tetra(4-n-methylpyridyl)porphine and Heart-Diseases

Local control of Ca(2+)-induced Ca(2+) release (CICR) depends on the spatial organization of L-type Ca(2+) channels and ryanodine receptors (RyR) in the dyad. Analogously, Ca(2+) uptake by mitochondria is facilitated by their close proximity to the Ca(2+) release sites, a process required for stimulating oxidative phosphorylation during changes in work. Mitochondrial feedback on CICR is less well understood. Since mitochondria are a primary source of reactive oxygen species (ROS), they could potentially influence the cytosolic redox state, in turn altering RyR open probability. We have shown that self-sustained oscillations in mitochondrial inner membrane potential (ΔΨ(m)), NADH, ROS, and reduced glutathione (GSH) can be triggered by a laser flash in cardiomyocytes. Here, we employ this method to directly examine how acute changes in energy state dynamically influence resting Ca(2+) spark occurrence and properties. Two-photon laser scanning microscopy was used to monitor cytosolic Ca(2+) (or ROS), ΔΨ(m), and NADH (or GSH) simultaneously in isolated guinea pig cardiomyocytes. Resting Ca(2+) spark frequency increased with each ΔΨ(m) depolarization and decreased with ΔΨ(m) repolarization without affecting Ca(2+) spark amplitude or time-to-peak. Stabilization of mitochondrial energetics by pretreatment with the superoxide scavenger TMPyP, or by acute addition of 4'-chlorodiazepam, a mitochondrial benzodiazepine receptor antagonist that blocks the inner membrane anion channel, prevented or reversed, respectively, the increased spark frequency. Cyclosporine A did not block the ΔΨ(m) oscillations or prevent Ca(2+) spark modulation by ΔΨ(m). The results support the hypothesis that mitochondria exert an influential role on the redox environment of the Ca(2+) handling subsystem, with mechanistic implications for the pathophysiology of cardiac disease.

Topics: Animals; Benzodiazepinones; Calcium; Calcium Channels, L-Type; Free Radical Scavengers; GABA-A Receptor Antagonists; Glutathione; Guinea Pigs; Heart Diseases; Ion Channel Gating; Lasers; Membrane Potential, Mitochondrial; Microscopy, Confocal; Mitochondria, Heart; Myocytes, Cardiac; NAD; Oxidation-Reduction; Oxidative Stress; Photons; Porphyrins; Reactive Oxygen Species; Receptors, GABA-A; Ryanodine Receptor Calcium Release Channel; Ultraviolet Rays