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Volume 272, Number 45, Issue of November 7, 1997 pp. 28360-28367
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

Store-operated Ca²⁺ Influx and Stimulation of Exocytosis in HL-60 Granulocytes

(Received for publication, December 10, 1996, and in revised form, September 5, 1997)

From the Division of Infectious Diseases, University Hospital Geneva, 1211 Geneva 14, Switzerland

This study addresses the role of store-operated Ca²⁺ influx in the regulation of exocytosis in inflammatory cells. In HL-60 granulocytes, which do not possess voltage-operated Ca²⁺ channels, the chemotactic peptide fMet-Leu-Phe (fMLP) was able to stimulate store-operated Ca²⁺ influx and to trigger exocytosis of primary granules. An efficient triggering of exocytosis by fMLP required the presence of extracellular Ca²⁺ and was inhibited by blockers of store-operated Ca²⁺ influx. However, receptor-independent activation of store-operated Ca²⁺ influx through thapsigargin did not trigger exocytosis. fMLP was unable to stimulate exocytosis in the absence of cytosolic free Ca²⁺ concentration [Ca²⁺]_c elevations. However, a second signal generated by fMLP synergized with store-operated Ca²⁺ influx to trigger exocytosis and led to a left shift of the exocytosis/[Ca²⁺]_c relationship in ionomycin-stimulated cells. The synergistic fMLP-generated signaling cascade was long-lasting, involved a pertussis toxin-sensitive G protein and a phosphatidylinositol 3-kinase. In summary, store-operated Ca²⁺ influx is crucial for the efficient triggering of exocytosis in HL-60 granulocytes, but, as opposed to Ca²⁺ influx through voltage-operated Ca²⁺ channels in neurons, it is not a sufficient stimulus by itself and requires synergistic receptor-generated signals.

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