Anion permeability and erythrocyte swelling

Abstract

Permeability of cell membranes to cations may increase as a result of membrane oxidation or in certain pathologies. We studied the effects of nonselective increases in cell membrane permeability to univalent cations on the volume of erythrocytes incubated in phosphate-buffered saline (PBS) using amphotericin B (5–10 mg/l suspension) or gramicidin D (10–100 μg/l suspension) as the membrane permeabilizing agents. Both antibiotics caused K + to leak, Na + to accumulate intracellularly, and cell volume to increase. The interval needed to reach the equilibrium between the intracellular and extracellular ion concentrations ranged from 30 min to several hours, depending on the antibiotic concentration. In spite of a rapid disappearance of cation transmembrane gradients, cell volume increased relatively slow. Even 24 h after the membrane permeability was changed, the volume of most erythrocytes did not increase to the lytic values (about 1.6 times the normal volume). The slow increase in erythrocyte volume was accounted for by slow changes in the transmembrane Cl − gradient. 4,4′-Diisothiocyanatostilbene-2,2′-disulfonic acid (DIDS), a specific inhibitor of anion transport, while producing no effect on the transmembrane Na + and K + fluxes induced by the antibiotics, significantly inhibited the decrease in the transmembrane Cl − gradient and the increase in erythrocyte volume. Analysis of these data by means of mathematical modeling showed that it failed to satisfactorily describe the experimental kinetics of erythrocyte swelling in response to increases in the membrane permeability to univalent cations if its permeability to Cl − was set to be constant. The satisfactory description of this kinetics could be achieved by assuming that the membrane permeability to anions decreased with increasing erythrocyte volume. The results obtained demonstrate that transmembrane anion transport may be considered to be a component of the mechanism responsible for the erythrocyte volume stabilization, because a significant decrease in the swelling rate allows the erythrocytes with damaged membranes to activate a relatively slow (metabolic) mechanisms of cell volume stabilization and/or repair their damaged membranes.