Deformation of swollen erythrocytes provides a model of sickling-induced leak pathways, including a novel bromide-sensitive component.
Deoxygenation-induced red blood cell (RBC) sickling probably activates multiple cation leak pathways. In an attempt to model this, we examined the net passive K efflux ("K leak") from normal and sickle RBCs undergoing elliptical deformation in hypotonic media (200 mOsmol/L). This hypotonic deformation activates two deformation-dependent K leak pathways that are not detectable during the balanced leak (Kefflux = Nainflux) resulting from deformation of RBCs in isotonic medium. These are (1) a calcium-dependent leak component and (2) a novel leak pathway that is inhibited by substitution of bromide (but not sulfamate) for chloride, which converts the unbalanced K leak (Kefflux > Nainflux) of hypotonic deformation to a residual balanced leak. This dramatic effect of hypotonic deformation is reversible, is detected in both normal and sickle RBCs, and is inhibited significantly by 4,4'-diisothiocyano-2,2'-stilbene disulfonate. Remarkably, bromide also inhibits by 55% the K leak resulting from authentic deoxygenation-induced RBC sickling and, thereby, blunts the imbalance of accompanying monovalent cation leaks. The unique effect of bromide is not readily explainable on the basis of known behaviors of known ion leak/transport pathways. The mechanical threshold for triggering K leak during hypotonic deformation is at applied shear stress of 164 dyne/cm2, a value similar to the abnormal susceptibility we previously found for oxygenated sickle RBCs during isotonic deformation. These data suggest that membrane stretch accompanying hypotonic deformation activates the same multiple leak pathways that contribute to net K leak during authentic RBC sickling, including a previously unknown bromide-sensitive leak.
Sugihara
T
T
Department of Medicine, Kawasaki Medical School, Kurashiki City, Japan.
Yawata
Y
Y
Hebbel
R P
RP
D000755:Anemia, Sickle Cell / Q000097:blood*; D001696:Biomechanical Phenomena; D001965:Bromides / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D018999:Charybdotoxin; D004907:Erythrocyte Deformability; D004910:Erythrocyte Membrane / Q000502:physiology; D006801:Humans; D007038:Hypotonic Solutions; D008954:Models, Biological; D010100:Oxygen / Q000097:blood; D011188:Potassium / Q000097:blood; D012604:Scorpion Venoms / Q000494:pharmacology; D012964:Sodium / Q000097:blood; D012965:Sodium Chloride
D001965:Bromides; D007038:Hypotonic Solutions; D012604:Scorpion Venoms; D018999:Charybdotoxin; D012965:Sodium Chloride; D012964:Sodium; D011188:Potassium; D010100:Oxygen; D002118:Calcium