Optical mapping of sarcoplasmic reticulum Ca2+ in the intact heart: ryanodine receptor refractoriness during alternans and fibrillation

Circ Res. 2014 Apr 25;114(9):1410-21. doi: 10.1161/CIRCRESAHA.114.302505. Epub 2014 Feb 25.

Abstract

Rationale: Sarcoplasmic reticulum (SR) Ca(2+) cycling is key to normal excitation-contraction coupling but may also contribute to pathological cardiac alternans and arrhythmia.

Objective: To measure intra-SR free [Ca(2+)] ([Ca(2+)]SR) changes in intact hearts during alternans and ventricular fibrillation (VF).

Methods and results: Simultaneous optical mapping of Vm (with RH237) and [Ca(2+)]SR (with Fluo-5N AM) was performed in Langendorff-perfused rabbit hearts. Alternans and VF were induced by rapid pacing. SR Ca(2+) and action potential duration (APD) alternans occurred in-phase, but SR Ca(2+) alternans emerged first as cycle length was progressively reduced (217±10 versus 190±13 ms; P<0.05). Ryanodine receptor (RyR) refractoriness played a key role in the onset of SR Ca(2+) alternans, with SR Ca(2+) release alternans routinely occurring without changes in diastolic [Ca(2+)]SR. Sensitizing RyR with caffeine (200 μmol/L) significantly reduced the pacing threshold for both SR Ca(2+) and APD alternans (188±15 and 173±12 ms; P<0.05 versus baseline). Caffeine also reduced the magnitude of spatially discordant SR Ca(2+) alternans, but not APD alternans, the pacing threshold for discordance, or threshold for VF. During VF, [Ca(2+)]SR was high, but RyR remained nearly continuously refractory, resulting in minimal SR Ca(2+) release throughout VF.

Conclusions: In intact hearts, RyR refractoriness initiates SR Ca(2+) release alternans that can be amplified by diastolic [Ca(2+)]SR alternans and lead to APD alternans. Sensitizing RyR suppresses spatially concordant but not discordant SR Ca(2+) and APD alternans. Despite increased [Ca(2+)]SR during VF, SR Ca(2+) release was nearly continuously refractory. This novel method provides insight into SR Ca(2+) handling during cardiac alternans and arrhythmia.

Keywords: arrhythmias, cardiac; ventricular fibrillation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Action Potentials
  • Adrenergic beta-Agonists / pharmacology
  • Animals
  • Caffeine / pharmacology
  • Calcium / metabolism*
  • Calcium Signaling* / drug effects
  • Cardiac Pacing, Artificial
  • Excitation Contraction Coupling
  • In Vitro Techniques
  • Isoproterenol / pharmacology
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / metabolism*
  • Perfusion
  • Rabbits
  • Receptors, Adrenergic, beta / drug effects
  • Receptors, Adrenergic, beta / metabolism
  • Refractory Period, Electrophysiological*
  • Ryanodine Receptor Calcium Release Channel / drug effects
  • Ryanodine Receptor Calcium Release Channel / metabolism*
  • Sarcoplasmic Reticulum / drug effects
  • Sarcoplasmic Reticulum / metabolism*
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases / metabolism
  • Time Factors
  • Ventricular Fibrillation / metabolism*
  • Ventricular Fibrillation / physiopathology
  • Voltage-Sensitive Dye Imaging*

Substances

  • Adrenergic beta-Agonists
  • Receptors, Adrenergic, beta
  • Ryanodine Receptor Calcium Release Channel
  • Caffeine
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases
  • Isoproterenol
  • Calcium