Nitric oxide-dependent activation of CaMKII increases diastolic sarcoplasmic reticulum calcium release in cardiac myocytes in response to adrenergic stimulation

Jerry Curran, Lifei Tang, Steve R. Roof, Sathya Velmurugan, Ashley Millard, Stephen Shonts, Honglan Wang, Demetrio Santiago, Usama Ahmad, Matthew Perryman, Donald M Bers, Peter J. Mohler, Mark T. Ziolo, Thomas R. Shannon

Research output: Contribution to journalArticle

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Abstract

Spontaneous calcium waves in cardiac myocytes are caused by diastolic sarcoplasmic reticulum release (SR Ca2+ leak) through ryanodine receptors. Beta-adrenergic (β-AR) tone is known to increase this leak through the activation of Ca-calmodulin-dependent protein kinase (CaMKII) and the subsequent phosphorylation of the ryanodine receptor. When b-AR drive is chronic, as observed in heart failure, this CaMKII-dependent effect is exaggerated and becomes potentially arrhythmogenic. Recent evidence has indicated that CaMKII activation can be regulated by cellular oxidizing agents, such as reactive oxygen species. Here, we investigate how the cellular second messenger, nitric oxide, mediates CaMKII activity downstream of the adrenergic signaling cascade and promotes the generation of arrhythmogenic spontaneous C a2+ waves in intact cardiomyocytes. Both SCaWs and SR Ca2+ leak were measured in intact rabbit and mouse ventricular myocytes loaded with the Ca-dependent fluorescent dye, fluo-4. CaMKII activity in vitro and immunoblotting for phosphorylated residues on CaMKII, nitric oxide synthase, and Akt were measured to confirm activity of these enzymes as part of the adrenergic cascade. We demonstrate that stimulation of the β-AR pathway by isoproterenol increased the CaMKII-dependent SR Ca2+ leak. This increased leak was prevented by inhibition of nitric oxide synthase 1 but not nitric oxide synthase 3. In ventricular myocytes isolated from wild-type mice, isoproterenol stimulation also increased the CaMKII-dependent leak. Critically, in myocytes isolated from nitric oxide synthase 1 knock-out mice this effect is ablated. We show that isoproterenol stimulation leads to an increase in nitric oxide production, and nitric oxide alone is sufficient to activate CaMKII and increase SR Ca2+ leak. Mechanistically, our data links Akt to nitric oxide synthase 1 activation downstream of β-AR stimulation. Collectively, this evidence supports the hypothesis that CaMKII is regulated by nitric oxide as part of the adrenergic cascade leading to arrhythmogenesis.

Original languageEnglish (US)
Article numbere87495
JournalPLoS One
Volume9
Issue number2
DOIs
StatePublished - Feb 3 2014

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Calcium-Calmodulin-Dependent Protein Kinase Type 2
sarcoplasmic reticulum
Sarcoplasmic Reticulum
Cardiac Myocytes
Adrenergic Agents
nitric oxide
Nitric Oxide
Chemical activation
Calcium
calcium
myocytes
ryanodine receptors
Nitric Oxide Synthase
mice
Isoproterenol
Muscle Cells
Ryanodine Receptor Calcium Release Channel
fluorescent dyes
second messengers
calmodulin

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Nitric oxide-dependent activation of CaMKII increases diastolic sarcoplasmic reticulum calcium release in cardiac myocytes in response to adrenergic stimulation. / Curran, Jerry; Tang, Lifei; Roof, Steve R.; Velmurugan, Sathya; Millard, Ashley; Shonts, Stephen; Wang, Honglan; Santiago, Demetrio; Ahmad, Usama; Perryman, Matthew; Bers, Donald M; Mohler, Peter J.; Ziolo, Mark T.; Shannon, Thomas R.

In: PLoS One, Vol. 9, No. 2, e87495, 03.02.2014.

Research output: Contribution to journalArticle

Curran, J, Tang, L, Roof, SR, Velmurugan, S, Millard, A, Shonts, S, Wang, H, Santiago, D, Ahmad, U, Perryman, M, Bers, DM, Mohler, PJ, Ziolo, MT & Shannon, TR 2014, 'Nitric oxide-dependent activation of CaMKII increases diastolic sarcoplasmic reticulum calcium release in cardiac myocytes in response to adrenergic stimulation', PLoS One, vol. 9, no. 2, e87495. https://doi.org/10.1371/journal.pone.0087495
Curran, Jerry ; Tang, Lifei ; Roof, Steve R. ; Velmurugan, Sathya ; Millard, Ashley ; Shonts, Stephen ; Wang, Honglan ; Santiago, Demetrio ; Ahmad, Usama ; Perryman, Matthew ; Bers, Donald M ; Mohler, Peter J. ; Ziolo, Mark T. ; Shannon, Thomas R. / Nitric oxide-dependent activation of CaMKII increases diastolic sarcoplasmic reticulum calcium release in cardiac myocytes in response to adrenergic stimulation. In: PLoS One. 2014 ; Vol. 9, No. 2.
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AU - Millard, Ashley

AU - Shonts, Stephen

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AU - Santiago, Demetrio

AU - Ahmad, Usama

AU - Perryman, Matthew

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