Evidence for apoptosis after intercerebral hemorrhage in rat striatum

J Cereb Blood Flow Metab. 2000 Feb;20(2):396-404. doi: 10.1097/00004647-200002000-00022.


The overall hypothesis that cell death after intracerebral hemorrhage is mediated in part by apoptotic mechanisms was tested. Intracerebral hemorrhage was induced in rats using stereotactic infusions of 0.5 U of collagenase (1-microL volume) into the striatum. After 24 hours, large numbers of TUNEL-positive stained cells with morphologies suggestive of apoptosis were present in the center and periphery of the hemorrhage. Double staining with Nissl and immunocytochemical labeling with antibodies against neuronal nuclei and glial fibrillary acidic protein suggested that these TUNEL-positive cells were mostly neurons and astrocytes. Electrophoresis of hemorrhagic brain extracts showed evidence of DNA laddering into approximately 200-bp fragments. Western blots showed cleavage of the cytosolic caspase substrate gelsolin. The density of TUNEL-positive cells at 24 and 48 hours after hemorrhage was significantly reduced by treatment with the broad-spectrum caspase inhibitor zVADfmk. It was unlikely that apoptotic changes were due to neurotoxicity of injected collagenase because TUNEL-positive cells and DNA laddering were also obtained in an alternative model of hemorrhage where autologous blood was infused into the striatum. Furthermore, equivalent doses of collagenase did not induce cell death in primary neuronal cultures. These results provide initial evidence that apoptotic mechanisms may mediate some of the injury in brain after intracerebral hemorrhage.

Publication types

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

MeSH terms

  • Amino Acid Chloromethyl Ketones / pharmacology
  • Animals
  • Apoptosis*
  • Astrocytes / chemistry
  • Basal Ganglia / blood supply*
  • Basal Ganglia / enzymology
  • Basal Ganglia / pathology*
  • Basal Ganglia Hemorrhage / metabolism
  • Basal Ganglia Hemorrhage / pathology*
  • Caspases / analysis
  • Caspases / metabolism
  • Cell Count
  • Cells, Cultured
  • Collagenases / metabolism
  • DNA / analysis
  • Gelsolin / analysis
  • Gelsolin / metabolism
  • Glial Fibrillary Acidic Protein / analysis
  • In Situ Nick-End Labeling
  • Male
  • Neurons / cytology
  • Neurons / drug effects
  • Neurons / enzymology
  • Neuroprotective Agents / pharmacology
  • Rats
  • Rats, Sprague-Dawley


  • Amino Acid Chloromethyl Ketones
  • Gelsolin
  • Glial Fibrillary Acidic Protein
  • Neuroprotective Agents
  • benzyloxycarbonylvalyl-alanyl-aspartyl fluoromethyl ketone
  • DNA
  • Caspases
  • Collagenases