To elucidate the vascular actions of lysophosphatidylcholine, we examined its effects on the concentration of cytosolic free calcium ([Ca2+]i), plasma membrane fluidity and release of lactate dehydrogenase in vascular endothelial cells cultured from bovine aortas. The [Ca2+]i of the endothelial cells was measured by a dual-wavelength fluorospectrophotometer using a fluorescent, calcium-specific indicator, Fura-2. Membrane fluidity was monitored by measuring changes in the steady-state fluorescence anisotropies, using 1,6-diphenyl-1,3,5-hexatriene as a fluorescence probe. In the presence of 1 mmol/L extracellular calcium, lysophosphatidylcholine caused a biphasic elevation of [Ca2+]i in Fura-2-loaded vascular endothelial cells, consisting of a large transient component followed by a relatively low, but more sustained component. At concentrations of lysophosphatidylcholine equal to or greater than 10 mumol/L, [Ca2+]i increased in a dose-dependent manner in the presence or absence of external calcium. In the absence of extracellular calcium, only an initial transient increment in the [Ca2+]i of endothelial cells was generated, the sustained component being eliminated. The sustained component was greatly depressed or almost abolished by the addition of the calcium influx blocker, NiCl2. Plasma membrane fluidity was greatly increased by incubation with lysophosphatidylcholine (30 and 50 mumol/L) concomitant with significant increases in the release of lactate dehydrogenase from the cells. At 50 mumol/L, lysophosphatidylcholine increased lactate dehydrogenase release and membrane fluidity in a time-related way.(ABSTRACT TRUNCATED AT 250 WORDS)