Compartmentalization of Cardiac &bgr;-Adrenergic Inotropy Modulation by Phosphodiesterase Type 5

Background— Recent cell-based studies have found that cGMP synthesis and hydrolysis by phosphodiesterase (PDE) appear compartmentalized, with nitric oxide synthase–derived and/or PDE type 5 (PDE-5)–hydrolyzable cGMP undetected at the sarcolemmal membrane in contrast to cGMP stimulated by natriuretic peptide. In the present study, we determine the functional significance of such compartments with a comparison of &bgr;-adrenergic modulation by PDE-5 inhibition to that of natriuretic peptide stimulation in both cardiomyocytes and intact hearts. The potential role of differential cGMP and protein kinase G stimulation by these 2 modulators was also studied. Methods and Results— Intact C57/BL6 mouse hearts were studied with pressure-volume analysis, and adult isolated myocytes were studied with fluorescence microscopy. PDE-5 inhibition with 0.1 to 1 &mgr;mol/L sildenafil (SIL) suppressed isoproterenol (ISO)-stimulated contractility, whereas 10 &mgr;mol/L atrial natriuretic peptide (ANP) had no effect. ISO suppression by SIL was prevented in cells pretreated with a protein kinase G inhibitor. Surprisingly, myocardial cGMP changed little with SIL+ISO yet rose nearly 5-fold with ANP, whereas protein kinase G activation (vasodilator-stimulated protein phosphorylation; ELISA assay) displayed the opposite: increased with SIL+ISO but unaltered by ANP+ISO. PDE-5 and ANP compartments were functionally separated, as inhibition of nitric oxide synthase by Nw-nitro-L-arginine methyl ester eliminated antiadrenergic effects of SIL, yet this was not restorable by co-stimulation with ANP. Conclusions— Regulation of cardiac &bgr;-adrenergic response by cGMP is specifically linked to a nitric oxide–synthesis/PDE-5–hydrolyzed pool signaling via protein kinase G. Natriuretic peptide stimulation achieves greater detectable increases in cGMP but not protein kinase G activity and does not modulate &bgr;-adrenergic response. Such disparities likely contribute to differential cardiac regulation by drugs that modulate cGMP synthesis and hydrolysis.

[1]  Manuela Zaccolo,et al.  Phosphodiesterases and compartmentalized cAMP signalling in the heart. , 2006, European journal of cell biology.

[2]  M. Silberbach,et al.  Natriuretic Peptides and Nitric Oxide Stimulate cGMP Synthesis in Different Cellular Compartments , 2006, The Journal of general physiology.

[3]  L. Langeberg,et al.  AKAP signaling complexes: getting to the heart of the matter. , 2006, Trends in molecular medicine.

[4]  Jacques Turgeon,et al.  The ATP-Binding Cassette Transporters and Their Implication in Drug Disposition: A Special Look at the Heart , 2006, Pharmacological Reviews.

[5]  D. Cooper,et al.  Cyclic Guanosine Monophosphate Compartmentation in Rat Cardiac Myocytes , 2006, Circulation.

[6]  L. Langeberg,et al.  Compartmentation of Cyclic Nucleotide Signaling in the Heart The Role of A-Kinase Anchoring Proteins , 2006 .

[7]  M. Zaccolo,et al.  Compartmentalized Phosphodiesterase-2 Activity Blunts &bgr;-Adrenergic Cardiac Inotropy via an NO/cGMP-Dependent Pathway , 2006, Circulation research.

[8]  D. Kass,et al.  Sildenafil Inhibits β-Adrenergic–Stimulated Cardiac Contractility in Humans , 2005 .

[9]  M. Semigran.  Type 5 phosphodiesterase inhibition: the focus shifts to the heart. , 2005, Circulation.

[10]  S. Reiken,et al.  Phosphodiesterase 4D Deficiency in the Ryanodine-Receptor Complex Promotes Heart Failure and Arrhythmias , 2005, Cell.

[11]  H. Weiss,et al.  Differential Effects of cGMP Produced by Soluble and Particulate Guanylyl Cyclase on Mouse Ventricular Myocytes , 2005, Experimental biology and medicine.

[12]  Anindita Das,et al.  Phosphodiesterase-5 Inhibitor Sildenafil Preconditions Adult Cardiac Myocytes against Necrosis and Apoptosis , 2005, Journal of Biological Chemistry.

[13]  G. Booz.  Putting the brakes on cardiac hypertrophy: exploiting the NO-cGMP counter-regulatory system. , 2005, Hypertension.

[14]  D. Kass,et al.  Chronic inhibition of cyclic GMP phosphodiesterase 5A prevents and reverses cardiac hypertrophy , 2005, Nature Medicine.

[15]  M. Zaccolo,et al.  cGMP Catabolism by Phosphodiesterase 5A Regulates Cardiac Adrenergic Stimulation by NOS3-Dependent Mechanism , 2004, Circulation research.

[16]  Sharon M. Cawley,et al.  Cygnets: in vivo characterization of novel cGMP indicators and in vivo imaging of intracellular cGMP. , 2005, Methods in molecular biology.

[17]  D. Kass,et al.  Sildenafil inhibits beta-adrenergic-stimulated cardiac contractility in humans. , 2005, Circulation.

[18]  Stepan Gambaryan,et al.  Fluorescent sensors for rapid monitoring of intracellular cGMP , 2005, Nature Methods.

[19]  J. Balligand,et al.  Cardiomyocyte-Restricted Overexpression of Endothelial Nitric Oxide Synthase (NOS3) Attenuates &bgr;-Adrenergic Stimulation and Reinforces Vagal Inhibition of Cardiac Contraction , 2004, Circulation.

[20]  D. Kass,et al.  Modulation of In Vivo Cardiac Function by Myocyte-Specific Nitric Oxide Synthase-3 , 2004, Circulation research.

[21]  R. Pilz,et al.  This Review Is Part of a Thematic Series on Cyclic Gmp–generating Enzymes and Cyclic Gmp–dependent Signaling, Which Includes the following Articles: Regulation of Nitric Oxide–sensitive Guanylyl Cyclase Cyclic Gmp Phosphodiesterases and Regulation of Smooth Muscle Function Structure, Regulation, and , 2022 .

[22]  U. Walter,et al.  Increased effects of C‐type natriuretic peptide on contractility and calcium regulation in murine hearts overexpressing cyclic GMP‐dependent protein kinase I , 2003, British journal of pharmacology.

[23]  F. Hofmann,et al.  Cyclic GMP-dependent protein kinases and the cardiovascular system: insights from genetically modified mice. , 2003, Circulation research.

[24]  M. Kuhn,et al.  Structure, regulation, and function of mammalian membrane guanylyl cyclase receptors, with a focus on guanylyl cyclase-A. , 2003, Circulation research.

[25]  U. Brinkmann,et al.  Expression and localization of the multidrug resistance protein 5 (MRP5/ABCC5), a cellular export pump for cyclic nucleotides, in human heart. , 2003, The American journal of pathology.

[26]  J. Balligand,et al.  Nitric Oxide and Cardiac Function: Ten Years After, and Continuing , 2003, Circulation research.

[27]  R. Kloner,et al.  Therapeutic potential of phosphodiesterase 5 inhibition for cardiovascular disease. , 2003, Circulation.

[28]  Michael D. Schneider,et al.  Pressure-independent cardiac hypertrophy in mice with cardiomyocyte-restricted inactivation of the atrial natriuretic peptide receptor guanylyl cyclase-A. , 2003, The Journal of clinical investigation.

[29]  A. Shah,et al.  Role of cyclic GMP‐dependent protein kinase in the contractile response to exogenous nitric oxide in rat cardiac myocytes , 2002, The Journal of physiology.

[30]  F. Hofmann,et al.  cGMP-Dependent Protein Kinase I Mediates the Negative Inotropic Effect of cGMP in the Murine Myocardium , 2002, Circulation research.

[31]  D. Kass,et al.  Cardiac phosphodiesterase 5 (cGMP‐specific) modulates β‐adrenergic signaling in vivo and is down‐regulated in heart failure , 2001, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[32]  D. Kass,et al.  Minimal force‐frequency modulation of inotropy and relaxation of in situ murine heart , 2001, The Journal of physiology.

[33]  M. Redfield,et al.  Differential effects of natriuretic peptides and NO on LV function in heart failure and normal dogs. , 2001, American journal of physiology. Heart and circulatory physiology.

[34]  J. Knowles,et al.  Pressure-independent enhancement of cardiac hypertrophy in natriuretic peptide receptor A-deficient mice. , 2001, The Journal of clinical investigation.

[35]  T. Meinertz,et al.  Vasodilator-Stimulated Phosphoprotein Serine 239 Phosphorylation as a Sensitive Monitor of Defective Nitric Oxide/cGMP Signaling and Endothelial Dysfunction , 2000, Circulation research.

[36]  A. Lawrie,et al.  Activation of the Particulate and Not the Soluble Guanylate Cyclase Leads to the Inhibition of Ca2+ Extrusion through Localized Elevation of cGMP* , 2000, The Journal of Biological Chemistry.

[37]  E. Lakatta,et al.  Activation of distinct cAMP-dependent and cGMP-dependent pathways by nitric oxide in cardiac myocytes. , 1999, Circulation research.

[38]  B. Prendergast,et al.  Nitric oxide enhances the inotropic response to β-adrenergic stimulation in the isolated guinea-pig heart , 1998, Basic Research in Cardiology.

[39]  M. Creager,et al.  Increased sensitivity to nitric oxide synthase inhibition in patients with heart failure: potentiation of beta-adrenergic inotropic responsiveness. , 1998, Circulation.

[40]  M. Creager,et al.  Nitric Oxide Inhibits the Positive Inotropic Response to β-Adrenergic Stimulation in Humans With Left Ventricular Dysfunction , 1995 .

[41]  M. Creager,et al.  Nitric oxide inhibits the positive inotropic response to beta-adrenergic stimulation in humans with left ventricular dysfunction. , 1995, Circulation.

[42]  R. Fischmeister,et al.  Atrial Natriuretic Factor Regulates the Calcium Current in Frog Isolated Cardiac Cells , 1988, Circulation research.

[43]  S. Shaughnessy,et al.  Do No Harm: Health Systems’ Duty to Promote Clinician Well-Being , 2022, American Journal of Hospital Medicine.