Temporal profile of cytochrome c and caspase-3 immunoreactivities and TUNEL staining after permanent middle cerebral artery occlusion in rats

Chihoko Sasaki, Hisashi Kitagawa, Wen Ri Zhang, Hitoshi Warita, Kenichi Sakai, Abe Koji

Research output: Contribution to journalArticlepeer-review

60 Citations (Scopus)

Abstract

Although apoptotic pathways play important roles in ischemic neuronal injury, exact mechanism of apoptotic enzyme cascade has not been fully studied. Immunohistochemical stainings for cytochrome c and caspase-3, and histochemical staining for a terminal deoxynucleotidyl-transferase (TdT)- mediated dUTP-biotin nick end-labeling method (TUNEL) were examined in a rat model of permanent middle cerebral artery (MCA) occlusion. Cytochrome c was strongly induced in neurons of the ischemic penumbra from 3 h after MCA occlusion, and caspase-3 began to be induced in the same area from 3 h with a peak at 8 h. Neuronal cells in MCA area became TUNEL positive at delayed time, reaching a peak at 24 h. Thus, the peak of induction of cytochrome c preceded that of caspase-3, and these two peaks were also precedence of the peak of DNA-fragmentation. Western blot analysis showed cytosolic expression of cytochrome c from mitochondria. This study demonstrated 1. Rapid release of cytochrome c from mitochondria to the cytosol, mainly in neurons of the cortex at 3 h after ischemia. 2. Subsequent peaks of caspase-3 and TUNEL in this order. These temporal profiles suggest a serial cascadic activation of apoptotic pathways in neuronal death after permanent MCA occlusion of rats.

Original languageEnglish
Pages (from-to)223-228
Number of pages6
JournalNeurological Research
Volume22
Issue number2
DOIs
Publication statusPublished - Jan 1 2000

Keywords

  • Apoptosis
  • Caspase-3
  • Cytochrome c
  • Permanent MCA occlusion
  • TUNEL
  • Western blotting

ASJC Scopus subject areas

  • Neurology
  • Clinical Neurology

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