Osmotic regulation of amino acids and system A transport in Madin-Darby canine kidney cells

Abstract

The effects of hypertonicity on the intracellular amino acid content and system A transport activity were studied in Madin-Darby canine kidney (MDCK) cells. Total content of 20 amino acids increased from 274 to 689 nmol/mg protein after 8 h of hypertonicity (500 mosmol/ kg), remaining almost constant until after 6 days of hypertonicity. The content of neutral amino acids increased from 77 to 307 and 395 nmol/mg protein after 8 h and 6 days of hypertonicity, respectively, accounting for 73% of the increased amount of total amino acids. In the hypertonic MDCK cells, system A transport activity, measured by Na+-dependent 2-(methylamino)isobutyric acid (MeAIB) uptake, increased approximately 60-fold relative to the uptake in isotonic cells. MeAIB was taken up primarily on the basal side in the isotonic MDCK cells cultured on permeable supports. Extracellular hypertonicity stimulated the MeAIB uptake predominantly on the basal side. These results indicated that amino acids, especially neutral amino acids, can function as volume-regulating osmolytes and that the stimulation of system A activity appears to contribute to the accumulation of neutral amino acids in hypertonic MDCK cells.