discussions and financial support made possible by an European Research Council (ERC) Advanced Grant.

Author information

Authors and Affiliations

  1. Department of Neurogenetics, Max Planck Institute of Experimental Medicine, Goettingen, Germany

    Michelle S. Erwig, Ramona B. Jung, Kathrin Kusch & Hauke B. Werner

  2. Proteomics Group, Max Planck Institute of Experimental Medicine, Goettingen, Germany

    Dörte Hesse, Marina Uecker & Olaf Jahn

  3. Institute of Immunology, University Medical Center, Johannes Gutenberg University, Mainz, Germany

    Stefan Tenzer

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  7. Olaf Jahn
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  8. Hauke B. Werner
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Correspondence to Olaf Jahn or Hauke B. Werner .

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Editors and Affiliations

  1. Centre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, UK

    Dr. David A. Lyons

  2. Centre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, UK

    Dr. Linde Kegel

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Erwig, M.S. et al. (2019). Myelin: Methods for Purification and Proteome Analysis. In: Lyons, D., Kegel, L. (eds) Oligodendrocytes. Methods in Molecular Biology, vol 1936. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9072-6_3

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  • DOI: https://doi.org/10.1007/978-1-4939-9072-6_3

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