Apoptosis after traumatic human spinal cord injury

View More View Less
  • 1 Department of Neurological Surgery and the Miami Project to Cure Paralysis, and Department of Physiology and Biophysics, University of Miami School of Medicine, Miami, Florida; and IDUN Pharmaceuticals, Inc., La Jolla, California
Restricted access

Purchase Now

USD  $45.00

JNS + Pediatrics - 1 year subscription bundle (Individuals Only)

USD  $505.00

JNS + Pediatrics + Spine - 1 year subscription bundle (Individuals Only)

USD  $600.00
Print or Print + Online

Object. Apoptosis is a form of programmed cell death seen in a variety of developmental and disease states, including traumatic injuries. The main objective of this study was to determine whether apoptosis is observed after human spinal cord injury (SCI). The spatial and temporal expression of apoptotic cells as well as the nature of the cells involved in programmed cell death were also investigated.

Methods. The authors examined the spinal cords of 15 patients who died between 3 hours and 2 months after a traumatic SCI. Apoptotic cells were found at the edges of the lesion epicenter and in the adjacent white matter, particularly in the ascending tracts, by using histological (cresyl violet, hematoxylin and eosin) and nuclear staining (Hoechst 33342). The presence of apoptotic cells was supported by staining with the terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick-end labeling technique and confirmed by immunostaining for the processed form of caspase-3 (CPP-32), a member of the interleukin-1β-converting enzyme/Caenorhabditis elegans D 3 (ICE/CED-3) family of proteases that plays an essential role in programmed cell death. Apoptosis in this series of human SCIs was a prominent pathological finding in 14 of the 15 spinal cords examined when compared with five uninjured control spinal cords. To determine the type of cells undergoing apoptosis, the authors immunostained specimens with a variety of antibodies, including glial fibrillary acidic protein, 2′,3′-cyclic nucleotide 3′-phosphohydrolase (CNPase), and CD45/68. Oligodendrocytes stained with CNPase and a number of apoptotic nuclei colocalized with positive staining for this antibody.

Conclusions. These results support the hypothesis that apoptosis occurs in human SCIs and is accompanied by the activation of caspase-3 of the cysteine protease family. This mechanism of cell death contributes to the secondary injury processes seen after human SCI and may have important clinical implications for the further development of protease inhibitors to prevent programmed cell death.

JNS + Pediatrics - 1 year subscription bundle (Individuals Only)

USD  $505.00

JNS + Pediatrics + Spine - 1 year subscription bundle (Individuals Only)

USD  $600.00

Contributor Notes

Address for Dr. Emery: Hôpital Beaujon, Clichy, France.Address reprint requests to: Allan D. O. Levi, M.D., The Miami Project to Cure Paralysis, 1600 NW 10th Avenue, R-48, Miami, Florida 33136. email: alevi@mednet.med.miami.edu.
  • 1.

    Adams JC: Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains. J Histochem Cytochem 40:14571463, 1992 Adams JC: Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains. J Histochem Cytochem 40:1457–1463, 1992

    • Search Google Scholar
    • Export Citation
  • 2.

    Alnemri ES, , Livingston DJ, & Nicholson DW, et al: Human ICE/CED-3 protease nomenclature. Cell 87:171, 1996 (Letter) Alnemri ES, Livingston DJ, Nicholson DW, et al: Human ICE/CED-3 protease nomenclature. Cell 87:171, 1996 (Letter)

    • Search Google Scholar
    • Export Citation
  • 3.

    Arends MJ, & Wyllie AH: Apoptosis: mechanisms and roles in pathology. Int Rev Exp Pathol 32:223254, 1991 Arends MJ, Wyllie AH: Apoptosis: mechanisms and roles in pathology. Int Rev Exp Pathol 32:223–254, 1991

    • Search Google Scholar
    • Export Citation
  • 4.

    Armstrong RC, , Aja TJ, & Hoang KD, et al: Activation of the CED3/ICE-related protease CPP32 in cerebellar granule neurons undergoing apoptosis but not necrosis. J Neurosci 17:553562, 1997 Armstrong RC, Aja TJ, Hoang KD, et al: Activation of the CED3/ICE-related protease CPP32 in cerebellar granule neurons undergoing apoptosis but not necrosis. J Neurosci 17:553–562, 1997

    • Search Google Scholar
    • Export Citation
  • 5.

    Barres BA, , Jacobson MD, & Schmid R: Does oligodendrocyte survival depend on axons? Curr Biol 3:489497, 1993 Barres BA, Jacobson MD, Schmid R: Does oligodendrocyte survival depend on axons? Curr Biol 3:489–497, 1993

    • Search Google Scholar
    • Export Citation
  • 6.

    Bauer J, , Wekerle H, & Lassmann H: Apoptosis in brain-specific autoimmune disease. Curr Opin Immunol 7:839843, 1995 Bauer J, Wekerle H, Lassmann H: Apoptosis in brain-specific autoimmune disease. Curr Opin Immunol 7:839–843, 1995

    • Search Google Scholar
    • Export Citation
  • 7.

    Blight AR: Effect of 4-aminopyridine on axonal conduction-block in chronic spinal cord injury. Brain Res Bull 22:4752, 1989 Blight AR: Effect of 4-aminopyridine on axonal conduction-block in chronic spinal cord injury. Brain Res Bull 22:47–52, 1989

    • Search Google Scholar
    • Export Citation
  • 8.

    Bracken MB, , Shepard MJ, & Collins WF, et al: A randomized, controlled trial of methylprednisolone or naloxone in the treatment of acute spinal-cord injury. Results of the second National Acute Spinal Cord Injury Study. N Engl J Med 322:14051411, 1990 Bracken MB, Shepard MJ, Collins WF, et al: A randomized, controlled trial of methylprednisolone or naloxone in the treatment of acute spinal-cord injury. Results of the second National Acute Spinal Cord Injury Study. N Engl J Med 322:1405–1411, 1990

    • Search Google Scholar
    • Export Citation
  • 9.

    Bunge RP, , Puckett WR, & Becerra JL, et al: Observations on the pathology of human spinal cord injury. A review and classification of 22 new cases with details from a case of chronic cord compression with extensive focal demyelination. Adv Neurol 59:7589, 1993 Bunge RP, Puckett WR, Becerra JL, et al: Observations on the pathology of human spinal cord injury. A review and classification of 22 new cases with details from a case of chronic cord compression with extensive focal demyelination. Adv Neurol 59:75–89, 1993

    • Search Google Scholar
    • Export Citation
  • 10.

    Cohen JJ: Apoptosis. Immunol Today 14:126130, 1993 Cohen JJ: Apoptosis. Immunol Today 14:126–130, 1993

  • 11.

    Crowe MJ, , Bresnahan JC, & Shuman SL: Apoptosis and delayed degeneration after spinal cord injury in rats and monkeys. Nature Med 3:7376, 1997 Crowe MJ, Bresnahan JC, Shuman SL: Apoptosis and delayed degeneration after spinal cord injury in rats and monkeys. Nature Med 3:73–76, 1997

    • Search Google Scholar
    • Export Citation
  • 12.

    Crowe MJ, , Shuman SL, & Masters JN, et al: Morphological evidence suggesting apoptotic nuclei in spinal cord injury. Soc Neurosci Abstr 21:232, 1995 (Abstract) Crowe MJ, Shuman SL, Masters JN, et al: Morphological evidence suggesting apoptotic nuclei in spinal cord injury. Soc Neurosci Abstr 21:232, 1995 (Abstract)

    • Search Google Scholar
    • Export Citation
  • 13.

    Dusart I, & Schwab ME: Secondary cell death and the inflammatory reaction after dorsal hemisection of the rat spinal cord. Eur J Neurosci 6:712724, 1994 Dusart I, Schwab ME: Secondary cell death and the inflammatory reaction after dorsal hemisection of the rat spinal cord. Eur J Neurosci 6:712–724, 1994

    • Search Google Scholar
    • Export Citation
  • 14.

    Duvall E, , Wyllie AH, & Morris RG: Macrophage recognition of cells undergoing programmed cell death (apoptosis). Immunology 56:351358, 1985 Duvall E, Wyllie AH, Morris RG: Macrophage recognition of cells undergoing programmed cell death (apoptosis). Immunology 56:351–358, 1985

    • Search Google Scholar
    • Export Citation
  • 15.

    Ellis RE, , Yvan J, & Horvitz HR: Mechanisms and functions of cell death. Annu Rev Cell Biol 7:663698, 1991 Ellis RE, Yvan J, Horvitz HR: Mechanisms and functions of cell death. Annu Rev Cell Biol 7:663–698, 1991

    • Search Google Scholar
    • Export Citation
  • 16.

    Enari M, , Talanian RV, & Wong WW, et al: Sequential activation of ICE-like and CPP32-like proteases during Fas-mediated apoptosis. Nature 380:723726, 1996 (Letter) Enari M, Talanian RV, Wong WW, et al: Sequential activation of ICE-like and CPP32-like proteases during Fas-mediated apoptosis. Nature 380:723–726, 1996 (Letter)

    • Search Google Scholar
    • Export Citation
  • 17.

    Fraser A, & Evan G: A license to kill. Cell 85:781784, 1996 Fraser A, Evan G: A license to kill. Cell 85:781–784, 1996

  • 18.

    Furukawa K, , Estus S, & Fu WM, et al: Neuroprotective action of cycloheximide involves induction of Bcl-2 and antioxidant pathways. J Cell Biol 136:11371149, 1997 Furukawa K, Estus S, Fu WM, et al: Neuroprotective action of cycloheximide involves induction of Bcl-2 and antioxidant pathways. J Cell Biol 136:1137–1149, 1997

    • Search Google Scholar
    • Export Citation
  • 19.

    Gavrieli Y, , Sherman Y, & Ben-Sasson SA: Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119:493501, 1992 Gavrieli Y, Sherman Y, Ben-Sasson SA: Identification of programmed cell death in situ via specific labeling of nuclear DNA fragmentation. J Cell Biol 119:493–501, 1992

    • Search Google Scholar
    • Export Citation
  • 20.

    Gehrmann J, & Banati RB: Microglial turnover in the injured CNS: activated microglia undergo delayed DNA fragmentation following peripheral nerve injury. J Neuropathol Exp Neurol 54:680688, 1995 Gehrmann J, Banati RB: Microglial turnover in the injured CNS: activated microglia undergo delayed DNA fragmentation following peripheral nerve injury. J Neuropathol Exp Neurol 54:680–688, 1995

    • Search Google Scholar
    • Export Citation
  • 21.

    Hall ED, , Yonkers PA, & Andrus PK, et al: Biochemistry and pharmacology of lipid antioxidants in acute brain and spinal cord injury. J Neurotrauma 9 (Suppl):S425S442, 1992 Hall ED, Yonkers PA, Andrus PK, et al: Biochemistry and pharmacology of lipid antioxidants in acute brain and spinal cord injury. J Neurotrauma 9 (Suppl):S425–S442, 1992

    • Search Google Scholar
    • Export Citation
  • 22.

    Hamburger V: Cell death in the development of the lateral motor column of the chick embryo. J Comp Neurol 160:535546, 1975 Hamburger V: Cell death in the development of the lateral motor column of the chick embryo. J Comp Neurol 160:535–546, 1975

    • Search Google Scholar
    • Export Citation
  • 23.

    Hansebout RR, , Blight AR, & Fawcett S, et al: 4-Aminopyridine in chronic spinal cord injury: a controlled, double-blind, crossover study in eight patients. J Neurotrauma 10:118, 1993 Hansebout RR, Blight AR, Fawcett S, et al: 4-Aminopyridine in chronic spinal cord injury: a controlled, double-blind, crossover study in eight patients. J Neurotrauma 10:1–18, 1993

    • Search Google Scholar
    • Export Citation
  • 24.

    Hara H, , Friedlander RM, & Gagliardini V, et al: Inhibition of interleukin 1β converting enzyme family proteases reduce ischemic and excitotoxic neuronal damage. Proc Natl Acad Sci USA 94:20072012, 1997 Hara H, Friedlander RM, Gagliardini V, et al: Inhibition of interleukin 1β converting enzyme family proteases reduce ischemic and excitotoxic neuronal damage. Proc Natl Acad Sci USA 94:2007–2012, 1997

    • Search Google Scholar
    • Export Citation
  • 25.

    Henderson CE: Programmed cell death in the developing nervous system. Neuron 17:579585, 1997 Henderson CE: Programmed cell death in the developing nervous system. Neuron 17:579–585, 1997

    • Search Google Scholar
    • Export Citation
  • 26.

    Hisahara S, , Shoji S, & Okano H, et al: ICE/CED-3 family executes oligodendrocyte apoptosis by tumor necrosis factor. J Neurochem 69:1020, 1997 Hisahara S, Shoji S, Okano H, et al: ICE/CED-3 family executes oligodendrocyte apoptosis by tumor necrosis factor. J Neurochem 69:10–20, 1997

    • Search Google Scholar
    • Export Citation
  • 27.

    Hockenberry D: Defining apoptosis. Am J Pathol 146:1619, 1995 Hockenberry D: Defining apoptosis. Am J Pathol 146:16–19, 1995

  • 28.

    Hockenberry DM, , Oltvai ZN, & Yin XM, et al: Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell 75:241251, 1993 Hockenberry DM, Oltvai ZN, Yin XM, et al: Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell 75:241–251, 1993

    • Search Google Scholar
    • Export Citation
  • 29.

    Horwitz HR, , Ellis HM, & Sternberg DW: Programmed cell death in nematode development. Neurosci Comment 1:5665, 1982 Horwitz HR, Ellis HM, Sternberg DW: Programmed cell death in nematode development. Neurosci Comment 1:56–65, 1982

    • Search Google Scholar
    • Export Citation
  • 30.

    Jänicke RU, , Sprengart ML, & Wati MR, et al: Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. J Biol Chem 273:93579360, 1997 Jänicke RU, Sprengart ML, Wati MR, et al: Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis. J Biol Chem 273:9357–9360, 1997

    • Search Google Scholar
    • Export Citation
  • 31.

    Kane AB: Redefining cell death. Am J Pathol 146:12, 1995 Kane AB: Redefining cell death. Am J Pathol 146:1–2, 1995

  • 32.

    Katoh K, , Ikata T, & Katoh S, et al: Induction and its spread of apoptosis in rat spinal cord after mechanical trauma. Neurosci Lett 216:912, 1996 Katoh K, Ikata T, Katoh S, et al: Induction and its spread of apoptosis in rat spinal cord after mechanical trauma. Neurosci Lett 216:9–12, 1996

    • Search Google Scholar
    • Export Citation
  • 33.

    Keane RW, , Srinivasan A, & Foster LM, et al: Activation of CPP32 during apoptosis of neurons and astrocytes. J Neurosci Res 48:168180, 1997 Keane RW, Srinivasan A, Foster LM, et al: Activation of CPP32 during apoptosis of neurons and astrocytes. J Neurosci Res 48:168–180, 1997

    • Search Google Scholar
    • Export Citation
  • 34.

    Kuida K, , Zheng TS, & Na S, et al: Decreased apoptosis in the brain and premature lethality in CPP32-deficient mice. Nature 384:368372, 1996 Kuida K, Zheng TS, Na S, et al: Decreased apoptosis in the brain and premature lethality in CPP32-deficient mice. Nature 384:368–372, 1996

    • Search Google Scholar
    • Export Citation
  • 35.

    Li GI, , Brodin G, & Farooque M, et al: Apoptosis and expression of Bcl-2 after compression trauma to rat spinal cord. J Neuropathol Exp Neurol 55:280289, 1996 Li GI, Brodin G, Farooque M, et al: Apoptosis and expression of Bcl-2 after compression trauma to rat spinal cord. J Neuropathol Exp Neurol 55:280–289, 1996

    • Search Google Scholar
    • Export Citation
  • 36.

    Liu XZ, , Xu XM, & Hu R, et al: Neuronal and glial apoptosis after traumatic spinal cord injury. J Neurosci 17:53955406, 1997 Liu XZ, Xu XM, Hu R, et al: Neuronal and glial apoptosis after traumatic spinal cord injury. J Neurosci 17:5395–5406, 1997

    • Search Google Scholar
    • Export Citation
  • 37.

    Majno G, & Joris I: Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol 146:315, 1995 Majno G, Joris I: Apoptosis, oncosis, and necrosis. An overview of cell death. Am J Pathol 146:3–15, 1995

    • Search Google Scholar
    • Export Citation
  • 38.

    Martin SJ, & Green DR: Protease activation during apoptosis: death by a thousand cuts. Cell 82:349352, 1995 Martin SJ, Green DR: Protease activation during apoptosis: death by a thousand cuts. Cell 82:349–352, 1995

    • Search Google Scholar
    • Export Citation
  • 39.

    Milligan CE, , Prevette D, & Yaginuma H, et al: Peptide inhibitors of the ICE protease family arrest programmed cell death of motoneurons in vivo and in vitro. Neuron 15:385393, 1995 Milligan CE, Prevette D, Yaginuma H, et al: Peptide inhibitors of the ICE protease family arrest programmed cell death of motoneurons in vivo and in vitro. Neuron 15:385–393, 1995

    • Search Google Scholar
    • Export Citation
  • 40.

    Nagata S: Apoptosis by death factor. Cell 88:355365, 1997 Nagata S: Apoptosis by death factor. Cell 88:355–365, 1997

  • 41.

    Namura S, , Zhu J, & Fink K, et al: Activation and cleavage of caspase-3 in apoptosis induced by experimental cerebral ischemia. J Neurosci 18:36593668, 1998 Namura S, Zhu J, Fink K, et al: Activation and cleavage of caspase-3 in apoptosis induced by experimental cerebral ischemia. J Neurosci 18:3659–3668, 1998

    • Search Google Scholar
    • Export Citation
  • 42.

    Nguyen KB, , McCombe PA, & Pender MP: Macrophage apoptosis in the central nervous system in experimental autoimmune encephalomyelitis. J Autoimmun 17:145152, 1994 Nguyen KB, McCombe PA, Pender MP: Macrophage apoptosis in the central nervous system in experimental autoimmune encephalomyelitis. J Autoimmun 17:145–152, 1994

    • Search Google Scholar
    • Export Citation
  • 43.

    Nicotera P, , Ankarcrona M, & Bonfoco E, et al: Neuronal necrosis and apoptosis: two distinct events induced by exposure to glutamate or oxidative stress. Adv Neurol 72:95101, 1997 Nicotera P, Ankarcrona M, Bonfoco E, et al: Neuronal necrosis and apoptosis: two distinct events induced by exposure to glutamate or oxidative stress. Adv Neurol 72:95–101, 1997

    • Search Google Scholar
    • Export Citation
  • 44.

    Pender MP, , Nguyen KB, & McCombe PA, et al: Apoptosis in the nervous system in experimental allergic encephalomyelitis. J Neurol Sci 104:8187, 1991 Pender MP, Nguyen KB, McCombe PA, et al: Apoptosis in the nervous system in experimental allergic encephalomyelitis. J Neurol Sci 104:81–87, 1991

    • Search Google Scholar
    • Export Citation
  • 45.

    Prineas JW, , Kwon EE, & Goldenberg PZ, et al: Multiple sclerosis. Oligodendrocyte proliferation and differentiation in fresh lesions. Lab Invest 61:489503, 1989 Prineas JW, Kwon EE, Goldenberg PZ, et al: Multiple sclerosis. Oligodendrocyte proliferation and differentiation in fresh lesions. Lab Invest 61:489–503, 1989

    • Search Google Scholar
    • Export Citation
  • 46.

    Rink A, , Fung KM, & Trojanowski JQ, et al: Evidence of apoptotic cell death after experimental traumatic brain injury in the rat. Am J Pathol 147:15751583, 1995 Rink A, Fung KM, Trojanowski JQ, et al: Evidence of apoptotic cell death after experimental traumatic brain injury in the rat. Am J Pathol 147:1575–1583, 1995

    • Search Google Scholar
    • Export Citation
  • 47.

    Savitz SI, & Rosenbaum DM: Apoptosis in neurological disease. Neurosurgery 42:555574, 1998 Savitz SI, Rosenbaum DM: Apoptosis in neurological disease. Neurosurgery 42:555–574, 1998

    • Search Google Scholar
    • Export Citation
  • 48.

    Shuman SL, , Bresnahan JC, & Beattie NS: Apoptosis of microglia and oligodendrocytes after spinal cord injury in rats. J Neurosci Res 50:798808, 1997 Shuman SL, Bresnahan JC, Beattie NS: Apoptosis of microglia and oligodendrocytes after spinal cord injury in rats. J Neurosci Res 50:798–808, 1997

    • Search Google Scholar
    • Export Citation
  • 49.

    Tator CH, & Fehlings MG: Review of the secondary injury theory of acute spinal cord trauma with emphasis on vascular mechanisms. J Neurosurg 75:1526, 1991 Tator CH, Fehlings MG: Review of the secondary injury theory of acute spinal cord trauma with emphasis on vascular mechanisms. J Neurosurg 75:15–26, 1991

    • Search Google Scholar
    • Export Citation
  • 50.

    Thompson CB: Apoptosis in the pathogenesis and treatment of disease. Science 267:14561462, 1995 Thompson CB: Apoptosis in the pathogenesis and treatment of disease. Science 267:1456–1462, 1995

    • Search Google Scholar
    • Export Citation
  • 51.

    Vela JM, , Dalmau I, & Gonzáles B, et al: The microglial reaction in spinal cords of jimpy mice is related to apoptotic oligodendrocytes. Brain Res 712:134142, 1996 Vela JM, Dalmau I, Gonzáles B, et al: The microglial reaction in spinal cords of jimpy mice is related to apoptotic oligodendrocytes. Brain Res 712:134–142, 1996

    • Search Google Scholar
    • Export Citation
  • 52.

    Weedon D, , Searle J, & Kerr JF: Apoptosis. Its nature and implications for dermatopathology. Am J Dermatopathol 1:133144, 1979 Weedon D, Searle J, Kerr JF: Apoptosis. Its nature and implications for dermatopathology. Am J Dermatopathol 1:133–144, 1979

    • Search Google Scholar
    • Export Citation
  • 53.

    Wood PL: Microglia as a unique cellular target in the treatment of stroke: potential neurotoxic mediators produced by activated microglia. Neurol Res 17:242248, 1995 Wood PL: Microglia as a unique cellular target in the treatment of stroke: potential neurotoxic mediators produced by activated microglia. Neurol Res 17:242–248, 1995

    • Search Google Scholar
    • Export Citation
  • 54.

    Yakovlev AG, , Knoblach SM, & Fan L, et al: Activation of CPP32-like caspases contributes to neuronal apoptosis and neurological dysfunction after traumatic brain injury. J Neurosci 17:74157424, 1997 Yakovlev AG, Knoblach SM, Fan L, et al: Activation of CPP32-like caspases contributes to neuronal apoptosis and neurological dysfunction after traumatic brain injury. J Neurosci 17:7415–7424, 1997

    • Search Google Scholar
    • Export Citation
  • 55.

    Young W: Secondary injury mechanisms in acute spinal cord injury. J Emerg Med 11 (Suppl 1):1322, 1993 Young W: Secondary injury mechanisms in acute spinal cord injury. J Emerg Med 11 (Suppl 1):13–22, 1993

    • Search Google Scholar
    • Export Citation

Metrics

All Time Past Year Past 30 Days
Abstract Views 712 464 70
Full Text Views 254 42 1
PDF Downloads 81 17 0
EPUB Downloads 0 0 0