Sodium/proton transport by apical membranes of type-II pneumocytes

Recent studies fail to confirm the coexistence of Na+ channels and Na+/H+ exchange at the apical membranes of lower airway epithelia. Availability of plasma membrane vesicles simplifies the investigation of membrane transport processes. Apical and basolateral plasma membrane vesicles of disrupted type-II pneumocytes were fractionated upon nonlinear, continuous sucrose gradients. To investigate sodium transport, 22Na+ uptake by apical membrane vesicles was assayed in the presence and absence of transmembrane sodium diffusion potentials. Interior-negative sodium diffusion potentials promoted 22Na+ uptake 1.5-fold. Internally-directed H+ gradients or NH+4 gradients inhibited 22Na+ uptake 40-50%. Amiloride (1-1000 microM) inhibited uptake 10-79%. To investigate H+ transport, decay of transmembrane pH gradients was monitored with pH probe acridine orange. In the presence or absence of externally-directed H+ gradients, external sodium promoted internal alkalinization, except in the presence of external amiloride. These observations of amiloride-sensitive, electrogenic Na+ uptake and amiloride-sensitive, electroneutral, Na+/H+ coupling indicate coexistence of Na+ channels and Na+/H+ exchange at the apical membrane of type-II pneumocytes.