Calpain-induced proteolysis after transient global cerebral ischemia and ischemic tolerance in a rat model.
The activation of the [Ca(2+)]-dependent cysteine protease calpain plays an important role in ischemic injury. Here, the levels of two calpain-specific substrates, p35 protein and eukaryotic initiation factor 4G (eIF4G), as well as its physiological regulator calpastatin, were investigated in a rat model of transient global cerebral ischemia with or without ischemic tolerance (IT). Extracts of the cerebral cortex, whole hippocampus and hippocampal subregions after 30 min of ischemia and different reperfusion times (30 min and 4 h) were used. In rats without IT, the p35 levels slightly decreased after ischemia or reperfusion, whereas the levels of p25 (the truncated form of p35) were much higher than those in sham control rats after ischemia and remained elevated during reperfusion. The eIF4G levels deeply diminished after reperfusion and the decrease was significantly greater in CA1 and the rest of the hippocampus than in the cortex. By contrast, the calpastatin levels did not significantly decrease during ischemia or early reperfusion, but were upregulated after 4 h of reperfusion in the cortex. Although IT did not promote significant changes in p35 and p25 levels, it induced a slight increase in calpastatin and eIF4G levels in the hippocampal subregions after 4 h of reperfusion.
García-Bonilla
L
L
Servicio de Bioquímica, Departamento de Investigación, Hospital Ramón y Cajal, Ctra Colmenar Km 9, 28034, Madrid, Spain.
Burda
J
J
Piñeiro
D
D
Ayuso
I
I
Gómez-Calcerrada
M
M
Salinas
M
M
D000818:Animals; D001921:Brain / Q000473:pathology; D001923:Brain Chemistry / Q000502:physiology; D002135:Calcium-Binding Proteins / Q000378:metabolism; D002154:Calpain / Q000378:metabolism*; D002540:Cerebral Cortex / Q000378:metabolism / Q000473:pathology; D039603:Eukaryotic Initiation Factor-4G / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D006624:Hippocampus / Q000201:enzymology / Q000378:metabolism; D002546:Ischemic Attack, Transient / Q000378:metabolism* / Q000473:pathology; D019194:Ischemic Preconditioning; D051379:Mice; D010770:Phosphotransferases / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015427:Reperfusion Injury / Q000378:metabolism / Q000473:pathology; D014020:Tissue Extracts / Q000494:pharmacology
D002135:Calcium-Binding Proteins; C492236:Cdk5r1 protein, mouse; D039603:Eukaryotic Initiation Factor-4G; D018840:HSP70 Heat-Shock Proteins; D014020:Tissue Extracts; C033668:calpastatin; D010770:Phosphotransferases; D002154:Calpain