Constitutive beta(2)-adrenergic signalling enhances sarcoplasmic reticulumCa2+ cycling to augment contraction in mouse heart

1. Transgenic overexpression of the beta(2)-adrenergic receptor (beta(2)AR) in mouse heart augments baseline cardiac function in a ligand-independent manner, due to the presence of spontaneously active beta(2)AR (beta(2)AR*). This study aims to elucidate the mechanism of beta(2)AR*-mediated modulati on of cardiac excitation-contraction (EC) coupling. 2. Confocal imaging was used to analyse Ca2+ sparks and spatially resolve C a2+ transients in single ventricular myocytes from transgenic (TG4) and non -transgenic (NTG) littermates. Whole-cell voltage- and current-clamp techni ques were used to record L-type Ca2+ currents (I-Ca) and action potentials, respectively. 3. In the absence of any beta(2)AR ligand, TG4 myocytes had greater contrac tion amplitudes, larger Ca2+ transients and faster relaxation times than di d NTG cells. 4. The action potentials of TG4 and NTG myocytes were similar, except for a prolonged end-stage repolarization in TG4 cells; the I-Ca density and kine tics were nearly identical. The relationship between peak Ca2+ and contract ion, which reflects myofilament Ca2+ sensitivity, was similar. 5. In TG4 cells, the frequency of Ca2+ sparks (spontaneous or evoked at -40 mV) was 2-7 times greater, despite the absence of change in the resting Ca 2+, sarcoplasmic reticulum (SR) Ca2+ content, and I-Ca. Individual sparks w ere brighter, broader and lasted longer, leading to a 2.3-fold greater sign al mass. Thus, changes in both spark frequency and size underlie the greate r Ca2+ transient in TG4 cells. 6. The inverse agonist ICI 118,551 (ICI, 5 x 10(-7) M), which blocks sponta neous beta(2)AR activation, reversed the aforementioned beta(2)AR* effects on cardiac EC coupling without affecting the sarcolemmal I-Ca. However, ICI failed to detect significant constitutive beta(2)AR activity in NTG cells. 7. We conclude that beta(2)AR*-mediated signalling enhances SR release chan nel activity and Ca2+-induced Ca2+ release in TG4 cardiac myocytes, and tha t beta(2)AR* enhances EC coupling by reinforcing SR Ca2+ cycling (release a nd reuptake), but bypassing the sarcolemmal I-Ca.