Ionic effects on firefly bioluminescence assay of red blood cell ATP

Aledort, Louis M.; Weed, Robert I.; Troup, Stanley B.

doi:10.1016/0003-2697(66)90205-3

Cited by 71 publications

(26 citation statements)

References 12 publications

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“…ATP was measured by the firefly bioluminescence method of Aledort et al (16). Intracellular ATP was expressed as micromoles per milliliter of erythrocytes as derived from the calibration curve and after correction for hematocrit and dilution.…”

Section: Methodsmentioning

confidence: 99%

Separate Mechanisms of Deformability Loss in ATP-depleted and Ca-loaded Erythrocytes

Clark¹,

Mohandas²,

Féo³

et al. 1981

J. Clin. Invest.

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A B S T R A C T Membrane rigidity has been widely accepted as the dominant cause of reduced deformability both of ATP-depleted erythrocytes and erythrocytes containing excess calcium (Ca). However, recent studies have shown normal membrane deformability in ATP-depleted erythrocytes. In addition, Ca accumulation causes massive ion and water loss, and it has been shown that extensive dehydration causes an increase in intracellular viscosity with attendant loss of whole cell deformability. To obtain a detailed understanding of the processes accompanying ATP depletion and/or Ca accumulation that limit cell deformability, we have used a viscodiffractometric method to identify the cellular factors contributing to reduced whole cell deformability.Analysis of the influence of the suspending medium osmolality on deformability showed the presence of two independent processes. One was a Ca-independent reduction in cell surface area/volume ratio, resulting from the spheroechinocyte formation that follows total ATP consumption. The other was a Cadependent increase in intracellular viscosity resulting from a Ca-induced loss of intracellular potassium and water. This deformability loss due to increased intracellular viscosity was found for cells depleted of ATP in the presence of Ca and in cells treated with Ca and A23187 without prior depletion. lonophoretreated cells at high Ca concentration (>500 ,uM) formed spheroechinocytes with reduced surface area and a further loss of whole cell deformability. The rate of deformability loss associated with Ca-induced spheroechinocytosis was much more rapid than that A portion of this material has been published in abstract form in 1979. Blood. 54(Suppl. 1): 25a. This is publication number 21 from the MacMillan-Cargill Hematology Research Laboratory.Address reprint requests to Dr. Clark in California.

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Section: Methodsmentioning

confidence: 99%

Separate Mechanisms of Deformability Loss in ATP-depleted and Ca-loaded Erythrocytes

Clark¹,

Mohandas²,

Féo³

et al. 1981

J. Clin. Invest.

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“…Red cell A TP measurements were carried out by the luciferin-luciferase assay method utilizing a liquid scintillation counter, as previously described (10).…”

Section: Methodsmentioning

confidence: 99%

Metabolic dependence of red cell deformability

Weed¹,

LaCelle²,

Merrill³

1969

J. Clin. Invest.

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787

251

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“…Light emis sion was counted in a Nuclear-Chicago liquid scintillation counter and concentrations calculated from internal standards, using known concentrations of disodium-ATP added to tissue extracts [2].…”

Section: Methodsmentioning

confidence: 99%

Effect of Extracellular Volume Expansion on Renal Na-K-ATPase and Cell Metabolism

Kramer

Gonick²

1974

Nephron

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In the present study the reduction in renal fractional reabsorption of sodium from 99.6 ± 0.2 to 86.0 ± 3.3 % following ECV expansion with isotonic saline in the rat was associated with a small decrease in renal cortical Na-K-ATPase activity from 12.2 ± 0.5 to 10.8 ± 0.6 µmol Pi/mg protein/h and an increase in medullary enzyme activity from 23.3 ± 1.3 to 28.6 ± 1.8 µmol Pi/mg protein/h. In addition, an increase in cortical ATP concentration from 1.21 ± 0.12 to 1.55 ± 0.10 µmol/g wet weight was observed. These alterations resemble renal metabolic changes induced by low dose ouabain and favor the hypothesis that decreased tubular reabsorption of sodium following ECV expansion may be associated with alterations in active sodium transport.

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Ionic effects on firefly bioluminescence assay of red blood cell ATP

Cited by 71 publications

References 12 publications

Separate Mechanisms of Deformability Loss in ATP-depleted and Ca-loaded Erythrocytes

Separate Mechanisms of Deformability Loss in ATP-depleted and Ca-loaded Erythrocytes

Metabolic dependence of red cell deformability

Effect of Extracellular Volume Expansion on Renal Na-K-ATPase and Cell Metabolism

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