Summary
Injection of ethidium bromide into X-irradiated spinal cord white matter produces a lesion in which demyelinated axons reside in an environment that is permanently depleted of glial cells. By transplanting defined populations of glial cells into this lesion it is possible to recreate normal or novel glial environments. In this study we have transplanted cultures of astrocytes into the X-irradiated ethidium bromide lesion in order to (1) assess the ability of these cells to relate to components within the lesion environment and thereby contribute to tissue reconstruction and (2) establish an astrocytic environment around demyelinated axons that resembles pathological states such as the chronic demyelinated plaques of multiple sclerosis. In order to focus attention on the interactions between astrocytes and demyelinated axons we developed a protocol for depleting astrocyte cultures of oligodendrocyte lineage cells and Schwann cells based on complement-mediated immunocytolysis andin vitro X-irradiation. In addition to establishing the ability of transplanted astrocytes to form an astrocytic matrix around demyelinated axons, this study has also revealed the diversity of cell types present within neonatal forebrain cultures.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Barbarese, E. &Barry, C. (1989) Radiation sensitivity of glial cells in primary culture.Journal of the Neurological Sciences 91, 97–107.
Blakemore, W. F. (1969) The ultrastructure of the subependymal plate in the rat.Journal of Anatomy 104, 423–33.
Blakemore, W. F. (1977) Remyelination of CNS axons by Schwann cells transplanted from the sciatic nerve.Nature 266, 68–9.
Blakemore, W. F. &Crang, A. J. (1985) The use of cultured autologous Schwann cells to remyelinate areas of persistent demyelination in the central nervous system.Journal of the Neurological Sciences 70, 207–23.
Blakemore, W. F. &Crang, A. J. (1988) Extensive oligodendrocyte remyelination following injection of cultured central nervous system cells into demyelinating lesions in adult central nervous system.Developmental Neuroscience 10, 1–10.
Blakemore, W. F. &Crang, A. J. (1992) Transplantation of glial cells into areas of demyelination in the adult rat spinal cord. InNeural Transplantation: A Practical Approach (edited byDunnett, S. B. &Björklund, A.) pp. 105–22. Oxford: Oxford University Press.
Blakemore, W. F. &Franklin, R. J. M. (1991) Transplantation of glial cells into the CNS.Trends in Neurosciences 14, 323–7.
Cavanagh, J. B. &Hopewell, J. W. (1972) Mitotic activity in the subependymal plate of rats and the long-term consequences of X-irradiation.Journal of the Neurological Sciences 15, 471–82.
Crang, A. J. &Blakemore, W. F. (1989) The effect of the number of oligodendrocytes transplanted into X-irradiated, glial-free lesions on the extent of oligodendrocyte remyelination.Neuroscience Letters 193, 269–74.
Crang, A. J., Franklin, R. J. M., Blakemore, W. F., Noble, M., Barnett, S., Groves, A., Trotter, J. &Schachner, M. (1992) The differentiation of glial cell progenitor populations following transplantation into non-repairing central nervous system glial lesions in adult animals.Journal of Neuroimmunology 40, 243–54.
Eisenbarth, G. S., Walsh, F. S. &Nirenberg, M. (1979) Monoclonal antibody to plasma membrane antigen of neurons.Proceedings of the National Academy of Sciences (USA) 76, 4913–17.
Emmett, C. J., Lawrence, J. M., Raisman, G. &Seeley, P. J. (1991) Cultured epitheloid astrocytes migrate after transplantation into the adult rat brain.Journal of Comparative Neurology 3, 330–41.
Franklin, R. J. M., Crang, A. J. &Blakemore, W. F. (1991) Transplanted type-1 astrocytes facilitate repair of demyelinating lesions by host oligodendrocytes in the adult rat spinal cord.Journal of Neurocytology 20, 420–30.
Franklin, R. J. M., Crang, A. J. &Blakemore, W. F. (1992) Type-1 astrocytes fail to inhibit Schwann cell remyelination of CNS axons in the absence of cells of the O-2A lineage.Developmental Neuroscience 14, 85–92.
Gard, A. L. &Pfeiffer, S. E. (1989) Oligodendrocyte progenitors isolated directly from developing telencephalon at a specific phenotypic stage: myelinogenic potential in a defined environment.Development 106, 119–32.
Goldberg, W. J. &Bernstein, J. J. (1988) Migration of cultured fetal spinal cord astrocytes into adult host cervical cord and medulla following transplantation into thoracic spinal cord.Journal of Neuroscience Research 19, 34–42.
Graça, D. L. &Blakemore, W. F. (1986) Delayed remyelination in rat spinal cord following ethidium bromide injection.Neuropathology and Applied Neurobiology 12, 593–605.
Grinspan, J. B., Stern, J. L., Pustilnik, S. M. &Pleasure, D. (1990) Cerebral white matter contains PDGF-responsive precursors to O-2A cells.Journal of Neuroscience 10, 1866–73.
Hardy, R. &Reynolds, R. (1991) Proliferation and differentiation potential of rat forebrain oligodendroglial progenitors bothin vitro andin vivo.Development 111, 1061–80.
Hatton, J. D., Garcia, R. &U, H. S. (1992) Migration of grafted rat astrocytes: dependence on source/target organ.Glia 5, 251–8.
Ignacio, V., Collins, V. P., Suard, I. M. &Jacque, C. M. (1989) Survival of astroglial cell lineage from adult brain transplant.Developmental Neuroscience 11, 174–8.
Ignacio, V., Gansmuller, A., Collins, V. P., Suard, I. &Jacque, C. (1990) Short-term post-grafting morphological alterations of glia from an adult brain transplant.Glia 3, 140–50.
Levine, S. M. &Goldman, J. E. (1988) Ultrastructural characteristics of GD3 ganglioside-positive immature glia in the rat forebrain white matter.Journal of Comparative Neurology 277, 456–64.
McDermott, K. W. &Lantos, P. L. (1991) Distribution and fine structural analysis of undifferentiated cells in the primate subependymal layer.Journal of Anatomy 178, 45–63.
Miller, R. H. &Szigeti, V. (1991) Clonal analysis of astrocyte diversity in neonatal rat spinal cord.Development 113, 353–62.
Mirsky, R., Jessen, K. R., Schachner, M. &Goridis, C. (1986) Distribution of the adhesion molecules N-CAM and L1 on peripheral neurons and glia in adult rats.Journal of Neurocytology 15, 799–815.
Mirsky, R., Dubois, C., Morgan, L. &Jessen, K. R. (1990) O4 and A007-sulfatide antibodies bind to embryonic Schwann cells prior to the appearance of galactocerebroside; regulation of the antigen by axons-Schwann cell signals and cyclic AMP.Development 109, 105–16.
Privat, A. &Leblond, C. P. (1972) The subependymal layer and neighbouring region in the brain of the young rat.Journal of Comparative Neurology 148, 277–302.
Raff, M. C. (1990) Subclasses of astrocytes in culture: what should we call them? InDifferentiation and functions of glial cells (edited byLevi, G.) pp. 17–23. New York: Alan R. Liss, inc.
Raff, M. C., Abney, E. R., Cohen, J., Lindsay, R. &Noble, M. (1983a) Two types of astrocyte in cultures of developing white matter: differences in morphology, surface gangliosides, and growth characteristics.Journal of Neuroscience 6, 1289–300.
Raff, M. C., Miller, R. H. &Noble, M. (1983b) A glial progenitor that developsin vitro into an astrocyte or an oligodendrocyte depending on tissue culture medium.Nature 303, 390–6.
Reyners, H., Gianfelici De Reyners, E., Regniers, L. &Maisin, J.-R. (1986) A glial progenitor cell in the cerebral cortex of the adult rat.Journal of Neurocytology 15, 53–61.
Smith, G. M. &Miller, R. H. (1991) Immature type-1 astrocytes suppress glial scar formation, are motile and interact with blood vessels.Brain Research 543, 111–22.
Sommer, I. &Schachner, M. (1981) Monoclonal antibodies (O1 to O4) to oligodendrocyte surfaces. An immunocytological study in the central nervous system.Developmental Biology 83, 3121–7.
Van Der Maazen, R. W. M., Verhagen, I. &Van Der Kogel, A. J. (1990) Anin vitro clonogenic assay to assess radiation damage in rat CNS glial progenitor cells.International Journal of Radiation Biology 58, 835–44.
Van Der Maazen, R. W. M., Kleiboer, B. J., Verhagen, I. &Van Der Kogel, A. J. (1991) Irradiationin vitro discriminates between different O-2A progenitor cell populations in the perinatal central nervous system of rats.Radiation Research 128, 64–72.
Vaysse, P. J.-J. &Goldman, J. E. (1992) A distinct type of GD3+, flat astrocyte in rat CNS cultures.Journal of Neuroscience 12, 330–37.
Yeh, H.-J., Ruit, K. G., Wang, Y.-X., Parks, W. C., Snider, W. D. &Deuel, T. F. (1991) PDGF A-chain gene is expressed by mammalian neurons during development and in maturity.Cell 64, 209–16.
Zhou, H. F., Lee, L. H.-C. &Lund, R. D. (1990) Timing and patterns of astrocyte migration from xenogeneic transplants of the cortex and corpus callosum.Journal of Comparative Neurology 292, 320–30.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Franklin, R.J.M., Crang, A.J. & Blakemore, W.F. The reconstruction of an astrocytic environment in glia-deficient areas of white matter. J Neurocytol 22, 382–396 (1993). https://doi.org/10.1007/BF01195559
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01195559