1 To investigate the role of endothelium in vascular spasm, we studied the in¯uence of endothelin-1 (ET-1) on the contracting and spasmogenic e ect of the K + -channel blocker, tetraethylammonium (TEA), in aorta rings of reserpine-treated guinea-pigs, perfused with either control (5.5 mM) or elevated (50 mM) glucose concentration. 2 Endothelium-dependent relaxation induced by acetylcholine was lost in rings contracted by noradrenaline in the presence of elevated glucose. In control medium, TEA (1 ± 20 mM) induced a sustained tonic contraction, followed by a phasic spasm, characterized by rhythmic contractions. Elevated glucose, ET-1 (3 nM), or both, reduced the EC 50 of TEA-induced tonic contraction, without modifying the maximum contractile e ect. 3 In control medium, ET-1 reduced the time before TEA-induced spasm and increased the rate of rhythmic contractions. TEA-induced spasm was abolished by elevated glucose, and restored by ET-1. The spasm induced by TEA and ET-1 was ampli®ed by the ET A antagonist, EMD 94246 , and suppressed by the ET A -ET B antagonist, bosentan. In endothelium-denuded vessels incubated with high glucose and ET-1, TEA evoked only a tonic contraction. 4 In control medium, L-NAME (N G -nitro-L-arginine methyl ester) abolished TEA-induced rhythmic contractions. L-arginine, but not D-arginine, prevented the e ect of L-NAME. In the presence of elevated glucose and ET-1, TEA-induced spasm was not a ected by L-NAME, whereas verapamil, indomethacin, metyrapone, glybenclamide or apamin abolished the phasic spasm, unmasking the tonic contracture. 5 In conclusion, endothelium plays a regulatory role in the genesis and maintenance of TEAinduced rhythmic contractions, through the release endothelium derived relaxing factor and vasodilating eicosanoids.