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The Journal of Neuroscience, July 1, 1998, 18(13):4914-4928

Induction of Caspase-3-Like Protease May Mediate Delayed Neuronal Death in the Hippocampus after Transient Cerebral Ischemia

Jun Chen1, Tetsuya Nagayama1, Kunlin Jin1, R. Anne Stetler1, 2, Raymond L. Zhu1, Steven H. Graham1, 2, and Roger P. Simon1

1 Department of Neurology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania 15213, and 2 Neurology Service, Department of Veterans Affairs Medical Center, Pittsburgh, Pennsylvania 15261

Delayed neuronal death after transient cerebral ischemia may be mediated, in part, by the induction of apoptosis-regulatory gene products. Caspase-3 is a newly characterized mammalian cysteine protease that promotes cell death during brain development, in neuronal cultures, and in other cell types under many different conditions. To determine whether caspase-3 serves to regulate neuronal death after cerebral ischemia, we have (1) cloned a cDNA encoding the rat brain caspase-3; (2) examined caspase-3 mRNA and protein expression in the brain using in situ hybridization, Northern and Western blot analyses, and double-labeled immunohistochemistry; (3) determined caspase-3-like activity in brain cell extracts; and (4) studied the effect of caspase-3 inhibition on cell survival and DNA fragmentation in the hippocampus in a rat model of transient global ischemia. At 8-72 hr after ischemia, caspase-3 mRNA and protein were induced in the hippocampus and caudate-putamen (CPu), accompanied by increased caspase-3-like protease activity. In the hippocampus, caspase-3 mRNA and protein were predominantly increased in degenerating CA1 pyramidal neurons. Proteolytic activation of the caspase-3 precursor was detected in hippocampus and CPu but not in cortex at 4-72 hr after ischemia. Double-label experiments detected DNA fragmentation in the majority of CA1 neurons and selective CPu neurons that overexpressed caspase-3. Furthermore, ventricular infusion of Z-DEVD-FMK, a caspase-3 inhibitor, decreased caspase-3 activity in the hippocampus and significantly reduced cell death and DNA fragmentation in the CA1 sector up to 7 d after ischemia. These data strongly suggest that caspase-3 activity contributes to delayed neuronal death after transient ischemia.

Key words: cerebral ischemia; caspase-3; apoptosis; cysteine protease; neuronal death; neuron

Copyright © 1998 Society for Neuroscience  0270-6474/98/18134914-15$05.00/0


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Caspase-8 and Caspase-3 Are Expressed by Different Populations of Cortical Neurons Undergoing Delayed Cell Death after Focal Stroke in the Rat
J. Neurosci., July 15, 1999; 19(14): 5932 - 5941.
[Abstract] [Full Text] [PDF]

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 J. Neurosci.Home page
D. A. Shackelford, T. Tobaru, S. Zhang, and J. A. Zivin
Changes in Expression of the DNA Repair Protein Complex DNA-Dependent Protein Kinase after Ischemia and Reperfusion
J. Neurosci., June 15, 1999; 19(12): 4727 - 4738.
[Abstract] [Full Text] [PDF]

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 J. Neurosci.Home page
D. Xu, Y. Bureau, D. C. McIntyre, D. W. Nicholson, P. Liston, Y. Zhu, W. G. Fong, S. J. Crocker, R. G. Korneluk, and G. S. Robertson
Attenuation of Ischemia-Induced Cellular and Behavioral Deficits by X Chromosome-Linked Inhibitor of Apoptosis Protein Overexpression in the Rat Hippocampus
J. Neurosci., June 15, 1999; 19(12): 5026 - 5033.
[Abstract] [Full Text] [PDF]

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 J. Neurosci.Home page
F. Colbourne, G. R. Sutherland, and R. N. Auer
Electron Microscopic Evidence against Apoptosis as the Mechanism of Neuronal Death in Global Ischemia
J. Neurosci., June 1, 1999; 19(11): 4200 - 4210.
[Abstract] [Full Text] [PDF]

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 FASEB J.Home page
R. S. B. CLARK, P. M. KOCHANEK, M. CHEN, S. C. WATKINS, D. W. MARION, J. CHEN, R. L. HAMILTON, J. E. LOEFFERT, and S. H. GRAHAM
Increases in Bcl-2 and cleavage of caspase-1 and caspase-3 in human brain after head injury
FASEB J, May 1, 1999; 13(8): 813 - 821.
[Abstract] [Full Text]

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 J. Neurosci.Home page
Z. F. Cheema, S. B. Wade, M. Sata, K. Walsh, F. Sohrabji, and R. C. Miranda
Fas/Apo [Apoptosis]-1 and Associated Proteins in the Differentiating Cerebral Cortex: Induction of Caspase-Dependent Cell Death and Activation of NF-kappa B
J. Neurosci., March 1, 1999; 19(5): 1754 - 1770.
[Abstract] [Full Text] [PDF]

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 StrokeHome page
F. Colbourne, H. Li, A. M. Buchan, and J. A. Clemens
Continuing Postischemic Neuronal Death in CA1 : Influence of Ischemia Duration and Cytoprotective Doses of NBQX and SNX-111 in Rats • Editorial Comment: Influence of Ischemia Duration and Cytoprotective Doses of NBQX and SNX-111 in Rats
Stroke, March 1, 1999; 30(3): 662 - 668.
[Abstract] [Full Text] [PDF]

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 JEMHome page
C. Iadecola, C. A. Salkowski, F. Zhang, T. Aber, M. Nagayama, S. N. Vogel, and M. Elizabeth Ross
The Transcription Factor Interferon Regulatory Factor 1 Is Expressed after Cerebral Ischemia and Contributes to Ischemic Brain Injury
J. Exp. Med., February 15, 1999; 189(4): 719 - 727.
[Abstract] [Full Text] [PDF]

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 J. Neurosci.Home page
S. Cho, B. T. Volpe, Y. Bae, O. Hwang, H. J. Choi, J. Gal, L. C. H. Park, C. K. Chu, J. Du, and T. H. Joh
Blockade of Tetrahydrobiopterin Synthesis Protects Neurons after Transient Forebrain Ischemia in Rat: A Novel Role for the Cofactor
J. Neurosci., February 1, 1999; 19(3): 878 - 889.
[Abstract] [Full Text] [PDF]

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 StrokeHome page
M. Kawase, M. Fujimura, Y. Morita-Fujimura, P. H. Chan, and C. Iadecola
Reduction of Apurinic/Apyrimidinic Endonuclease Expression After Transient Global Cerebral Ischemia in Rats : Implication of the Failure of DNA Repair in Neuronal Apoptosis • Editorial Comment: Implication of the Failure of DNA Repair in Neuronal Apoptosis
Stroke, February 1, 1999; 30(2): 441 - 449.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
E. Vereker, V. Campbell, E. Roche, E. McEntee, and M. A. Lynch
Lipopolysaccharide Inhibits Long Term Potentiation in the Rat Dentate Gyrus by Activating Caspase-1
J. Biol. Chem., August 18, 2000; 275(34): 26252 - 26258.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
D. Chen, R. A. Stetler, G. Cao, W. Pei, C. O'Horo, X.-M. Yin, and J. Chen
Characterization of the Rat DNA Fragmentation Factor 35/Inhibitor of Caspase-activated DNase (Short Form). THE ENDOGENOUS INHIBITOR OF CASPASE-DEPENDENT DNA FRAGMENTATION IN NEURONAL APOPTOSIS
J. Biol. Chem., December 1, 2000; 275(49): 38508 - 38517.
[Abstract] [Full Text] [PDF]

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 J. Biol. Chem.Home page
K. Blomgren, C. Zhu, X. Wang, J.-O. Karlsson, A.-L. Leverin, B. A. Bahr, C. Mallard, and H. Hagberg
Synergistic Activation of Caspase-3 by m-Calpain after Neonatal Hypoxia-Ischemia. A MECHANISM OF "PATHOLOGICAL APOPTOSIS"?
J. Biol. Chem., March 23, 2001; 276(13): 10191 - 10198.
[Abstract] [Full Text] [PDF]

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 J. Neurosci.Home page
T. Sugawara, M. Fujimura, Y. Morita-Fujimura, M. Kawase, and P. H. Chan
Mitochondrial Release of Cytochrome c Corresponds to the Selective Vulnerability of Hippocampal CA1 Neurons in Rats after Transient Global Cerebral Ischemia
J. Neurosci., November 15, 1999; 19(22): RC39 - RC39.
[Abstract] [Full Text] [PDF]


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