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Ca²⁺ sparks and Ca²⁺ waves in saponin-permeabilized rat ventricular myocytes

Valeriy Lukyanenko and Sandor Györke

1. We carried out confocal Ca²⁺ imaging in myocytes permeabilized with saponin in 'internal' solutions containing: MgATP, EGTA and fluo-3 potassium salt.

2. Permeabilized myocytes exhibited spontaneous Ca²⁺ sparks and waves similar to those observed in intact myocytes loaded with fluo-3 AM.

3. In the presence of 'low' [EGTA] (0·05 mM), Ca²⁺ waves arose regularly, even at relatively low [Ca²⁺] (50-100 nM, free). Increasing [EGTA] resulted in decreased frequency and propagation velocity of Ca²⁺ waves. Propagating waves were completely abolished at [EGTA] > 0·3 mM.

4. The frequency of sparks increased as a function of [Ca²⁺] (50-400 nM range) with no sign of a high affinity Ca²⁺-dependent inactivation process.

5. The rate of occurrence of Ca²⁺ sparks was increased by calmodulin and cyclic adenosine diphosphate-ribose (cADPR).

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