Detection of a Na+-H+ antiporter in cultured rat renal papillary collecting duct cells

Abstract

To examine whether Na+-dependent H+ transport is present in the papillary collecting duct, changes in intracellular pH (pHi) were evaluated in cultured papillary collecting duct cells acidified to a pHi of 6.3 and then placed into Na+-free or Na+-containing solutions. pHi was determined from changes in the fluorescent signal of the pH-sensitive dye BCECF. pHi did not change significantly when cells were placed in tetramethylammonium chloride- or KCl-containing solutions; however, a significant rise in pHi occurred when acid-loaded cells were placed in solutions containing 140 mM NaCl. The Na+-dependent rise in pHi was blocked by high concentrations of amiloride, but was not affected by alterations in membrane potential across the cell. The rate of rise in pHi was a function of extracellular sodium concentration with a Km for Na+ of 30 +/- 12 mM (n = 6). The properties of this Na+-dependent H+ efflux supports the presence of a Na+-H+ antiporter in the papillary collecting duct.