Increased Ca2+ sensitivity of the ryanodine receptor mutant RyR2R4496C underlies catecholaminergic polymorphic ventricular tachycardia

María Fernández-Velasco, Angélica Rueda, Nicoletta Rizzi, Jean Pierre Benitah, Barbara Colombi, Carlo Napolitano, Silvia G. Priori, Sylvain Richard, Ana María Gómez

Research output: Contribution to journalArticle

Abstract

Cardiac ryanodine receptor (RyR2) mutations are associated with autosomal dominant catecholaminergic polymorphic ventricular tachycardia, suggesting that alterations in Ca handling underlie this disease. Here we analyze the underlying Ca release defect that leads to arrhythmia in cardiomyocytes isolated from heterozygous knock-in mice carrying the RyR2 mutation. RyR2 littermates (wild type) were used as controls. [Ca]i transients were obtained by field stimulation in fluo-3-loaded cardiomyocytes and viewed using confocal microscopy. In our basal recording conditions (2-Hz stimulation rate), [Ca]i transients and sarcoplasmic reticulum Ca load were similar in wild-type and RyR2 cells. However, paced RyR2 ventricular myocytes presented abnormal Ca release during the diastolic period, viewed as Ca waves, consistent with the occurrence of delayed afterdepolarizations. The occurrence of this abnormal Ca release was enhanced at faster stimulation rates and by β-adrenergic stimulation, which also induced triggered activity. Spontaneous Ca sparks were more frequent in RyR2 myocytes, indicating increased RyR2 activity. When permeabilized cells were exposed to different cytosolic [Ca]i, RyR2 showed a dramatic increase in Ca sensitivity. Isoproterenol increased [Ca]i transient amplitude and Ca spark frequency to the same extent in wild-type and RyR2 cells, indicating that the β-adrenergic sensitivity of RyR2 cells remained unaltered. This effect was independent of protein expression variations because no difference was found in the total or phosphorylated RyR2 expression levels. In conclusion, the arrhythmogenic potential of the RyR2 mutation is attributable to the increased Ca sensitivity of RyR2, which induces diastolic Ca release and lowers the threshold for triggered activity.

Original languageEnglish
Pages (from-to)201-209
Number of pages9
JournalCirculation Research
Volume104
Issue number2
DOIs
Publication statusPublished - Jan 30 2009

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Keywords

  • [Ca2+]i transients
  • Ca2+ sparks
  • CPVT
  • Excitation contraction coupling
  • Ryanodine receptor

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

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