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. 1994 Apr;144(4):702–710.

Amyloid beta precursor protein and ubiquitin epitopes in human and experimental dystrophic axons. Ultrastructural localization.

B Bacci 1, E Cochran 1, M G Nunzi 1, E Izeki 1, T Mizutani 1, A Patton 1, S Hite 1, L M Sayre 1, L Autilio-Gambetti 1, P Gambetti 1
PMCID: PMC1887249  PMID: 7512790

Abstract

Dystrophic axons (DA) represent a major pathological feature of several neurodegenerative disorders, including infantile neuroaxonal dystrophy (INAD) and Alzheimer disease. We have previously presented evidence that amyloid beta precursor protein (BPP) and ubiquitin (Ub) are present in DA of different origin. We have now characterized the immunoreactivity of DA experimentally induced in rat by the administration of parabromophenylacetylurea (BPAU) and examined the subcellular localization of Ub and BPP in BPAU-induced DA and in DA present in subjects affected by INAD. BPAU-induced DA strongly immunoreacted with antisera to Ub and to COOH- and NH2-terminal regions of BPP. Immunoblots of DA-enriched brain regions were consistent with an increase in the amount of Ub and BPP in DA. Moreover, BPAU-induced DA immunoreacted with antibodies to PGP 9.5, a neuronal-specific Ub COOH-terminal hydrolase, and to the inducible heat shock protein 70. Antigenic characterization also indicated that the tubulovesicular membranes within DA derived largely from the smooth endoplasmic reticulum rather than from the Golgi system or the synaptic vesicles. Subcellular immunolocalization of Ub and BPP in both INAD- and BPAU-induced DA revealed that Ub and BPP colocalize in granulovesicular material in both conditions. In INAD DA intense Ub immunoreactivity was also detected in nonmembranous electron dense structures that were present only in these DA, probably because of the chronic course of INAD. Although BPP immunostaining may be related to accumulation of BPP-containing membranes in DA, Ub immunostaining is likely to result from activation of the Ub system by the neuron in the attempt to remove excessive and possibly abnormal proteins. A similar pathogenesis can be postulated for DA of Alzheimer disease.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aicardi J., Castelein P. Infantile neuroaxonal dystrophy. Brain. 1979 Dec;102(4):727–748. doi: 10.1093/brain/102.4.727. [DOI] [PubMed] [Google Scholar]
  2. Ananthan J., Goldberg A. L., Voellmy R. Abnormal proteins serve as eukaryotic stress signals and trigger the activation of heat shock genes. Science. 1986 Apr 25;232(4749):522–524. doi: 10.1126/science.3083508. [DOI] [PubMed] [Google Scholar]
  3. Bizzi A., Schaetzle B., Patton A., Gambetti P., Autilio-Gambetti L. Axonal transport of two major components of the ubiquitin system: free ubiquitin and ubiquitin carboxyl-terminal hydrolase PGP 9.5. Brain Res. 1991 May 10;548(1-2):292–299. doi: 10.1016/0006-8993(91)91135-n. [DOI] [PubMed] [Google Scholar]
  4. Blakemore W. F., Cavanagh J. B. "Neuroaxonal dystrophy" occurring in an experimental "dying back" process in the rat. Brain. 1969;92(4):789–804. doi: 10.1093/brain/92.4.789. [DOI] [PubMed] [Google Scholar]
  5. COWEN D., OLMSTEAD E. V. Infantile neuroaxonal dystrophy. J Neuropathol Exp Neurol. 1963 Apr;22:175–236. doi: 10.1097/00005072-196304000-00001. [DOI] [PubMed] [Google Scholar]
  6. Cavanagh J. B., Chen F. C., Kyu M. H., Ridley A. The experimental neuropathy in rats caused by p-bromophenylacetylurea. J Neurol Neurosurg Psychiatry. 1968 Oct;31(5):471–478. doi: 10.1136/jnnp.31.5.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cochran E., Bacci B., Chen Y., Patton A., Gambetti P., Autilio-Gambetti L. Amyloid precursor protein and ubiquitin immunoreactivity in dystrophic axons is not unique to Alzheimer's disease. Am J Pathol. 1991 Sep;139(3):485–489. [PMC free article] [PubMed] [Google Scholar]
  8. Cole G., Masliah E., Huynh T. V., DeTeresa R., Terry R. D., Okuda C., Saitoh T. An antiserum against amyloid beta-protein precursor detects a unique peptide in Alzheimer brain. Neurosci Lett. 1989 May 22;100(1-3):340–346. doi: 10.1016/0304-3940(89)90710-6. [DOI] [PubMed] [Google Scholar]
  9. Dickson D. W., Wertkin A., Mattiace L. A., Fier E., Kress Y., Davies P., Yen S. H. Ubiquitin immunoelectron microscopy of dystrophic neurites in cerebellar senile plaques of Alzheimer's disease. Acta Neuropathol. 1990;79(5):486–493. doi: 10.1007/BF00296107. [DOI] [PubMed] [Google Scholar]
  10. Haas A. L., Bright P. M. The immunochemical detection and quantitation of intracellular ubiquitin-protein conjugates. J Biol Chem. 1985 Oct 15;260(23):12464–12473. [PubMed] [Google Scholar]
  11. Herman M. M., Huttenlocher P. R., Bensch K. G. Electron microscopic observations in infantile neuroaxonal dystrophy. Report of a cortical biopsy and review of the recent literature. Arch Neurol. 1969 Jan;20(1):19–34. doi: 10.1001/archneur.1969.00480070029004. [DOI] [PubMed] [Google Scholar]
  12. Ivy G. O., Kitani K., Ihara Y. Anomalous accumulation of tau and ubiquitin immunoreactivities in rat brain caused by protease inhibition and by normal aging: a clue to PHF pathogenesis? Brain Res. 1989 Oct 2;498(2):360–365. doi: 10.1016/0006-8993(89)91117-7. [DOI] [PubMed] [Google Scholar]
  13. Jakobsen J., Lambert E. H., Carlson G., Brimijoin S. Clinical and electrophysiological characteristics of the experimental neuropathy caused by p-bromophenylacetylurea. Exp Neurol. 1982 Jan;75(1):158–172. doi: 10.1016/0014-4886(82)90015-2. [DOI] [PubMed] [Google Scholar]
  14. Kawarabayashi T., Shoji M., Yamaguchi H., Tanaka M., Harigaya Y., Ishiguro K., Hirai S. Amyloid beta protein precursor accumulates in swollen neurites throughout rat brain with aging. Neurosci Lett. 1993 Apr 16;153(1):73–76. doi: 10.1016/0304-3940(93)90080-5. [DOI] [PubMed] [Google Scholar]
  15. Koo E. H., Sisodia S. S., Archer D. R., Martin L. J., Weidemann A., Beyreuther K., Fischer P., Masters C. L., Price D. L. Precursor of amyloid protein in Alzheimer disease undergoes fast anterograde axonal transport. Proc Natl Acad Sci U S A. 1990 Feb;87(4):1561–1565. doi: 10.1073/pnas.87.4.1561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lowe J., McDermott H., Landon M., Mayer R. J., Wilkinson K. D. Ubiquitin carboxyl-terminal hydrolase (PGP 9.5) is selectively present in ubiquitinated inclusion bodies characteristic of human neurodegenerative diseases. J Pathol. 1990 Jun;161(2):153–160. doi: 10.1002/path.1711610210. [DOI] [PubMed] [Google Scholar]
  17. McLean I. W., Nakane P. K. Periodate-lysine-paraformaldehyde fixative. A new fixation for immunoelectron microscopy. J Histochem Cytochem. 1974 Dec;22(12):1077–1083. doi: 10.1177/22.12.1077. [DOI] [PubMed] [Google Scholar]
  18. Mignery G. A., Südhof T. C., Takei K., De Camilli P. Putative receptor for inositol 1,4,5-trisphosphate similar to ryanodine receptor. Nature. 1989 Nov 9;342(6246):192–195. doi: 10.1038/342192a0. [DOI] [PubMed] [Google Scholar]
  19. Moretto G., Sparaco M., Monaco S., Bonetti B., Rizzuto N. Cytoskeletal changes and ubiquitin expression in dystrophic axons of Seitelberger's disease. Clin Neuropathol. 1993 Jan-Feb;12(1):34–37. [PubMed] [Google Scholar]
  20. Nagata H., Brimijoin S. Axonal transport in the motor neurons of rats with neuropathy induced by p-bromophenylacetylurea. Ann Neurol. 1986 May;19(5):458–464. doi: 10.1002/ana.410190506. [DOI] [PubMed] [Google Scholar]
  21. Oka N., Brimijoin W. S. Tubulomembranous lesions in p-bromophenylacetylurea neuropathy reflect local stasis of fast axonal transport: evidence from electron microscopic autoradiography. Mayo Clin Proc. 1992 Apr;67(4):341–348. doi: 10.1016/s0025-6196(12)61550-x. [DOI] [PubMed] [Google Scholar]
  22. Parag H. A., Raboy B., Kulka R. G. Effect of heat shock on protein degradation in mammalian cells: involvement of the ubiquitin system. EMBO J. 1987 Jan;6(1):55–61. doi: 10.1002/j.1460-2075.1987.tb04718.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sahenk Z., Lasek R. J. Inhibition of proteolysis blocks anterograde-retrograde conversion of axonally transported vesicles. Brain Res. 1988 Sep 13;460(1):199–203. doi: 10.1016/0006-8993(88)91224-3. [DOI] [PubMed] [Google Scholar]
  24. Sandbank U., Lerman P., Geifman M. Infantile neuroaxonal dystrophy: cortical axonic and presynaptic changes. Acta Neuropathol. 1970;16(4):342–352. doi: 10.1007/BF00686897. [DOI] [PubMed] [Google Scholar]
  25. Schwartz A. L., Ciechanover A., Brandt R. A., Geuze H. J. Immunoelectron microscopic localization of ubiquitin in hepatoma cells. EMBO J. 1988 Oct;7(10):2961–2966. doi: 10.1002/j.1460-2075.1988.tb03158.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Selkoe D. J., Podlisny M. B., Joachim C. L., Vickers E. A., Lee G., Fritz L. C., Oltersdorf T. Beta-amyloid precursor protein of Alzheimer disease occurs as 110- to 135-kilodalton membrane-associated proteins in neural and nonneural tissues. Proc Natl Acad Sci U S A. 1988 Oct;85(19):7341–7345. doi: 10.1073/pnas.85.19.7341. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Suzuki K., Terry R. D. Fine structural localization of acid phosphatase in senile plaques in Alzheimer's presenile dementia. Acta Neuropathol. 1967 May 5;8(3):276–284. doi: 10.1007/BF00688828. [DOI] [PubMed] [Google Scholar]
  28. Takauchi S., Miyoshi K. Degeneration of neuronal processes in rats induced by a protease inhibitor, leupeptin. Acta Neuropathol. 1989;78(4):380–387. doi: 10.1007/BF00688174. [DOI] [PubMed] [Google Scholar]
  29. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Troncoso J. C., Griffin J. W., Price D. L., Hess-Kozlow K. M. Pathology of the peripheral neuropathy induced by p-bromophenylacetylurea. Lab Invest. 1982 Feb;46(2):215–223. [PubMed] [Google Scholar]
  31. Tsukita S., Ishikawa H. The movement of membranous organelles in axons. Electron microscopic identification of anterogradely and retrogradely transported organelles. J Cell Biol. 1980 Mar;84(3):513–530. doi: 10.1083/jcb.84.3.513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wiedenmann B., Franke W. W. Identification and localization of synaptophysin, an integral membrane glycoprotein of Mr 38,000 characteristic of presynaptic vesicles. Cell. 1985 Jul;41(3):1017–1028. doi: 10.1016/s0092-8674(85)80082-9. [DOI] [PubMed] [Google Scholar]
  33. Wilkinson K. D., Lee K. M., Deshpande S., Duerksen-Hughes P., Boss J. M., Pohl J. The neuron-specific protein PGP 9.5 is a ubiquitin carboxyl-terminal hydrolase. Science. 1989 Nov 3;246(4930):670–673. doi: 10.1126/science.2530630. [DOI] [PubMed] [Google Scholar]

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