Phenamil inhibits electrogenic sodium absorption in rabbit ileum

Electrogenic Na absorption, independent of either nutrients or other ions, occurs in the rabbit ileum. However, unlike electrogenic Na absorption in the distal colon and other tight epithelia, this ileal transport system is not inhibited by amiloride. Because of this amiloride insensitivity, ileal electrogenic Na absorption has been poorly characterized. To more clearly delineate the underlying mechanisms of this pathway, we examined the effects of phenamil, an amiloride analogue, on ion fluxes and electrical parameters in rabbit ileum in vitro under short-circuit conditions. Phenamil has been shown to have a high affinity for Na channels, but minimal effect on Na-H exchange. Amiloride (10(-8) through 10(-4) M) had a minimal effect on short-circuit current. In contrast, phenamil induced a significant decrease in short-circuit current; the maximal effect was seen at 10(-4) M phenamil. There was an associated decrease in conductance at 10(-4) M phenamil. Ion flux studies were performed in normal, chloride-free and bicarbonate-free Ringer's solution; under each condition, 10(-4) M phenamil inhibited mucosal-to-serosal Na flux, net Na flux, and short-circuit current without significantly altering other fluxes. Phenamil did not inhibit the electrical response to either 10 mM glucose or 1 mM theophylline, indicating that the drug did not block either nutrient-coupled electrogenic Na absorption or electrogenic Cl secretion, and did not inhibit sodium-potassium-stimulated adenosine triphosphatase. These results demonstrate that electrogenic Na absorption in rabbit ileum may be blocked by the amiloride analogue phenamil, suggesting that, in this epithelium, Na absorption may occur via Na channels in which the amiloride-binding site has been significantly altered.