In vivo introduction of transgenes into mouse sciatic nerve cells in situ using viral vectors.
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.
INSERM U1051, Institut des Neurosciences de Montpellier (INM), Université de Montpellier 1 and 2, Montpellier, France.
Sergio
S
Gonzalez
INSERM U1051, Institut des Neurosciences de Montpellier (INM), Université de Montpellier 1 and 2, Montpellier, France.
Ruani N
RN
Fernando
INSERM U1051, Institut des Neurosciences de Montpellier (INM), Université de Montpellier 1 and 2, Montpellier, France.
Claire
C
Perrin-Tricaud
INSERM U1051, Institut des Neurosciences de Montpellier (INM), Université de Montpellier 1 and 2, Montpellier, France.
Nicolas
N
Tricaud
D000256:Adenoviridae; D000818:Animals; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D016086:Lentivirus; D051379:Mice; D034741:RNA, Small Interfering / Q000235:genetics; D012583:Schwann Cells / Q000166:cytology / Q000378:metabolism*; D012584:Sciatic Nerve / Q000166:cytology*; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes / Q000235:genetics
D034741:RNA, Small Interfering