Temporal relationship of peroxynitrite-induced oxidative damage, calpain-mediated cytoskeletal degradation and neurodegeneration after traumatic brain injury

Exp Neurol. 2007 May;205(1):154-65. doi: 10.1016/j.expneurol.2007.01.023. Epub 2007 Feb 3.


We assessed the temporal and spatial characteristics of PN-induced oxidative damage and its relationship to calpain-mediated cytoskeletal degradation and neurodegeneration in a severe unilateral controlled cortical impact (CCI) traumatic brain injury (TBI) model. Quantitative temporal time course studies were performed to measure two oxidative damage markers: 3-nitrotyrosine (3NT) and 4-hydroxynonenal (4HNE) at 30 min, 1, 3, 6, 12, 24, 48, 72 h and 7 days after injury in ipsilateral cortex of young adult male CF-1 mice. Secondly, the time course of Ca(++)-activated, calpain-mediated proteolysis was also analyzed using quantitative western-blot measurement of breakdown products of the cytoskeletal protein alpha-spectrin. Finally, the time course of neurodegeneration was examined using de Olmos silver staining. Both oxidative damage markers increased in cortical tissue immediately after injury (30 min) and elevated for the first 3-6 h before returning to baseline. In the immunostaining study, the PN-selective marker, 3NT, and the lipid peroxidation marker, 4HNE, were intense and overlapping in the injured cortical tissue. alpha-Spectrin breakdown products, which were used as biomarker for calpain-mediated cytoskeletal degradation, were also increased after injury, but the time course lagged behind the peak of oxidative damage and did not reach its maximum until 24 h post-injury. In turn, cytoskeletal degradation preceded the peak of neurodegeneration which occurred at 48 h post-injury. These studies have led us to the hypothesis that PN-mediated oxidative damage is an early event that contributes to a compromise of Ca(++) homeostatic mechanisms which causes a massive Ca(++) overload and calpain activation which is a final common pathway that results in post-traumatic neurodegeneration.

MeSH terms

  • Aldehydes / metabolism
  • Animals
  • Brain / metabolism
  • Brain / pathology
  • Brain Injuries / complications*
  • Brain Injuries / metabolism*
  • Brain Injuries / pathology
  • Calcium / metabolism
  • Calpain / metabolism*
  • Cerebral Cortex / injuries
  • Cerebral Cortex / metabolism
  • Cerebral Cortex / pathology
  • Cytoskeleton / pathology*
  • Lipid Peroxidation
  • Male
  • Mice
  • Mice, Inbred Strains
  • Nerve Degeneration / etiology*
  • Nerve Degeneration / metabolism
  • Nerve Degeneration / pathology
  • Nerve Tissue Proteins / metabolism
  • Nitrates / metabolism
  • Oxidative Stress*
  • Peroxynitrous Acid / metabolism*
  • Spectrin / metabolism
  • Time Factors
  • Tissue Distribution
  • Tyrosine / analogs & derivatives
  • Tyrosine / metabolism


  • Aldehydes
  • Nerve Tissue Proteins
  • Nitrates
  • Spectrin
  • Peroxynitrous Acid
  • 3-nitrotyrosine
  • Tyrosine
  • Calpain
  • 4-hydroxy-2-nonenal
  • Calcium