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Deoxygenation inhibits the volume-stimulated, Cl(-)-dependent K+ efflux in
SS and young AA cells: a cytosolic Mg2+ modulation
M Canessa, ME Fabry and RL Nagel
Brigham and Women's Hospital, Department of Medicine, Harvard University,
Boston.
We recently reported that the Cl(-)-dependent K+ (K:Cl) efflux, which can
be stimulated by cell swelling in the presence of inhibitors of the Na+
pump (ouabain) and of the Na-K-Cl cotransport (bumetanide), is highly
active in young AA and SS RBCs. We report here that deoxygenation inhibits
volume-stimulated K:Cl efflux in SS and reticulocyte-enriched
density-separated SS and AA RBCs. In SS whole blood, the K:Cl efflux
stimulated by hypotonic (220 mOsm) swelling was reduced from 9.2 +/- 2
(mean +/- SE) in oxygen to 2.7 +/- 1.9 (mmol/L cell/h = flux units = FU) (n
= 4) under deoxygenated conditions (P less than .005). Deoxygenation also
decreased the acid pH-stimulated K:Cl efflux from 5.9 +/- 1.5 to 3.7 +/-
1.1 FU (n = 3) (P less than .025) but did not inhibit NEM-stimulated K:Cl
transport. The effect of deoxygenation on density-separated SS cells is
similar: When fraction SS2 (reversible discocytes) is deoxygenated under
hypotonic conditions, the K:Cl efflux is reduced by 50%. In
reticulocyte-enriched AA cells obtained from anemic patients, deoxygenation
under hypotonic conditions also reduces K+ efflux by 50%. In SS cells only,
deoxygenation under isotonic conditions results in an increased
Cl(-)-independent K+ efflux. Because ionized Mg2+ in the cytosol increases
during deoxygenation, we investigated the effect of external and internal
Mg2+ on the volume-stimulated K:Cl efflux. Removal of external Mg2+ did not
influence the rate of transport in oxygenated cells. When internal Mg2+ was
clamped at 0.15 mmol/L with A23187 and EDTA at ionized cytosolic Ca2+ = O,
however, the inhibitory effect of deoxygenation on the K:Cl efflux was
eliminated. We conclude that deoxygenation inhibits the volume-stimulated,
Cl(-)-dependent K+ efflux in AA and SS young red cells by concomitantly
increasing ionized cytosolic Mg2+.
Volume 70,
Issue 6,
pp. 1861-1866,
12/01/1987
Copyright © 1987 by The American Society of Hematology
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