Early processing of Bid and caspase-6, -8, -10, -14 in the canine brain during cardiac arrest and resuscitation

Exp Neurol. 2004 Oct;189(2):261-79. doi: 10.1016/j.expneurol.2004.05.020.


A clinically relevant model of transient global brain ischemia involving cardiac arrest followed by resuscitation in dogs was utilized to study the expression and proteolytic processing of apoptosis-regulatory proteins. In the hippocampus, an increase in pro-apoptotic Bcl-2 family proteins Bcl-XS and Bak was detected, concomitant with proteolysis of Bcl-XL and Bcl-2, following ischemia-reperfusion injury. Also, biphasic cleavage of Bid was found in this region of the brain, with early generation of tBid-p11 within 10 min of cardiac arrest, followed by generation of tBid-p15 within 30-min reperfusion, consistent with activation of this pro-apoptotic protein. In addition, cardiac arrest and resuscitation induced early, reperfusion-dependent proteolytic processing of pro-caspase-6, -8, -10, and -14, which preceded caspase-3 activation. Immunohistochemical analysis using antibodies, which preferentially recognize processed caspase-3, -6, -8, and -10, provided evidence of time-dependent activation of these proteases in both neurons and glia in ischemia-sensitive regions of the brain. In conclusion, extremely rapid, cell-selective processing of apoptosis-regulatory proteins occurs in a clinically relevant model of ischemic brain injury caused by cardiac arrest and resuscitation. The early cleavage of Bid and rapid depletion of 32-kDa pro-caspase-14 from the canine hippocampus after induction of ischemia suggests the involvement of calpains in the processing of these proteins. Demonstration of in vitro cleavage of recombinant mouse caspase-14 by calpain I in the present study lends support to this hypothesis, further implicating cross-talk between different protease families in the pathophysiology of ischemic neural cell death.

Publication types

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

MeSH terms

  • Animals
  • Apoptosis / physiology*
  • BH3 Interacting Domain Death Agonist Protein
  • Brain Ischemia / enzymology*
  • Brain Ischemia / physiopathology
  • Calpain / metabolism
  • Carrier Proteins / metabolism*
  • Caspase 14
  • Caspases / metabolism*
  • Disease Models, Animal
  • Dogs
  • Female
  • Heart Arrest, Induced
  • Hippocampus / enzymology*
  • Hippocampus / physiopathology
  • Membrane Proteins / metabolism
  • Nerve Degeneration / enzymology
  • Nerve Degeneration / physiopathology
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • Reaction Time / physiology
  • Reperfusion Injury / enzymology*
  • Reperfusion Injury / physiopathology
  • Resuscitation
  • bcl-2 Homologous Antagonist-Killer Protein
  • bcl-X Protein


  • BH3 Interacting Domain Death Agonist Protein
  • Carrier Proteins
  • Membrane Proteins
  • Proto-Oncogene Proteins c-bcl-2
  • bcl-2 Homologous Antagonist-Killer Protein
  • bcl-X Protein
  • Calpain
  • Casp14 protein, mouse
  • Caspase 14
  • Caspases