Sodium fluxes in human fibroblasts: effect of serum, Ca+2, and amiloride

Journal of Cellular Physiology
M L Villereal

Abstract

Human fibroblasts that have been serum deprived for 4 hours have a digitoxin-insensitive Na influx of 9.5 +/- 1.0 (n = 4) mumol/g prot/min which is not significantly different from the influx of 9.4 +/- 0.6 (n = 3) mumol/g prot/min measured in cells arrested in the G1/G0 state by serum-deprivation for a period of four days. The Na influx in serum-deprived cells is rapidly stimulated (within one minute) simply by assaying the cells in medium containing 10% fetal bovine serum (FBS). The digitoxin-insensitive NA influx for cells in the presence of 10% FBS is 22.9 +/- 1.1 (n = 6) mumol/g prot/min. The stimulation of Na influx in serum-deprived cells can also be achieved by the addition of the purified mitogen, epidermal growth factor (EGF). Addition of EGF to serum-deprived cells gives a maximal stimulation of Na influx of approximately 1.6-fold, with the concentration for half-maximal stimulation being 7.5 ng/ml. The stimulation of Na influx results from the activation of an amiloride-sensitive pathway, which appears to be minimally active in serum-deprived cells. Kinetic analysis of Na influx experiments in the presence of 10% FBS and varying concentrations of amiloride indicate that at infinite concentrations of amiloride the Na...Continue Reading

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