We tested the hypothesis that in the rat duodenum and intestine the mesenteric hyperemia due to beta-adrenoceptor stimulation is mediated by capsaicin-sensitive afferent nerves. Superior mesenteric artery blood flow was measured by pulsed Doppler flowmetry in the anesthetized rat. Functional ablation of afferent nerves was accomplished by subcutaneous 125 mg/kg capsaicin pretreatment 10 to 14 days before blood flow studies. Blockade of capsaicin-sensitive cation-selective ion channels of the duodenal and intestinal mucosal afferent nerves was achieved by intraduodenal 0.1% ruthenium red given 15 min prior to the intraduodenal administration of 5 mg/kg isoproterenol. Functional ablation of the afferent nerves and blockade of the capsaicin-sensitive cation-selective ion channels alone or in combination resulted in a significant reduction of mesenteric hyperemia induced by intraduodenal isoproterenol. These data support the hypothesis that beta-adrenoceptor stimulation by intraduodenal isoproterenol induces mesenteric hyperemia in part through afferent nerves in the rat duodenal and intestinal mucosa. The results suggest for the first time a link between beta-adrenoceptor function and peripheral capsaicin-sensitive afferent nerve-mediated mechanism in the rat gut.