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Journal of Neurocytology

, Volume 16, Issue 2, pp 239–248 | Cite as

The distribution of (Na+ + K+)ATPase is continuous along the axolemma of unensheathed axons from spinal roots of ‘dystrophic’ mice

  • Reginald G. Ariyasu
  • Mark H. Ellisman
Article

Summary

(Na++K+)ATPase-like immunoreactivity along the axolemma of sensory and motor neurons and the plasmalemma of Schwann cells from spinal roots of dystrophic mice (129 ReJ Dy/Dy) was determined using polyclonal antibodies specific for guinea pig renal (Na++K+)ATPase (GP-17), along with polyclonal (439-2) and monoclonal (9A5) antibodies specific for rat renal (Na++K+)ATPase. In normal and dystrophic mice, (Na++K+)ATPase-like immunoreactivity was observed along the axolemma at nodes of Ranvier using GP-17 and 439-2, each of which binds to isozymes of (Na++K+)ATPase composed of the α and α+ forms of the catalytic subunit. Staining was not seen along the nodal axolemma with 9A5, a preparation that binds to the α form of the catalytic subunit. The terminal processes and microvilli of Schwann cells were stained using all three antibody probes. The axolemma of unensheathed axons in dystrophic mice was continuously and uniformly labelled with GP-17 and 439-2, but not 9A5. Concentrations of (Na++K+)ATPase-like immunoreactivity along Schwann cell processes were observed most often in areas adjacent to unensheathed axolemma. At heminodes, staining abruptly decreased along Schwann cell processes in areas that were separated from the unensheathed axolemma by other intervening Schwann cell processes. It was concluded from these data that in dystrophic mice (Na++K+)ATPase is uniformly distributed along unensheathed portions of axons without evidence of detectable focal concentations of the enzyme, and that the catalytic subunit of (Na++K+)ATPase along unensheathed axons is distinct from the α form found in Schwann cells and other organs. In addition, (Na++K+)ATPase is concentrated along the plasmalemma of Schwann cells in regions of close apposition to axolemmal areas associated with large ionic fluxes.

Keywords

Polyclonal Antibody Motor Neuron Schwann Cell Catalytic Subunit Cell Process 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Abbott, N. J. (1986) The neuronal microenvironment.Trends in Neurosciences 9, 3–6.Google Scholar
  2. Aguayo, A. J., Bray, G. M. &Perkins, C. S. (1979) Axon Schwann cell relationships in neuropathies of mutant mice.Annals of the New York Academy of Sciences 317, 512–31.Google Scholar
  3. Ariyasu, R. G., Nichol, J. A. &Ellisman, M. H. (1985) Localization of sodium/potassium adenosine triphosphatase in multiple cell types of the murine nervous system with antibodies raised against the enzyme from kidney.Journal of Neuroscience 5, 2581–96.Google Scholar
  4. Bostock, H., Hall, S. M. &Smith, K. J. (1980) Demyelinated axons can form ‘nodes’ prior to remyelination.Journal of Physiology 308, 21P-23P.Google Scholar
  5. Bradley, W. G. &Jenkison, M. (1973) Abnormalities of peripheral nerves in murine muscular dystrophy.Journal of the Neurological Sciences 18, 227–47.Google Scholar
  6. Bray, G. M. (1973) A comparison of the ouabain-sensitive (Na++K+)-adenosine triphosphatase of normal and dystrophic skeletal muscle.Biochimica et Biophysica Acta 298, 239–45.Google Scholar
  7. Bray, G. M. &Aguayo, A. J. (1975) Quantitative ultrastructural studies of the axon-Schwann cell abnormality in spinal nerve roots from dystrophic mice.Journal of Neuropathology and Experimental Neurology 34, 517–30.Google Scholar
  8. Bray, G. M., Cullen, M. J., Aguayo, A. J. &Rasminsky, M. (1979) Node-like areas of intramembraneous particles in the unensheathed axons of dystrophic mice.Neuroscience Letters 13, 203–8.Google Scholar
  9. Ellisman, M. H. (1976) The distribution of membrane molecular specializations characteristic of the node of Ranvier is not dependent upon myelination.Society for Neuroscience Abstracts 2, 410.Google Scholar
  10. Ellisman, M. H. (1979) Molecular specializations of the axon membrane at nodes of Ranvier are not dependent upon myelination.Journal of Neurocytology 8, 719–35.Google Scholar
  11. Ellisman, M. H. &Levinson, S. R. (1982) Immunocytochemical localization of sodium channel distributions in the excitable membranes ofElectrophorus electricus.Proceedings of the National Academy of Sciences USA 79, 6707–11.Google Scholar
  12. Ellisman, M. H., Miller, J. A. &Agnew, W. S. (1983) Molecular morphology of the tetrodotoxin-binding sodium channel protein fromElectrophorus electricus in solubilized and reconstituted preparations.Journal of Cell Biology 97, 1834–40.Google Scholar
  13. Grisar, T., Franck, G. &Schoffeniels, E. (1980) Glial control of neuronal excitability in mammals. II. Enzymatic evidence: two molecular forms of the (Na+,K+)-ATPase in brain.Neurochemistry International 2, 311–20.Google Scholar
  14. Hayat, M. A. (1972)Basic Electron Microscopy Techniques. New York: Van Nostrand Reinhold Company.Google Scholar
  15. Hildebrand, C. (1971) Ultrastructural and light-microscopic studies of the nodal region in large myelinated fibres of the adult feline spinal cord white matter.Acta physiologica scandinavica 364S, 43–79.Google Scholar
  16. Hubert, J. J., Schenk, D. B., Skelly, H. &Leffert, H. L. (1986) Rat hepatic [Na+,K+]-ATPase: alpha-subunit isolation by immunoaffinity chromatography and structural analysis by peptide mapping.Biochemistry 25, 4156–63.Google Scholar
  17. Huizar, P., Kuno, M. &Miyata, Y. (1975) Electrophysiological properties of spinal motoneurones of normal and dystrophic mice.Journal of Physiology 248, 231–46.Google Scholar
  18. Leffert, H. L., Schenk, D. B., Hubert, J. J., Skelly, H., Schumacher, M., Ariyasu, R., Ellisman, M., Koch, K. S. &Keller, G. A. (1985) Hepatic (Na+,K+)-ATPase: A current view of its structure, function and localization in rat liver as revealed by studies with monoclonal antibodies.Hepatology 5, 501–7.Google Scholar
  19. Madrid, R. E., Jaros, E., Cullen, M. J. &Bradley, W. G. (1975) Genetically determined defect of Schwann cell basement membrane in dystrophic mouse.Nature 257, 319–21.Google Scholar
  20. McDonough, A., Hiatt, A. &Edelman, I. S. (1982) Characteristics of antibodies to guinea pig (Na++K+)-adenosine triphosphatase and their use in cell-free synthesis studies.Journal of Membrane Biology 69, 13–22.Google Scholar
  21. McDonough, A. &Schmitt, C. (1985) Comparison of subunits of cardiac, brain, and kidney Na+-K+-ATPase.American Journal of Physiology 248, C247-C251.Google Scholar
  22. McGrail, K. M. &Sweadner, K. J. (1986) Immunofluorescent localization of two different Na,K-ATPases in the rat retina and in identified dissociated retinal cells.Journal of Neuroscience 6, 1272–83.Google Scholar
  23. McLean, I. W. &Nakane, P. K. (1974) Periodate-lysine-paraformaldehyde fixative. A new fixative for immunoelectron microscopy.Journal of Histochemistry and Cytochemistry 22, 1077–83.Google Scholar
  24. Moonen, G., Franck, G. &Schoffeniels, E. (1980) Glial control of neuronal excitability in mammals: I. Electrophysiological and isotopic evidence in culture.Neurochemistry International 2, 299–310.Google Scholar
  25. Perkins, C. S., Bray, G. M. &Aguayo, A. J. (1981) Ongoing block of Schwann cell differentiation and deployment in dystrophic mouse spinal roots.Developmental Brain Research 1, 213–20.Google Scholar
  26. Rasminsky, M. (1978) Ecotopic generation of impulses and cross-talk in spinal nerve roots of ‘dystrophic’ mice.Annals of Neurology 3, 351–7.Google Scholar
  27. Rasminsky, M., Kearney, R. E., Aguayo, A. J. &Bray, G. M. (1978) Conduction of nervous impulses in spinal roots and peripheral nerves of dystrophic mice.Brain Research 143, 71–85.Google Scholar
  28. Sanes, J. R. &Hall, Z. W. (1979) Antibodies that bind specifically to synaptic sites on muscle fiber basal lamina.Journal of Cell Biology 38, 347–70.Google Scholar
  29. Schenk, D. &Leffert, H. L. (1983) Monoclonal antibodies to rat (Na++K+) ATPase block enzymatic activity.Proceedings of the National Academy of Sciences USA 80, 5281–5.Google Scholar
  30. Schwartz, M., Ernst, S. A., Siegel, G. J. &Agranoff, B. W. (1981) Immunocytochemical localization of (Na++K+)-ATPase in the goldfish optic nerve.Journal of Neurochemistry 36, 107–15.Google Scholar
  31. Skriver, E., Maunsbach, A. B. &Jorgensen, P. L. (1980) Ultrastructure of Na,K-transport vesicles reconstituted with purified renal Na,K-ATPase.Journal of Cell Biology 86, 746–54.Google Scholar
  32. Specht, S. C. &Sweadner, K. J. (1984) Two different Na,K-ATPases in the optic nerve: cells of origin and axonal transport.Proceedings of the National Academy of Sciences USA 81, 1234–8.Google Scholar
  33. Sternberger, L. A. (1979)Immunocytochemistry, 2nd edn. New York: John Wiley and Sons, Inc.Google Scholar
  34. Stirling, C. A. (1975) Abnormalities in Schwann cell sheaths in spinal nerve roots of dystrophic mice.Journal of Anatomy 119, 169–80.Google Scholar
  35. Sweadner, K. J. (1979) Two molecular forms of (Na++K+)-stimulated ATPase in brain. Separation, and difference in affinity for strophanthidin.Journal of Biological Chemistry 254, 6060–7.Google Scholar
  36. Walz, W., Wuttke, W. &Hertz, L. (1984) Astrocytes in primary cultures: membrane potential characteristics reveal exclusive potassium conductance and potassium accumulator properties.Brain Research 292, 367–74.Google Scholar
  37. Waxman, S. G., Black, J. A. &Foster, R. E. (1982) Freeze-fracture heterogeneity of the axolemma of premyelinated fibers in the CNS.Neurology 32, 418–21.Google Scholar
  38. Wiley-Livingston, C. A. &Ellisman, M. H. (1980) Development of axonal membrane specializations defines nodes of Ranvier and precedes Schwann cell myelin elaboration.Developmental Biology 79, 334–55.Google Scholar
  39. Wood, J. G., Jean, D. H., Whitaker, J. N., McLaughlin, B. J. &Albers, R. W. (1977) Immunocytochemical localization of the sodium, potassium activated ATPase in knifefish brain.Journal of Neurocytology 6, 571–81.Google Scholar

Copyright information

© Chapman and Hall Ltd 1987

Authors and Affiliations

  • Reginald G. Ariyasu
    • 1
  • Mark H. Ellisman
    • 1
  1. 1.Laboratory for Neurocytology, Department of Neurosciences, School of MedicineUniversity of CaliforniaLajollaUSA

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