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Am J Physiol Heart Circ Physiol 277: H1771-H1785, 1999;
0363-6135/99 $5.00
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Vol. 277, Issue 5, H1771-H1785, November 1999

PKC-dependent activation of p46/p54 JNKs during ischemic preconditioning in conscious rabbits

Peipei Ping1,2, Jun Zhang2, Shuang Huang3, Xinan Cao2, Xian-Liang Tang1, Richard C. X. Li1, Yu-Ting Zheng2, Yumin Qiu1, Angela Clerk4, Peter Sugden4, Jiahuai Han3, and Roberto Bolli1,2

1 Experimental Research Laboratory, Division of Cardiology, and 2 Department of Physiology and Biophysics, University of Louisville, Louisville, Kentucky 40202; 4 Division of Cardiac Medicine, National Heart and Lung Institute, London SW3 6LY, United Kingdom; and 3 Division of Immunology, Scripps Institute of Molecular Medicine, La Jolla, California 92037

A conscious rabbit model was used to study the effect of ischemic preconditioning (PC) on stress-activated kinases [c-Jun NH2-terminal kinases (JNKs) and p38 mitogen-activated protein kinase (MAPK)] in an environment free of surgical trauma and attending external stress. Ischemic PC (6 cycles of 4-min ischemia/4-min reperfusion) induced significant activation of protein kinase C (PKC)-epsilon in the particulate fraction, which was associated with activation of p46 JNK in the nuclear fraction and p54 JNK in the cytosolic fraction; all of these changes were completely abolised by the PKC inhibitor chelerythrine. Selective enhancement of PKC-epsilon activity in adult rabbit cardiac myocytes resulted in enhanced activity of p46/p54 JNKs, providing direct in vitro evidence that PKC-epsilon is coupled to both kinases. Studies in rabbits showed that the activation of p46 JNK occurred during ischemia, whereas that of p54 JNK occurred after reperfusion. A single 4-min period of ischemia induced a robust activation of the p38 MAPK cascade, which, however, was attenuated after 5 min of reperfusion and disappeared after six cycles of 4-min ischemia/reperfusion. Overexpression of PKC-epsilon in cardiac myocytes failed to increase the p38 MAPK activity. These results demonstrate that ischemic PC activates p46 and p54 JNKs via a PKC-epsilon -dependent signaling pathway and that there are important differences between p46 and p54 JNKs with respect to the subcellular compartment (cytosolic vs. nuclear) and the mechanism (ischemia vs. reperfusion) of their activation after ischemic PC.

stress-activated protein kinases; p38 mitogen-activated protein kinases; c-Jun NH2-terminal kinases; ischemia-reperfusion; protein kinase C


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