Severe global cerebral ischemia-induced programmed necrosis of hippocampal CA1 neurons in rat is prevented by 3-methyladenine: a widely used inhibitor of autophagy

J Neuropathol Exp Neurol. 2011 Apr;70(4):314-22. doi: 10.1097/NEN.0b013e31821352bd.

Abstract

The true programmed mechanisms of delayed neuronal death induced by global cerebral ischemia/reperfusion injury remain incompletely characterized. Autophagic cell death and programmed necrosis are 2 kinds of programmed cell death distinct from apoptosis. Here, we studied the death mechanisms of hippocampal cornu ammonis 1 neuronal death after a 20-minute severe global ischemia/reperfusion injury in young adult rats and the effects of 3-methyladenine (3-MA), a widely used inhibitor of autophagy. The morphological changes detected by electron microscopy, together with the activation of autophagy, transferase-mediated UTP nick end-labeling-positive neurons, and delayed death, demonstrated that cornu ammonis 1 neuronal death induced in this paradigm was programmed necrosis. No significant activation of caspase-3 after injury was detected by Western blot and immunohistochemistry. Treatment with 3-MA provided time-dependent protection against cornu ammonis 1 neuronal death at 7 days of reperfusion when it was administered before ischemia; administration 60 minutes after reperfusion was not beneficial. The redistribution of the lysosomal enzyme cathepsin B after injury was inhibited by 3-MA administered before ischemia, suggesting that this might be another important mechanism for the protective effect of 3-MA in ischemic neuronal injury.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenine / analogs & derivatives*
  • Adenine / pharmacology
  • Animals
  • Autophagy / drug effects
  • Blotting, Western
  • Brain Ischemia / pathology*
  • Brain Ischemia / prevention & control*
  • CA1 Region, Hippocampal / pathology*
  • Caspase 3 / metabolism
  • Cathepsin B / metabolism
  • Cell Count
  • Cell Death / drug effects
  • Immunohistochemistry
  • In Situ Nick-End Labeling
  • Male
  • Microscopy, Electron, Transmission
  • Necrosis / pathology
  • Necrosis / prevention & control
  • Neurons / pathology*
  • Rats
  • Rats, Sprague-Dawley

Substances

  • 3-methyladenine
  • Caspase 3
  • Cathepsin B
  • Adenine