Summary
In previous studies two neurite growth inhibiting protein fractions of 35 and 250 kDa were identified in myelin preparations of the rat CNS. These activities were not found in the myelin of PNS. A monoclonal antibody (mAb IN-1) was raised against the 250 kDa protein fraction and selected for its ability to neutralize the inhibitory effect of CNS myelin and of both isolated protein fractions. IN-1 has been shown bothin vitro andin vivo to neutralize the inhibitory effect of differentiated oligodendrocytes and CNS white matter. In the present study, the antigens of IN-1 were localized by immunohistochemistry on cryostat sections of the adult rat nervous system. The staining pattern of IN-1 was compared to that of mAbs specific for proteins found in CNS and PNS myelin. These proteins include myelin basic protein, myelin oligodendrocyte glycoprotein, and myelin associated glycoprotein. IN-1 stained white matter and myelinated fibre tracts in the CNS on sections of fresh frozen tissue fixed with 95% ethanol: 5% acetic acid (Clark's solution). Sciatic nerve myelin and spinal roots remained unstained. The staining pattern of IN-1 corresponded most closely to that of a mAb against myelin oligodendrocyte glycoprotein, a protein which occurs exclusively in CNS myelin and on differentiated oligodendrocytes.
Similar content being viewed by others
References
Agrawal, H. C., Hartmann, B. K., Shearer, W. T., Kalmbach, S. &Margolis, F. L. (1977) Purification and immunohistochemical localization of rat brain myelin proteolipid protein.Journal of Neurochemistry 28, 495–508.
Bandtlow, C., Zachleder, T. &Schwab, M. E. (1990) Oligodendrocytes arrest neurite growth by contact inhibition.Journal of Neuroscience 10, 3837–48.
Bandtlow, C. E., Schmidt, M. F., Hassinger, T. D., Schwab, M. E. &Kater, S. B. (1993) Role of intracellular calcium in NI-35-evoked collapse of neuronal growth cones.Science 259, 80–3.
Bedi, K. S., Winter, J., Berry, M. &Cohen, J. (1992) Adult rat dorsal root ganglion neurons extend neurites on predegenerated but not on normal peripheral nervesin vitro.European Journal of Neuroscience 4, 193–200.
Berthold, C. H., Carlstedt, T. &Corneliusson, O. (1984) Anatomy of the nerve root at the central-peripheral transitional region. InPeripheral Neuropathy, Vol. 1, 2nd ed. (edited byDyck, P. J., Thomas, P. K., Lambert, E. H., Bunge, R. P.) pp. 156–70. Philadelphia: Saunders.
Brown, M. C., Lunn, E. R. &Perry, V. H. (1991) Poor growth of mammalian motor and sensory axons into intact proximal nerve stumps.European Journal of Neuroscience 3, 1366–9.
Cadelli, D. &Schwab, M. E. (1991) Regeneration of lesioned septohippocampal acetylcholinesterase-positive axons is improved by antibodies against the myelin-associated neurite growth inhibitors NI-35/250.European Journal of Neuroscience 3, 825–32.
Carbonetto, S., Evans, D. &Cochard, P. (1987) Nerve fiber growth in culture on tissue substrata from central and peripheral nervous systems.Journal of Neuroscience 7, 610–20.
Caroni, P. &Schwab, M. E. (1988a) Two membrane protein fractions from rat central myelin with inhibitory properties for neurite growth and fibroblast spreading.Journal of Cell Biology 106, 1281–8.
Caroni, P. &Schwab, M. E. (1988b) Antibody against myelin-associated inhibitor of neurite growth neutralizes nonpermissive substrate properties of CNS white matter.Neuron 1, 85–96.
Crutcher, K. A. (1989) Tissue sections from the mature rat brain and spinal cord as substrates for neurite outgrowthin vitro: extensive growth on gray matter but little growth on white matter.Experimental Neurology 104, 39–54.
Igarashi, M., Strittmatter, S. M., Vartanian, T. &Fishman, M. C. (1993) Mediation by G proteins of signals that cause collapse of growth cones.Science 259, 77–9.
Jacobson, S. (1963) Sequence of myelination in the brain of the albino rat.Journal of Comparative Neurology 121, 5–29.
Kapfhammer, J. P., Schwab, M. E. &Schneider, G. E. (1992) Antibody neutralization of neurite growth inhibitors from oligodendrocytes results in expanded pattern of postnatally sprouting retinocollicular axons.Journal of Neuroscience 12, 2112–19.
Keller, F., Rubin, B., Bandtlow, C. &Schwab, M. E. (1992) Purification and characterization of inhibitory proteins from rat CNS myelin.Society for Neuroscience Abstracts 18, Part 2, 945.
Linington, C., Webb, M. &Woodhams, P. L. (1984) A novel myelin-associated glycoprotein defined by a mouse monoclonal antibody.Journal of Neuroimmunology 6, 387–96.
Omlin, F. X., Def. Webster, H., Palkovits, C. A. &Cohen, S. R. (1982) Immunocytochemical localization of basic protein in major dense line regions of central and peripheral myelin.Journal of Cell Biology 95, 242–8.
Poltorak, M., Sadoul, R., Keilhauer, G., Landa, C., Fahrig, T. &Schachner, M. (1987) Myelin-associated glycoprotein, a member of the L2/HNK-1 family of neural cell adhesion molecules, is involved in neuron-oligodendrocyte and oligodendrocyte-oligodendrocyte interaction.Journal of Cell Biology 105, 1893–9.
Sagot, Y., Swerts, J.-P. &Cochard, P. (1991) Changes in permissivity for neuronal attachment and neurite outgrowth of spinal cord grey and white matter during development: a study with the ‘cryoculture’ bioassay.Brain Research 543, 25–35.
Sandrock, A. W. &Matthew, W. D. (1987) Identification of a peripheral nerve neurite growth-promoting activity by development and use of anin vitro bioassay.Proceedings of the National Academy of Sciences (USA) 84, 6934–8.
Savio, T. &Schwab, M. E. (1989) Rat CNS white matter, but not gray matter, is nonpermissive for neuronal cell adhesion and fiber outgrowth.Journal of Neuroscience 9, 1126–33.
Schnell, L. &Schwab, M. E. (1990) Axonal regeneration in the rat spinal cord produced by an antibody against myelin-associated neurite growth inhibitors.Nature 343, 269–72.
Schnell, L. &Schwab, M. E. (1993) Sprouting and regeneration of lesioned corticospinal tract fibers in the adult rat spinal cord.European Journal of Neuroscience 5, 1156–71.
Schwab, M. E. &Caroni, P. (1988) Oligodendrocytes and CNS myelin are nonpermissive substrates for neurite growth and fibroblast spreadingin vitro.Journal of Neuroscience 8, 2381–93.
Schwab, M. E., Kapfhammer, J. P. &Bandtlow, C. E. (1993) Inhibitors of neurite growth.Annual Review of Neuroscience 16, 565–95.
Schwab, M. E. &Thoenen, H. (1985) Dissociated neurons regenerate into sciatic but not optic nerve explants in culture irrespective of neurotrophic factors.Journal of Neuroscience 5, 2415–23.
Sires, L. R., Hruby, S., Alvord, Jr, E. C., Hellström, I., Hellström, K.-E., Kies, M. W., Martenson, R., Deibler, G. E., Beckman, E. D. &Casnellie, J. E. (1981) Species restriction of a monoclonal antibody reacting with residues 130 to 137 in encephalitogenic myelin basic protein.Science 214, 87–9.
Trapp, B. D. &Quarles, R. H. (1982) Presence of myelinassociated glycoprotein correlates with alterations in the periodicity of peripheral myelin.Journal of Cell Biology 92, 877–82.
Tuttle, R. &Matthew, W. D. (1991) Anin vitro bioassay for neurite growth using cryostat sections of nervous tissue as a substratum.Journal of Neuroscience Methods 39, 193–202.
Watanabe, E. &Murakami, F. (1990) Cell attachment to and neurite outgrowth on tissue sections of developing, mature and lesioned brain: the role of inhibitory factor(s) in the CNS white matter.Neuroscience Research 8, 83–99.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rubin, B.P., Dusart, I. & Schwab, M.E. A monoclonal antibody (IN-1) which neutralizes neurite growth inhibitory proteins in the rat CNS recognizes antigens localized in CNS myelin. J Neurocytol 23, 209–217 (1994). https://doi.org/10.1007/BF01275525
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01275525