BACKGROUND:Endothelin (ET) modulates motility of the internal anal sphincter through unclear receptor subtypes. METHODS:We measured relaxation of guinea pig internal anal sphincter strips caused by ET-related peptides and binding of (125)I-ET-1 to cell membranes prepared from the internal anal sphincter muscle. Visualization of (125)I-ET-1 binding sites in tissue was performed by autoradiography. KEY RESULTS:In the guinea pig internal anal sphincter, ET-1 caused a marked relaxation insensitive to tetrodotoxin, atropine, or omega-conotoxin GVIA. ET-2 was as potent as ET-1. ET-3 caused a mild relaxation. The relative potencies for ETs to cause relaxation were ET-1 = ET-2 > ET-3. The ET-1-induced relaxation was inhibited by BQ-123, an ET(A) antagonist, but not by BQ-788, an ET(B) antagonist. These indicate that ET(A) receptors mediate the relaxation. The relaxant response of ET-1 was attenuated by LY 83583, KT 5823, Rp-8CPT-cGMPS, tetraethyl ammonium, 4-aminopyridine and N(omega)-nitro-L-arginine, but not significantly affected by N(G)-nitro-L-arginine methyl ester, N(G)-methyl-L-arginine, charybdotoxin, apamin, KT 5720, and Rp-cAMPS. These suggest the involvement of cyclic guanosine 3',5'-cyclic monophosphate (cGMP), and potassium channels. Autoradiography localized (125)I-ET-1 binding to the internal anal sphincter. Binding of (125)I-ET-1 to the cell membranes prepared from the internal anal sphincter revealed the presence of two subtypes of ET receptors, ET(A) and ET(B) receptors. CONCLUSIONS & INFERENCES:Taken together, these results demonstrate that ET(A) receptors mediate relaxation of guinea pig internal anal sphincter through the cGMP pathway.