Furosemide-sensitive Na+ and K+ transport and human erythrocyte volume

The relationship between cation transport and cell volume in human erythrocytes was investigated by measuring ouabain-sensitive K+ influx, ouabain-resistant, furosemide-sensitive K+ influx, and ouabain + furosemide-resistant K+ influx, and maximal ouabain binding in microcytic, normocytic and macrocytic red cells. A significant correlation was found between the mean corpuscular volume and furosemide-sensitive K+ influx normalized either to cell number (r = 0.636, P less than 0.001) or to cell volume (r = 0.488, P less than 0.001). No relationship was seen between mean corpuscular volume and ouabain-sensitive K+ influx, and the number of ouabain-binding sites per cell was only weakly correlated with mean corpuscular volume (r = 0.337, P less than 0.05). A slight, negative relationship existed between mean corpuscular volume and ouabain + furosemide-resistant K+ influx expressed per volume of cells (r = -0.359, P less than 0.01), and an apparent relationship between furosemide-sensitive K+ influx and mean corpuscular hemoglobin concentration (r = 0.446, P less than 0.01) disappeared when microcytic samples were excluded from analysis. Furosemide-sensitive transport, including Na+ influx and K+ and Na+ efflux, was completely absent in microcytic cells from one patient with alpha-thalassemia minor. In addition, these cells exhibited a furosemide-resistant, Cl(-)-dependent K+ influx. Exposure of normal erythrocytes to hypotonic conditions (196 mosM) increased furosemide-sensitive K+ influx by a mean of 45% (P less than 0.05), while exposure to hypertonic conditions (386 mosM) had no significant effect. The results indicate that furosemide-sensitive transport and cell volume are interrelated in human erythrocytes. However, the inability to fully recreate this relationship with in vitro manipulation of cell volume suggest that this relationship is established prior to red cell maturation.