Abstract

2,2'-dithiodipyridine (2,2'-DTDP), a reactive disulphide that mobilizes Ca(2+) in muscle, induced an increase in cytoplasmic free Ca(2+)concentrations ([Ca(2+)](i)) in MG63 human osteosarcoma cells loaded with the Ca(2+)-sensitive dye fura-2. 2,2'-DTDP acted in a concentration-independent manner with an EC(50) of 50 microM. The Ca(2+) signal comprised an initial spike and a prolonged increase. Removing extracellular Ca(2+) did not alter the Ca(2+) signal, suggesting that the Ca(2+) signal was due to store Ca(2+) release. In Ca(2+)-free medium, the 2,2'-DTDP-induced [Ca(2+)](i) increase was not changed by depleting store Ca(2+) with 50 microM bredfeldin A (a Golgi apparatus permeabilizer), 2 microM carbonylcyanide m-chlorophenylhydrazone (CCCP, a mitochondrial uncoupler), 1 microM thapsigargin (an endoplasmic reticulum Ca(2+)pump inhibitor) or 5 microM ryanodine. Conversely, 2,2'-DTDP pretreatment abolished CCCP and thapsigargin-induced [Ca(2+)](i) increases. 2,2'-DTDP-induced Ca(2+) signals in Ca(2+)-containing medium were not affected by modulation of protein kinase C activity or suppression of phospholipase C activity. However, 2,2'-DTDP-induced Ca(2+) release was inhibited by a thiol-selective reducing reagent, dithiothreitol (5-25 microM) in a concentration-dependent manner. Collectively, this study shows that 2,2'-DTDP induced [Ca(2+)](i) increases in human osteosarcoma cells via releasing store Ca(2+)from multiple stores in a manner independent of protein kinase C or phospholipase C activity. The 2,2'-DTDP-induced store Ca(2+) release appeared to be dependent on oxidation of membranes.

Copyright 2002 Elsevier Science Inc.