cAMP-dependent secretagogues stimulate the NaHCO3 cotransporter in the villous epithelium of the brushtail possum, Trichosurus vulpecula

Abstract

In the ileum of the brushtail possum, Trichosurus vulpecula, fluid secretion appears to be driven by electrogenic HCO secretion. Consistent with this, the cystic fibrosis transmembrane conductance regulator is expressed in the apical membrane of the ileal epithelial cells and the pancreatic or secretory variant of the NaHCO cotransporter in the basolateral membrane. This suggests that in the possum ileum, electrogenic HCO secretion is driven by basolateral NaHCO cotransporter (NBC) activity. To determine if the NBC contributes to HCO secretion in the possum ileum, intracellular pH (pHi) measurements in isolated villi were used to demonstrate NBC activity in the ileal epithelial cells and investigate the effect of cAMP-dependent secretagogues. In CO/HCO-free solutions, recovery of the epithelial cells from an acid load was Na-dependent and ≈80% inhibited by ethyl-isopropyl-amiloride (EIPA, 10 µmol L), indicative of the presence of an Na/H exchanger, most likely NHE1. However, in the presence of CO/HCO, EIPA only inhibited ≈ 50% of the recovery, the remainder was inhibited by 4,4'-diisothiocyano-2,2'-stilbenedisulfonic acid (DIDS, 500 µmol L), indicative of NBC activity. Under steady-state conditions, NHE1 inhibition by EIPA had little effect on pHi in the presence or absence of secretagogues, but NBC inhibition with DIDS resulted in a rapid acidification of the cells, which was increased fivefold by secretagogues. These data demonstrate the functional activity of an NaHCO cotransporter in the ileal epithelial cells. Furthermore, the stimulation of NBC activity by secretagogues is consistent with the involvement of an NaHCO cotransporter in electrogenic HCO secretion.