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Markers of axonal injury in post mortem human brain

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Abstract

β-Amyloid precursor protein (βAPP) can be detected immunocytochemically at sites of axonal injury in the brain, and has recently been found to be a useful marker for injured axons in patients who survived for only 3 h after head trauma. It is transported by last axonal transport and is thought to accumulate in detectable levels where the cytoskeleton breaks down. If this theory is correct, other substances should accumulate here in the same way, so we have used antibodies to other neuronal proteins to compare their efficacy as markers of axonal injury. SNAP-25, chromogranin A and cathepsin D also marked injured axons at all survival times studied (2.5h–2 weeks), although they were not as sensitive or specific as βAPP. Immunolabelling for the 68-kDa neurofilament subunit (NF68) was present in most uninjured axons, and allowed axonal swellings to be seen in some cases. Synaptophysin, GAP-43, ubiquitin or tau did not label any normal or injured axons in this study. We, therefore, suggest that βAPP should be the immunocytochemical marker of choice for the detection of injured axons. This study also showed that microwave antigen retrieval significantly enhances the immunoreactivity of SNAP-25, chromogranin A, synaptophysin, GAP-43, ubiquitin and tau, in addition to that of βAPP, in formalin-fixed, paraffin-embedded tissue, and reveals NF68 antigenicity where it was not previously detectable.

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References

  1. Adams JH, Graham DI, Murray LS, Scott G (1982) Diffuse axonal injury due to non-missile head injury in humans. Ann Neurol 12: 557–563

    PubMed  Google Scholar 

  2. Adams JH, Graham DI, Gennarelli TA, Maxwell WI (1991) Diffuse axonal injury in non-missile head injury. J Neurol Neurosurg Psychiatry 54: 481–483

    PubMed  Google Scholar 

  3. Adams LA, Ang L-C, Munoz DG (1993) Chromogranin A, a soluble synaptic vesicle protein, is found in cortical neurons other than previously defined peptidergic neurons in the human neocortex. Brain Res 602: 336–341

    Article  PubMed  Google Scholar 

  4. Barrett AJ (1980) Cathepsin D: the lysosomal aspartic proteinase. In: Barrett AJ (ed) Protein degradation in health and disease, Ciba Foundation symposium, 1979. Excerpta Medica, Amsterdam, pp 37–50

    Google Scholar 

  5. Benowitz LI, Routtenberg A (1987) A membrane phosphoprotein associated with neural development, axonal regeneration, phospholipid metabolism, and synaptic plasticity. Trends Neurosci 10: 527–532

    Article  Google Scholar 

  6. Benowitz LI, Rodriguez W, Pakevich P, Mufson EJ, Schenk D, Neve RL (1989) The amyloid precursor protein is concentrated in neuronal lysosomes in normal and Alzheimer's subjects. Exp Neurol 106: 237–250

    Article  PubMed  Google Scholar 

  7. Benowitz LI, Perrone-Bizzozero NI, Finkelstein SP, Bird ED (1989) Localisation of the growth-associated phosphoprotein GAP-43 (B-50, F1) in the human cerebral cortex. J Neurosci 9: 990–995

    PubMed  Google Scholar 

  8. Catteruccia N, Willingdale-Theune J, Bunke D, Prior R, Masters CL, Crisanti A, Beyreuther K (1990) Ultrastructural localisation of the putative precursors of the A4 amyloid protein associated with Alzheimer's disease. Am J Pathol 137: 19–26

    PubMed  Google Scholar 

  9. Cattoretti G, Pileri S, Parravicini C, Becker MHG, Poggi S, Bifulco C, Key G, D'Amato L, Sabattini E, Feudale E, Reynold F, Gerdes J, Rilke F (1993) Antigen unmasking on formalin-fixed, paraffin-embedded tissue sections. J Pathol 171: 83–98

    PubMed  Google Scholar 

  10. Cochran E, Bacci B, Chen Y, Patton A, Gambetti P, Autilio-Gambetti L (1991) Amyloid precursor protein and ubiquitin immunoreactivity in dystrophic axons is not unique to Alzheimer's Disease. Am J Pathol 139: 485–489

    PubMed  Google Scholar 

  11. Crooks DA, Scoltz CL, Vowles G, Greenwald S (1992) Axonal injury in closed head injury by assault: a quantitative study. Med Sci Law 32: 109–117

    PubMed  Google Scholar 

  12. De Camilli P, Jahn R (1990) Pathways to regulated exocytosis in humans. Annu Rev Physiol 52: 625–645

    Article  PubMed  Google Scholar 

  13. Ferreira A, Caceras A, Kosik KS (1993) Intraneuronal compartments of the amyloid precursor protein. J Neurosci 13: 3112–3123

    PubMed  Google Scholar 

  14. Fischer-Colbrie R, Hagn C, Schober M (1987) Chromogranins A, B and C: widespread constituents of secretory vesicles. Ann NY Acad Sci 493: 120–134

    PubMed  Google Scholar 

  15. Geinisman Y, Bondareff W, Telser A (1977) Transport of [3H]fucose-labeled glycoproteins in the septo-hippocampal pathway of young adult and senescent rats. Brain Res 125: 182–186

    Article  PubMed  Google Scholar 

  16. Gentleman SM, Nash MJ, Sweeting CJ, Graham DI, Roberts GW (1993) β-amyloid precursor protein (βAPP) as a marker for axonal injury after head injury. Neurosci Lett 160: 139–144

    PubMed  Google Scholar 

  17. Golde TE, Estus S, Younkin LH, Selkoe DJ, Younkin SG (1992) Processing of the amyloid precursor to potentially amyloidogenic derivatives. Science 255: 728–730

    PubMed  Google Scholar 

  18. Grady MS, McLaughlin MR, Christman CW, Valadka AB, Fligner CL, Povlishock JT (1993) The use of antibodies targeted against the neurofilament subunits for the detection of diffuse axonal injury in humans. J Neuropathol Exp Neurol 52: 143–152

    PubMed  Google Scholar 

  19. Gultekin SH, Smith TW (1994) Diffuse axonal injury in craniocerebral trauma: a comparative histologic and immunocytochemical study. Arch Pathol Lab Med 118: 168–171

    PubMed  Google Scholar 

  20. Haass N, Koo EH, Mellon A, Hung AY, Selkoe DJ (1992) Targeting of cell-surface β-amyloid precursor protein to lysosomes: alternative processing into amyloid-bearing fragments. Nature 357: 500–503

    PubMed  Google Scholar 

  21. Hershko A (1991) The ubiquitin pathway for protein degradation. Trends Biochem Sci 16: 265–268

    Article  PubMed  Google Scholar 

  22. Joachim C, Games D, Morris J, Ward P, Frenkel D, Selkoe D (1991) Antibodies to non-beta regions of the beta amyloid precursor protein detect a subset of senile plaques. Am J Pathol 138: 373–384

    PubMed  Google Scholar 

  23. Jortner BS, Scarratt WK, Modransky PD, Perkins SK (1993) Ubiquitin expression in degenerating axons of equine cervical stenotic myelopathy. Soc Neurosci Abstr 765.2

  24. Kawarabayashi T, Shoji M, Yamaguchi H, Tanaka M, Harigaya Y, Ishiguro K, Hirai S (1993) Amyloid β-protein precursor accumulates in swollen neurites throughout rat brain with aging. Neurosci Lett 153: 73–76

    PubMed  Google Scholar 

  25. Kidd M (1963) Paired helical filaments in electron microscopy of Alzheimer's disease. Nature 197: 192–193

    Google Scholar 

  26. Koo EH, Sisodia SS, Archer DR, Martin LJ, Weidemann A, Beyruther K, Fischer P, Masters CL, Price DL (1990) Precursor of amyloid protein in Alzheimer's disease undergoes fast anterograde axonal transport. Proc Natl Acad Sci USA 87: 1561–1565

    PubMed  Google Scholar 

  27. Lee VM-Y, Balin BJ, Otvos L Jr, Trojanowski JQ (1991) A major subunit of paired helical filaments and derivatised forms of normal tau. Science 251: 675–678

    PubMed  Google Scholar 

  28. Li J-Y, Kling-Petersen A, Dahlstrom A (1993) GAP 43-like immunoreactivity in normal adult rat sciatic nerve, spinal cord, and motoneurons: axonal transport and effect of spinal cord transection. Neuroscience 57: 759–776

    Article  PubMed  Google Scholar 

  29. Martin JE, Mather KS, Swash M, Garofalo O, Dale GE, Leigh PN, Anderton BBH (1990) Spinal cord trauma in man: expression. Brain 113: 1553–1562

    PubMed  Google Scholar 

  30. Munoz D (1991) Chromogranin A-like immunoreactive neurites are major constituents of senile plaques. Lab Invest 64: 826–832

    PubMed  Google Scholar 

  31. Ohgami T, Kitamoto T, Tateishi J (1992) Alzheimer's amyloid precursor protein accumulates within axonal swellings in human brain lesions. Neurosci Lett 136: 75–78

    PubMed  Google Scholar 

  32. Oyler GA, Higgins GA, Hart RA, Battenberg E, Billingsby M, Bloom FE, Wilson MC (1989) The identification of a novel synaptosomal-associated protein, SNAP-25, differentially expressed by neuronal subpopulations. J Cell Biol 109: 3039–3052

    Article  PubMed  Google Scholar 

  33. Pappola MA, Omar R, Saran B (1989) The ‘normal’ brain. ‘Abnormal’ ubiquitiated deposits highlight an age-related protein change. Am J Pathol 135: 585–591

    PubMed  Google Scholar 

  34. Povlishock JT (1992) Traumatically induced axonal injury: pathogenesis and pathobiological implications. Brain Pathol 2: 1–12

    PubMed  Google Scholar 

  35. Reid WA, Valler MJ, Kay J (1986) Immunolocalisation of cathepsin D in normal and neoplastic human tissues. J Clin Pathol 39: 1323–1330

    PubMed  Google Scholar 

  36. Roberts GW, Gentleman SM, Lynch A, Murray L, Landon M, Graham DI (1994) β-Amyloid protein deposition in the brain after severe head injury: implications for the pathogenesis of Alzheimer's disease. J Neurol Neurosurg Psychiatry 57: 419–425

    PubMed  Google Scholar 

  37. Schubert W, Prior R, Weidemann A, Dircksen H, Multhaup G, Masters C, Beyreuther K (1991) Localisation of Alzheimer βA4 amyloid precursor protein at central and peripheral synaptic sites. Brain Res 563: 184–194

    Article  PubMed  Google Scholar 

  38. Schweitzer JB, Park MR, Einhaus SL, Robertson JT (1993) Ubiquitin marks the reactive swellings of diffuse axonal injury. Acta Neuropathol 85: 503–507

    PubMed  Google Scholar 

  39. Selkoe DJ (1993) Physiological production of the b-amyloid protein and the mechanism of Alzheimer's disease. Trends Neurosci 16: 403–409

    Article  PubMed  Google Scholar 

  40. Sherriff FE, Bridges LR, Sivaloganathan S (1994) Early detection of axonal injury after human head trauma using immunocytochemistry for β-amyloid precursor protein. Acta Neuropathol 87: 55–62

    Article  PubMed  Google Scholar 

  41. Sherriff FE, Bridges LR, Jackson P (1994) Microwave antigen retrieval enhances β-amyloid precursor protein immunoreactivity in formalin-fixed brain in Alzheimer's disease and after head injury. Neuroreport 5: 1085–1088

    PubMed  Google Scholar 

  42. Skene JHP, Jacobson RD, Snipes GJ, McGuire CB, Norden JJ, Freeman JA (1987) A protein induced during nerve growth (GAP-43) is a major component of growth cone membranes. Science 223: 783–786

    Google Scholar 

  43. Snipes GJ, Chan SY, McGuire CB, Costello BR, Norden JJ, Freeman JA, Routtenberg A (1987) Evidence for the coidentification of GAP-43, a growth-associated protein, and, F1, a plasticity-associated protein. J Neurosci 7: 4066–4075

    PubMed  Google Scholar 

  44. Sollner T, Whiteheart SW, Brunner M, Erdjument-Bromage H, Germanos S, Tempst P, Rothman JE (1993) SNAP receptors implicated in vesicle targeting and fusion. Nature 362: 318–324

    Article  PubMed  Google Scholar 

  45. Somogyi P, Hodgson AJ, DePotter RW, Fischer-Colbrie R, Schober M, Winkler H, Chubb IW (1984) Chromogranin immunoreactivity in the central nervous system, immunocytochemical characterisation and relationship to catecholamine and enkephalin pathways. Brain Res Rev 8: 193–230

    Article  Google Scholar 

  46. Tomlinson DR (1975) Two populations of granular vesicles in constricted post-gangloinic sympathetic nerves. J Physiol (Lond) 245: 727–735

    Google Scholar 

  47. Tsukita S, Ishikawa H (1980) The movement of membraneous organelles in axons, electron microscopic identification of anterogradely and retrogradely transported organelles. J Cell Biol 84: 513–530

    Article  PubMed  Google Scholar 

  48. Viancour TA, Kreiter NA (1993) Vesicular fast axonal transport rates in young and old rat axons. Brain Res 628: 209–217

    Article  PubMed  Google Scholar 

  49. Weidenmann B, Franke WW (1985) Identification and localisation of synaptophysin, an integral membrane glycoprotein of Mr 38,000 characteristic of presynaptic vesicles. Cell 41: 1017–1028

    PubMed  Google Scholar 

  50. Winkler H, Fischer-Colbrie R (1990) Common membrane proteins of chromaffin granules, endocrine and synaptic vesicles: properties, tissue distribution, membrane topography and regulation of synthesis. Neurochem Int 17: 245–262

    Article  Google Scholar 

  51. Wisniewski HM, Narang HK, Corsellis JAN, Terry RD (1976) Ultrastructural studies of the neuropil and neurofibrillary tangles in Alzheimer's disease and post-traumatic dementia. J Neuropathol Exp Neurol 35: 367

    Google Scholar 

  52. Yaghmai A, Povlishock JT (1992) Traumatically induced reactive change as visualised through the use of monoclonal antibodies targeted to neurofilament subunits. J Neuropathol Exp Neurobiol 51: 158–176

    Google Scholar 

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Authors and Affiliations

  1. Department of Clinical Neurology (Neuropathology), Radeliffe Infirmary, University of Oxford, Woodstock Road, OX2 6HE, Oxford, UK

    F. E. Sherriff

  2. Neuropathology Laboratory, Academic Unit of Pathology, University of Leeds, LS2 9JT, Leeds, UK

    F. E. Sherriff, L. R. Bridges, S. Sivaloganathan & S. Wilson

  3. Department of Anatomy and Psychiatry, Charing Cross and Westminster Medical School, St. Dunstan's Road, W6 8RP, London, UK

    S. M. Gentleman

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  1. F. E. Sherriff
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  2. L. R. Bridges
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  3. S. M. Gentleman
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  4. S. Sivaloganathan
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  5. S. Wilson
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Supported by the Home Office Policy Advisory Board for Forensic Pathology (UK)

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Sherriff, F.E., Bridges, L.R., Gentleman, S.M. et al. Markers of axonal injury in post mortem human brain. Acta Neuropathol 88, 433–439 (1994). https://doi.org/10.1007/BF00389495

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  • DOI: https://doi.org/10.1007/BF00389495

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