Abstract
Current methods for studying central nervous system myelination necessitate permissive axonal substrates conducive to myelin wrapping by oligodendrocytes. We have developed a neuron-free culture system in which electron-spun nanofibers of varying sizes substitute for axons as a substrate for oligodendrocyte myelination, thereby allowing manipulation of the biophysical elements of axonal-oligodendroglial interactions. To investigate axonal regulation of myelination, this system effectively uncouples the role of molecular (inductive) cues from that of biophysical properties of the axon. We use this method to uncover the causation and sufficiency of fiber diameter in the initiation of concentric wrapping by rat oligodendrocytes. We also show that oligodendrocyte precursor cells display sensitivity to the biophysical properties of fiber diameter and initiate membrane ensheathment before differentiation. The use of nanofiber scaffolds will enable screening for potential therapeutic agents that promote oligodendrocyte differentiation and myelination and will also provide valuable insight into the processes involved in remyelination.
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Acknowledgements
We thank W. Stallcup for the rabbit anti-PDGFRα antibody; M.L. Wong for sectioning nanofiber cultures for electron microscopy analysis at the W.M. Keck Foundation Advanced Microscopy Laboratory at UCSF; R. Langen and M. Isas for assistance, advice and support with the electron microscopy; the other members of the Chan laboratory and the MS Research Group at UCSF for encouragement, advice and insightful discussions. This work was supported by the US National Multiple Sclerosis Society Career Transition Award (TA 3008A2/T), the Harry Weaver Neuroscience Scholar Award (JF 2142-A2/T) and the US National Institutes of Health/National Institute of Neurological Disorders and Stroke (NS062796-02) to J.R.C.
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S.L., M.K.L., S.A.R., S.Y.C.C. and J.R.C. performed experiments. S.L., M.K.L., S.H.M., S.J.T., Z.-Q.F., J.M.C. and J.R.C. provided reagents. S.L., M.K.L., S.A.R., S.Y.C.C., S.H.M., S.J.T., Z.-Q.F., J.M.C. and J.R.C. provided intellectual contributions. S.L. and J.R.C. analyzed the data and wrote the paper.
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Lee, S., Leach, M., Redmond, S. et al. A culture system to study oligodendrocyte myelination processes using engineered nanofibers. Nat Methods 9, 917–922 (2012). https://doi.org/10.1038/nmeth.2105
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DOI: https://doi.org/10.1038/nmeth.2105
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