Acetylcholine and cholinomimetics such as carbachol are potent stimulants of epithelial Cl- secretion in the small and large intestines of several mammalian species. In this study, the effects of carbachol were characterized in vitro on active ion transport in sheets of submucosa-mucosa from the distal jejunum of swine. Carbachol (10 microM) produced an increase in serosa-positive short-circuit current (Isc) in this tissue after its serosal, but not luminal administration. The Na(+)-K(+)-Cl- transport blocker bumetanide (10 microM) produced a 50% decrease in the carbachol-induced Isc elevation after its serosal administration. Peak increases in Isc evoked by carbachol were significantly reduced by 60-85% in tissues bathed in media lacking Cl-, HCO3-, or both anions. The initial drug-induced increase in net charge transfer from serosa to lumen was dependent upon both HCO3- and Cl-, whereas sustained elevations in charge transfer were dependent upon extracellular Cl- only. Radiotracer flux analyses revealed that the drug decreased net Na+ absorption and increased Cl- secretion. In the absence of HCO3-, carbachol decreased Cl- absorption. The effects of carbachol on HCO3- transport were examined by pH-stat titration. The drug rapidly alkalinized the luminal medium immediately after its serosal administration. These results suggest that carbachol stimulates electrogenic anion secretion in the mucosa of the porcine distal jejunum. Furthermore, the ability of carbachol to inhibit spontaneous Na+ absorption is dependent upon extracellular HCO3-.