Effects of intra- and extracellular H+ and Na+ concentrations on Na(+)-H+ antiport activity in the lacrimal gland acinar cells

Abstract

Kinetic properties of the Na(+)-H+ antiport in the acinar cells of the isolated, superfused mouse lacrimal gland were studied by measuring intracellular pH (pHi) and Na+ activity (aNai) with the aid of double-barreled H(+)- and Na(+)-selective microelectrodes, respectively. Bicarbonate-free solutions were used throughout. Under untreated control conditions, pHi was 7.12 +/- 0.01 and aNai was 6.7 +/- 0.6 mmol/l. The cells were acid-loaded by exposure to an NH4+ solution followed by an Na(+)-free N-methyl-D-glucamine (NMDG+) solution. Intracellular Na+ and H+ concentrations were manipulated by changing the duration of exposure to the above solutions. Subsequent addition of the standard Na+ solution rapidly increased pHi. This Na(+)-induced increase in pHi was almost completely inhibited by 0.5 mmol/l amiloride and was associated with a rapid, amiloride-sensitive increase in aNai. The rate of pHi recovery induced by the standard Na+ solution increased in a saturable manner as pHi decreased, and was negligible at pHi 7.2-7.3, indicating an inactivation of the Na(+)-H+ antiport. The apparent Km for intracellular H+ concentration was 105 nmol/l (pH 6.98). The rate of acid extrusion from the acid-loaded cells increased proportionally to the increase in extracellular pH. Depletion of aNai to less than 1 mmol/l by prolonged exposure to NMDG+ solution significantly increased the rate of Na(+)-dependent acid extrusion. The rate of acid extrusion increased as the extracellular Na+ concentration increased following Michaelis-Menten kinetics (Vmax was 0.55 pH/min and the apparent Km was 75 mmol/l at pHi 6.88).(ABSTRACT TRUNCATED AT 250 WORDS)