Gene profiling in spinal cord injury shows role of cell cycle in neuronal death

Ann Neurol. 2003 Apr;53(4):454-68. doi: 10.1002/ana.10472.

Abstract

Spinal cord injury causes secondary biochemical changes leading to neuronal cell death. To clarify the molecular basis of this delayed injury, we subjected rats to spinal cord injury and identified gene expression patterns by high-density oligonucleotide arrays (8,800 genes studied) at 30 minutes, 4 hours, 24 hours, or 7 days after injury (total of 26 U34A profiles). Detailed analyses were limited to 4,300 genes consistently expressed above background. Temporal clustering showed rapid expression of immediate early genes (30 minutes), followed by genes associated with inflammation, oxidative stress, DNA damage, and cell cycle (4 and 24 hours). Functional clustering showed a novel pattern of cell cycle mRNAs at 4 and 24 hours after trauma. Quantitative reverse transcription polymerase chain reaction verified mRNA changes in this group, which included gadd45a, c-myc, cyclin D1 and cdk4, pcna, cyclin G, Rb, and E2F5. Changes in their protein products were quantified by Western blot, and cell-specific expression was determined by immunocytochemistry. Cell cycle proteins showed an increased expression 24 hours after injury and were, in part, colocalized in neurons showing morphological evidence of apoptosis. These findings suggest that cell cycle-related genes, induced after spinal cord injury, are involved in neuronal damage and subsequent cell death.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Cell Cycle / genetics
  • Cell Death / genetics*
  • Gene Expression Profiling*
  • Genes, Immediate-Early / genetics
  • Immunoblotting
  • Immunohistochemistry
  • In Situ Nick-End Labeling
  • Male
  • Myelitis / genetics
  • Myelitis / pathology
  • Myelitis / physiopathology
  • Neurons / pathology*
  • Neurons / physiology
  • Oxidative Stress / genetics
  • Rats
  • Rats, Sprague-Dawley
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Spinal Cord Injuries / genetics*
  • Spinal Cord Injuries / pathology*
  • Spinal Cord Injuries / physiopathology
  • Up-Regulation / genetics