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Nitric oxide regulates cardiac intracellular Na(+) and Ca(2 +) by modulating Na/K ATPase via PKCε and phospholemman-dependent mechanism()

In the heart, Na/K-ATPase regulates intracellular Na(+) and Ca(2 +) (via NCX), thereby preventing Na(+) and Ca(2 +) overload and arrhythmias. Here, we test the hypothesis that nitric oxide (NO) regulates cardiac intracellular Na(+) and Ca(2 +) and investigate mechanisms and physiological consequence...

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Autores principales: Pavlovic, Davor, Hall, Andrew R., Kennington, Erika J., Aughton, Karen, Boguslavskyi, Andrii, Fuller, William, Despa, Sanda, Bers, Donald M., Shattock, Michael J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Academic Press 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3981027/
https://www.ncbi.nlm.nih.gov/pubmed/23612119
http://dx.doi.org/10.1016/j.yjmcc.2013.04.013
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author Pavlovic, Davor
Hall, Andrew R.
Kennington, Erika J.
Aughton, Karen
Boguslavskyi, Andrii
Fuller, William
Despa, Sanda
Bers, Donald M.
Shattock, Michael J.
author_facet Pavlovic, Davor
Hall, Andrew R.
Kennington, Erika J.
Aughton, Karen
Boguslavskyi, Andrii
Fuller, William
Despa, Sanda
Bers, Donald M.
Shattock, Michael J.
author_sort Pavlovic, Davor
collection PubMed
description In the heart, Na/K-ATPase regulates intracellular Na(+) and Ca(2 +) (via NCX), thereby preventing Na(+) and Ca(2 +) overload and arrhythmias. Here, we test the hypothesis that nitric oxide (NO) regulates cardiac intracellular Na(+) and Ca(2 +) and investigate mechanisms and physiological consequences involved. Effects of both exogenous NO (via NO-donors) and endogenously synthesized NO (via field-stimulation of ventricular myocytes) were assessed in this study. Field stimulation of rat ventricular myocytes significantly increased endogenous NO (18 ± 2 μM), PKCε activation (82 ± 12%), phospholemman phosphorylation (at Ser-63 and Ser-68) and Na/K-ATPase activity (measured by DAF-FM dye, western-blotting and biochemical assay, respectively; p < 0.05, n = 6) and all were abolished by Ca(2 +)-chelation (EGTA 10 mM) or NOS inhibition l-NAME (1 mM). Exogenously added NO (spermine-NONO-ate) stimulated Na/K-ATPase (EC50 = 3.8 μM; n = 6/grp), via decrease in K(m), in PLM(WT) but not PLM(KO) or PLM(3SA) myocytes (where phospholemman cannot be phosphorylated) as measured by whole-cell perforated-patch clamp. Field-stimulation with l-NAME or PKC-inhibitor (2 μM Bis) resulted in elevated intracellular Na(+) (22 ± 1.5 and 24 ± 2 respectively, vs. 14 ± 0.6 mM in controls) in SBFI-AM-loaded rat myocytes. Arrhythmia incidence was significantly increased in rat hearts paced in the presence of l-NAME (and this was reversed by l-arginine), as well as in PLM(3SA) mouse hearts but not PLM(WT) and PLM(KO). We provide physiological and biochemical evidence for a novel regulatory pathway whereby NO activates Na/K-ATPase via phospholemman phosphorylation and thereby limits Na(+) and Ca(2 +) overload and arrhythmias. This article is part of a Special Issue entitled “Na(+) Regulation in Cardiac Myocytes”.
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spelling pubmed-39810272014-04-10 Nitric oxide regulates cardiac intracellular Na(+) and Ca(2 +) by modulating Na/K ATPase via PKCε and phospholemman-dependent mechanism() Pavlovic, Davor Hall, Andrew R. Kennington, Erika J. Aughton, Karen Boguslavskyi, Andrii Fuller, William Despa, Sanda Bers, Donald M. Shattock, Michael J. J Mol Cell Cardiol Original Article In the heart, Na/K-ATPase regulates intracellular Na(+) and Ca(2 +) (via NCX), thereby preventing Na(+) and Ca(2 +) overload and arrhythmias. Here, we test the hypothesis that nitric oxide (NO) regulates cardiac intracellular Na(+) and Ca(2 +) and investigate mechanisms and physiological consequences involved. Effects of both exogenous NO (via NO-donors) and endogenously synthesized NO (via field-stimulation of ventricular myocytes) were assessed in this study. Field stimulation of rat ventricular myocytes significantly increased endogenous NO (18 ± 2 μM), PKCε activation (82 ± 12%), phospholemman phosphorylation (at Ser-63 and Ser-68) and Na/K-ATPase activity (measured by DAF-FM dye, western-blotting and biochemical assay, respectively; p < 0.05, n = 6) and all were abolished by Ca(2 +)-chelation (EGTA 10 mM) or NOS inhibition l-NAME (1 mM). Exogenously added NO (spermine-NONO-ate) stimulated Na/K-ATPase (EC50 = 3.8 μM; n = 6/grp), via decrease in K(m), in PLM(WT) but not PLM(KO) or PLM(3SA) myocytes (where phospholemman cannot be phosphorylated) as measured by whole-cell perforated-patch clamp. Field-stimulation with l-NAME or PKC-inhibitor (2 μM Bis) resulted in elevated intracellular Na(+) (22 ± 1.5 and 24 ± 2 respectively, vs. 14 ± 0.6 mM in controls) in SBFI-AM-loaded rat myocytes. Arrhythmia incidence was significantly increased in rat hearts paced in the presence of l-NAME (and this was reversed by l-arginine), as well as in PLM(3SA) mouse hearts but not PLM(WT) and PLM(KO). We provide physiological and biochemical evidence for a novel regulatory pathway whereby NO activates Na/K-ATPase via phospholemman phosphorylation and thereby limits Na(+) and Ca(2 +) overload and arrhythmias. This article is part of a Special Issue entitled “Na(+) Regulation in Cardiac Myocytes”. Academic Press 2013-08 /pmc/articles/PMC3981027/ /pubmed/23612119 http://dx.doi.org/10.1016/j.yjmcc.2013.04.013 Text en © 2013 The Authors https://creativecommons.org/licenses/by/3.0/ Open Access under CC BY 3.0 (https://creativecommons.org/licenses/by/3.0/) license
spellingShingle Original Article
Pavlovic, Davor
Hall, Andrew R.
Kennington, Erika J.
Aughton, Karen
Boguslavskyi, Andrii
Fuller, William
Despa, Sanda
Bers, Donald M.
Shattock, Michael J.
Nitric oxide regulates cardiac intracellular Na(+) and Ca(2 +) by modulating Na/K ATPase via PKCε and phospholemman-dependent mechanism()
title Nitric oxide regulates cardiac intracellular Na(+) and Ca(2 +) by modulating Na/K ATPase via PKCε and phospholemman-dependent mechanism()
title_full Nitric oxide regulates cardiac intracellular Na(+) and Ca(2 +) by modulating Na/K ATPase via PKCε and phospholemman-dependent mechanism()
title_fullStr Nitric oxide regulates cardiac intracellular Na(+) and Ca(2 +) by modulating Na/K ATPase via PKCε and phospholemman-dependent mechanism()
title_full_unstemmed Nitric oxide regulates cardiac intracellular Na(+) and Ca(2 +) by modulating Na/K ATPase via PKCε and phospholemman-dependent mechanism()
title_short Nitric oxide regulates cardiac intracellular Na(+) and Ca(2 +) by modulating Na/K ATPase via PKCε and phospholemman-dependent mechanism()
title_sort nitric oxide regulates cardiac intracellular na(+) and ca(2 +) by modulating na/k atpase via pkcε and phospholemman-dependent mechanism()
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3981027/
https://www.ncbi.nlm.nih.gov/pubmed/23612119
http://dx.doi.org/10.1016/j.yjmcc.2013.04.013
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