Ae4 (Slc4a9) belongs to the Slc4a family of Cl−/HCO3− exchangers and Na+-HCO3− cotransporters, but its ion transport cycle is poorly understood. In this study, we find that native Ae4 activity in mouse salivary gland acinar cells supports Na+-dependent Cl−/HCO3− exchange that is comparable with that obtained upon heterologous expression of mouse Ae4 and human AE4 in CHO-K1 cells. Additionally, whole cell recordings and ion concentration measurements demonstrate that Na+ is transported by Ae4 in the same direction as HCO3− (and opposite to that of Cl−) and that ion transport is not associated with changes in membrane potential. We also find that Ae4 can mediate Na+-HCO3− cotransport–like activity under Cl−-free conditions. However, whole cell recordings show that this apparent Na+-HCO3− cotransport activity is in fact electroneutral HCO3−/Na+-HCO3− exchange. Although the Ae4 anion exchanger is thought to regulate intracellular Cl− concentration in exocrine gland acinar cells, our thermodynamic calculations predict that the intracellular Na+, Cl−, and HCO3− concentrations required for Ae4-mediated Cl− influx differ markedly from those reported for acinar secretory cells at rest or under sustained stimulation. Given that K+ ions share many properties with Na+ ions and reach intracellular concentrations of 140–150 mM (essentially the same as extracellular [Na+]), we hypothesize that Ae4 could mediate K+-dependent Cl−/HCO3− exchange. Indeed, we find that Ae4 mediates Cl−/HCO3− exchange activity in the presence of K+ as well as Cs+, Li+, and Rb+. In summary, our results strongly suggest that Ae4 is an electroneutral Cl−/nonselective cation–HCO3− exchanger. We postulate that the physiological role of Ae4 in secretory cells is to promote Cl− influx in exchange for K+(Na+) and HCO3− ions.
Ae4 (Slc4a9) is an electroneutral monovalent cation-dependent Cl−/HCO3− exchanger