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The Journal of Neuroscience, May 15, 1998, 18(10):3659-3668
Activation and Cleavage of Caspase-3 in Apoptosis Induced by
Experimental Cerebral Ischemia
Shobu
Namura1,
Jinmin
Zhu1,
Klaus
Fink1,
Matthias
Endres1,
Anu
Srinivasan2,
Kevin J.
Tomaselli2,
Junying
Yuan3, and
Michael A.
Moskowitz1
1 Stroke and Neurovascular Regulation, Neurosurgical
Service, Departments of Surgery and Neurology, Massachusetts General
Hospital, Harvard Medical School, Charlestown, Massachusetts 02129, 2 IDUN Pharmaceuticals, Inc., La Jolla, California 92037, and 3 Department of Cell Biology, Harvard Medical School,
Boston, Massachusetts 02115
We examined the expression, activation, and cellular localization
of caspase-3 (CPP32) using immunohistochemistry, immunoblots, and
cleavage of the fluorogenic substrate
N-benzyloxycarbonyl-Asp-Glu-Val-Asp-7-amino-4-trifluoromethyl coumarin (zDEVD-afc) in adult mouse brain after temporary (2 hr) middle
cerebral artery occlusion produced by filament insertion into the
carotid artery. Immunoreactive caspase-3p32 but not its cleavage
product caspase-3p20 was constitutively expressed in neurons throughout
brain and was most prominent in neuronal perikarya within piriform
cortex. Caspase-like enzyme activity was elevated in brain homogenate
0-3 hr after reperfusion and reached a peak within 30 to 60 min.
Caspase-3p20 immunoreactivity became prominent in neuronal perikarya
within the middle cerebral artery territory at the time of reperfusion
and on immunoblots 1-12 hr later. DNA laddering (agarose gels) and
terminal deoxynucleotidyl transferase-mediated dUTP biotin nick-end
labeling (TUNEL)-stained cells were detected 6-24 hr after
reperfusion. At 12-24 hr, immunoreactive p20 was visualized in
TUNEL-positive cells, a finding also observed in apoptotic mouse
cerebellar granule cells on postnatal day 5. Together, these
observations suggest the existence of a time-dependent evolution of
ischemic injury characterized by the close correspondence between caspase-like enzyme activation and an associated increase in
immunoreactive product (caspase-3p20) beginning at or before
reperfusion and followed several hours later by morphological and
biochemical features of apoptosis.
Key words:
ischemia; caspase; CED-3/ICE proteases; mouse brain; apoptosis; caspase-3 cleavage; caspase-like enzyme activity; TUNEL; CNS
development
Copyright © 1998 Society for Neuroscience 0270-6474/98/18103659-10$05.00/0
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T. Nakagawa and J. Yuan
Cross-talk between Two Cysteine Protease Families: Activation of Caspase-12 by Calpain in Apoptosis
J. Cell Biol.,
August 21, 2000;
150(4):
887 - 894.
[Abstract]
[Full Text]
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D. C. Henshall, R. S. B. Clark, P. D. Adelson, M. Chen, S. C. Watkins, and R. P. Simon
Alterations in bcl-2 and caspase gene family protein expression in human temporal lobe epilepsy
Neurology,
July 25, 2000;
55(2):
250 - 257.
[Abstract]
[Full Text]
[PDF]
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P. M. Kang, A. Haunstetter, H. Aoki, A. Usheva, and S. Izumo
Morphological and Molecular Characterization of Adult Cardiomyocyte Apoptosis During Hypoxia and Reoxygenation
Circ. Res.,
July 21, 2000;
87(2):
118 - 125.
[Abstract]
[Full Text]
[PDF]
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