Spontaneous β2-Adrenergic Signaling Fails To Modulate L-Type Ca2+ Current in Mouse Ventricular Myocytes

A receptor can be activated either by specific ligand-directed changes in conformation or by intrinsic, spontaneous conformational change. In the β2-adrenergic receptor (AR) overexpression transgenic (TG4) murine heart, spontaneously activated β2AR (β2-R*) in the absence of ligands has been evidenced by elevated basal adenylyl cyclase activity and cardiac function. In the present study, we determined whether the signaling mediated by β2-R* differs from that of a ligand-elicited β2AR activation (β2-LR*). In ventricular myocytes from TG4 mice, the properties of L-type Ca2+ current (ICa), a major effector of β2-LR* signaling, was unaltered, despite a 2.5-fold increase in the basal cAMP level and a 1.9-fold increase in baseline contraction amplitude as compared with that of wild-type (WT) cells. Although the contractile response to β2-R* in TG4 cells was abolished by a β2AR inverse agonist, ICI118,551 (5 × 10−7 M), or an inhibitory cAMP analog, Rp-CPT-cAMPS (10−4 M), no change was detected in the simultaneously recorded ICa. These results suggest that the increase in basal cAMP due to β2-R*, while increasing contraction amplitude, does not affect ICa characteristics. In contrast, the β2AR agonist, zinterol elicited a substantial augmentation of ICa in both TG4 and WT cells (pertussis toxin-treated), indicating that L-type Ca2+channel in these cells can respond to ligand-directed signaling. Furthermore, forskolin, an adenylyl cyclase activator, elicited similar dose-dependent increase in ICa amplitude in WT and TG4 cells, suggesting that the sensitivity of L-type Ca2+channel to cAMP-dependent modulation remains intact in TG4 cells. Thus, we conclude that β2-R* bypasses ICa to modulate contraction, and that β2-LR* and β2-R* exhibit different intracellular signaling and target protein specificity.