Mechanism of action of leukotriene D4 on guinea pig tracheal smooth muscle cells: roles of Ca++ influx and intracellular Ca++ release.

The effects of leukotriene D4 (LTD4) on the concentration of intracellular cytosolic free calcium ([Ca++]i) and on phosphoinositide hydrolysis were studied in cultured guinea pig tracheal smooth muscle cells. In Fura-2-loaded cells, LTD4 (10(-9)-10(-6) M) induced concentration-dependent changes in [Ca++]i consisting of a slow, transient increase followed by a sustained phase. Preincubation of cells with LTD4 receptor antagonist MK-571 (10(-6) M) blocked the increase in [Ca++]i. Similarly, LTD4-induced inositol phosphate ([3H]InsP(s) synthesis was transient, concentration-dependent and inhibited by the LTD4 antagonist. In the absence of extracellular Ca++, LTD4 failed to induce [Ca++]i increases and [3H]InsP(s) formation. Accordingly, NiCl2 completely inhibited the LTD4-stimulated [3H]InsP(s) synthesis. Nifedipine (10(-5) M) had a slight inhibitory effect on [Ca++]i increase but significantly reduced (40-50%) the [3H]lnsP(s) accumulation. These findings indicate that LTD4-stimulated inositol phosphate synthesis and [Ca++]i increases in tracheal smooth muscle cells are receptor-mediated events and are dependent on the availability of extracellular Ca++. It is suggested that Ca++ influx plays a major role in the LTD4 signal transduction mechanism.