The effect of endothelin (ET) on cyclic GMP levels in cultured porcine kidney epithelial cells, LLC-PK1, was investigated. ET-1 or ET-3, but not big ET-1 or ET C-terminal hexapeptide 16-21, elevated cyclic GMP levels in a concentration-dependent manner with an EC50 value of about 5 x 10(-10) M. This effect of ET-1 was enhanced with superoxide dismutase, diminished with oxyhemoglobin, inhibited with methylene blue, totally dependent on extracellular calcium and unaffected by indomethacin. L-Arginine derivatives, NG-methyl-L-arginine and NG-nitro-L-arginine also inhibited cyclic GMP responses to 10(-8) M ET-1 with IC50 values of 1.2 x 10(-6) M and 7.6 x 10(-8) M, respectively, and the inhibition was prevented with L-arginine. These data strongly suggest that ET-1 stimulates formation of an endothelium-derived relaxing factor-like substance from L-arginine or a related endogenous material(s) in a Ca(++)-dependent fashion, which in turn activates soluble guanylate cyclase to elevate cellular cyclic GMP levels. The concentrations required for these effects were 10 times lower than those required for atrial natriuretic factor. Thus, the effects of ET on cyclic GMP accumulation may be related to the natriuretic effects of ET in vivo.