Protein Heterogeneity in Rat CNS Myelin Subfractions

  • T. V. Waehneldt
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 100)

Summary

Microsomal fraction-free myelin from forebrain and spinal cord of young and mature rats, when subjected to hypo-osmotic shock and slow speed centrifugation, yielded a myelin pellet and a supernatant fraction (SN 4). Fraction SN 4 consisted of small vesicular profiles in which the major myelin proteins were reduced whereas high molecular weight material such as Wolfgram protein, myelin-associated glycoprotein and CNP were substantially increased over myelin. A close correlation of the SN 4 fraction to the myelin-like fraction of Davison and coworkers was suggested.

The myelin pellets were subfractioned on zonal sucrose gradients to yield bell-shaped particle distributions. Besides shifts in densities of the maxima between myelin of young and mature forebrain and spinal cord, a decrease was observed from the light to the heavy gradient end in basic proteins, and an increase in Wolfgram protein and other high molecular weight proteins. Proteolipid protein took an intermediate position. Light fractions from adult spinal cord displayed CNP activities below those of the total homogenate. This result, together with the very high CNP activities in fraction SN 4 casts some doubt on CNP being a marker for compact myelin; rather it appears that CNP is a marker for the process of myelin formation.

Keywords

Cholesterol Sucrose Phosphorus Electrophoresis Fractionation 

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References

  1. 1.
    Adams, D.H. and Fox, M.E., The homogeneity and protein composition of rat brain myelin, Brain Res. 14 (1969) 647–661.PubMedCrossRefGoogle Scholar
  2. 2.
    Agrawal, H.C., Banik, N.L., Bone, S., Davison, A.N., Mitchell, R.F. and Spohn, M., The identity of myelin-like fraction isolated from developing brain, Biochem. J. 120 (1970) 635–642.Google Scholar
  3. 3.
    Agrawal, H.C., Burton, R.M., Fishman, M.A., Mitchell, R.F. and Prensky, A.L., Partial characterization of a new myelin protein component, J. Neurochem. 19 (1972) 2083–2089.PubMedCrossRefGoogle Scholar
  4. 4.
    Agrawal, H.C., Trotter, J.L., Mitchell, R.F. and Burton, R.M., Criteria for identifying a myelin-like fraction from developing brain, Biochem. J. 136 (1973) 1117–1119.PubMedGoogle Scholar
  5. 5.
    Agrawal, H.C., Trotter, J.L., Burton, R.M. and Mitchell, R., Metabolic studies on myelin: evidence for a precursor role of a myelin subfraction, Biochem. J. 140 (1974) 99–109.PubMedGoogle Scholar
  6. 6.
    Autilio, L.A., Norton, W.T. and Terry, R.D., The preparation and some properties of purified myelin from the central nervous system, J. Neurochem. 11 (1964) 17–22.PubMedCrossRefGoogle Scholar
  7. 7.
    Benjamins, J.A., Miller, K. and McKhann, G.M., Myelin sub-fractions in developing rat brain: characterization and sulpha-tide metabolism, J. Neurochem. 20 (1973) 1589–1603.PubMedCrossRefGoogle Scholar
  8. 8.
    Benjamins, J.A., Miller, S.L. and Morell, P., Metabolic relationships between myelin subfractions: entry of galactolipids and phospholipids, J. Neurochem. 27 (1976) 565–570.PubMedCrossRefGoogle Scholar
  9. 9.
    Benjamins, J.A., Gray, M. and Morell, P., Metabolic relationships between myelin subfractions: entry of proteins, J. Neurochem. 27 (1976) 571–575.PubMedCrossRefGoogle Scholar
  10. 10.
    Cammer, W. and Norton, W.T., Disc gel electrophoresis of myelin proteins: new observations on development of the intermediate proteins (DM-20), Brain Res. 109 (1976) 643–648.PubMedCrossRefGoogle Scholar
  11. 11.
    Fujimoto, K., Roots, B.I., Burton, R.M. and Agrawal, H.C., Morphological and biochemical characterization of light and heavy myelin isolated from developing rat brain, Biochim. Biophys. Acta 426 (1976) 659–668.CrossRefGoogle Scholar
  12. 12.
    Igarashi, M. and Suzuki, K., Solubilization and characterization of the rat brain cholesterol ester hydrolase localized in the myelin sheath, J. Neurochem. 28 (1977) 729–738.PubMedCrossRefGoogle Scholar
  13. 13.
    Kurihara, T. and Tsukada, Y., The regional and subcellular distribution of 2’,3’-cyclic nucleotide 3’-phosphohydrolase in the central nervous system, J. Neurochem. 14 (1967) 1167–1174.PubMedCrossRefGoogle Scholar
  14. 14.
    Kurihara, T. and Tsukada, Y., 2′,3′-cyclic nucleotide 3′phosphohydrolase in the developing chick brain and spinal cord, J. Neurochem. 15 (1968) 827–832.PubMedCrossRefGoogle Scholar
  15. 15.
    Martenson, R.E., Deibler, G.E. and Kies, M.W., The occurrence of two myelin basic proteins in the central nervous system of rodents in the suborders Myomorpha and Sciuromorpha, J. Neurochem. 18 (1971) 2427–2433.PubMedCrossRefGoogle Scholar
  16. 16.
    Matthieu, J.-M., Quarles, R.H., Brady, R.O. and Webster, H. deF., Variations of proteins, enzyme markers and gangliosides in myelin subfractions, Biochim. Biophys. Acta329 (1973) 305–317.CrossRefGoogle Scholar
  17. 17.
    Matthieu, J.-M., Brady, R.O. and Quarles, R.H., Change in a myelin-associated glycoprotein in rat brain during development: metabolic aspects, Brain Res. 86 (1975) 55–65.PubMedCrossRefGoogle Scholar
  18. 18.
    McIntyre, R.J., Quarles, R.H., Webster, H. deF. and Brady, R.O., Isolation and characterization of myelin-related membranes, Trans. Am. Soc. Neurochem. 8 (1977) 159.Google Scholar
  19. 19.
    Mehl, E. and Wolfgram, F., Myelin types with different protein components in the same species, J. Neurochem. 16 (1969) 1091–1097.PubMedCrossRefGoogle Scholar
  20. 20.
    Mehl, E., Separation and characterization of myelin proteins, Adv. Exp. Med. Biol. 32 (1972) 157–170.PubMedCrossRefGoogle Scholar
  21. 21.
    Mokrasch, L.C., Biophysical chemistry and dynamics of myelin, in Myelin (L.C. Mokrasch, R.S. Bear and F.O. Schmitt, eds.) Neurosciences Res. Progr. Bull. 9 (1971) 452–506.Google Scholar
  22. 22.
    Moore, W.J., Smith, R. and Chapman, B.E., Conformation and function of myelin basic proteins, Trans. Am. Soc. Neurochem. 8 (1977) 67.Google Scholar
  23. 23.
    Morell, P., Greenfield, S., Costantino-Ceccarini, E. and Wisniewski, H., Changes in the protein composition of mouse brain myelin during development, J. Neurochem. 19 (1972) 2545–2554.Google Scholar
  24. 24.
    Norton, W.T. and Poduslo, S.E., Myelination in rat brain: method of myelin isolation, J. Neurochem. 21 (1973) 749–758.PubMedCrossRefGoogle Scholar
  25. 25.
    Poduslo, J.F. and Braun, P., Topographical arrangement of membrane proteins in the intact myelin sheath, J. biol. Chem. 250 (1975) 1099–1105.PubMedGoogle Scholar
  26. 26.
    Poduslo, J.F., Quarles, R.H. and Brady, R.O., External labeling of galactose in surface membrane glycoproteins of the intact myelin sheath, J. biol. Chem. 251 (1976) 153–158.PubMedGoogle Scholar
  27. 27.
    Quarles, R.H., Everly, J.L. and Brady, R.O., Evidence for the close association of a glycoprotein with myelin in rat brain, J. Neurochem. 21 (1973) 1177–1191.PubMedCrossRefGoogle Scholar
  28. 28.
    Quarles, R.H., Everly, J.L. and Brady, R.O., Myelin-associated glycoprotein: a developmental change, Brain Res. 58 (1973) 506–509.PubMedCrossRefGoogle Scholar
  29. 29.
    Quarles, R.H., The biochemical and morphological heterogeneity of myelin and myelin-related membranes, in Biochemistry of Brain (S. Kumar, ed.) Pergamon Press Ltd., in press.Google Scholar
  30. 30.
    Sabri, M.I., Tremblay, C., Banik, N.L., Scott, T., Gohil, K. and Davison, A.N., Biochemical and morphological changes in the subcellular fractions during myelination of rat brain, Biochem. Soc. Trans. (London) 3 (1975) 275–276.Google Scholar
  31. 31.
    Smith, M.E. and Sedgewick, L.M., Studies of the mechanism of demyelination. Regional differences in myelin stability in vitro, J. Neurochem. 24 (1975) 763–770.PubMedGoogle Scholar
  32. 32.
    Storm-Mathisen, J., Localization of transmitter candidates in the brain: the hippocampal formation as a model, Prog. Neurobio1.8 (1977) 119–181.CrossRefGoogle Scholar
  33. 33.
    Toews, A.D., Horrocks, L.A. and King, J.S., Simultaneous isolation of purified microsomal and myelin fractions from rat spinal cord, J. Neurochem. 27 (1976) 25–31.PubMedCrossRefGoogle Scholar
  34. 34.
    Waehneldt, T.V. and Mandel, P., Proteins of rat brain myelin. Extraction with sodium dodecylsulphate and electrophoresis on analytical and preparative scale, FEBS Lett. 9 (1970) 209–212.PubMedCrossRefGoogle Scholar
  35. 35.
    Waehneldt, T.V. and Neuhoff, V., Membrane proteins of rat brain: compositional changes during postnatal development, J. Neurochem. 23 (1974) 71–77.PubMedCrossRefGoogle Scholar
  36. 36.
    Waehneldt, T.V., Ontogenetic study of a myelin-derived fraction with 2’, 3’-cyclic nucleotide 3’-phosphohydrolase activity higher than that of myelin, Biochem. J. 151 (1975) 435–437.PubMedGoogle Scholar
  37. 37.
    Waehneldt, T.V., Matthieu, J.-M. and Neuhoff, V., Characteriza- tion of a myelin-related fraction (SN 4) isolated from rat forebrain at two developmental stages, Brain Res., in press.Google Scholar
  38. 38.
    Wolfgram, F., A new proteolipid fraction of the nervous system. I. Isolation and amino acid analysis, J. Neurochem. 13 (1966) 461–470.PubMedCrossRefGoogle Scholar
  39. 39.
    Zgorzalewicz, B., Neuhoff, V. and Waehneldt, T.V., Rat myelin proteins. Compositional changes in various regions of the nervous system during ontogenetic development, Neurobiology 4 (1974) 265–276.Google Scholar
  40. 40.
    Zimmermann, A.W., Quarles, R.H., Webster, H. deF., Matthieu, J.-M. and Brady, R.O., Characterization and protein analysis of myelin subfractions in rat brain: developmental and regional comparison, J. Neurochem. 25 (1975) 749–757.CrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1978

Authors and Affiliations

  • T. V. Waehneldt
    • 1
  1. 1.Forschungsstelle NeurochemieMax-Planck-Institut für experimentelle MedizinGöttingenGermany

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