Journal of Neurocytology

, Volume 29, Issue 9, pp 633–643 | Cite as

Anatomical repair of nerve membranes in crushed mammalian spinal cord with polyethylene glycol

  • R. Shi
  • R. B. Borgens
Article

Abstract

Acute damage to axons is manifested as a breach in their membranes, ion exchange across the compromised region, local depolarization, and sometimes conduction block. This condition can worsen leading to axotomy. Using a novel recording chamber, we demonstrate immediate arrest of this process by application of polyethylene glycol (PEG) to a severe compression of guinea pig spinal cord. Variable magnitudes of compound actions potentials (CAPs) were rapidly restored in 100% of the PEG-treated spinal cords. Using a dye exclusion test, in which horseradish peroxidase is imbibed by damaged axons, we have shown that the physiological recovery produced by polyethylene glycol was associated with sealing of compromised axolemmas. Injured axons readily imbibe horseradish peroxidase—but not following sealing of their membranes. The density of nerve fibers taking up the marker is significantly reduced following polyethylene glycol treatment compared to a control group. We further show that all axons—independent of their caliber—are equally susceptible to the compression injury and equally susceptible to polyethylene glycol mediated repair. Thus, polyethylene glycol—induced reversal of permeabilization by rapid membrane sealing is likely the basis for physiological recovery in crushed spinal cords. We discuss the clinical importance of these findings.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. AHKONG, Q. F., DESMAZES, J. P., GEORGESCAULD, D. & LUCY, J. A. (1987) Movements of fluorescent probes in the mechanism of cell fusion induced by polyethylene glycol. Journal of Cell Science 88, 389–98.PubMedGoogle Scholar
  2. ASANO, T., SHI, R. & BLIGHT, A. R. (1995) Horseradish proxidase used to examine the distribution of axonal damage in spinal cord compression injury in vitro. Journal of Neurotrauma 12, 993, Abstract.Google Scholar
  3. BLIGHT, A. R., MCGINNIS, M. E. & BORGENS, R. B. (1990) Cutaneus trunci muscle reflex of the guinea pig. Journal of Comparative Neurology 296, 614–633.PubMedGoogle Scholar
  4. BORGENS, R. B., BLIGHT, A. R. & MURPHY, D. J. (1986) Axonal regeneration in spinal cord injury: A perspective and new technique. Journal of Comparative Neurology 250, 157–167.PubMedGoogle Scholar
  5. BORGENS, R. B., TOOMBS, J. P., BLIGHT, A. R., MCGINNIS, M. E., BAUER, M. S., WIDMER, W. R. & COOK, J. R. Jr. (1993) Effects of applied electric fields on clinical cases of complete paraplegia in dogs. J. Restorative Neurology and Neurosci. 5, 305–322.Google Scholar
  6. BORGENS, R. B. & SHI, R. (2000) Immediate recovery from spinal cord injury through molecular repair of nerve membranes with polyethylene glycol. FASEB Journal 14, 27–35.PubMedGoogle Scholar
  7. BORGENS, R. B., TOOMBS, J. P., BREUR, G., WIDMER, W. R., WATER, D., HARBATH, A. M., MARCH, P. & ADAMS, L. G. (1999) An imposed oscillating electrical field improves the recovery of function in neurologically complete paraplegic dogs. Journal of Neurotrama 16, 639–657.Google Scholar
  8. DAVIDSON, R. L., O'MALLEY, K. A. & WHEELER, T. B. (1976) Induction of mammalian somatic cell hybridization by polyethylene glycol. Somatic Cell and Molecular Genetics 2, 271–280.Google Scholar
  9. DIMITRIJEVIC, M. R. (1995) Clinical aspects of traumatic injury to central nervous system axons. In The Axon (edited by WAXMAN, S. G., KOCSIS, J. D. & STYS, P. K.) pp. 669–679. New York: Oxford UP.Google Scholar
  10. FEHLINGS, M. & TATOR, C. (1995) The relationships among the severity of spinal cord injury, residual neurological function, axon counts, and counts of retrogradely labeled neurons after experimental spinal cord injury. Experimental Neurology 132, 123–134.PubMedGoogle Scholar
  11. GRIFFIN, J. W., GEORGE, E. B., HSIEH, S. & GLASS, J. D. (1995) Axonal degeneration and disorders of the axonal cytoskeleton. In The Axon (edited by WAXMAN, S. G., KOCSIS, J. D. & STYS, P. K.) pp. 375–390. New York: Oxford UP.Google Scholar
  12. GUTH, L. (1969) Trophic effects of vertebrate neurons. Neuroscience Research Program Bulletin 7, 1–73.Google Scholar
  13. HANNIG, J., YU, J., BECKETT, M., WEICHSELBAUM, R. & LEE, R. C. (1999) Poloxamine 1107 sealing of radiopermeabilized erythrocyte membranes. International Journal of Radiation Biology 75, 379–385.PubMedGoogle Scholar
  14. HONMOU, O. & YOUNG, W. (1995) Traumatic injury to the spinal axons. In The Axon (edited by WAXMAN, S. G., KOCSIS, J. D. & STYS, P. K), pp. 480–503. New York: Oxford UP.Google Scholar
  15. LEE, J. & LENTZ, B. R. (1997) Evolution of lipid structures during model membrane fusion and the relation of this process to cell membrane fusion. Biochemistry 36, 6251–6259.PubMedGoogle Scholar
  16. LENTZ, B. R. (1994) Induced membrane fusion; Potential mechanism and relation to cell fusion events. Chemistry and Physics of Lipids 73, 91–106.PubMedGoogle Scholar
  17. MALMGREN, L. & OLSSON, L. (1977) A sensitive histochemical method for light and electron microscopic demonstration of horseradish peroxidase. Journal of Histochemistry and Cytochemistry 25, 1280–1283.PubMedGoogle Scholar
  18. MAXWELL, W. L. (1996) Histopathological changes at central nodes of ranvier after stretch-injury. Microscopy Research and Technique 34, 522–535.PubMedGoogle Scholar
  19. MAXWELL, W. L. & GRAHAM, D. I. (1997) Loss of axonal microtubules and neurofilaments after stretch-injury to guinea pig optic nerve fibers. Journal of Neurotrauma 14, 603–614.PubMedGoogle Scholar
  20. MAXWELL, W. L., WATT, C., GRAHAM, D. I. & GENNARELLI, T. A. (1993) Ultrastructural evidence of axonal shearing as a result of lateral acceleration of the head in non-human primates. Acta Neuropathologica 86, 136–144.PubMedGoogle Scholar
  21. MORIARTY, L. J., DUERSTOCK, B. S., BAJAJ, C. L., LIN, K. & BORGENS, R. B. (1998) Two and three dimensional computer graphic evaluation of the subacute spinal cord injury. Journal of The Neurological Sciences 155, 121–137.PubMedGoogle Scholar
  22. NAKAJIMA, N. & IKADA, Y. (1994) Fusogenic activity of various water-soluble polymers. Journal of Biomaterials Science, Polymer Edition 6, 751–9.Google Scholar
  23. O'LAGUE, P. H. & HUNTTER, S. L. (1980) Physiological and morphological studies of rat pheochromocytoma cells (PC12) chemically fused and grown in culture. Proceedings of the National Academy of Sciences USA 77, 1701–1705.Google Scholar
  24. PADANLAM, J. T., BISCHOF, J. C., CRAVALHO, E. G., TOMPKINS, R. G., YARMUSH, M. L. & TONER, M. (1994) Effectiveness of Poloxamer 188 in arresting calcein leakage from thermally damaged isolated skeletal muscle cells. Annals of the New York Academy of Sciences 92, 111–123.Google Scholar
  25. PALMER, J. S., CROMIE, W. J. & LEE, R. C. (1998) Surfactant administration reduces testicular ischemiareprefusion injury. Journal of Urology 159, 2136–2139.PubMedGoogle Scholar
  26. SHI, R., ASANO, T. & BLIGHT, A. R. (1996) Sucrose gap recording of membranes resealing in mammalian spinal cord axons. Society for Neuroscience Abstracts 22, 1185.Google Scholar
  27. SHI, R. & BLIGHT, A. R. (1996) Compression injury of mammalian spinal cord in vitro and the dynamics of action potential conduction failure. Journal of Neurophysiology 76, 1572–1580.PubMedGoogle Scholar
  28. SHI, R., ASANO, T., VINING, N. C. & BLIGHT, A. R. (1997) m-Calpain dependence of membrane sealing in mammalian spinal cord axons. Society for Neuroscience Abstracts 108, 16.Google Scholar
  29. SHI, R. & BLIGHT, A. R. (1997) Differential effects of low and high concentrations of 4-Aminopyridine on axonal conduction in normal and injured spinal cord. Neuroscience 77, 553–562.PubMedGoogle Scholar
  30. SHI, R. & BORGENS, R. B. (1999) Acute repair of crushed guinea pig spinal cord by polyethylene glycol. Journal of Neurophysiology 81, 2406–2414.PubMedGoogle Scholar
  31. SHI, R., BORGENS, R. B. & BLIGHT, A. R. (1999) Functional reconnection of severed mammalian spinal cord axons with polyethylene glycol. Journal of Neurotrauma 16, 727–738.PubMedGoogle Scholar
  32. XIE, X. & BARRETT, J. N. (1991) Membrane resealing in cultured rat septal neurons after neurite transection: evidence for enhancement by Ca2+-triggered protease activity and cytoskeletal disassembly. Journal of Neuroscience 11, 3257–3267.PubMedGoogle Scholar
  33. YAWO, H. & KUNO, M. (1985) Calcium dependence of membrane sealing at the cut end of the cockroach giant axon. Journal of Neuroscience 5, 1626–1632.PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • R. Shi
    • 1
  • R. B. Borgens
    • 1
  1. 1.Institute for Applied Neurology, Center for Paralysis Research, Department of Basic Medical SciencesPurdue UniversityWest LafayetteUSA

Personalised recommendations