Summary
Rabbit antisera to the rat myelin proteins P0 and P1 were used to assay for the presence of these components by both immunochemical and immunofluorescence methods. The antiserum to P0 did not react detectably with polyacrylamide gels containing central myelin, or with P1 and P2 in peripheral myelin; it did react with P0 in peripheral myelin, and in extracts of adult and neonatal sciatic nerve. When reacted with frozen tissue sections using indirect immunofluorescence, it did not stain central myelin but did stain myelin in adult sciatic nerve, the myelinated fibres in cervical sympathetic trunk and occasional areas in neonatal sciatic nerve where Schwann cells had presumably begun to form myelin. Antiserum to basic protein reacted with both of the basic protein bands in central and peripheral myelin, but not P0; P1 and P2 were detectable in adult and neonatal sciatic nerve. In indirect immunofluorescence assays, the antiserum stained both central and peripheral myelin, the few myelinated fibres of sympathetic trunk and myelinating regions of neonatal sciatic nerve.
Cultured secondary rat Schwann cells showed no detectable reaction with either reagent, using either technique. We conclude that these three proteins are probably expressed as a consequence of the neuron-Schwann cell interaction that initiates myelination.
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References
Brockes, J. P., Fields, K. L. &Raff, M. C. (1977) A surface antigenic marker for rat Schwann cells.Nature 266, 364–6.
Brockes, J. P., Fields, K. L. &Raff, M. C. (1979) Studies on cultured rat Schwann cells. I. Derivation of purified populations from cultures of peripheral nerve.Brain Research 165, 105–18.
Brockes, J. P. &Hall, Z. W. (1975) Acetylcholine receptors in normal and denervated rat diaphragm muscle. I. Purification and interaction with125I-α-bungarotoxin.Biochemistry 14, 2092–9.
Brockes, J. P. &Raff, M. C. (1979) Studies on cultured rat Schwann cells. II. Comparison with a rat Schwann cell line.In vitro (in press).
Brostoff, S. W. &Eylar, E. H. (1972) The proposed amino acid sequence of P1 protein of rabbit sciatic nerve myelin.Archives in Biochemistry and Biophysics 153, 590–8.
Brostoff, S. W., Karkhanis, Y. D., Carlo, D. J., Reuter, W. &Eylar, E. H. (1975) Isolation and partial characterization of the major proteins of rabbit sciatic nerve myelin.Brain Research 86, 449–58.
Burridge, K. (1976) Changes in cellular glycoproteins after transformation: identification of specific glycoproteins and antigens in sodium dodecyl sulfate gels.Proceedings of the National Academy of Sciences (U.S.A.) 73, 4457–61.
Dunkley, P. R. &Carnegie, P. R. (1974) Amino acid sequence of the smaller basic protein from rat brain myelin.Biochemical Journal 141, 243–55.
Everly, J. L., Brady, R. O. &Quarles, R. H. (1974) Evidence that the major protein in rat sciatic nerve myelin is a glycoprotein.Journal of Neurochemistry 21, 329–34.
Fairbanks, G., Steck, T. L. &Wallach, D. H. F. (1971) Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane.Biochemistry 10, 2606–16.
Greenfield, S., Brostoff, S., Eylar, E. H. &Morell, P. (1973) Protein composition of myelin of the peripheral nervous system.Journal of Neurochemistry 20, 1207–16.
Kronvall, G., Seal, U. S., Finstad, J. &Williams, R. C. (1970) Phylogenetic insight into evolution of mammalian Fc fragments of globulins usingStaphylococcal protein A.Journal of Immunology 104, 140–7.
Laemmli, U. K. &Favre, M. (1974) Maturation of the head of bacteriophage T4. I. DNA packaging events.Journal of Molecular Biology 80, 575–99.
Martenson, R. E., Deibler, G. E. &Kies, M. W. (1971) The occurrence of two myelin basic proteins in the central nervous system of rodents in the subordersMyomorpha andSciurmorpha.Journal of Neurochemistry 18, 2427–33.
Mendell, J. R. &Whittaker, J. N. (1978) Immunocytochemical localisation studies of myelin basic protein.Journal of Cell Biology 76, 502–11.
Mirsky, R., Winter, J., Abney, E., Pruss, R., Gavrilovic, J. &Raff, M. (1980) Myelin-specific proteins and glycolipids in rat Schwann cells and oligodendrocytes in culture.Journal of Cell Biology (in press).
Raff, M. C., Hornby-Smith, A. &Brockes, J. P. (1978a) Cyclic AMP as a mitogenic signal for cultured rat Schwann cells.Nature 273, 672–3.
Raff, M. C., Abney, E. R., Hornby-Smith, A. &Brockes, J. P. (1978b) Schwann cell growth factors.Cell 15, 813–22.
Spencer, P. S. &Weinberg, H. J. (1978) Axonal specification of Schwann cell expression and myelination. InPhysiology and Pathobiology of Axons (edited byWaxman, S. G.), pp. 389–405. New York: Raven Press.
Standring, R. &Williams, A. F. (1978) Glycoproteins and antigens of membranes prepared from rat thymocytes after lysis by shearing or with the detergent Tween 40.Biochemica et Biophysica Acta 508, 85–96.
Sternberger, H. H., Itoyama, Y., Kies, M. W. &Webster, H. deF. (1978) Immunochemical method to identify basic protein in myelin-forming oligodendrocytes of newborn rat C.N.S.Journal of Neurocytology 7, 251–63.
Uyemura, K., Tobani, C., Hirano, S. &Tsukada, Y. (1972) Comparative studies on the myelin proteins of bovine peripheral nerve and spinal cord.Journal of Neurochemistry 19, 2607–14.
Uyemura, L., Kato-Yamanaka, T. &Kitamura, K. (1977) Distribution and optical activity of the basic protein in bovine peripheral nerve myelin.Journal of Neurochemistry 29, 61–8.
Webster, H. deF. (1971) The geometry of peripheral myelin sheaths during their formation and growth in rat sciatic nerves.Journal of Cell Biology 48, 348–67.
Wood, J. G. &Dawson, R. M. C. (1974) Some properties of a major structural glycoprotein of sciatic nerve.Journal of Neurochemistry 22, 631–5.
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Brockes, J.P., Raff, M.C., Nishiguchi, D.J. et al. Studies on cultured rat Schwann cells. III. Assays for peripheral myelin proteins. J Neurocytol 9, 67–77 (1980). https://doi.org/10.1007/BF01205227
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DOI: https://doi.org/10.1007/BF01205227