Calpain activity in the rat brain after transient forebrain ischemia.
Activity of the Ca(2+)-dependent protease calpain is increased in neurons after global and focal brain ischemia, and may contribute to postischemic injury cascades. Understanding the time course and location of calpain activity in the post-ischemic brain is essential to establishing causality and optimizing therapeutic interventions. This study examined the temporal and spatial characteristics of brain calpain activity after transient forebrain ischemia (TFI) in rats. Male Long Evans rats underwent 10 min of normothermic TFI induced by bilateral carotid occlusion with hypovolemic hypotension (MABP 30 mm Hg). Brain calpain activity was examined between 1 and 72 h after reperfusion. Western blot analysis of regional brain homogenates demonstrated a bimodal pattern of calpain-mediated alpha-spectrin degradation in the hippocampus, cortex, and striatum with an initial increase at 1 h followed by a more prominent secondary increase at 36 h after reperfusion. Immunohistochemical analysis revealed that calpain activity was primarily localized to dendritic fields of selectively vulnerable neurons at one hour after reperfusion. Between 24 and 48 h after reperfusion neuronal calpain activity progressed from the dorsal to ventral striatum, medial to lateral CA1 hippocampus, and centripetally expanded from watershed foci in the cerebral cortex. This progression was associated with fragmentation of dendritic processes, calpain activation in the neuronal soma and subsequent neuronal degeneration. These observations demonstrate a clear association between calpain activation and subsequent delayed neuronal death and suggest broad therapeutic window for interventions aimed at preventing delayed intracellular Ca(2+) overload and pathologic calpain activation.
Neumar
R W
RW
Department of Emergency Medicine, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-4283, USA.
Meng
F H
FH
Mills
A M
AM
Xu
Y A
YA
Zhang
C
C
Welsh
F A
FA
Siman
R
R
D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000201:enzymology* / Q000473:pathology; D002154:Calpain / Q000378:metabolism*; D002540:Cerebral Cortex / Q000201:enzymology; D003342:Corpus Striatum / Q000201:enzymology; D004195:Disease Models, Animal; D004789:Enzyme Activation; D006624:Hippocampus / Q000201:enzymology; D007150:Immunohistochemistry; D002546:Ischemic Attack, Transient / Q000201:enzymology* / Q000473:pathology; D008297:Male; D009474:Neurons / Q000473:pathology; D009928:Organ Specificity; D051381:Rats; D020318:Rats, Long-Evans; D013049:Spectrin / Q000378:metabolism
D013049:Spectrin; D002154:Calpain