Advertisement

Myelin pp 263-276 | Cite as

In Vivo Introduction of Transgenes into Mouse Sciatic Nerve Cells Using Viral Vectors

  • Gerben Van Hameren
  • Sergio Gonzalez
  • Ruani N. Fernando
  • Claire Perrin-Tricaud
  • Nicolas Tricaud
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1791)

Abstract

Myelinated fibers are essential for the rapid and efficient propagation of nerve information throughout the body. These fibers result from an intimate crosstalk between myelinating glia and the myelinated axons and, because it is difficult to fully reproduce these interactions in vitro, the basic molecular mechanisms that regulate myelination, demyelination, and remyelination remain unclear. Schwann cells produce myelin in the peripheral nervous system (PNS) and remain associated with the axons of peripheral neurons throughout axonal migration to the target. In order to investigate more closely the biology of myelinated fibers, we developed a local transgenesis approach based on the injection of engineered viral vectors in the sciatic nerve of mice to locally transduce peripheral nerve cells. This approach represents an alternative to germline modifications as it facilitates and speed up the investigation of peripheral nerve biology in vivo. Indeed the protocol we describe here requires just 3 weeks to complete. The injection of engineered viral vectors in the sciatic nerve of mice is a reproducible and straightforward method for introducing exogenous factors into myelinating Schwann cells and myelinated axons in vivo in order to investigate specific molecular mechanisms.

Key words

Peripheral nerve Viral transduction Schwann cells Axons Transgenesis Sciatic nerve 

Notes

Acknowledgements

N.T. is grateful for the support of European research council (FP7-IDEAS-ERC 311610) and ATIP-Avenir program. R.F. work was supported by Fondation pour la Recherche Médicale and the Marie-Curie fellowship program. G.H. work has benefited from support by the Labex EpiGenMed.

References

  1. 1.
    Suter U, Scherer SS (2003) Disease mechanisms in inherited neuropathies. Nat Rev Neurosci 4:714–726. https://doi.org/10.1038/nrn1196 CrossRefPubMedGoogle Scholar
  2. 2.
    McGoldrick P, Joyce PI, Fisher EMC, Greensmith L (2013) Rodent models of amyotrophic lateral sclerosis. Biochim Biophys Acta 1832:1421–1436. https://doi.org/10.1016/j.bbadis.2013.03.012 CrossRefPubMedGoogle Scholar
  3. 3.
    Höke A (2012) Animal models of peripheral neuropathies. Neurotherapeutics 9:262–269. https://doi.org/10.1007/s13311-012-0116-y CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Davey RA, MacLean HE (2006) Current and future approaches using genetically modified mice in endocrine research. Am J Physiol Endocrinol Metab 291:E429. https://doi.org/10.1152/ajpendo.00124.2006 CrossRefPubMedGoogle Scholar
  5. 5.
    Joung JK, Sander JD (2013) TALENs: a widely applicable technology for targeted genome editing. Nat Rev Mol Cell Biol 14:49–55. https://doi.org/10.1038/nrm3486 CrossRefPubMedGoogle Scholar
  6. 6.
    Sung YH, Baek I-J, Seong JK et al (2012) Mouse genetics: catalogue and scissors. BMB Rep 45:686–692CrossRefGoogle Scholar
  7. 7.
    Sherman DL, Brophy PJ (2005) Mechanisms of axon ensheathment and myelin growth. Nat Rev Neurosci 6:683–690. https://doi.org/10.1038/nrn1743 CrossRefPubMedGoogle Scholar
  8. 8.
    Viader A, Sasaki Y, Kim S et al (2013) Aberrant Schwann cell lipid metabolism linked to mitochondrial deficits leads to axon degeneration and neuropathy. Neuron 77:886–898. https://doi.org/10.1016/j.neuron.2013.01.012 CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Nave K-A (2010) Myelination and support of axonal integrity by glia. Nature 468:244–252. https://doi.org/10.1038/nature09614 CrossRefPubMedGoogle Scholar
  10. 10.
    Cotter L, Ozçelik M, Jacob C et al (2010) Dlg1-PTEN interaction regulates myelin thickness to prevent damaging peripheral nerve overmyelination. Science 328:1415–1418. https://doi.org/10.1126/science.1187735 CrossRefPubMedGoogle Scholar
  11. 11.
    Ozçelik M, Cotter L, Jacob C et al (2010) Pals1 is a major regulator of the epithelial-like polarization and the extension of the myelin sheath in peripheral nerves. J Neurosci 30:4120–4131. https://doi.org/10.1523/JNEUROSCI.5185-09.2010 CrossRefPubMedGoogle Scholar
  12. 12.
    Fernando RN, Cotter L, Perrin-Tricaud C et al (2016) Optimal myelin elongation relies on YAP activation by axonal growth and inhibition by Crb3/hippo pathway. Nat Commun 7:12186. https://doi.org/10.1038/ncomms12186 CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Perrin-Tricaud C, Rutishauser U, Tricaud N (2007) P120 catenin is required for thickening of Schwann cell myelin. Mol Cell Neurosci 35:120–129. https://doi.org/10.1016/j.mcn.2007.02.010 CrossRefPubMedGoogle Scholar
  14. 14.
    Tricaud N, Perrin-Tricaud C, Brusés JL, Rutishauser U (2005) Adherens junctions in myelinating Schwann cells stabilize Schmidt-Lanterman incisures via recruitment of p120 catenin to E-cadherin. J Neurosci 25:3259–3269. https://doi.org/10.1523/JNEUROSCI.5168-04.2005 CrossRefPubMedGoogle Scholar
  15. 15.
    Glatzel M, Flechsig E, Navarro B et al (2000) Adenoviral and adeno-associated viral transfer of genes to the peripheral nervous system. Proc Natl Acad Sci U S A 97:442–447CrossRefGoogle Scholar
  16. 16.
    Guénard V, Schweitzer B, Flechsig E et al (1999) Effective gene transfer of lacZ and P0 into Schwann cells of P0-deficient mice. Glia 25:165–178CrossRefGoogle Scholar
  17. 17.
    He T-C, Zhou S, da Costa LT et al (1998) A simplified system for generating recombinant adenoviruses. Proc Natl Acad Sci 95:2509–2514CrossRefGoogle Scholar

Copyright information

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

Authors and Affiliations

  • Gerben Van Hameren
    • 1
  • Sergio Gonzalez
    • 1
  • Ruani N. Fernando
    • 1
  • Claire Perrin-Tricaud
    • 1
  • Nicolas Tricaud
    • 1
  1. 1.Institut des Neurosciences de Montpellier, INSERM U1051Université de MontpellierMontpellierFrance

Personalised recommendations