1 The effects of high K+ ion concentration on the isometric tension in dilator muscle strips of the rat and porcine iris were examined. A high K+ solution, prepared by the replacement of Na+ in the medium with equimolar K+, was applied in the presence of 1 UM phentolamine, 1 UM propranolol and 1 or lOAuM atropine. High K+ (>20 mM) induced a biphasic response; an initial phasic contraction followed by relaxation rather than tonic contraction. 2 An additional application of a Ca2+ antagonist, 1 HM nifedipine or nicardipine, almost completely blocked the K+-induced initial contraction and enhanced the following relaxation. The effect of K+ under these conditions was concentration-dependent in the range 20 to 80 mm. The maximum amplitude of the atropine-resistant relaxation induced by high K+ corresponds to 50-75% of that produced by acetylcholine in the absence of atropine. A similar K+-induced relaxation was observed in the porcine iris dilator. 3 The atropine-resistant relaxation in the rat iris dilator was not affected by pretreatment with 10pM ouabain. The relaxation induced by 40 or 80mM K+ in the porcine dilator was slightly enhanced or not affected, respectively, in the presence of 1 FM ouabain. Application of 10 uM ouabain per se induced relaxation in the porcine iris dilator.4 The low Na+ ion concentration present in high K+ solutions was not responsible for the K+-induced relaxation since the complete replacement of Na in the medium with Tris did not affect significantly the relaxation produced by high K+-containing solutions.5 Neither 1 yM tetrodotoxin, 10 JM indomethacin, 10 JM nordihydroguaiaretic acid nor hypoxic conditions affected the high K+-induced relaxation. 6 The inherent tone of the rat iris dilator was not affected by either 8-bromo cyclic GMP, dibutyryl cyclic GMP (0.1-0.3 mM) or nitroprusside (1-100 JM). 7 These results may suggest that the atropine-resistant relaxation induced by high K+ is not due to either activation of the Na-K pump or release of a relaxing factor produced by oxidative metabolism. Although the relaxation mechanism has not been elucidated, it is probably not mediated by an increase in cellular cyclic GMP levels.