© 1999 American Heart Association, Inc.
Original Contribution |
Coupling of ß2-Adrenoceptor to Gi Proteins and Its Physiological Relevance in Murine Cardiac Myocytes
From the Laboratory of Cardiovascular Science, Gerontology Research Center, National Institute on Aging (R.-P.X., Y.-Y.Z., H.C., E.G.L.), Baltimore, Md; Institute of Developmental Biology (P.A.), Russian Academy of Sciences, Moscow, Russia; Department of Surgery (S.A.A., W.J.K.), Department of Medicine and Howard Hughes Medical Institute (R.J.L.), Duke University Medical Center, Durham, NC; and Abteilung Allgemeine Pharmakologie, Universitats-Krankenhaus Eppendorf (T.E.), Hamburg, Germany.
Correspondence to Rui-Ping Xiao, MD, PhD, Laboratory of Cardiovascular Science, Gerontology Research Center, National Institute on Aging, 5600 Nathan Shock Dr, Baltimore, MD 21224. E-mail Xiaor{at}GRC.NIA.NIH.Gov
Abstract—Transgenic mouse models
have been developed to manipulate ß-adrenergic receptor (ßAR)
signal transduction. Although several of these models have altered
ßAR subtypes, the specific functional sequelae of ßAR stimulation
in murine heart, particularly those of ß2-adrenergic
receptor (ß2AR) stimulation, have not been characterized.
In the present study, we investigated effects of ß2AR
stimulation on contraction, [Ca2+]i
transient, and L-type Ca2+ currents
(ICa) in single ventricular
myocytes isolated from transgenic mice overexpressing human
ß2AR (TG4 mice) and wild-type (WT) littermates. Baseline
contractility of TG4 heart cells was increased by
3-fold relative to WT controls as a result of the presence of
spontaneous ß2AR activation. In contrast,
ß2AR stimulation by zinterol or isoproterenol plus a
selective ß1-adrenergic receptor (ß1AR)
antagonist CGP 20712A failed to enhance the
contractility in TG4 myocytes, and more surprisingly,
ß2AR stimulation was also ineffective in increasing
contractility in WT myocytes. Pertussis toxin (PTX)
treatment fully rescued the ICa,
[Ca2+]i, and contractile responses to
ß2AR agonists in both WT and TG4 cells. The PTX-rescued
murine cardiac ß2AR response is mediated by
cAMP-dependent mechanisms, because it was totally blocked by the
inhibitory cAMP analog Rp-cAMPS. These results suggest that
PTX-sensitive G proteins are responsible for the unresponsiveness of
mouse heart to agonist-induced ß2AR stimulation. This was
further corroborated by an increased incorporation of the photoreactive
GTP analog [
-32P]GTP azidoanilide into 
subunits of
Gi2 and Gi3 after ß2AR
stimulation by zinterol or isoproterenol plus the ß1AR
blocker CGP 20712A. This effect to activate Gi
proteins was abolished by a selective ß2AR blocker ICI
118,551 or by PTX treatment. Thus, we conclude that (1)
ß2ARs in murine cardiac myocytes couple to concurrent
Gs and Gi signaling, resulting in null
inotropic response, unless the Gi signaling is inhibited;
(2) as a special case, the lack of cardiac contractile response to
ß2AR agonists in TG4 mice is not due to a saturation of
cell contractility or of the cAMP signaling cascade but
rather to an activation of ß2AR-coupled Gi
proteins; and (3) spontaneous ß2AR activation may differ
from agonist-stimulated ß2AR signaling.
Key Words: ß2-adrenergic receptor • inhibitory G protein • cardiac contractility • L-type Ca2+ current • mice, transgenic
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