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Ryanodol action on calcium sparks in ventricular myocytes

  • Ion Channels, Receptors and Transporters
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Abstract

The action of ryanodol on single cardiac ryanodine receptor (RyR2) channels in bilayers and local RyR2-mediated Ca2+ release events (Ca2+ sparks) in ventricular myocytes was defined. At the single-channel level, ryanodol intermittently modified single channels into a long-lived subconductance state with an average duration of 3.8 ± 0.2 s. Unlike ryanodine, ryanodol did not change the open probability (Po) of unmodified channels, and high concentrations did not promote full-channel closure. Ryanodol action was Po dependent with the K D varying roughly from 20 to 80 μM as Po changed from ∼0.2 to 1, respectively. Ryanodol preferentially bound during long channel openings. In intact and permeabilized rat myocytes, ryanodol evoked trains of sparks at active release sites resulting in a significant increase in overall spark frequency. Ryanodol did not increase the number of active release sites. Long-lived Ca2+ release events were observed but infrequently, and ryanodol action was readily reversed upon drug washout. We propose that ryanodol modifies a few channels during a Ca2+ spark. These modified channels mediate a sustained low-intensity Ca2+ release that repeatedly triggers sparks at the same release site. We conclude that ryanodol is an easily generated reversible probe that can be effectively used to explore RyR2-mediated Ca2+ release in cells.

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Acknowledgements

This work was supported by National Institutes of Health Grants HL057832 and HL064210 to M.F., HL071741 to J.R.F., GM078665 to J.A.C., and Agence Nationale de la Recherche Grants ANR-09-GENO-012 and ANR-09-GENO-034 to A.M.G..

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Authors and Affiliations

  1. Department of Molecular Biophysics & Physiology, Rush University Medical Center, 1750 West Harrison St., Chicago, IL, 60612, USA

    Josefina Ramos-Franco, Alma Nani, Yiwei Liu & Michael Fill

  2. INSERM U-637, CHU A. de Villeneuve, 34295, Montpellier, France

    Ana M. Gomez

  3. Department of Pharmacology, Southern Illinois University, 801 North Rutledge St., Springfield, IL, 62794, USA

    Julio A. Copello

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  1. Josefina Ramos-Franco
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  2. Ana M. Gomez
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  3. Alma Nani
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  4. Yiwei Liu
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  5. Julio A. Copello
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  6. Michael Fill
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Correspondence to Michael Fill.

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Ramos-Franco, J., Gomez, A.M., Nani, A. et al. Ryanodol action on calcium sparks in ventricular myocytes. Pflugers Arch - Eur J Physiol 460, 767–776 (2010). https://doi.org/10.1007/s00424-010-0839-8

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  • DOI: https://doi.org/10.1007/s00424-010-0839-8

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