Perfusion of the kidney with methylene blue, a soluble guanylate cyclase inhibitor, significantly enhanced the vasoconstrictor effects of angiotensin II, noradrenaline and phenylephrine but significantly reduced the vasodilator effect of acetylcholine without altering that of iloprost. 2. In the kidneys, which were perfused with Triton X-100 to remove endothelium, acetylcholine-induced vasodilation was completely abolished and angiotensin II-, noradrenaline- and phenylephrine-induced vasoconstriction was greatly reduced. 3. The vasodilator effect of iloprost was unchanged after perfusion of kidney with Triton X-100. 4. Neither methylene blue nor Triton X-100 significantly altered urine volume form normal and angiotensin II induced increase of urine volume. 5. These results were taken as evidence for the involvement of renal vascular endothelium originated EDRF in the responses of various vasoactive agents in the rabbit isolated perfused kidney.