Abstract
Modeling myelination in vitro allows mechanistic study of developmental myelination and short-term myelin maintenance, but analyses possible to carry out using currently available models are usually limited because of high cell density and the lack of separation between neurons and myelinating cells. Furthermore, regeneration studies of myelinated systems after lesion require compartmentalization of neuronal cell bodies, axons, and myelinating cells. Here we describe a compartmentalized method using microfluidics that allows live-cell imaging at the single-cell level to follow short- and long-term dynamic interactions of neurons and myelinating cells and large-scale analyses, e.g., RNA sequencing on pure or highly enriched neurons or myelinating cells, separately.
Key words
- Microfluidics
- Myelinated models
- Schwann cells
- Oligodendrocytes
- Myelin maintenance
- Regeneration
- Single-cell live imaging
This is a preview of subscription content, access via your institution.
Buying options
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Einheber S, Hannocks M-J, Metz CN, Rifkin DB, Salzer JL (1995) TGF-β1 regulates axon-Schwann cell interactions. J Cell Biol 129:443–458
Stendel C, Roos A, Kleine H, Arnaud E, Ozçelik M, Sidiropoulos PN et al (2010) SH3TC2, a protein mutant in Charcot-Marie-tooth neuropathy, links peripheral nerve myelination to endosomal recycling. Brain 133:2462–2474
Zanazzi G, Einheber S, Westreich R, Hannocks M-J, Bedell-Hogan D, Marchionni MA et al (2001) Glial growth factor/neuregulin inhibits Schwann cell myelination and induces demyelination. J Cell Biol 152:1289–1300
Brügger V, Engler S, Pereira JA, Ruff S, Horn M, Welzl H et al (2015) HDAC1/2-dependent P0 expression maintains paranodal and nodal integrity independently of myelin stability through interactions with neurofascins. PLoS Biol 13:e1002258
Campenot RB (1994) NGF and the local control of nerve terminal growth. Dev Neurobiol 25:599–611
Jadhav AD, Wei L, Shi P (2016) Compartmentalized platforms for neuro-pharmacological research. Curr Neuropharmacol 14:72–86
Taylor AM, Blurton-Jones M, Rhee SW, Cribbs DH, Cotman CW, Jeon NL (2005) A microfluidic culture platform for CNS axonal injury, regeneration and transport. Nat Methods 2:599–605
Chen Y, Balasubramaniyan V, Peng J, Hurlock EC, Tallquist M, Li J et al (2007) Isolation and culture of rat and mouse oligodendrocyte precursor cells. Nat Protoc 2:1044–1051
Acknowledgments
We thank Dr. Roman Chrast and Dr. Luca Bartesaghi for advices on microfluidic device dimensions and cell culture, and Dr. Noo Li Jeon for producing Master molds for the microfluidic devices. This work was supported by Swiss National Science Foundation grants PP00P3_1139163, PP00P3_163759, and 31003A_173072 to C.J.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2018 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Vaquié, A., Sauvain, A., Jacob, C. (2018). Modeling PNS and CNS Myelination Using Microfluidic Chambers. In: Woodhoo, A. (eds) Myelin. Methods in Molecular Biology, vol 1791. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7862-5_12
Download citation
DOI: https://doi.org/10.1007/978-1-4939-7862-5_12
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-7861-8
Online ISBN: 978-1-4939-7862-5
eBook Packages: Springer Protocols