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Oligodendrocytes pp 169–183Cite as

Ex Vivo Slice Cultures to Study Myelination, Demyelination, and Remyelination in Mouse Brain and Spinal Cord

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1936))

Abstract

In vitro culture systems have been invaluable in understanding the cell biology of oligodendrocytes; the monoculture of primary oligodendroglia has helped characterize different stages of oligodendrocyte maturation in the absence of neurons. However, oligodendrocyte monocultures do not model the interaction of oligodendrocytes with neurons where they form myelin wraps. To circumvent this problem, coculture systems were developed; oligodendrocytes and neurons are cultured together, facilitating the study of myelin wraps and the interaction between the two cell types. However, this coculture system also has limitations, as other cells are not present and it does not represent the three-dimensional multicellular structure seen in vivo. Some of these limitations are resolved by using ex vivo slice cultures to serve as a three-dimensional culture system that is more similar to in vivo and can be used to study myelination, demyelination, and remyelination, over extended periods of time. Slice cultures are economical compared to in vivo studies and live imaging using them is less challenging. The focus of this chapter is to describe how to culture brain and spinal cord slices of mice and use them to study myelination, demyelination, and remyelination.

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References

  1. Baumann N, Pham-Dinh D (2001) Biology of oligodendrocyte and myelin in the mammalian central nervous system. Physiol Rev 81:871–927

    Article  CAS  Google Scholar 

  2. Szuchet S, Polak PE, Yim SH (1986) Mature oligodendrocytes cultured in the absence of neurons recapitulate the ontogenic development of myelin membranes. Dev Neurosci 8:208–221

    Article  CAS  Google Scholar 

  3. Wood P, Okada E, Bunge R (1980) The use of networks of dissociated rat dorsal root ganglion neurons to induce myelination by oligodencrocytes in culture. Brain Res 196:247–252

    Article  CAS  Google Scholar 

  4. Wood PM, Bunge RP (1986b) Myelination of cultured dorsal root ganglion neurons by oligodendrocytes obtained from adult rats. J Neurol Sci 74:153–169

    Article  CAS  Google Scholar 

  5. Chan JR et al (2004) NGF controls axonal receptivity to myelination by Schwann cells or oligodendrocytes. Neuron 43:183–191

    Article  CAS  Google Scholar 

  6. Huang JK, Jarjour AA et al (2011) Retinoid X receptor gamma signalling accelerates CNS remyelination. Nat Neurosci 14:45–53

    Article  CAS  Google Scholar 

  7. Bornstein MB (1972) Immunopathology of the central and peripheral nervous systems. Verh Dtsch Ges Inn Med 78:777–790

    CAS  PubMed  Google Scholar 

  8. Yavin E, Yavin Z (1974) Attachment and culture of dissociated cells from rat embryo cerebral hemispheres on polylysine-coated surface. J Cell Biol 62:540–546

    Article  CAS  Google Scholar 

  9. Jarjour AA, Zhang H, Bauer N, ffrench-constant C, Williams A (2012) In vitro modelling of central nervous system myelination and remyelination. Glia 60:1–12

    Article  Google Scholar 

  10. Avossa D, Rosato-Siri MD, Mazzarol F, Ballerini L (2003) Spinal circuits formation: a study of developmentally regulated markers in organotypic cultures of emb1ryonic mouse spinal cord. Neuroscience 122:391–405

    Article  CAS  Google Scholar 

  11. Schnadelbach O, Ozen I, Blaschuk OW, Gour BJ, Meyer RL, Fawcett JW (2001) N-cadherin is involved in axon-oligodendrocyte contact and myelination. Mol Cell Neurosci 17:1084–1093

    Article  CAS  Google Scholar 

  12. Hild W (1956) Myelin formation in central nervous system tissue cultures. Verh Anat Ges 53:315–317

    Google Scholar 

  13. Bornstein MB, Murray MR (1958) Serial observations on patterns of growth, myelin formation, maintenance and degeneration in cultures of newborn rat and kitten cerebellum. J Biophys Biochem Cytol 4:499–504

    Article  CAS  Google Scholar 

  14. Field EJ, Hughes D, Raine CS (1969) Electron microscopic observations on the development of myelin in cultures of neonatal rat cerebellum. J Neurol Sci 8:49–60

    Article  CAS  Google Scholar 

  15. Kim SU (1971) Electron microscope study of mouse cerebellum in tissue culture. Exp Neurol 33:30–44

    Article  CAS  Google Scholar 

  16. Ross LL, Bornstein MB, Lehrer GM (1962) Electron microscopic observation of rat and mouse cerebellum in tissue culture. J Cell Biol 14:19–30

    Article  CAS  Google Scholar 

  17. Bunge RP, Wood P (1973) Studies on the transplantation of spinal cord tissue in the rat. I. The development of a culture system for hemisections of embryonic spinal cord. Brain Res 57:261–276

    Article  CAS  Google Scholar 

  18. Zhang H, Jarjour AA, Boyd A, Williams A (2011) Central nervous system remyelination in culture—a tool for multiple sclerosis research. Exp Neurol 230:138–148

    Article  Google Scholar 

  19. Jarjour AA, Bull SJ, Almasieh M, Rajasekharan S, Baker KA, Mui J, Antel JP, Di Polo A, Kennedy TE (2008) Maintenance of axo-oligodendroglial paranodal junctions requires DCC, netrin-1. J Neurosci 28:11003–11014

    Article  CAS  Google Scholar 

  20. Birgbauer E, Rao TS, Webb M (2004) Lysolecithin induced demyelination in vitro in a cerebellar slice culture system. J Neurosci Res 78:157–166

    Article  CAS  Google Scholar 

  21. Huang JK, Jarjour AA, Nait-Oumesmar B, Kerninon C, Williams A, Krezel W, Kagechika H, Bauer J, Zhao C, Baron-Van Evercooren A, Chambon P, ffrench-Constant C, Franklin RJM (2011) Retinoid X receptor gamma signaling accelerates CNS remyelination. Nat Neurosci 14(1):45–53

    Article  CAS  Google Scholar 

  22. Mi S, Miller RH, Tang W, Lee X, Hu B, Wu W, Zhang Y, Shields CB, Zhang Y, Miklasz S, Shea D, Mason J, Franklin RJ, Ji B, Shao Z, Chédotal A, Bernard F, Roulois A, Xu J, Jung V, Pepinsky B (2009) Promotion of central nervous system remyelination by induced differentiation of oligodendrocyte precursor cells. Ann Neurol 65:304–315

    Article  CAS  Google Scholar 

  23. Miron VE, Ludwin SK, Darlington PJ, Jarjour AA, Soliven B, Kennedy TE, Antel JP (2010) Fingolimod (FTY720) enhances remyelination following demyelination of organotypic cerebellar slices. Am J Pathol 176:2682–2694

    Article  CAS  Google Scholar 

  24. Hill RA, Medved J, Patel KD, Nishiyama A (2014) Organotypic slice cultures to study oligodendrocyte dynamics and myelination. J Vis Exp 90:e51835. https://doi.org/10.3791/51835

  25. Seidl AH, Rubel EW (2010) A simple method for multiday imaging of slice cultures. Microsc Res Tech 73(1):37–44

    PubMed  PubMed Central  Google Scholar 

  26. Miron VE, Boyd A, Zhao JW, Yuen TJ, Ruckh JM, Shadrach JL, van Wijngaarden P, Wagers AJ, Williams A, Franklin RJ, ffrench-Constant C (2013) M2 microglia and macrophages drive oligodendrocyte differentiation during CNS remyelination. Nat Neurosci 16:1211–1218

    Article  CAS  Google Scholar 

  27. Yuen TJ, Johnson KR, Miron VE, Zhao C, Quandt J, Harrisingh MC, Swire M, Williams A, McFarland HF, Franklin RJ, ffrench-Constant C (2013) Identification of endothelin 2 as an inflammatory factor that promotes central nervous system remyelination. Brain 136:1035–1047

    Article  Google Scholar 

  28. Foran DR, Peterson AC (1992) Myelin acquisition in the central nervous system of the mouse revealed by an MBP-LacZ transgene. J Neurosci 12(12):4890–4897

    Article  CAS  Google Scholar 

  29. Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez JY, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682

    Article  CAS  Google Scholar 

  30. Ghoumari AM, Ibanez C, El-Etr M, Leclerc P, Eychenne B, O’Malley BW, Baulieu EE, Schumacher M (2003) Progesterone and its metabolites increase myelin basic protein expression in organotypic slice cultures of rat cerebellum. J Neurochem 86:848–859

    Article  CAS  Google Scholar 

  31. Eisenbach M, Kartvelishvily E, Eshed-Eisenbach Y, Watkins T, Sorensen A, Thomson C, Ranscht B, Barnett SC, Brophy P, Peles E (2009) Differential clustering of Caspr by oligodendrocytes and Schwann cells. J Neurosci Res 87:3492–3501

    Article  CAS  Google Scholar 

  32. Neumann H, Kotter MR, Franklin RJ (2009) Debris clearance by microglia: an essential link between degeneration and regeneration. Brain 132(Pt 2):288–295

    CAS  PubMed  Google Scholar 

  33. Marksteiner J, Humpel C (2008) Beta-amyloid expression, release and extracellular deposition in aged rat brain slices. Mol Psychiatry 13:939–952

    Article  CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. MRC Centre for Regenerative Medicine and MS Society Edinburgh Centre, Edinburgh bioQuarter, The University of Edinburgh, Edinburgh, UK

    Sowmya Sekizar & Anna Williams

Authors
  1. Sowmya Sekizar
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  2. Anna Williams
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Corresponding author

Correspondence to Anna Williams .

Editor information

Editors and Affiliations

  1. Centre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, UK

    David A. Lyons

  2. Centre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, UK

    Linde Kegel

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Sekizar, S., Williams, A. (2019). Ex Vivo Slice Cultures to Study Myelination, Demyelination, and Remyelination in Mouse Brain and Spinal Cord. 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_10

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  • DOI: https://doi.org/10.1007/978-1-4939-9072-6_10

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  • 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

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