The sphingosine derivatives sphingosylphosphorylcholine (SPC) and sphingosine-1-phosphate (S1P) caused a similar elevation of the intracellular Ca2+ concentration ([Ca2+]i) in an immortalized airway epithelial cell line (CFNP9o-) incubated in Ca(2+)-free medium. The maximal effect was obtained with 2 microM SPC and 0.1 microM S1P and was sensitive to pre-incubation with pertussis toxin, indicating the involvement of a Gi/G(o) type of G protein. In Ca2+ containing medium, [Ca2+]i elevation by SPC was significantly higher than that by S1P, due to the fact that SPC was able to stimulate Mn2+ entry, whereas S1P was ineffective. SPC, but not S1P, caused a dose-dependent production of total inositol phosphates. Conversely, S1P, but not SPC, increased the level of phosphatidic acid. These findings suggest the presence of two distinct receptors, specific for SPC and S1P, respectively. Depletion of intracellular Ca2+ stores by SPC makes cells unable to respond to a subsequent addition of S1P. Conversely, cells do respond to SPC after a challenge with S1P, suggesting that the two receptors likely share one or more intracellular signalling component(s).