Histamine-induced inositol phosphate accumulation in HeLa cells: lithium sensitivity

Abstract

1. In the presence of 10 mM Li+ the histamine-stimulated accumulation of [3H]-inositol monophosphates [( 3H]-IP1) in HeLa cells prelabelled with [3H]-inositol increased over 10-20 min to a plateau level, which was normally maintained up to 60 min. Levels of [3H]-inositol bis- and trisphosphates [( 3H]-IP2 and [3H]-IP3) initially increased rapidly but declined to near basal levels by 20 min. 2. The same pattern of histamine-induced [3H]-IP1 accumulation was observed in cells in which [3H]-inositol was present 30 min before and during the incubation with histamine. Concentration-response curves for histamine measured in the presence of 10 mM Li+ were closely similar in cells prelabelled for 24 h with [3H]-inositol and in cells exposed to [3H]-inositol for only 30 min before addition of histamine, without removing the [3H]-inositol. The EC50 for histamine was 1.6 +/- 0.2 microM. 3. [3H]-IP1 accumulation induced by a sub-maximal concentration of histamine, 1 microM, also reached a plateau, but at a lower level than with 1 mM histamine. 4. Addition of 10 mM NaF at the plateau phase of [3H]-IP1 accumulation induced by 1 mM histamine resulted in a further increase in the level of [3H]-IP1. The level of [3H]-IP1 in the presence of histamine + NaF was 1.4 +/- 0.2 fold of that of the sum of the responses to histamine and NaF acting alone (basal levels subtracted). 5. Addition of 1 microM mepyramine at the plateau phase of [3H]-IP1 accumulation induced by 1 mM histamine in the presence of 10mM Li + resulted in a decline in the level of [3H]-IP1, implying that the metabolism of [3H]-IP1 is not completely blocked by 10mM Li' in HeLa cells. 6. Omission of the 15 min preincubation period with 10mM Li+ before stimulation with 100 microM histamine for 15 min resulted in an approximate halving of the level of [3H]-IP1, without any significant change in basal accumulation. Periods of preincubation with 10mM Li' longer than 15min did not produce any further increase in the level of [3H]-IP1 induced by histamine. 7. Basal and histamine-induced levels of [3H]-IP, increased as the concentration of Li+ was increased from 0 to 60mm, but the effect on the histamine-induced response was greater than on the basal level. Increasing the concentration of Li+ from 0 to 60 mm had only a small and mostly statistically insignificant effects on the levels of [3H]-1P2 and [3H]-1P3. The EC50 for histamine-induced [3H]-4P1 accumulation in the presence of 30mm Li + was 2.4 +/- 0.4 microM. 8. The results indicate that IP1 metabolism in HeLa cells is much less sensitive to Li+ than in mammalian brain. The plateau phase of histamine-induced [3H]-IP1 accumulation in the presence of 10mM Li+ thus represents a steady-state level. On a simple model the plateau level will be proportional to the rate of [3H]-IP1 formation at that time. The lower the concentration of Li' present, the better the approximation will be. The information obtained is thus not the same as when [3H]-1P, metabolism is completely blocked, in which case [3H]-IP1 accumulated can be used to calculate a mean rate of formation over the whole of the incubation period.