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Circulation Research. 1999;84:43-52

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(Circulation Research. 1999;84:43-52.)
© 1999 American Heart Association, Inc.


Original Contribution

Coupling of ß2-Adrenoceptor to Gi Proteins and Its Physiological Relevance in Murine Cardiac Myocytes

Rui-Ping Xiao, Pavel Avdonin, Ying-Ying Zhou, Heping Cheng, Shahab A. Akhter, Thomas Eschenhagen, Robert J. Lefkowitz, Walter J. Koch, Edward G. Lakatta

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 2AR) 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 [{gamma}-32P]GTP azidoanilide into {alpha} 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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