Na +/K +/Cl − cotransport in resealed ghosts from erythrocytes of the Milan hypertensive rats

Abstract

The erythrocytes (RBC) of the Milan hypertensive rats (MHS) have a smaller volume and faster Na +/K +/Cl cotransport than RBC from normotensive controls (MNS). The difference in Na +/K +/Cl cotransport is no longer present in inside-out Vesicles (IOV) of RBC membrane. To differentiate between cytoplasmic or membrane skeleton abnormalities as possible causes of these differences. Resealed ghosts (RG) were used to measure ion transport systems. The following results have been obtained: (1) RG from MHS have a smaller volume than MNS (mean ± S.E. 20.7 ± 0.45 vs. 22.09 ± 0.42 fl, P < 0.05). (2) RG showed a bumetanide-sensitive Na efflux that retains the characteristics of the Na +/K +/Cl cotransport of the original RBC: it is K +- and Cl −-sensitive and dependent on the intracellular Na + concentration. (3) The Na +/K +/Cl − cotransport was faster in RG from MHS than in those from MNS (mean ± S.E. 0.095 ± 0.01 vs. 0.066 ± 0.01 rate constant h 1, P < 0.01). These results, together with those of IOV, support the hypothesis that an abnormality in the membrane skeletal proteins may play a role in the different Na +/K +/Cl cotransport modulation between MHS and MNS erythrocytes.