Sodium 22⁺ washout from cultured rat cells

Abstract

The washout of Na+ isotopes from tissues and cells is quite complex and not well defined. To further gain insight into this process, we have studied 22Na+ washout from cultured Wistar rat skin fibroblasts and vascular smooth muscle cells (VSMCs). In these preparations, 22Na+ washout is described by a general three-exponential function. The exponential factor of the fastest component (k1) and the initial exchange rate constant (kie) of cultured fibroblasts decrease in magnitude in response to incubation in K+-deficient medium or in the presence of ouabain and increase in magnitude when the cells are incubated in a Ca++-deficient medium. As the magnitude of the kie declines (in the presence of ouabain) to the level of the exponential factor of the middle component (k2), 22Na+ washout is adequately described by a two-exponential function. When the kie is further diminished (in the presence of both ouabain and phloretin) to the range of the exponential factor of the slowest component (k3), the washout of 22Na+ is apparently monoexponential. Calculations of the cellular Na+ concentrations, based on the 22Na+ activity in the cells at the initiation of the washout experiments, and the medium specific activity agree with atomic absorption spectrometry measurements of the cellular concentration of this ion. Thus, all three components of 22Na+ washout from cultured rat cells are of cellular origin. Using the exponential parameters, compartmental analyses of two models (in parallel and in series) with three cellular Na+ pools were performed. The results indicate that, independent of the model chosen, the relative size of the largest Na+ pool is 92-93% in fibroblasts and approximately 96% in VSMCs. This pool is most likely to represent the cytosol.

Effect of acute serum depletion on Na+ ‐K+ homeostasis in cultured human skin fibroblasts

“…The details for the 2zNaf washout experiments are given elsewhere (Kino et al, 1986). Briefly, cells were loaded with 25 pCi/ml "Na+ for 2 hours.…”

A higher cellular sodium turnover rate in cultured skin fibroblasts from blacks.

“…5 Briefly, cells in each well were washed twice with 5-ml aliquots of DMEM plus 10% FBS and loaded with 22 Na + in fresh medium for 20 minutes (20 /iCi/ml; Amersham, Arlington Heights, IL, US A) in the absence of ouabain and for 3 hours (8 /i.Ci/ml) in the presence of 10" 4 M ouabain. In preliminary studies we found that steady states with respect tô a" 1 " and Na + were attained at the indicated intervals.…”

Refined Evaluation of the Exponential Curve Parameters and Initial Exchange Rate Constant for 22Na+ Washout in Cultured Human Skin Fibroblasts

“…Hence, the largest and fastest of the three exponential components is likely to represent cellular Na+. This assertion is further substantiated by subsequent studies demonstrating that the exponential factor of the largest component is reduced by ouabain and is increased in a Ca2+-deficient medium (27). The exponential factor of the middle component was also sensitive to ouabain (27).…”