Relations between intracellular Ca2+ stores and store-operated Ca2+ entry in primary cultured human glioblastoma...

1. In primary cultured human glioblastoma cells extracellular application of ATP triggered elevation in cytoplasmic calcium concentration ([Ca2+]i) mediated entirely by generation of inositol 1,4,5-trisphosphate (InsP3)-dependent Ca2+ release from endoplasmic reticulum Ca2+ stores followed by the activation of store-operated Ca2+ entry into the cells. 2. Sensitivity of P2Y purinoceptors to extracellular ATP was regulated by extracellular Ca2+: in Ca2+-free extracellular solution the threshold concentration of ATP that induced an increase in [Ca2+]i was reduced by one order of magnitude. 3. Activation of Ca2+ release and store-operated Ca2+ entry was dissociated: low concentrations of ATP induced substantial Ca2+ release without activation of Ca2+ entry; activation of the latter required higher ATP concentrations. 4. Mitochondria participated in buffering Ca2+ loads that resulted from store-operated Ca2+ influx; in contrast Ca2+ released from intracellular stores was not accumulated by the mitochondrial depot. 5. We conclude that ATP-induced Ca2+ responses are governed by several pathways with different sensitivities to the agonist. This enables cells to respond either with pure Ca2+ release from intracellular stores (at low ATP concentrations) or (at high ATP concentrations) the response is amplified by plasmalemmal Ca2+ influx. Store-operated Ca2+ entry increases mitochondrial Ca2+ content providing a link between cellular activation and mitochondrial function.