The pathway for refilling intracellular Ca2+ stores passes through the cytosol in human leukaemia cells

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Abstract
The pathway for refilling the intracellular Ca2+ stores of HL60 and U937 human leukaemia cells loaded with fura-2 has been investigated. On addition of external Ca2+ to cells with empty stores there was an increase in the cytosolic Ca2+ concentration ([Ca2+]i) which preceded the refilling of the stores. The increase in [Ca2+]i was faster than the refilling, by 3-to 15-fold, depending on the cell type. In measurements in single HL60 cells we found that the refilling of the stores correlated with the extent of the [Ca2+]i increase on addition of external Ca2+. The cells showing no [Ca2+]i increase were unable to refill their stores. The addition of Ni2+ to the extracellular medium prevented both the [Ca2+]i increase and the refilling of the stores. These results indicate that the limiting step for store refilling is the entry of Ca2+ from the extracellular medium to the cytosol. Hence, we conclude that extracellular Ca2+ cannot gain access directly to the intracellular Ca2+ stores in these cells, but must first enter the cytosol and be taken up from there into the stores.

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