The relationships between arachidonic acid (AA) metabolism and chloride secretion were investigated in mucosal preparations of rabbit distal colon. Tissues displayed a significant cyclooxygenenase activity already in nonstimulated conditions and incubation with exogenous AA and calcium ionophore A23187 produced a predominant prostaglandin F2-alpha (PGF2-alpha) profile [PGF2-alpha > PGE2 > thromboxane B2 (TxB2) > 6-keto-PGF1-alpha] as assessed by HPLC of tissue homogenates, whereas 5-hydroxy-6,8,11,14-eicosatetraenoic acid (5-HETE) was not detected in AA- or A23187-stimulated tissues. Radioimmunological assays showed that PGE2 synthesis was time dependent, plateaued at 10 min, and proceeded at rates 15-20 times over TxB2 and 6-keto-PGF1-alpha. Among the PGs produced by colonic mucosa, only PGE2 and, to a lower extent, PGF2-alpha, were found to stimulate chloride secretion and cAMP synthesis. Pretreatment with 10-mu-M 5,8,11,14-eicosatetraynoic acid, a cyclo- and lipoxygenase inhibitor, prevented AA-induced chloride secretion and PG and cAMP synthesis with the same strength as the cyclooxygenase inhibitor indomethacin. No effects were found after preincubation with nordihydroguaiaretic acid, a lipoxygenase blocker with moderate cyclooxygenase inhibitory properties, and caffeic acid, a lipoxygenase inhibitor. 5-HETE (5-mu-M) had no effect on short-circuit currents (I(sc)) and chloride transport, but it significantly reduced the increase in I(sc), chloride secretion, and PGE2 synthesis elicited by AA or A23187. Platelet-activating factor, reported to stimulate rabbit colon I(sc) through an indomethacin-sensitive pathway, was not detected at concentrations as low as 10(-10) M.