1. Effects of arachidonic acid (AA) on voltage-dependent Ca channel currents were investigated by whole-cell-clamp methods in single smooth muscle cells freshly isolated from vas deferens of the guinea-pig. 2. Ca channel current was decreased by application of 1-30 microM AA in a concentration-dependent manner. When Ca2+ or Ba2+ was the charge carrier, Ca channel current (ICa or IBa) was reduced by AA to a similar extent (IC50 = 10 and 6 microM, respectively). Addition of 15 mM BAPTA to the pipette solution did not affect the reduction of IBa by 10 microM AA. 3. The effect of AA on IBa was not prevented by internal application of 1 mM nordihydroguaiaretic acid (NDGA) and 1 mM indomethacin (Indo). When the pipette solution contained 0.1 mM guanosine-5'-triphosphate (GTP), IBa was decreased slightly but significantly by application of 30 microM prostaglandin F2 alpha (PGF2 alpha) but not by PGE2. This effect of PGF2 alpha was irreversible or not observed when the pipette solution contained 0.3 mM guanosine-5'-(3-thiotriphosphate) (GTP gamma S) or both GTP or guanosine-5'-O-(2-thiodiphosphate) (GDP beta S), respectively. 4. External application of 100 units ml-1 superoxide dismutase slightly but significantly attenuated the inhibition of IBa by 1-30 microM AA. Intracellular application of 1 mM GDP beta S or 0.3 mM GTP gamma S did not significantly change the effect of AA. Intracellular application of 0.1 mM 1-(5-isoquinolinesulphonyl)-2-methylepiperazine (H-7) also did not change the effect of AA. 5. These results indicate that the decrease in Ca channel currents in vas deferens smooth muscle cells is mainly due to AA itself, as opposed to its metabolites. The effect of AA may be due to AA itself, as opposed to its metabolites. The effect of AA may be due to its direct action on Ca channels or membrane phospholipids, but may not be mediated by activation of GTP binding proteins or protein kinase C. The inhibition of Ca channel current by AA may be partly induced by superoxide radicals derived from AA oxidation. PGF2A also reduces Ca channel currents but probably by a separate mechanism via activation of a GTP binding protein.