Biophys J, November 2000, p. 2547-2556, Vol. 79, No. 5

G_i-Dependent Localization of ₂-Adrenergic Receptor Signaling to L-Type Ca²⁺ Channels

Ye Chen-Izu,^* Rui-Ping Xiao,^* Leighton T. Izu, Heping Cheng,^* Meike Kuschel,^* Harold Spurgeon,^* and Edward G. Lakatta^*

 ^*Laboratory of Cardiovascular Sciences, Gerontology Research Center, National Institute on Aging, National Institutes of Health, Baltimore, Maryland 21224-6823;  Department of Physiology/Cardiology, University of Maryland School of Medicine, Baltimore, Maryland 21201 USA

A plausible determinant of the specificity of receptor signaling is the cellular compartment over which the signal is broadcast. In rat heart, stimulation of ₁-adrenergic receptor (₁-AR), coupled to G_s-protein, or ₂-AR, coupled to G_s- and G_i-proteins, both increase L-type Ca²⁺ current, causing enhanced contractile strength. But only ₁-AR stimulation increases the phosphorylation of phospholamban, troponin-I, and C-protein, causing accelerated muscle relaxation and reduced myofilament sensitivity to Ca²⁺. ₂-AR stimulation does not affect any of these intracellular proteins. We hypothesized that ₂-AR signaling might be localized to the cell membrane. Thus we examined the spatial range and characteristics of ₁-AR and ₂-AR signaling on their common effector, L-type Ca²⁺ channels. Using the cell-attached patch-clamp technique, we show that stimulation of ₁-AR or ₂-AR in the patch membrane, by adding agonist into patch pipette, both activated the channels in the patch. But when the agonist was applied to the membrane outside the patch pipette, only ₁-AR stimulation activated the channels. Thus, ₁-AR signaling to the channels is diffusive through cytosol, whereas ₂-AR signaling is localized to the cell membrane. Furthermore, activation of G_i is essential to the localization of ₂-AR signaling because in pertussis toxin-treated cells, ₂-AR signaling becomes diffusive. Our results suggest that the dual coupling of ₂-AR to both G_s- and G_i-proteins leads to a highly localized ₂-AR signaling pathway to modulate sarcolemmal L-type Ca²⁺ channels in rat ventricular myocytes.

Biophys J, November 2000, p. 2547-2556, Vol. 79, No. 5
© 2000 by the Biophysical Society   0006-3495/00/11/2547/10  $2.00

This article has been cited by other articles:

				G. S. Lynch and J. G. Ryall Role of {beta}-Adrenoceptor Signaling in Skeletal Muscle: Implications for Muscle Wasting and Disease Physiol Rev, April 1, 2008; 88(2): 729 - 767. [Abstract] [Full Text] [PDF]

				Y. Ikeda, M. Hoshijima, and K. R. Chien Toward Biologically Targeted Therapy of Calcium Cycling Defects in Heart Failure Physiology, February 1, 2008; 23(1): 6 - 16. [Abstract] [Full Text] [PDF]

				F. A. Faucher, F. E. Gannier, J. M. Lignon, P. Cosnay, and C. O. Malecot Roles of PKA, PI3K, and cPLA2 in the NO-mediated negative inotropic effect of {beta}2-adrenoceptor agonists in guinea pig right papillary muscles Am J Physiol Cell Physiol, January 1, 2008; 294(1): C106 - C117. [Abstract] [Full Text] [PDF]

				V. O. Nikolaev, M. Bunemann, E. Schmitteckert, M. J. Lohse, and S. Engelhardt Cyclic AMP Imaging in Adult Cardiac Myocytes Reveals Far-Reaching {beta}1-Adrenergic but Locally Confined {beta}2-Adrenergic Receptor-Mediated Signaling Circ. Res., November 10, 2006; 99(10): 1084 - 1091. [Abstract] [Full Text] [PDF]

				J. T. Hulme, R. E. Westenbroek, T. Scheuer, and W. A. Catterall Phosphorylation of serine 1928 in the distal C-terminal domain of cardiac CaV1.2 channels during beta1-adrenergic regulation PNAS, October 31, 2006; 103(44): 16574 - 16579. [Abstract] [Full Text] [PDF]

				R. Fischmeister, L. R.V. Castro, A. Abi-Gerges, F. Rochais, J. Jurevicius, J. Leroy, and G. Vandecasteele Compartmentation of Cyclic Nucleotide Signaling in the Heart: The Role of Cyclic Nucleotide Phosphodiesterases Circ. Res., October 13, 2006; 99(8): 816 - 828. [Abstract] [Full Text] [PDF]

				M. A.G. van der Heyden, T. J.M. Wijnhoven, and T. Opthof Molecular aspects of adrenergic modulation of the transient outward current Cardiovasc Res, August 1, 2006; 71(3): 430 - 442. [Abstract] [Full Text] [PDF]

				M. S. George and G. S. Pitt The real estate of cardiac signaling: Location, location, location PNAS, May 16, 2006; 103(20): 7535 - 7536. [Full Text] [PDF]

				W. Zhu, X. Zeng, M. Zheng, and R.-P. Xiao The Enigma of {beta}2-Adrenergic Receptor Gi Signaling in the Heart: The Good, the Bad, and the Ugly Circ. Res., September 16, 2005; 97(6): 507 - 509. [Full Text] [PDF]

				A. P. Barnes, G. Livera, P. Huang, C. Sun, W. K. O'Neal, M. Conti, M. J. Stutts, and S. L. Milgram Phosphodiesterase 4D Forms a cAMP Diffusion Barrier at the Apical Membrane of the Airway Epithelium J. Biol. Chem., March 4, 2005; 280(9): 7997 - 8003. [Abstract] [Full Text] [PDF]

				R. L Winslow, S. Cortassa, and J. L Greenstein Using models of the myocyte for functional interpretation of cardiac proteomic data J. Physiol., February 15, 2005; 563(1): 73 - 81. [Abstract] [Full Text] [PDF]

				A. J. Tanskanen, J. L. Greenstein, B. O'Rourke, and R. L. Winslow The Role of Stochastic and Modal Gating of Cardiac L-Type Ca2+ Channels on Early After-Depolarizations Biophys. J., January 1, 2005; 88(1): 85 - 95. [Abstract] [Full Text] [PDF]

				M. A.G. van der Heyden, T. J.M. Wijnhoven, and T. Opthof Molecular aspects of adrenergic modulation of cardiac L-type Ca2+ channels Cardiovasc Res, January 1, 2005; 65(1): 28 - 39. [Abstract] [Full Text] [PDF]

				V. Leblais, S.-H. Jo, K. Chakir, V. Maltsev, M. Zheng, M. T. Crow, W. Wang, E. G. Lakatta, and R.-P. Xiao Phosphatidylinositol 3-Kinase Offsets cAMP-Mediated Positive Inotropic Effect via Inhibiting Ca2+ Influx in Cardiomyocytes Circ. Res., December 10, 2004; 95(12): 1183 - 1190. [Abstract] [Full Text] [PDF]

				K. W. Dilly, J. Kurokawa, C. Terrenoire, S. Reiken, W. J. Lederer, A. R. Marks, and R. S. Kass Overexpression of {beta}2-Adrenergic Receptors cAMP-dependent Protein Kinase Phosphorylates and Modulates Slow Delayed Rectifier Potassium Channels Expressed in Murine Heart: EVIDENCE FOR RECEPTOR/CHANNEL CO-LOCALIZATION J. Biol. Chem., September 24, 2004; 279(39): 40778 - 40787. [Abstract] [Full Text] [PDF]

				A. Breit, M. Lagace, and M. Bouvier Hetero-oligomerization between {beta}2- and {beta}3-Adrenergic Receptors Generates a {beta}-Adrenergic Signaling Unit with Distinct Functional Properties J. Biol. Chem., July 2, 2004; 279(27): 28756 - 28765. [Abstract] [Full Text] [PDF]

				E. N. Nikolov and T. T. Ivanova-Nikolova Coordination of Membrane Excitability through a GIRK1 Signaling Complex in the Atria J. Biol. Chem., May 28, 2004; 279(22): 23630 - 23636. [Abstract] [Full Text] [PDF]

				J. L. GREENSTEIN, A. J. TANSKANEN, and R. L. WINSLOW Modeling the Actions of {beta}-Adrenergic Signaling on Excitation--Contraction Coupling Processes Ann. N.Y. Acad. Sci., May 1, 2004; 1015(1): 16 - 27. [Abstract] [Full Text] [PDF]

				R. A. Rose, A. E. Lomax, C. S. Kondo, M. B. Anand-Srivastava, and W. R. Giles Effects of C-type natriuretic peptide on ionic currents in mouse sinoatrial node: a role for the NPR-C receptor Am J Physiol Heart Circ Physiol, May 1, 2004; 286(5): H1970 - H1977. [Abstract] [Full Text] [PDF]

				R.-P. Xiao, S.-J. Zhang, K. Chakir, P. Avdonin, W. Zhu, R. A. Bond, C. W. Balke, E. G. Lakatta, and H. Cheng Enhanced Gi Signaling Selectively Negates {beta}2-Adrenergic Receptor (AR)- but Not {beta}1-AR-Mediated Positive Inotropic Effect in Myocytes From Failing Rat Hearts Circulation, September 30, 2003; 108(13): 1633 - 1639. [Abstract] [Full Text] [PDF]

				S. Bartel, E.-G. Krause, G. Wallukat, and P. Karczewski New insights into {beta}2-adrenoceptor signaling in the adult rat heart Cardiovasc Res, March 1, 2003; 57(3): 694 - 703. [Abstract] [Full Text] [PDF]

				T. Cesetti, J. M. Hernandez-Guijo, P. Baldelli, V. Carabelli, and E. Carbone Opposite Action of beta 1- and beta 2-Adrenergic Receptors on CaV1 L-Channel Current in Rat Adrenal Chromaffin Cells J. Neurosci., January 1, 2003; 23(1): 73 - 83. [Abstract] [Full Text] [PDF]

				Y. Xiang, E. Devic, and B. Kobilka The PDZ Binding Motif of the beta 1 Adrenergic Receptor Modulates Receptor Trafficking and Signaling in Cardiac Myocytes J. Biol. Chem., September 6, 2002; 277(37): 33783 - 33790. [Abstract] [Full Text] [PDF]

				S.-H. Jo, V. Leblais, P. H. Wang, M. T. Crow, and R.-P. Xiao Phosphatidylinositol 3-Kinase Functionally Compartmentalizes the Concurrent Gs Signaling During {beta}2-Adrenergic Stimulation Circ. Res., July 12, 2002; 91(1): 46 - 53. [Abstract] [Full Text] [PDF]

				K. D. Keef, J. R. Hume, and J. Zhong Regulation of cardiac and smooth muscle Ca2+ channels (CaV1.2a,b) by protein kinases Am J Physiol Cell Physiol, December 1, 2001; 281(6): C1743 - C1756. [Abstract] [Full Text] [PDF]

				R.-P. Xiao {beta}-Adrenergic Signaling in the Heart: Dual Coupling of the {beta}2-Adrenergic Receptor to Gs and Gi Proteins Sci. Signal., October 16, 2001; 2001(104): re15 - re15. [Abstract] [Full Text] [PDF]

				T. C. Rich, K. A. Fagan, T. E. Tse, J. Schaack, D. M. F. Cooper, and J. W. Karpen A uniform extracellular stimulus triggers distinct cAMP signals in different compartments of a simple cell PNAS, October 12, 2001; (2001) 221381398. [Abstract] [Full Text] [PDF]

				D. M. Bers and M. T. Ziolo When Is cAMP Not cAMP?: Effects of Compartmentalization Circ. Res., August 31, 2001; 89(5): 373 - 375. [Full Text] [PDF]

				M. A. Davare, V. Avdonin, D. D. Hall, E. M. Peden, A. Burette, R. J. Weinberg, M. C. Horne, T. Hoshi, and J. W. Hell A beta 2 Adrenergic Receptor Signaling Complex Assembled with the Ca2+ Channel Cav1.2 Science, July 6, 2001; 293(5527): 98 - 101. [Abstract] [Full Text] [PDF]

				T. C. Rich, K. A. Fagan, T. E. Tse, J. Schaack, D. M. F. Cooper, and J. W. Karpen A uniform extracellular stimulus triggers distinct cAMP signals in different compartments of a simple cell PNAS, November 6, 2001; 98(23): 13049 - 13054. [Abstract] [Full Text] [PDF]

				L.-S. Song, S.-Q. Wang, R.-P. Xiao, H. Spurgeon, E. G. Lakatta, and H. Cheng {beta}-Adrenergic Stimulation Synchronizes Intracellular Ca2+ Release During Excitation-Contraction Coupling in Cardiac Myocytes Circ. Res., April 27, 2001; 88(8): 794 - 801. [Abstract] [Full Text] [PDF]

				M. G. Vila Petroff, J. M. Egan, X. Wang, and S. J. Sollott Glucagon-Like Peptide-1 Increases cAMP but Fails to Augment Contraction in Adult Rat Cardiac Myocytes Circ. Res., August 31, 2001; 89(5): 445 - 452. [Abstract] [Full Text] [PDF]