Intracellular Ca2+ Pools and Fluxes in Cardiac Muscle-Derived H9c2 Cells

Abstract

Relevant Ca(2+) pools and fluxes in H9c2 cells have been studied using fluorescent indicators and Ca(2+)-mobilizing agents. Vasopressin produced a cytoplasmic Ca(2+) peak with half-maximal effective concentration of 6 nM, whereas thapsigargin-induced Ca(2+) increase showed half-maximal effect at 3 nM. Depolarization of the mitochondrial inner membrane by protonophore was also associated with an increase in cytoplasmic Ca(2+). Ionomycin induced a small and sustained depolarization, while thapsigargin had a small but transient effect. The thapsigargin-sensitive Ca(2+) pool was also sensitive to ionomycin, whereas the protonophore-sensitive Ca(2+) pool was not. The vasopressin-induced cytoplasmic Ca(2+) signal, which caused a reversible discharge of the sarco-endoplasmic reticulum Ca(2+) pool, was sensed as a mitochondrial Ca(2+) peak but was unaffected by the permeability transition pore inhibitor cyclosporin A. The mitochondrial Ca(2+) peak was affected by cyclosporin A when the Ca(2+) signal was induced by irreversible discharge of the intracellular Ca(2+) pool, i.e., adding thapsigargin. These observations indicate that the mitochondria interpret the cytoplasmic Ca(2+) signals generated in the reticular store.