The effect of acetylcholine (ACh), dopamine (DA), and bradykinin (BK) on vascular tone was examined in interlobular arteries and superficial afferent and efferent arterioles isolated from rabbit kidney. A single microvessel was dissected and cannulated, and lumen diameter was measured at a fixed intraluminal pressure. ACh caused a dose-dependent relaxation of norepinephrine-induced tone in all three vessel types. Significant relaxation (10-20%) was observed with 10(-8) M ACh and higher concentrations caused complete relaxation. In afferent and efferent arterioles DA caused a dose-dependent relaxation that was indistinguishable from the one caused by ACh. However, DA was much less effective on interlobular arteries. Significant relaxation was not observed until 10(-5) M DA, and 10(-4) M caused only a 30-40% relaxation. In afferent arterioles atropine blocked the effect of ACh, and metoclopramide selectively inhibited DA-induced relaxation. BK (10(-9) to 10(-5) M) caused a dose-dependent relaxation of norepinephrine-induced tone only in efferent arterioles. BK, either in the bath or lumen, had no effect on the preglomerular microvessels. ACh and DA also caused relaxation of afferent arterioles with spontaneous tone while all three vasodilators relaxed efferent arterioles with spontaneous tone. The results demonstrate segmental heterogeneity for these vasodilators in the rabbit renal microvasculature, with ACh causing relaxation in all three vessel types, DA acting primarily on the glomerular arterioles, and BK affecting only the efferent arteriole.