Diverse effects of sphingosine on calcium mobilization and influx in differentiated HL-60 cells.
CELL CALCIUM(2000)
Abstract
Sphingosine induces a biphasic increase in cytosolic-free Ca2+ ([Ca2+](i)) with an initial peak followed by a sustained increase in HL-60 cells differentiated into neutrophil-like cells. The initial peak is not affected by the presence of ethylene glycol bis (beta-aminoethyl ether) N, N, N', N-tetraacetic acid (EGTA) in the buffer and appears to be dependent on conversion of sphingosine to sphingosine -1-phosphate (S1P) by sphingosine kinase, since it is blocked by the presence of N, N-dimethylsphingosine (DMS), which, like sphingosine, causes a sustained increase itself. The sustained increase that is elicited by sphingosine or DMS is abolished by the presence of EGTA in the buffer. The sustained sphingosine-induced Ca2+ influx does not appear due to Ca2+ influx through store-operated Ca2+ (SOC) channels, since the influx is not inhibited by SKF 96365, nor is it augmented by loperamide. In addition, sphingosine and DMS attenuate the Ca2+ influx through SOC channels that occurs after depletion of intracellular stores by ATP or thapsigargin. Both the initial peak and the sustained increase in [Ca2+](i) elicited by sphingosine can be blocked by phorbol 12-myristate 13-acetate (PMA)-elicited activation of protein kinase C. Thus, in HL-60 cells sphingosine causes a mobilization of Ca2+ from intracellular Ca2+ stores, which requires conversion to S1P, while both sphingosine and DMS elicit a Ca2+ influx through an undefined Ca2+ channel and cause a blockade of SOC channels. (C) Harcourt Publishers Ltd 2000.
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Key words
protein kinase c,cell differentiation
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