title: Constitutive beta2-adrenergic signalling enhances sarcoplasmic reticulum Ca2+ cycling to augment contraction in mouse heart.
abstract: 1. Transgenic overexpression of the beta2-adrenergic receptor (beta2AR) in mouse heart augments baseline cardiac function in a ligand-independent manner, due to the presence of spontaneously active beta2AR (beta2AR*). This study aims to elucidate the mechanism of beta2AR*-mediated modulation of cardiac excitation-contraction (EC) coupling. 2. Confocal imaging was used to analyse Ca2+ sparks and spatially resolve Ca2+ transients in single ventricular myocytes from transgenic (TG4) and non-transgenic (NTG) littermates. Whole-cell voltage- and current-clamp techniques were used to record L-type Ca2+ currents (ICa) and action potentials, respectively. 3. In the absence of any beta2AR ligand, TG4 myocytes had greater contraction amplitudes, larger Ca2+ transients and faster relaxation times than did NTG cells. 4. The action potentials of TG4 and NTG myocytes were similar, except for a prolonged end-stage repolarization in TG4 cells; the ICa density and kinetics were nearly identical. The relationship between peak Ca2+ and contraction, 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 Ca2+, sarcoplasmic reticulum (SR) Ca2+ content, and ICa. Individual sparks were brighter, broader and lasted longer, leading to a 2.3-fold greater signal mass. Thus, changes in both spark frequency and size underlie the greater Ca2+ transient in TG4 cells. 6. The inverse agonist ICI 118,551 (ICI, 5 x 10-7 M), which blocks spontaneous beta2AR activation, reversed the aforementioned beta2AR* effects on cardiac EC coupling without affecting the sarcolemmal ICa. However, ICI failed to detect significant constitutive beta2AR activity in NTG cells. 7. We conclude that beta2AR*-mediated signalling enhances SR release channel activity and Ca2+-induced Ca2+ release in TG4 cardiac myocytes, and that beta2AR* enhances EC coupling by reinforcing SR Ca2+ cycling (release and reuptake), but bypassing the sarcolemmal ICa.
authors.affiliation: Laboratory of Cardiovascular Science, Gerontology Research Center, National Institute on Aging, National Institutes of Health, Baltimore, MD 21224, USA.
authors.forename: Y Y
authors.initials: YY
authors.lastname: Zhou
authors.forename: L S
authors.initials: LS
authors.lastname: Song
authors.forename: E G
authors.initials: EG
authors.lastname: Lakatta
authors.forename: R P
authors.initials: RP
authors.lastname: Xiao
authors.forename: H
authors.initials: H
authors.lastname: Cheng
mesh_terms: D000200:Action Potentials / Q000502:physiology; D000814:Aniline Compounds; D000818:Animals; D002118:Calcium / Q000378:metabolism; D005456:Fluorescent Dyes; D006321:Heart / Q000502:physiology*; D006352:Heart Ventricles / Q000737:chemistry / Q000166:cytology; D051379:Mice; D008822:Mice, Transgenic; D018485:Muscle Fibers, Skeletal / Q000737:chemistry / Q000378:metabolism; D009200:Myocardial Contraction / Q000502:physiology*; D009206:Myocardium / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000502:physiology*; D012519:Sarcoplasmic Reticulum / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology*; D016276:Ventricular Function; D014966:Xanthenes
chemical_list: D000814:Aniline Compounds; D005456:Fluorescent Dyes; D018343:Receptors, Adrenergic, beta-2; D014966:Xanthenes; C059715:Fluo-3; D002118:Calcium