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Epac stimulation induces rapid increases in connexin43 phosphorylation and function without preconditioning effect.

Nicolas Duquesnes¹, Mickael Derangeon, Mélanie Métrich

¹Signalisation et Physiopathologie Cardiaque, Châtenay-Malabry, France.

Pflugers Archiv : European Journal of Physiology

|June 30, 2010

Summary

Beta-adrenergic receptors activate phospholipase C via Epac, influencing isoproterenol (Iso)-induced preconditioning. Epac activation increases PKCepsilon, phosphorylates connexin43 (Cx43), and enhances gap junctional intercellular communication (GJIC), but does not confer ischemic preconditioning.

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Area of Science:

Cardiology
Molecular Biology
Cell Signaling

Background:

Beta-adrenergic receptors can activate phospholipase C through the cyclic adenosine monophosphate-binding protein Epac.
This pathway may be involved in isoproterenol (Iso)-induced preconditioning, a phenomenon that protects the heart against ischemic injury.

Purpose of the Study:

To investigate if Epac activation can induce protein kinase C epsilon (PKCepsilon) activation.
To determine the role of Epac in ischemic preconditioning via connexin43 (Cx43) phosphorylation and gap junctional intercellular communication (GJIC) modulation.

Main Methods:

Cultured rat neonatal cardiomyocytes and adult rat hearts were used.
Stimulation with Iso and the Epac activator 8-CPT.
Analysis of PKCepsilon, Cx43 phosphorylation, and GJIC.
Inhibition of PKA and PKC.

Main Results:

Iso and 8-CPT increased PKCepsilon in particulate fractions independently of PKA, leading to enhanced Cx43 phosphorylation.
Both Iso and 8-CPT increased GJIC, which was blocked by a PKC inhibitor.
PKA inhibition partially reduced Iso-induced Cx43 phosphorylation and GJIC.
Adult rat hearts showed similar PKCepsilon-dependent Cx43 phosphorylation via Epac.

Conclusions:

Epac stimulation activates PKCepsilon, promotes Cx43 phosphorylation, and increases GJIC.
Despite these effects, Epac activation alone does not induce the preconditioning effect observed with beta-adrenergic stimulation.