Distribution of dehydration rates generated by maximal Gardos-channel activation in normal and sickle red blood cells

Blood ◽  
2005 ◽  
Vol 105 (1) ◽  
pp. 361-367 ◽  
Author(s):  
Virgilio L. Lew ◽  
Teresa Tiffert ◽  
Zipora Etzion ◽  
Deisy Perdomo ◽  
Nuala Daw ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Osmotic Fragility ◽  
Large Cell ◽  
Volume Distribution ◽  
Human Red Blood Cells ◽  
Gardos Channel ◽  
Cell Dehydration ◽  
Osmotic Lysis ◽  
Channel Variation

Abstract The Ca2+-activated K+ channels of human red blood cells (RBCs) (Gardos channels, hIK1, hSK4) can mediate rapid cell dehydration, of particular relevance to the pathophysiology of sickle cell disease. Previous investigations gave widely discrepant estimates of the number of Gardos channels per RBC, from as few as 1 to 3 to as many as 300, with large cell-to-cell differences, suggesting that RBCs could differ extensively in their susceptibility to dehydration by elevated Ca2+. Here we investigated the distribution of dehydration rates induced by maximal and uniform Ca2+ loads in normal (AA) and sickle (SS) RBCs by measuring the time-dependent changes in osmotic fragility and RBC volume distributions. We found a remarkable conservation of osmotic lysis and volume distribution profiles during Ca2+-induced dehydration, indicating overall uniformity of dehydration rates among AA and SS RBCs. In light of these results, alternative interpretations were suggested for the previously proposed low estimates and heterogeneity of channel numbers per cell. The results support the view that stochastic Ca2+ permeabilization rather than Gardos-channel variation is the main determinant selecting which SS cells dehydrate through Gardos channels in each sickling episode. (Blood. 2005;105:361-367)

scholarly journals Age Decline in the Activity of the Ca2+-sensitive K+ Channel of Human Red Blood Cells

2007 ◽  
Vol 129 (5) ◽  
pp. 429-436 ◽  
Author(s):  
Teresa Tiffert ◽  
Nuala Daw ◽  
Zipora Etzion ◽  
Robert M. Bookchin ◽  
Virgilio L. Lew
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
K Channel ◽  
Human Red Blood Cells ◽  
Gardos Channel ◽  
Cell Age ◽  
Cell Dehydration ◽  
The Mean ◽  
Density Boundary ◽  
Substantial Degree

The Ca2+-sensitive K+ channel of human red blood cells (RBCs) (Gardos channel, hIK1, hSK4) was implicated in the progressive densification of RBCs during normal senescence and in the mechanism of sickle cell dehydration. Saturating RBC Ca2+ loads were shown before to induce rapid and homogeneous dehydration, suggesting that Gardos channel capacity was uniform among the RBCs, regardless of age. Using glycated hemoglobin as a reliable RBC age marker, we investigated the age–activity relation of Gardos channels by measuring the mean age of RBC subpopulations exceeding a set high density boundary during dehydration. When K+ permeabilization was induced with valinomycin, the oldest and densest cells, which started nearest to the set density boundary, crossed it first, reflecting conservation of the normal age–density distribution pattern during dehydration. However, when Ca2+ loads were used to induce maximal K+ fluxes via Gardos channels in all RBCs (Fmax), the youngest RBCs passed the boundary first, ahead of the older RBCs, indicating that Gardos channel Fmax was highest in those young RBCs, and that the previously observed appearance of uniform dehydration concealed a substantial degree of age scrambling during the dehydration process. Further analysis of the Gardos channel age–activity relation revealed a monotonic decline in Fmax with cell age, with a broad quasi-Gaussian Fmax distribution among the RBCs.

The Effects of Medical Diagnostic Low Dose X-rays after in vitro Exposure of Human Red Blood Cells: Hemolysis and Osmotic Fragility

2019 ◽  
Vol 11 (3) ◽  
pp. 237-243 ◽  
Author(s):  
Montree Tungjai ◽  
Jetchada Sopapang ◽  
Natdanai Tasri ◽  
Chanatip Osothsongkroh ◽  
Attapon Jantarato ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Low Dose ◽  
Osmotic Fragility ◽  
X Rays ◽  
Human Red Blood Cells ◽  
In Vitro Exposure ◽  
Medical Diagnostic

Effects of elevated intracellular calcium on the osmotic fragility of human red blood cells

Cell Calcium ◽  
2010 ◽  
Vol 47 (1) ◽  
pp. 29-36 ◽  
Author(s):  
Anne Cueff ◽  
Rachel Seear ◽  
Agnieszka Dyrda ◽  
Guillaume Bouyer ◽  
Stéphane Egée ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Intracellular Calcium ◽  
Blood Cells ◽  
Osmotic Fragility ◽  
Human Red Blood Cells

Stabilization of Human Erythrocytes for Lyophilization: Recovery of Biochemical Activities and Membrane Filterability.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3670-3670
Author(s):  
Shang-Zhi Xu ◽  
Arthur P. Bode
Keyword(s):  
Red Blood Cells ◽  
Pore Size ◽  
Blood Cells ◽  
White Blood Cells ◽  
Osmotic Fragility ◽  
Homocysteic Acid ◽  
Human Red Blood Cells ◽  
Before And After ◽  
Pass Through ◽  
2,3 Dpg

Abstract It has much more advantages to preserve human red blood cells (RBC) in freezing and drying form than conventional blood storage, even though it is still under developing. In the present study, we successfully used 0.045% glutaraldehyde(GA) to fix RBCs under appropriate conditions prior to lyophilization and regained the bulk of RBCs membrane filterability. The phlebotomized whole blood was anti-coagulated with CPDA-1 and the plasma was removed by centrifugation. White blood cells were removed using Leucocytes filter (Pall Corporation). 1 x 10 9 cells/ml were separately incubated with 0.03%, 0.045% and 0.05% GA at 37 °C for 10 min; RBCs at concentrations 0.5 x 10 9, 1 x 10 9 and 2 x 10 9 cell/ml were incubated with 0.045% GA at 37 °C for 10 min; Bovine serum albumin (BSA), trehalose, hydroxyethyl starch and dextran were used as protective additive in freezing and drying RBCs, respectively; ethanolamine, glycine, lysine, BSA, glutamic acid and homocysteic acid were used to recover membrane deformability due to GA fixation. Recovery RBCs yield after reconstituion was determined by a cell counter. Fee hemoglobin , glutathione (GSH), 2,3-diphosphoglycerate (2,3-DPG), ATP, and Glucose-6-phosphate dehydrogenase (G-6-PDH) in RBCs were determined before and after lyophilization. Membrane flexibility was assayed by osmotic fragility test. RBCs filterability was determined using positive filter apparatus through 5 micrometer pore size nitrocellulose membrane. Our results suggested that RBCs at 1 x 10 9 cells/ml fixed by 0.045% GA for10 min at pH 7.8 were enough to resist freezing and drying damages and the bulk of RBCs’s membrane filterability were remained. The effective fixation of GA is dependent on GA’s concentration, RBCs concentration, incubation period and pH etc. BSA is the most potential additive in preserving RBCs. More interestingly, the following reagents can recover the reduced RBCs membrane filterability by GA fixation. The potency of recovery is in the order of : 10 mM ethanolamine > 5 mM lysine > 0.5%BSA > 10 mM glutamatic acid > 5 mM homcysteic acid > 2% glycine. After reconstitution, 85 ± 2.3% RBCs yield is achieved, 71 ± 4.1% of these RBCs can freely pass through 5 micrometer pore size filter membrane. Biochemical function indexes as GSH, 2,3-DPG, ATP, and G-6-PDH are saved by 20 ± 1.2%. Lyophilized human red blood cells yields from reconstitution by different treatment. 1x 109 cells/ml fixed at 0.03% glutaraldehyde 44.7 ± 4.1 0.045%glutaraldehyde 84.7 ± 5.8 0.05%glutaraldehyde 88.7 ± 7.6 0.045% glutaraldehyde fix: 0.5x 109 cells/ml 91 ± 4.3 1x 109 cells/ml 84.7 ± 5.8 2 x 109 cells/ml 35.2 ± 2.3 Effects of various treatment on lyophilized RBCs after reconstitution. Group Filterability(%) 1x9 cells/ml fixed by: N = 5. 0.03%glutaraldehyde 88.3 ± 1.2 0.045%glutaraldehyde 73.3 ± 1.8 0.05%glutaraldehyde 44.7 ± 2.8 0.045%glutaraldehyde fixation at: 0.5x109 cells/ml 0 1x109 cells/ml 73.3 ± 1.8 2x109 cells/ml 83.9 ± 4.6

Effect of interleukin-8 and RANTES on the Gardos channel activity in sickle human red blood cells: Role of the Duffy antigen receptor for chemokines

2010 ◽  
Vol 44 (4) ◽  
pp. 219-223 ◽  
Author(s):  
Marie-Claude Durpès ◽  
Danitza Nebor ◽  
Pierre Couespel du Mesnil ◽  
Danièle Mougenel ◽  
Monique Decastel ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Antigen Receptor ◽  
Interleukin 8 ◽  
Channel Activity ◽  
Duffy Antigen ◽  
Human Red Blood Cells ◽  
Gardos Channel

scholarly journals The hSK4 (KCNN4) isoform is the Ca2+-activated K+ channel (Gardos channel) in human red blood cells

2003 ◽  
Vol 100 (12) ◽  
pp. 7366-7371 ◽  
Author(s):  
J. F. Hoffman ◽  
W. Joiner ◽  
K. Nehrke ◽  
O. Potapova ◽  
K. Foye ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
K Channel ◽  
Human Red Blood Cells ◽  
Gardos Channel

Ion Transport Pathology in the Mechanism of Sickle Cell Dehydration

2005 ◽  
Vol 85 (1) ◽  
pp. 179-200 ◽  
Author(s):  
Virgilio L. Lew ◽  
Robert M. Bookchin
Keyword(s):  
Red Blood Cells ◽  
Sickle Cell ◽  
Blood Cells ◽  
Experimental Tests ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Open Questions ◽  
Gardos Channel ◽  
Cell Dehydration ◽  
Experimental Findings

Polymers of deoxyhemoglobin S deform sickle cell anemia red blood cells into sickle shapes, leading to the formation of dense, dehydrated red blood cells with a markedly shortened life-span. Nearly four decades of intense research in many laboratories has led to a mechanistic understanding of the complex events leading from sickling-induced permeabilization of the red cell membrane to small cations, to the generation of the heterogeneity of age and hydration condition of circulating sickle cells. This review follows chronologically the major experimental findings and the evolution of guiding ideas for research in this field. Predictions derived from mathematical models of red cell and reticulocyte homeostasis led to the formulation of an alternative to prevailing gradualist views: a multitrack dehydration model based on interactive influences between the red cell anion exchanger and two K+transporters, the Gardos channel (hSK4, hIK1) and the K-Cl cotransporter (KCC), with differential effects dependent on red cell age and variability of KCC expression among reticulocytes. The experimental tests of the model predictions and the amply supportive results are discussed. The review concludes with a brief survey of the therapeutic strategies aimed at preventing sickle cell dehydration and with an analysis of the main open questions in the field.

Sign in / Sign up

Export Citation Format

Share Document