3 research outputs found
H2O2-mediated modulation of cytosolic signaling and organelle function in rat hippocampus
Reactive oxygen species (ROS) released from (dys-)functioning mitochondria contribute to normal and pathophysiological cellular signaling by modulating cytosolic redox state and redox-sensitive proteins. To identify putative redox targets involved in such signaling, we exposed hippocampal neurons to hydrogen peroxide (H2O2). Redox-sensitive dyes indicated that externally applied H2O2 may oxidize intracellular targets in cell cultures and acute tissue slices. In cultured neurons, H2O2 (EC50 118 µM) induced an intracellular Ca2+ rise which could still be evoked upon Ca2+ withdrawal and mitochondrial uncoupling. It was, however, antagonized by thapsigargin, dantrolene, 2-aminoethoxydiphenyl borate, and high levels of ryanodine, which identifies the endoplasmic reticulum (ER) as the intracellular Ca2+ store involved. Intracellular accumulation of endogenously generated H2O2—provoked by inhibiting glutathione peroxidase—also released Ca2+ from the ER, as did extracellular generation of superoxide. Phospholipase C (PLC)-mediated metabotropic signaling was depressed in the presence of H2O2, but cytosolic cyclic adenosine-5′-monophosphate (cAMP) levels were not affected. H2O2 (0.2–5 mM) moderately depolarized mitochondria, halted their intracellular trafficking in a Ca2+- and cAMP-independent manner, and directly oxidized cellular nicotinamide adenine dinucleotide (NADH) and flavin adenine dinucleotide (FADH2). In part, the mitochondrial depolarization reflects uptake of Ca2+ previously released from the ER. We conclude that H2O2 releases Ca2+ from the ER via both ryanodine and inositol trisphosphate receptors. Mitochondrial function is not markedly impaired even by millimolar concentrations of H2O2. Such modulation of Ca2+ signaling and organelle interaction by ROS affects the efficacy of PLC-mediated metabotropic signaling and may contribute to the adjustment of neuronal function to redox conditions and metabolic supply