The ability of endothelin (ET) isopeptides to interact with ET receptor subtypes and stimulate phosphoinositide (PI) hydrolysis was examined in the rat lung. [125I]ET-1 and [125I]ET-3 binding to lung homogenates was saturable with maximal binding capacity values of 438 and 125 fmol/mg of protein and Kd values of 29 and 13 pM. The nonselective peptides, ET-1 and ET-2, produced steep inhibition of both [125I]ET-1 and [125I] ET-3 binding. The ETB-selective peptides, ET-3, sarafotoxin (SFX) S6a, SFX S6b and SFX S6c and the ETA-selective antagonist, BQ-123, generated shallow inhibition curves of [125I]ET-1 binding indicating the presence of both ETA and ETB receptors in the lung. Whereas the peptides exhibited similar potency in stimulating PI turnover in rat lung slices, the ability of ET-3 (1.6-fold) and SFX S6c (2-fold) to maximally stimulate [3H]inositol phosphate release was significantly different from the maximal response produced by ET-1 (4-fold) or SFX S6b (3.2-fold). The ETA-selective antagonist, BQ-123 [cyclo(L-Leu-D-Trp-D-Asp-L-Pro-D-Val)], inhibited PI hydrolysis induced by ET-1 or SFX S6b by approximately 80%, although having no effect on ET-3- or SFX S6c-induced PI turnover. Furthermore, ET-1- and SFX S6b-stimulated [3H]inositol phosphate release was significantly decreased in the presence of quinacrine and nordihydroguairetic acid, but not indomethacin. In contrast, these inhibitors had no effect on PI hydrolysis induced by SFX S6c.(ABSTRACT TRUNCATED AT 250 WORDS)