Activation of calcineurin expression in ischemia-reperfused rat heart and in human ischemic myocardium.
Calcineurin (CaN) has been reported as a critical mediator for cardiac hypertrophy and cardiac myocyte apoptosis. In the present study, we investigated the activity and expression of CaN and the effect of calpain in rat heart after ischemia and reperfusion. Rat ischemic heart showed significant increase in CaN activity. Western blot analysis of normal rat heart extract with a polyclonal antibody raised against bovine CaN indicated a prominent immunoreactive band of 60 kDa (CaN A). In ischemic-reperfused hearts, the expression of CaN A was significantly low and immunoreactivity was observed in proteolytic bands of 46 kDa. This may be due to the proteolytic degradation of CaN A in ischemic tissues by m-calpain. We also noticed in vitro proteolysis of bovine cardiac CaN A by m-calpain. Immunohistochemical studies showed strong staining of immunoreactivity in rat hearts that had gone under 30 min ischemia followed by 30 min reperfusion similar to that found in human ischemic heart. Ischemia is associated with multiple alterations in the extracellular and intracellular signaling of cardiomyocytes and may act as an inducer of apoptosis. The increase in CaN activity and strong immunostaining observed in ischemic/perfused rat heart may be due to the calpain-mediated proteolysis of this phosphatase.
Lakshmikuttyamma
Ashakumary
A
Department of Pathology, College of Medicine and Cancer Research Unit, Health Research Division, Saskatchewan Cancer Agency, University of Saskatchewan, Saskatoon, Canada S7N 4H4.
Selvakumar
Ponniah
P
Kakkar
Rakesh
R
Kanthan
Rani
R
Wang
Rui
R
Sharma
Rajendra K
RK
D000818:Animals; D015153:Blotting, Western; D019703:Calcineurin / Q000378:metabolism*; D002154:Calpain / Q000494:pharmacology; D002417:Cattle; D006321:Heart / Q000187:drug effects; D008297:Male; D017202:Myocardial Ischemia / Q000378:metabolism*; D015428:Myocardial Reperfusion Injury / Q000378:metabolism* / Q000473:pathology; D009206:Myocardium / Q000378:metabolism* / Q000473:pathology; D051381:Rats; D017207:Rats, Sprague-Dawley
D019703:Calcineurin; D002154:Calpain; C449000:m-calpain