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1 Surgery, University of Kentucky, Lexington, Kentucky, United States
* To whom correspondence should be addressed. E-mail: ccrof2{at}uky.edu.
Ischemia/reperfusion activates ERK and p38 MAPK in cardiac membranes, but the role of caveolae in MAPK signaling during this stress has not been studied. The purpose of this study was to determine the effect of in vivo myocardial ischemia/reperfusion on the level and distribution of caveolin-1/3 and cholesterol as well as MAPK activation in caveolin-enriched fractions. Adult male rats were submitted to in vivo regional myocardial ischemia induced by 25 minutes coronary artery occlusion and either 10 minutes (n=5) or 2 hours (n=4) reperfusion. Appropriate nonischemic time controls (n=4). Caveolae/lipid rafts were isolated from ischemic and nonischemic heart tissue using a discontinuous sucrose density gradient. Caveolin-1 and 3, as well as cholesterol, were enriched in the light fractions. A redistribution of caveolin-3, and a reduction in caveolin-1 and cholesterol levels in the light fractions occurred after 10 minutes reperfusion. The ERKs were activated in ischemic zone light and heavy fractions by short reperfusion. P44 ERK was activated after 2 hours reperfusion only in the light fractions, whereas p42 ERK phosphorylation was increased in both light and heavy fractions. Although no p38 MAPK activation occurred after 10 minutes reperfusion, longer reperfusion caused significant activation of p38 MAPK in nonischemic zone light and heavy fractions. These results show the importance of caveolar membrane/lipid rafts in MAPK signaling and suggest that subcellular compartmentation of p44/p42 ERKs and p38 MAPK may play distinct roles in the response to myocardial ischemia/reperfusion.
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