Advertisement

Schwann Cells pp 339-348 | Cite as

Whole Mount Immunostaining on Mouse Sciatic Nerves to Visualize Events of Peripheral Nerve Regeneration

  • Xin-Peng DunEmail author
  • David B. Parkinson
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1739)

Abstract

Injury to the peripheral nervous system triggers a series of well-defined events within both neurons and the Schwann cells to allow efficient axonal regeneration, remyelination, and functional repair. The study of these events has previously been done using sections of nerve material to analyze axonal regrowth, cell migration, and immune cell infiltration following injury. This approach, however, has the obvious disadvantage that it is not possible to follow, for instance, the path of regenerating axons in three dimensions within the nerve trunk or the nerve bridge. In order to provide a fuller picture of such events, we have developed a whole mount staining procedure to visualize blood vessel regeneration, Schwann cell migration, axonal regrowth, and remyelination in models of nerve injury.

Key words

Whole mount immunostaining Peripheral nerve injury Axon regeneration Schwann cell migration Blood vessel regeneration 

Notes

Acknowledgments

This work was supported by a Wellcome Trust grant (WT088228) and Medical Research Council grant (MR/J012785/1) to D.B.P. We are grateful to Prof. Wendy Macklin (University of Colorado Denver School of Medicine, USA) for the gift of the PLP-EGFP transgenic mouse line.

References

  1. 1.
    Klymkowsky MW, Hanken J (1991) Whole-mount staining of Xenopus and other vertebrates. Methods Cell Biol 36:419–441CrossRefPubMedGoogle Scholar
  2. 2.
    Kim HA, Mindos T, Parkinson DB (2013) Plastic fantastic: Schwann cells and repair of the peripheral nervous system. Stem Cells Transl Med 2:553–557CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Parrinello S, Napoli I, Ribeiro S et al (2010) EphB signaling directs peripheral nerve regeneration through Sox2-dependent Schwann cell sorting. Cell 143:145–155CrossRefPubMedGoogle Scholar
  4. 4.
    Cattin AL, Lloyd AC (2016) The multicellular complexity of peripheral nerve regeneration. Curr Opin Neurobiol 39:38–46CrossRefPubMedGoogle Scholar
  5. 5.
    Chen YY, McDonald D, Cheng C et al (2005) Axon and Schwann cell partnership during nerve regrowth. J Neuropathol Exp Neurol 64:613–622CrossRefPubMedGoogle Scholar
  6. 6.
    McDonald D, Cheng C, Chen Y et al (2006) Early events of peripheral nerve regeneration. Neuron Glia Biol 2:139–147CrossRefPubMedGoogle Scholar
  7. 7.
    Dun XP, Parkinson DB (2015) Visualizing peripheral nerve regeneration by whole mount staining. PLoS One 10:e0119168CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Painter MW, Brosius Lutz A, Cheng YC et al (2014) Diminished Schwann cell repair responses underlie age-associated impaired axonal regeneration. Neuron 83:331–343CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Tian F, Yang W, Mordes DA et al (2016) Monitoring peripheral nerve degeneration in ALS by label-free stimulated Raman scattering imaging. Nat Commun 7:13283CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Mallon BS, Shick HE, Kidd GJ et al (2002) Proteolipid promoter activity distinguishes two populations of NG2-positive cells throughout neonatal cortical development. J Neurosci 22:876–885PubMedGoogle Scholar
  11. 11.
    Cattin AL, Burden JJ, Van Emmenis L et al (2015) Macrophage-induced blood vessels guide Schwann cell-mediated regeneration of peripheral nerves. Cell 162:1127–1139CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.Plymouth University Peninsula Schools of Medicine and DentistryPlymouthUK

Personalised recommendations