Ischemic preconditioning promotes a transient, but not sustained translocation of protein kinase C and sensitization of adenylyl cyclase

Objective Brief periods of ischemia precondition the heart and reduce the size of infarction caused by a subsequent sustained ischemia. The molecular memory of preconditioning, i.e., the molecule persistently activated by the preconditioning ischemia exhibiting protection during the infarct-inducing event, is subject to debate. Protein kinase C, adenylyl cyclase and beta-adrenergic receptors are candidates for this memory, supposedly their activation persists during several cycles of ischemia and reperfusion. The goal of this study was, therefore, to determine the activation of those signaling molecules after 1 - 3 cycles of myocardial ischemia and reperfusion. Methods Rat hearts were perfused according to the method of Langendorff with 1 - 3 cycles of ischemia (5 min) and reperfusion (10 min). In the particulate fraction of these hearts, densities of beta-adrenergic receptors, activities of adenylyl. cyclase, and the activities and isozyme distributions of PKC-alpha, PKC-beta, and PKC-epsilon were determined. Results The ischemia-induced upregulation of beta-adrenergic receptors is not influenced by preconditioning. In contrast, the sensitization of adenylyl cyclase observed after 5 min of ischemia is lost after repetitive periods of ischemia and reperfusion. Translocation of protein kinase C to the particulate fraction could be shown after 1 and 2 periods of ischemia including all major cardiac isozymes, with a rapid relocation to the cytosol after every cycle of reperfusion. A third period of ischemia was unable to promote a repeated translocation of protein kinase C. Conclusion The ischemia-induced regulation process of beta-adrenergic receptors is not influenced in preconditioning. Moreover, a sustained translocation of protein kinase C and a sustained sensitization of adenylyl cyclase are obviously no prerequisite for preconditioning after various cycles of ischemia and reperfusion. Thus, those signaling molecules do not seem to be operative for the preconditioning's memory. It is suggested that the initial, synergistic burst of sensitization of the adenylyl cyclase and of protein kinase C translocation induces myocardial protection very early in ischemia and reperfusion.