Selective alteration of erythrocyte deformability by SH-reagents. Evidence for an involvement of spectrin in membrane shear elasticity

Fischer, Thomas M.; Haest, C.W.M.; Stöhr, M.; Kamp, D.; Deüticke, B.

doi:10.1016/0005-2736(78)90027-5

Cited by 167 publications

(77 citation statements)

References 19 publications

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“…13,15 This effect is pronounced in RBCs where G6PD is the sole producer of NADPH, an essential cofactor in the antioxidant defense mechanism. In type 2 diabetes, RBCs are under high oxidative stress and are believed to be less deformable, leading to lowered levels of deformation-induced ATP release.…”

Section: Discussionmentioning

confidence: 99%

“…[10][11][12] In its reduced form glutathione (GSH), a small tripeptide, is the main non-enzymatic antioxidant in the RBC and readily undergoes oxidation to the dimer (GSSG) to protect such important cellular components as proteins from oxidant insults (figure 1). [13][14][15] Under normal conditions, cells are capable of regenerating GSH at the expense of the reducing factor nicotinamide adenine dinucleotide phosphate (NADPH). Though other cells have several methods to produce NADPH, RBCs rely solely on the enzyme glucose-6-phosphate dehydrogenase (G6PD) for regeneration of NADPH, thereby rendering G6PD activity important to the antioxidant status of the RBC.…”

Section: Introductionmentioning

confidence: 99%

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Simultaneous determination of cell aging and ATP release from erythrocytes and its implications in type 2 diabetes

Subasinghe¹,

Spence²

2008

Analytica Chimica Acta

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Glucose-6-phosphate dehydrogenase (G6PD) is a determinant in the antioxidant status of the red blood cell (RBC) and is also used as an indicator of cell age. However, it is unknown if the relationship between antioxidant status, cell age, and RBC-derived adenosine triphosphate (ATP) occurs immediately or over a period of time. Therefore, the development of a simultaneous determination of G6PD activity (via the determination of nicotinamide adenine dinucleotide phosphate (NADPH)) in RBCs and the determination of deformation-induced RBC-derived ATP is described. The NADPH and ATP were determined while undergoing a chemically-induced aging process via inhibition of G6PD with dehydroepiandroesterone (DHEA). Upon incubation with DHEA for 30 minutes, NADPH levels measured in a flow stream decreased to 7.96 ± 1.10 μM from an original value of 13.20 ± 1.80 μM in a 0.02% solution of RBCs. In order to demonstrate a direct relationship between G6PD activity and deformation-induced ATP release from RBCs, a simultaneous microflow determination of G6PD activity and ATP release was performed. Upon inhibition with DHEA, NADPH levels decreased to 8.62 ± 0.29 μM from its original value of 12.73 ± 0.50 μM while ATP release decreased from 0.21 ± 0.07 μM to 0.06 ± 0.02 μM. These values were validated by an examination of NADPH levels in, and ATP release from, RBC fractions containing younger and older cells (separated by cell density centrifugation). This determination provides evidence that antioxidant status in the RBC and its ability to release ATP, a known stimulus of nitric oxide production, are closely related.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Simultaneous determination of cell aging and ATP release from erythrocytes and its implications in type 2 diabetes

Subasinghe¹,

Spence²

2008

Analytica Chimica Acta

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“…2a, expressed by DI vs. shear stress). The DI increased with the applied shear stress but it began to saturate at a higher shear stress (FISCHER et al, 1978a;KON et al, 1987).…”

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confidence: 98%

“…5% of the total erythrocytes' was collected (presumably containing younger and more deformable cells). In some cases, the fractionated cells were treated with diamide (diazene dicarboxylic acid bis(N,Ndimethylamide)) to reduce their deformability artificially (FIsCHER et al, 1978a;MAEDA et al, 1983).…”

mentioning

confidence: 99%

“…Application to detect a subtle change in deformability: It is known that the diamide treatment of erythrocytes induces the cross-linking of membrane proteins and reduces the cellular deformability (FISCHER et al, 1978a), e.g., we have quantitatively demonstrated the decrease in DI as increasing the degree of spectrin cross-linking (MAEDA et al, 1983) In Fig. 2a, the influence of diamide (0.2 mM) treatment on the deformation in uniform shear stress is shown: no change in DI was detected for the diamide-treated cells under constant shear flow (Fig.…”

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A novel method for measuring the erythrocyte deformability, under oscillatory shear stress.

et al. 1987

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Summary A novel apparatus for detecting a subtle decrease in erythrocyte deformability was developed. The oscillatory shape change of erythrocytes was monitored under an oscillatory shear stress (modulating the oscillation frequency in the range of 9-90 dyn/cm2). The ellipsoidal deformation of intact erythrocytes perfectly followed the oscillatory force of up to about 2 Hz, but the diamide-treated cells showed a phase difference of the oscillatory deformation at 1.7 Hz.Key words erythrocytes, deformability, oscillatory flow.The circulating erythrocytes are repeatedly exposed to the physical stresses which induce cellular deformation. Therefore, it is important to clarify the timedependent process of deformation, with respect to the biochemical properties. Although several quantitative methods have been developed for the measurement of erythrocyte deformability (OKA, 1981), none can be used to observe the oscillatory shape change in the flowing condition.Modifying the "cone-plate rheoscope" (SCHMID-SCHONBEIN and WELLS, 1969;SuDA et al., 1982), we constructed a novel apparatus to apply the oscillatory shear stress to the erythrocyte suspension. The flowing cells exposed to the oscillatory shear stress change their shape alternately, as a function of both the frequency and the amplitude of the oscillatory force. In its sensitivity to a subtle change in deformability, this method is superior to the conventional methods with uniform shear stress.In this note, we describe the design of the apparatus and show a typical example demonstrating its usefulness.Apparatus: As shown in Fig. 1, a transparent 0.8° cone was mounted on an inverted microscope (Olympus Optics, model IMT, Tokyo) and rotated by a stepping motor. The rotation rate of the cone can be modified by modulating the frequency of the driving pulses. The human erythrocytes were immersed in an

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Erythrocyte (Ca⁺²+Mg⁺²)‐ATPase activity: Increased sensitivity to oxidative stress in glucose‐6‐phosphate dehydrogenase deficiency

et al. 1985

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The effect of the thiol-oxidizing agent diamide on erythrocyte (Ca+2 + Mg+2)-ATPase activity was measured in normal and glucose-6-phosphate-dehydrogenase-deficient (G6PD-) cells. Although the enzyme activity before the oxidative stress was similar in both groups, diamide induced a markedly greater inhibition in the enzyme activity in the G6PD- cells than in the normal controls. These data indicate dependence of erythrocyte (Ca+2 + Mg+2)-ATPase, in part, on the redox status of the cell. The increased vulnerability of (Ca+2 + Mg+2)-ATPase to oxidative stress in G6PD- may be of pathophysiological relevance to their premature destruction in oxidant-induced hemolysis.

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Selective alteration of erythrocyte deformability by SH-reagents. Evidence for an involvement of spectrin in membrane shear elasticity

Cited by 167 publications

References 19 publications

Simultaneous determination of cell aging and ATP release from erythrocytes and its implications in type 2 diabetes

Simultaneous determination of cell aging and ATP release from erythrocytes and its implications in type 2 diabetes

A novel method for measuring the erythrocyte deformability, under oscillatory shear stress.

Erythrocyte (Ca⁺²+Mg⁺²)‐ATPase activity: Increased sensitivity to oxidative stress in glucose‐6‐phosphate dehydrogenase deficiency

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Selective alteration of erythrocyte deformability by SH-reagents. Evidence for an involvement of spectrin in membrane shear elasticity

Cited by 167 publications

References 19 publications

Simultaneous determination of cell aging and ATP release from erythrocytes and its implications in type 2 diabetes

Simultaneous determination of cell aging and ATP release from erythrocytes and its implications in type 2 diabetes

A novel method for measuring the erythrocyte deformability, under oscillatory shear stress.

Erythrocyte (Ca+2+Mg+2)‐ATPase activity: Increased sensitivity to oxidative stress in glucose‐6‐phosphate dehydrogenase deficiency

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Erythrocyte (Ca⁺²+Mg⁺²)‐ATPase activity: Increased sensitivity to oxidative stress in glucose‐6‐phosphate dehydrogenase deficiency