Expression and Impact of Adenosine A₃ Receptors on Calcium Homeostasis in Human Right Atrium

Increased adenosine A_2A receptor (A_2AR) expression and activation underlies a higher incidence of spontaneous calcium release in atrial fibrillation (AF). Adenosine A₃ receptors (A₃R) could counteract excessive A_2AR activation, but their functional role in the atrium remains elusive, and we therefore aimed to address the impact of A₃Rs on intracellular calcium homeostasis. For this purpose, we analyzed right atrial samples or myocytes from 53 patients without AF, using quantitative PCR, patch-clamp technique, immunofluorescent labeling or confocal calcium imaging. A₃R mRNA accounted for 9% and A_2AR mRNA for 32%. At baseline, A₃R inhibition increased the transient inward current (I_TI) frequency from 0.28 to 0.81 events/min (p < 0.05). Simultaneous stimulation of A_2ARs and A₃Rs increased the calcium spark frequency seven-fold (p < 0.001) and the I_TI frequency from 0.14 to 0.64 events/min (p < 0.05). Subsequent A₃R inhibition caused a strong additional increase in the I_TI frequency (to 2.04 events/min; p < 0.01) and increased phosphorylation at s2808 1.7-fold (p < 0.001). These pharmacological treatments had no significant effects on L-type calcium current density or sarcoplasmic reticulum calcium load. In conclusion, A₃Rs are expressed and blunt spontaneous calcium release at baseline and upon A_2AR-stimulation in human atrial myocytes, pointing to A₃R activation as a means to attenuate physiological and pathological elevations of spontaneous calcium release events.