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J Membr Biol. 1994 Dec;142(3):349-62.

Rate of activation and deactivation of K:Cl cotransport by changes in cell volume in hemoglobin SS, CC and AA red cells.

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Endocrine-Hypertension Division, Brigham and Women's Hospital, Boston, Massachusetts 02115.


Red blood cells (RBC) of subjects homozygous for hemoglobin A (AA), C (CC) and S (SS) exhibit different cell volumes which might be related to differences in cell volume regulation. We have investigated how rapidly K:Cl cotransport is activated and deactivated to regulate the cell volume in these cells. We measured the time course of net K+ efflux after step changes in cell volume and determined two delay times: one for activation by cell swelling and a second for deactivation by cell shrinkage. Cell swelling induced by 220 mOsm media activated K+ efflux to high values (10-20 mmol/liter cell x hr) in CC and SS; normal AA had a threefold lower activity. The delay time for activation was very short in blood with a high percentage of reticulocytes (retics): (SS, 10% retics, 1.7 +/- 0.3 min delay, n = 8; AA, 10% retics, 4 +/- 1.5 min, n = 3; CC, 11.6% retics, 4 +/- 0.3, n = 3) and long in cells with a smaller percentage of reticulocytes: (AA, 1.5% retics, 10 +/- 1.4 min, n = 8; CC whole blood 6% retics, 10 +/- 2.0 min, n = 10, P < 0.02 vs. SS). The delay times for deactivation by cell shrinking were very short in SS (3.6 +/- 0.4 min, n = 8, P < 0.02) and AA cells with high retics (2.7 +/- 1 min, n = 3) and normal retics (2.8 +/- 1 min, n = 3), but 8-15-fold longer in CC cells (29 +/- 2.8 min, n = 9). Density fractionation of CC cells (n = 3) resulted in coenrichment of the top fraction in reticulocytes and in swelling-activated cotransport (fourfold) with short delay time for activation (4 +/- 0.3 min) and long delay for deactivation (14 +/- 4 min). The delay time for activation, but not for deactivation, increased markedly with increasing cell density. These findings indicate that all CC cells do not promptly shut off cotransport with cell shrinkage and high rates of cellular K+ loss persist after return to isotonic conditions. In summary, (i) K:Cl cotransport is not only very active in young cells but it is also very rapidly activated and deactivated in young AA and SS cells by changes in cell volume. (ii) Delay times for cotransport activation markedly increased with RBC age and in mature cells with low cotransport rates, long delay times for activation were observed.(ABSTRACT TRUNCATED AT 400 WORDS).

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