QUALITATIVE DIFFERENCES IN [CA-2+]I INCREASES AND INSP3 GENERATION FOLLOWING STIMULATION OF N1E-115 CELLS WITH MICROMOLAR AND MILLIMOLAR ATP

Addition of ATP (100 muM) to populations of the neuronal cell line N1E-115 caused a transient increase in intracellular calcium ([Ca2+]i) which rapidly reached a peak (maximum of 243 +/- 7 nM above basal) and returned to basal after approximately 50 sec. The response was concentration-dependent (EC50 21 +/- 4 muM) and was unchanged when calcium was omitted from the extracellular medium. Transient increases in D-myo-inositol-1,4,5-trisphosphate levels (InsP3) were also observed over a similar time period. Addition of millimolar ATP, however, produced characteristically different responses; [Ca2+]i again increased rapidly (reaching a maximum of 639 +/- 23 nM above basal) but returned to a new maintained plateau (274 +/- 34 nM) which was abolished by the inorganic calcium channel blocker, nickel (1 mM), and omission of calcium from the extracellular medium. InsP3 levels were also maintained but were, however, unaffected by nickel or removal of extracellular calcium, The qualitative difference in the mechanism of calcium elevation produced with millimolar ATP, compared with lower concentrations, suggests that the N1E-115 cells might also contain a low affinity P3 receptor coupled with a calcium channel.