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Received July 24, 2001
Accepted after revision November 30, 2001
1 Department of Physiology I, Gmelinstrasse 5, 72076 Tuebingen, Germany
2 Department of Physiology, University of Tuebingen, Gmelinstrasse 5, D 72076 Tuebingen, Germany
* To whom correspondence should be addressed. E-mail: christophe.duranton{at}uni-tuebingen.de.
Oxidative stress induces complex alterations of membrane proteins in red blood cells (RBCs) eventually leading to haemolysis. To study changes of membrane ion permeability induced by oxidative stress, whole-cell patch-clamp recordings and haemolysis experiments were performed in control and oxidised human RBCs. Control RBCs exhibited a small cation-selective whole-cell conductance (236 ± 38 pS; n = 8) which was highly sensitive to the external Cl- concentration: replacement of NaCl in the bath by sodium gluconate induced an increase of this cation conductance by about 85 %. Exposing RBCs to t-butylhydroxyperoxide (1 mm for 10 min) induced a twofold increase in this cation conductance which was further stimulated after replacement of extracellular Cl- by gluconate, Br-, I- or SCN-. In addition, lowering the ionic strength of the bath solution by isosmotic substitution of NaCl by sorbitol activated the cation conductance. The Cl--sensitive and oxidation-induced cation conductance was Ca2+ permeable, exhibited a permselectivity of Cs+ > K+ > Na+ = Li+ >> NMDG+, and was partially inhibited by amiloride (1 mm) and almost completely inhibited by GdCl3 (150 µm), but was insensitive to TEA, BaCl2, NPPB, flufenamic acid or quinidine. DIDS (100 µm) reversibly inhibited the activation of the cation conductance by removal of external Cl-. Oxidation induced haemolysis in NaCl-bathed human RBCs. This haemolysis was attenuated by amiloride (1 mm) and inhibited by replacement of bath Na+ by the impermeant cation NMDG+. The Na+- and Ca2+-permeable conductance might be involved in haemolytic diseases induced by elevated oxidative stress, such as glucose-6-phosphate dehydrogenase deficiency.
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