Schematic diagram highlighting Ca²⁺ and cAMP changes observed in this study under different experimental conditions (basal, ISO and forskolin stimulated cardiomyocytes with and without pacing).

Abstract

<p>Pacing leads to an increase in Ca²⁺ levels which is further augmented by forskolin>ISO via PKA-dependent phosphorylation of Ca²⁺ handling proteins. However, pacing has little effect on cAMP levels, apart from the case when it is combined with forskolin stimulation, together both lead to PDE1 activation. Increase of PDE1 activity affects presumably a discrete subcellular microdomain which constitutes a small percentage of the whole cellular AMP content and can be therefore revealed in the cytosol only by the use of a PDE1 inhibitor. Forskolin and ISO generate quantitatively comparable but differently shaped amounts of cAMP which may come from ISO-induced dissociation of PDE4D8 from the β₁-adrenergic receptor. This mechanism regulates local second messenger pool at the receptor and allows more rapid increase of cAMP in the cytosol, as compared to forskolin stimulation.</p