Abstract
Important advances in our understanding of oligodendrocyte precursor cell biology and differentiation have stemmed from in vitro experiments using cultures of isolated primary oligodendrocyte precursor cells. To examine the process of myelination in the final stages of oligodendrocyte development, experimental systems have previously been limited to models utilizing neurons. Recent advances in three-dimensional culture systems, however, have opened the possibility to observe myelin sheath formation with only one cell type, the oligodendrocyte precursor cell. In this chapter, such a method is described for examining oligodendrocyte myelin sheath formation with isolated oligodendrocytes in the absence of neurons. This assay is ideal for gaining mechanistic insight into oligodendrocyte-specific regulation of myelin sheath formation. Oligodendrocyte heterogeneity can be readily assessed, determining whether different oligodendrocyte sources influence myelin sheath formation. As well, the direct impact of both physical and molecular cues on oligodendrocytes can be determined in this defined system. This assay extends the capability of two-dimensional oligodendrocyte cultures, permitting post-differentiation analysis of myelinating oligodendrocytes, the number of sheaths formed by individual oligodendrocytes, as well as the lengths of myelin sheaths formed.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Gard AL, Pfeiffer SE (1989) Oligodendrocyte progenitors isolated directly from developing telencephalon at a specific phenotypic stage: myelinogenic potential in a defined environment. Development 106:119–132
Baumann N, Pham-Dinh D (2001) Biology of oligodendrocyte and myelin in the mammalian central nervous system. Physiol Rev 81:871–927
Pfeiffer SE, Warrington AE, Bansal R (1993) The oligodendrocyte and its many cellular processes. Trends Cell Biol 3:191–197
Aggarwal S, Yurlova L, Snaidero N et al (2011) A size barrier limits protein diffusion at the cell surface to generate lipid-rich myelin-membrane sheets. Dev Cell 21:445–456
Jarjour AA, Zhang H, Bauer N et al (2012) In vitro modeling of central nervous system myelination and remyelination. Glia 60:1–12
Nave K-A (2010) Myelination and support of axonal integrity by glia. Nature 468:244–252
Chernoff GF (1981) Shiverer: an autosomal recessive mutant mouse with myelin deficiency. J Hered 72:128
Klugmann M, Schwab MH, Pühlhofer A et al (1997) Assembly of CNS myelin in the absence of proteolipid protein. Neuron 18:59–70
Griffiths I, Klugmann M, Anderson T et al (1998) Axonal swellings and degeneration in mice lacking the major proteolipid of myelin. Science 280:1610–1613
Waxman SG (1980) Determinants of conduction velocity in myelinated nerve fibers. Muscle Nerve 3:141–150
Wood PM, Williams AK (1984) Oligodendrocyte proliferation and CNS myelination in cultures containing dissociated embryonic neuroglia and dorsal root ganglion neurons. Brain Res 314:225–241
Rosen CL, Bunge RP, Ard MD et al (1989) Type 1 astrocytes inhibit myelination by adult rat oligodendrocytes in vitro. J Neurosci 9:3371–3379
Chan JR, Watkins TA, Cosgaya JM et al (2004) NGF controls axonal receptivity to myelination by Schwann cells or oligodendrocytes. Neuron 43:183–191
Watkins TA, Emery B, Mulinyawe S et al (2008) Distinct stages of myelination regulated by γ-secretase and astrocytes in a rapidly myelinating CNS coculture system. Neuron 60:555–569
Althaus HH, Burgisser P, Klöppner S et al (1987) Oligodendrocytes ensheath carbon fibres and produce myelin in vitro. In: Althaus HH, Seifert W (eds) Glial-neuronal communication in development and regeneration. Springer Berlin Heidelberg, Berlin, Heidelberg, pp 779–798
Bullock PN, Rome LH (1990) Glass micro-fibers: a model system for study of early events in myelination. J Neurosci Res 27:383–393
Howe CL (2006) Coated glass and vicryl microfibers as artificial axons. Cells Tissues Organs 183:180–194
Lee S, Leach MK, Redmond SA et al (2012) A culture system to study oligodendrocyte myelination processes using engineered nanofibers. Nat Methods 9:917–922
Bechler ME, Byrne L, ffrench-Constant C (2015) CNS myelin sheath lengths are an intrinsic property of oligodendrocytes. Curr Biol 25:2411–2416
Zoupi L, Savvaki M, Karagogeos D (2011) Axons and myelinating glia: an intimate contact. IUBMB Life 63:730–735
Faivre-Sarrailh C, Devaux JJ (2013) Neuro-glial interactions at the nodes of Ranvier: implication in health and diseases. Front Cell Neurosci 7:196
Wang Z, Colognato H, ffrench-Constant C (2007) Contrasting effects of mitogenic growth factors on myelination in neuron–oligodendrocyte co-cultures. Glia 55:537–545
Bottenstein JE, Sato GH (1979) Growth of a rat neuroblastoma cell line in serum-free supplemented medium. Proceed Natl Acad Sci U S A 76:514–517
Schindelin J, Arganda-Carreras I, Frise E et al (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682
McCarthy KD, de Vellis J (1980) Preparation of separate astroglial and oligodendroglial cell cultures from rat cerebral tissue. J Cell Biol 85:890–902
Emery B, Dugas JC (2013) Purification of oligodendrocyte lineage cells from mouse cortices by immunopanning. Cold Spring Harb Protoc 2013:854–868
Lee S, Chong SYC, Tuck SJ et al (2013) A rapid and reproducible assay for modeling myelination by oligodendrocytes using engineered nanofibers. Nat Protoc 8:771–782
Chen Y, et al (2008) NS21: Re-defined and modified supplement B27 for neuronal cultures. J Neurosci Methods 171(2):239–247. https://doi.org/10.1016/j.jneumeth.2008.03.013
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Bechler, M.E. (2019). A Neuron-Free Microfiber Assay to Assess Myelin Sheath Formation. In: Lyons, D., Kegel, L. (eds) Oligodendrocytes. Methods in Molecular Biology, vol 1936. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9072-6_6
Download citation
DOI: https://doi.org/10.1007/978-1-4939-9072-6_6
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-9070-2
Online ISBN: 978-1-4939-9072-6
eBook Packages: Springer Protocols