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Myelin pp 193-206 | Cite as

Analyzing Autophagic Flux in Nerve Cultures

  • Miguel Tamayo Caro
  • Marta Palomo Irigoyen
  • Encarnación Pérez Andrés
  • Adrián Barreira Manrique
  • Marta Varela Rey
  • Ashwin Woodhoo
Part of the Methods in Molecular Biology book series (MIMB, volume 1791)

Abstract

Autophagy is a key cellular mechanism involved in the degradation of long-lived proteins and organelles. We and others have previously shown that Schwann cells are able to degrade their own myelin by a form of selective autophagy, or myelinophagy. There is now increasing evidence that myelinophagy could also be aberrantly activated in other demyelinating diseases, including hereditary or inflammatory neuropathies, implicating this pathway in the pathogenesis of these disorders. In this chapter, we describe our protocol to monitor autophagy in peripheral nerves, using the autophagy flux assay. This assay can be useful to compare basal and demyelination-induced autophagy in genetic mice models, or after treatment with specific compounds.

Key words

Myelinophagy Autophagy flux LC3 Western blot Schwann cell 

Notes

Acknowledgments

AW is grateful for the support of the Ministerio de Economía y Competitividad–Plan Nacional de I+D+I (Subprograma Ramón y Cajal RYC2010-06901; Proyectos Retos Investigación SAF2015-65360-R; Proyectos ExploraCiencia SAF2015-72416-EXP; Proyectos Europa Excelencia SAF2015-62588-ERC), the BBVA foundation and Ikerbasque Foundation. MP is grateful for the support of the Basque Government of Education fellowship. MT isgrateful for the support of the “Ayudas para contratos predoctorales para la formación de doctores” from the Ministerio de Economía y Competitividad. MVR is grateful for the support of a 2017 Leonardo Grant for Researchers and Cultural Creators, BBVA Foundation. CIBERehd is funded by the Instituto de Salud Carlos III. We thank MINECO for the Severo Ochoa Excellence Accreditation (SEV-2016-0644).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Miguel Tamayo Caro
    • 1
  • Marta Palomo Irigoyen
    • 1
  • Encarnación Pérez Andrés
    • 1
  • Adrián Barreira Manrique
    • 1
  • Marta Varela Rey
    • 2
  • Ashwin Woodhoo
    • 3
    • 4
  1. 1.Nerve Disorders Laboratory, CIC bioGUNEDerioSpain
  2. 2.Liver disease Laboratory, Liver metabolism LaboratoryCIC bioGUNE, Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas (CIBERehd)DerioSpain
  3. 3.Nerve Disorders LaboratoryCIC bioGUNEDerioSpain
  4. 4.IKERBASQUE, Basque Foundation for ScienceBilbaoSpain

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