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Rationale  In ventricular myocytes of large mammals, not all ryanodine receptor (RyR) clusters are associated with T-tubules (TTs); this fraction increases with cellular remodeling after myocardial infarction (MI).    Objective  To characterize RyR functional properties in relation to TT proximity, at baseline and after MI.    Methods  Myocytes were isolated from left ventricle of healthy pigs (CTRL) or from the area adjacent to a myocardial infarction (MI). Ca 2+  transients were measured under whole-cell voltage clamp during confocal linescan imaging (fluo-3) and segmented according to proximity of TTs (sites of early Ca 2+  release, F>F 50  within 20 ms) or their absence (delayed areas). Spontaneous Ca 2+  release events during diastole, Ca 2+  sparks, reflecting RyR activity and properties, were subsequently assigned to either category.    Results  In CTRL, spark frequency was higher in proximity of TTs, but spark duration was significantly shorter. Block of Na + /Ca 2+  exchanger (NCX) prolonged spark duration selectively near TTs, while block of Ca 2+  influx via Ca 2+  channels did not affect sparks properties. In MI, total spark mass was increased in line with higher SR Ca 2+  content. Extremely long sparks (>47.6 ms) occurred more frequently. The fraction of near-TT sparks was reduced; frequency increased mainly in delayed sites. Increased duration was seen in near-TT sparks only; Ca 2+  removal by NCX at the membrane was significantly lower in MI.    Conclusion  TT proximity modulates RyR cluster properties resulting in intracellular heterogeneity of diastolic spark activity. Remodeling in the area adjacent to MI differentially affects these RyR subpopulations. Reduction of the number of sparks near TTs and reduced local NCX removal limit cellular Ca 2+  loss and raise SR Ca 2+  content, but may promote Ca 2+  waves.

Subcellular Heterogeneity of Ryanodine Receptor Properties in Ventricular Myocytes with Low T-Tubule Density