Abstract
The myelin sheath is essential for the rapid and efficient propagation of action potentials. However, our understanding of the basic molecular mechanisms that regulate myelination, demyelination and remyelination is limited. Schwann cells produce myelin in the peripheral nervous system and remain associated with the axons of peripheral neurons throughout axonal migration to the target. Owing to the intimate relationship between these cell types it is difficult to fully reproduce their function in vitro. For this reason, we developed an approach based on the injection of an engineered virus into the sciatic nerve of mice to locally transduce peripheral nerve cells. This approach can be used as an alternative to germline transgenesis to facilitate the investigation of peripheral nerve biology in vivo. The detailed protocol, described here, requires 3 weeks to complete. In comparison with genetic modification strategies, this protocol is a fast, reproducible and straightforward method for introducing exogenous factors into myelinating Schwann cells and myelinated axons in vivo to investigate specific molecular mechanisms.
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Acknowledgements
We acknowledge Montpellier RIO Imaging Platform and the Animal Facility of the INM for help and technical assistance. This work was supported by the European Research Council (grant no. FP7-IDEAS-ERC 311610) and an INSERM-AVENIR grant. All animal experiments were conducted in accordance with institutional (approval no. CEEA-LR-11032) and French governmental regulations.
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S.G., R.F. and N.T. wrote the paper. C.P.-T., S.G., R.F. and N.T. performed experiments and analyzed data. N.T. developed the protocol and supervised the project.
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Gonzalez, S., Fernando, R., Perrin-Tricaud, C. et al. In vivo introduction of transgenes into mouse sciatic nerve cells in situ using viral vectors. Nat Protoc 9, 1160–1169 (2014). https://doi.org/10.1038/nprot.2014.073
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DOI: https://doi.org/10.1038/nprot.2014.073
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