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

Abstract

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. 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 μm. [3H]‐IP1 accumulation induced by a sub‐maximal concentration of histamine, 1 μm, also reached a plateau, but at a lower level than with 1 mm histamine. 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). Addition of 1 μm mepyramine at the plateau phase of [3H]‐IP1 accumulation induced by 1 mm histamine in the presence of 10 mm Li+ resulted in a decline in the level of [3H]‐IP1, implying that the metabolism of [3H]‐IP1 is not completely blocked by 10 mm Li+ in HeLa cells. Omission of the 15 min preincubation period with 10 mm Li+ before stimulation with 100 μm 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 10 mm Li+ longer than 15 min did not produce any further increase in the level of [3H]‐IP1 induced by histamine. Basal and histamine‐induced levels of [3H]‐IP1 increased as the concentration of Li+ was increased from 0 to 60 mm, 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]‐IP2 and [3H]‐IP3. The EC50 for histamine‐induced [3H]‐IP1 accumulation in the presence of 30 mm Li+ was 2.4 ± 0.4 μm. 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 10 mm 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]‐IP1 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. 1991 British Pharmacological Society