Anti-hypertrophic and anti-oxidant effect of beta3-adrenergic stimulation in myocytes requires differential neuronal NOS phosphorylation

Vabren L. Watts, Fernando M. Sepulveda, Oscar Cingolani, Alice S. Ho, Xiaolin Niu, Rosa Kim, Karen L. Miller, Koenraad Vandegaer, Djahida Bedja, Kathleen L Gabrielson, Gerald Rameau, Brian O'Rourke, David A Kass, Lili Barouch

Research output: Contribution to journalArticle

Abstract

Rationale: Stimulation of β3-adrenoreceptors (β3-AR) blunts contractility and improves chronic left ventricular function in hypertrophied and failing hearts in a neuronal nitric oxide synthase (nNOS) dependent manner. nNOS can be regulated by post-translational modification of stimulatory phosphorylation residue Ser1412 and inhibitory residue Ser847. However, the role of phosphorylation of these residues in cardiomyocytes and β3-AR protective signaling has yet to be explored. Objective: We tested the hypothesis that β3-AR regulation of myocyte stress requires changes in nNOS activation mediated by differential nNOS phosphorylation. Methods and results: Endothelin (ET-1) or norepinephrine induced hypertrophy in rat neonatal ventricular cardiomyocytes (NRVMs) was accompanied by increased β3-AR gene expression. Co-administration of the β3-AR agonist BRL-37433 (BRL) reduced cell size and reactive oxygen species (ROS) generation, while augmenting NOS activity. BRL-dependent augmentation of NOS activity and ROS suppression due to NE were blocked by inhibiting nNOS (L-VNIO). BRL augmented nNOS phosphorylation at Ser1412 and dephosphorylation at Ser847. Cells expressing constitutively dephosphorylated Ser1412A or phosphorylated Ser847D nNOS mutants displayed reduced nNOS activity and a lack of BRL modulation. BRL also failed to depress ROS from NE in cells with nNOS-Ser847D. Inhibiting Akt decreased BRL-induced nNOS-Ser1412 phosphorylation and NOS activation, whereas Gi/o blockade blocked BRL-regulation of both post-translational modifications, preventing enhancement of NOS activity and ROS reduction. BRL resulted in near complete dephosphorylation of Ser847 and a moderate rise in Ser1412 phosphorylation in mouse myocardium exposed to chronic pressure-overload. Conclusion: β3-AR regulates myocardial NOS activity and ROS via activation of nNOS involving reciprocal changes in phosphorylation at two regulatory sites. These data identify a novel and potent anti-oxidant and anti-hypertrophic pathway due to nNOS post-translational modification that is coupled to β3-AR receptor stimulation.

Original languageEnglish (US)
Pages (from-to)8-17
Number of pages10
JournalJournal of Molecular and Cellular Cardiology
Volume62
DOIs
StatePublished - Sep 2013

Fingerprint

Nitric Oxide Synthase Type I
Adrenergic Agents
Muscle Cells
Antioxidants
Phosphorylation
Reactive Oxygen Species
Post Translational Protein Processing
Cardiac Myocytes
Endothelin-1
Cell Size
Left Ventricular Function
Oxidants
Hypertrophy
Myocardium
Norepinephrine

Keywords

  • Beta3-adrenergic receptors
  • Heart failure
  • Hypertrophy
  • Neuronal nitric oxide synthase
  • Reactive oxygen species

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine
  • Medicine(all)

Cite this

Anti-hypertrophic and anti-oxidant effect of beta3-adrenergic stimulation in myocytes requires differential neuronal NOS phosphorylation. / Watts, Vabren L.; Sepulveda, Fernando M.; Cingolani, Oscar; Ho, Alice S.; Niu, Xiaolin; Kim, Rosa; Miller, Karen L.; Vandegaer, Koenraad; Bedja, Djahida; Gabrielson, Kathleen L; Rameau, Gerald; O'Rourke, Brian; Kass, David A; Barouch, Lili.

In: Journal of Molecular and Cellular Cardiology, Vol. 62, 09.2013, p. 8-17.

Research output: Contribution to journalArticle

Watts, Vabren L. ; Sepulveda, Fernando M. ; Cingolani, Oscar ; Ho, Alice S. ; Niu, Xiaolin ; Kim, Rosa ; Miller, Karen L. ; Vandegaer, Koenraad ; Bedja, Djahida ; Gabrielson, Kathleen L ; Rameau, Gerald ; O'Rourke, Brian ; Kass, David A ; Barouch, Lili. / Anti-hypertrophic and anti-oxidant effect of beta3-adrenergic stimulation in myocytes requires differential neuronal NOS phosphorylation. In: Journal of Molecular and Cellular Cardiology. 2013 ; Vol. 62. pp. 8-17.
@article{eaa9e0cfb9984a7ead1f3095a8b7a65d,
title = "Anti-hypertrophic and anti-oxidant effect of beta3-adrenergic stimulation in myocytes requires differential neuronal NOS phosphorylation",
abstract = "Rationale: Stimulation of β3-adrenoreceptors (β3-AR) blunts contractility and improves chronic left ventricular function in hypertrophied and failing hearts in a neuronal nitric oxide synthase (nNOS) dependent manner. nNOS can be regulated by post-translational modification of stimulatory phosphorylation residue Ser1412 and inhibitory residue Ser847. However, the role of phosphorylation of these residues in cardiomyocytes and β3-AR protective signaling has yet to be explored. Objective: We tested the hypothesis that β3-AR regulation of myocyte stress requires changes in nNOS activation mediated by differential nNOS phosphorylation. Methods and results: Endothelin (ET-1) or norepinephrine induced hypertrophy in rat neonatal ventricular cardiomyocytes (NRVMs) was accompanied by increased β3-AR gene expression. Co-administration of the β3-AR agonist BRL-37433 (BRL) reduced cell size and reactive oxygen species (ROS) generation, while augmenting NOS activity. BRL-dependent augmentation of NOS activity and ROS suppression due to NE were blocked by inhibiting nNOS (L-VNIO). BRL augmented nNOS phosphorylation at Ser1412 and dephosphorylation at Ser847. Cells expressing constitutively dephosphorylated Ser1412A or phosphorylated Ser847D nNOS mutants displayed reduced nNOS activity and a lack of BRL modulation. BRL also failed to depress ROS from NE in cells with nNOS-Ser847D. Inhibiting Akt decreased BRL-induced nNOS-Ser1412 phosphorylation and NOS activation, whereas Gi/o blockade blocked BRL-regulation of both post-translational modifications, preventing enhancement of NOS activity and ROS reduction. BRL resulted in near complete dephosphorylation of Ser847 and a moderate rise in Ser1412 phosphorylation in mouse myocardium exposed to chronic pressure-overload. Conclusion: β3-AR regulates myocardial NOS activity and ROS via activation of nNOS involving reciprocal changes in phosphorylation at two regulatory sites. These data identify a novel and potent anti-oxidant and anti-hypertrophic pathway due to nNOS post-translational modification that is coupled to β3-AR receptor stimulation.",
keywords = "Beta3-adrenergic receptors, Heart failure, Hypertrophy, Neuronal nitric oxide synthase, Reactive oxygen species",
author = "Watts, {Vabren L.} and Sepulveda, {Fernando M.} and Oscar Cingolani and Ho, {Alice S.} and Xiaolin Niu and Rosa Kim and Miller, {Karen L.} and Koenraad Vandegaer and Djahida Bedja and Gabrielson, {Kathleen L} and Gerald Rameau and Brian O'Rourke and Kass, {David A} and Lili Barouch",
year = "2013",
month = "9",
doi = "10.1016/j.yjmcc.2013.04.025",
language = "English (US)",
volume = "62",
pages = "8--17",
journal = "Journal of Molecular and Cellular Cardiology",
issn = "0022-2828",
publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - Anti-hypertrophic and anti-oxidant effect of beta3-adrenergic stimulation in myocytes requires differential neuronal NOS phosphorylation

AU - Watts, Vabren L.

AU - Sepulveda, Fernando M.

AU - Cingolani, Oscar

AU - Ho, Alice S.

AU - Niu, Xiaolin

AU - Kim, Rosa

AU - Miller, Karen L.

AU - Vandegaer, Koenraad

AU - Bedja, Djahida

AU - Gabrielson, Kathleen L

AU - Rameau, Gerald

AU - O'Rourke, Brian

AU - Kass, David A

AU - Barouch, Lili

PY - 2013/9

Y1 - 2013/9

N2 - Rationale: Stimulation of β3-adrenoreceptors (β3-AR) blunts contractility and improves chronic left ventricular function in hypertrophied and failing hearts in a neuronal nitric oxide synthase (nNOS) dependent manner. nNOS can be regulated by post-translational modification of stimulatory phosphorylation residue Ser1412 and inhibitory residue Ser847. However, the role of phosphorylation of these residues in cardiomyocytes and β3-AR protective signaling has yet to be explored. Objective: We tested the hypothesis that β3-AR regulation of myocyte stress requires changes in nNOS activation mediated by differential nNOS phosphorylation. Methods and results: Endothelin (ET-1) or norepinephrine induced hypertrophy in rat neonatal ventricular cardiomyocytes (NRVMs) was accompanied by increased β3-AR gene expression. Co-administration of the β3-AR agonist BRL-37433 (BRL) reduced cell size and reactive oxygen species (ROS) generation, while augmenting NOS activity. BRL-dependent augmentation of NOS activity and ROS suppression due to NE were blocked by inhibiting nNOS (L-VNIO). BRL augmented nNOS phosphorylation at Ser1412 and dephosphorylation at Ser847. Cells expressing constitutively dephosphorylated Ser1412A or phosphorylated Ser847D nNOS mutants displayed reduced nNOS activity and a lack of BRL modulation. BRL also failed to depress ROS from NE in cells with nNOS-Ser847D. Inhibiting Akt decreased BRL-induced nNOS-Ser1412 phosphorylation and NOS activation, whereas Gi/o blockade blocked BRL-regulation of both post-translational modifications, preventing enhancement of NOS activity and ROS reduction. BRL resulted in near complete dephosphorylation of Ser847 and a moderate rise in Ser1412 phosphorylation in mouse myocardium exposed to chronic pressure-overload. Conclusion: β3-AR regulates myocardial NOS activity and ROS via activation of nNOS involving reciprocal changes in phosphorylation at two regulatory sites. These data identify a novel and potent anti-oxidant and anti-hypertrophic pathway due to nNOS post-translational modification that is coupled to β3-AR receptor stimulation.

AB - Rationale: Stimulation of β3-adrenoreceptors (β3-AR) blunts contractility and improves chronic left ventricular function in hypertrophied and failing hearts in a neuronal nitric oxide synthase (nNOS) dependent manner. nNOS can be regulated by post-translational modification of stimulatory phosphorylation residue Ser1412 and inhibitory residue Ser847. However, the role of phosphorylation of these residues in cardiomyocytes and β3-AR protective signaling has yet to be explored. Objective: We tested the hypothesis that β3-AR regulation of myocyte stress requires changes in nNOS activation mediated by differential nNOS phosphorylation. Methods and results: Endothelin (ET-1) or norepinephrine induced hypertrophy in rat neonatal ventricular cardiomyocytes (NRVMs) was accompanied by increased β3-AR gene expression. Co-administration of the β3-AR agonist BRL-37433 (BRL) reduced cell size and reactive oxygen species (ROS) generation, while augmenting NOS activity. BRL-dependent augmentation of NOS activity and ROS suppression due to NE were blocked by inhibiting nNOS (L-VNIO). BRL augmented nNOS phosphorylation at Ser1412 and dephosphorylation at Ser847. Cells expressing constitutively dephosphorylated Ser1412A or phosphorylated Ser847D nNOS mutants displayed reduced nNOS activity and a lack of BRL modulation. BRL also failed to depress ROS from NE in cells with nNOS-Ser847D. Inhibiting Akt decreased BRL-induced nNOS-Ser1412 phosphorylation and NOS activation, whereas Gi/o blockade blocked BRL-regulation of both post-translational modifications, preventing enhancement of NOS activity and ROS reduction. BRL resulted in near complete dephosphorylation of Ser847 and a moderate rise in Ser1412 phosphorylation in mouse myocardium exposed to chronic pressure-overload. Conclusion: β3-AR regulates myocardial NOS activity and ROS via activation of nNOS involving reciprocal changes in phosphorylation at two regulatory sites. These data identify a novel and potent anti-oxidant and anti-hypertrophic pathway due to nNOS post-translational modification that is coupled to β3-AR receptor stimulation.

KW - Beta3-adrenergic receptors

KW - Heart failure

KW - Hypertrophy

KW - Neuronal nitric oxide synthase

KW - Reactive oxygen species

UR - http://www.scopus.com/inward/record.url?scp=84878696585&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84878696585&partnerID=8YFLogxK

U2 - 10.1016/j.yjmcc.2013.04.025

DO - 10.1016/j.yjmcc.2013.04.025

M3 - Article

C2 - 23643588

AN - SCOPUS:84878696585

VL - 62

SP - 8

EP - 17

JO - Journal of Molecular and Cellular Cardiology

JF - Journal of Molecular and Cellular Cardiology

SN - 0022-2828

ER -