| Connexin 43 (Cx43) is the basic unit in the composition of Gap junction channels but also of the non-junctional unapposed hemichannels (Hc). Gap junction channels play key roles in cardiac function by allowing conduction of electrical impulses and exchange of biologically important molecules between cells. The unapposed Hc, however, perform functions different from those achieved by Gap junction channels mainly by providing pathways between the cytosol and the extracellular space allowing movement of ions and other small metabolites. Although they are much less studied than Gap junction channels, Hc are believed to remain normally in a closed state and that phosphorylation is an important factor promoting their closure. Under ischemic stress, the amount of non-phosphorylated Cx43 increases resulting in increasing hemichannels opening, an effect that can lead to irreversible tissue injury and cell death.;Protein kinase C (PKC) possesses the largest number of phosphorylation sites on Cx43 and exerts significant control on Cx43 channels. Its function depends on the involvement of at least 12 distinct isoformes. Various PKC isoforms exert specific cellular and cardiovascular functions, nonetheless the functional role of PKC isoforms in the modulation of the unapposed Cx43 hemichannels has never been assessed, neither has the therapeutic potential of Cx43Hc modulation in the protection of ischemic heart. In this context, three studies have been performed, they form the body of this thesis.;In the first study, a unique set of synthetic PKC isoform-selective activator and inhibitor peptides was utilised. In combination with the patch-clamp technique, we have demonstrated that Cx43Hc conductance is strongly inhibited by, among many isoforms, epsilon PKC isoforme, known for its cardioprotective effect against ischemic injury.;In the second study, we characterized the effect of a synthetic structural mimetic peptide of Cx43. Using patch clamp technique, we have demonstrated that the peptide Gap26 inhibits directly and specifically Cx43Hc, we also showed that Gap26 can confer resistance to cardiomyocytes (in vitro ) and intact heart (ex vivo) against ischemia.;In the third study, we investigated for the first time in vivo the capability of a unique pair of structural Cx43 mimetic peptides, Gap26 and Gap27, to protect heart from ischemic injury when administered in single low-dose intravenous boluses. We demonstrated that administration of either one or both peptides, before or after the onset of ischemia renders heart more resistant to ischemia and reduces significantly the size of myocardial infarct.;Altogether, our results revealed salvatory effect of Cx43Hc inhibition during ischemia and uncovered therapeutic potential of the synthetic structural mimetic peptides of Cx43 in ischemic heart disease.;Keywords: Heart, Ischemia, Connexin 43, Protein kinase C, Synthetic peptides. |
