The use of a fluorescent probe to monitor alterations in trans-membrane potential in single cell suspensions

Bramhall, John; Morgan, James I.; Perris, A. D.; Britten, A. Z.

doi:10.1016/s0006-291x(76)80090-3

Cited by 32 publications

(6 citation statements)

References 13 publications

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“…In contrast, addition of 0.3 mM calcium ionophore A23187, caused =6 mV increase in membrane potential. The membrane potential changes induced by these compounds (ouabain, valinomycin, and A23187) in CEM cells, as measured by "F-NMR, and the direction of change of the membrane potential are similar to those induced by the same compounds in lymphocytes and many other types of cells using different methods for membrane potential determinations (Bramhall et al, 1976;Smith and Robinson, 1982;Wittenkeller et al 1992). The results of the experiments with ouabain, valinomycin, and A23187 are shown in Figure 5.…”

Section: Resultsmentioning

confidence: 93%

¹⁹F NMR studies of changes in membrane potential and intracellular volume during dexamethasone‐induced apoptosis in human leukemic cell lines

Adebodun

Post

1993

Journal Cellular Physiology

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The induction of apoptosis in leukemic cells by dexamethasone is well known, but the mechanism of this type of cell death and of dexamethasone resistance by some variants is still poorly understood. Apoptotic cell death is preceded by many changes in cellular properties, such as glucose metabolism, cell size, cell density, and others. In this study, 19F-NMR has been used to characterize changes in cell membrane potential and intracellular accessible volume during dexamethasone induced apoptosis. One dex-sensitive (CEM-C7) and three dex-resistant variants (CEM-C1, CEM-ICR27, and CEM-4R4) were examined. We have observed separate intracellular and extracellular resonances for trifluoroacetate and trifluoroacetamide added to suspended leukemic cells. From the equilibrium distribution of these fluoro-compounds between intra and extracellular spaces, the changes in membrane potential and intracellular accessible volume were calculated. The membrane potential for CEM-C7 cells was found to significantly decrease in the presence of dexamethasone (9-mV decrease within 18 h of dexamethasone treatment), while that of CEM-ICR27 was found in some samples to increase on dexamethasone incubation. The membrane potential for CEM-C1 decreased slightly, while that of CEM-4R4 was not appreciably affected by dexamethasone. The reduction of membrane potential seems to be an early step in the mechanism of dexamethasone induced apoptosis. Although the intracellular volume varied with cell type and dexamethasone incubation (for CEM-C7), the fractional intracellular volume (alpha = Vin/Vcell) was found to be the same (0.82 +/- 0.06) for all the cell lines in the presence and absence of dexamethasone.

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Section: Resultsmentioning

confidence: 93%

¹⁹F NMR studies of changes in membrane potential and intracellular volume during dexamethasone‐induced apoptosis in human leukemic cell lines

Adebodun

Post

1993

Journal Cellular Physiology

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“…The application of optical methods using dye 64 to the measurement of membrane potentials in thymic and splenic lymphocytes and in platelets was established by Bramhall et al (1976). The effects of extracellular calcium concentrations, as well as steroids, on the membrane potential of lymphocytes was then studied .…”

Section: Lymphocytes and Plateletsmentioning

confidence: 99%

Optical measurement of membrane potential

Cohen

Salzberg

1978

Reviews of Physiology, Biochemistry and Pharmacology

482

285

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“…Waggoner (34) has developed a series of carbocyanine dyes, the fluorescence response of which has been shown to correlate with independently measured changes in membrane potential (34)(35)(36)(37). Investigators using these dyes have indicated that platelets maintain a trans membrane potential, inside negative, in the range of -4 8 to -6 0 mV (9)(10)(11)(12).…”

Section: Effects O F Platelet Activation On Membrane Potentialmentioning

confidence: 99%

“…There is evidence that early events may involve alterations in ionic gradients which are maintained across the plasma membrane (5)(6)(7)(8). Investigators using fluorometric dye techniques have provided evidence for a platelet transmembrane potential, inside negative, that is perturbed by aggregating agents (9)(10)(11)(12). How ever, the suitability of fluorescent probes to measure membrane potential during platelet activation is uncertain due to the responsiveness of the dyes to changes in cytoskeletal structure as well as ion fluxes (14)(15).…”

Section: Introductionmentioning

confidence: 99%

ADP Induced Depolarization of Human Platelet Membrane Potential

Wencel-Drake

Feinberg

1985

Thromb Haemost

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SummaryThe transmembrane potential of human blood platelets suspended in plasma was investigated by studying the distribution of a radiolabeled permeant ion [14C] thiocyanate. The membrane potential of resting platelets was found to be -54.50 mV ± 9.23 S.D. with a range of -39 to -76 mV (n = 27). The possibility that platelet activation alters membrane potential or that changes in membrane potential serve as an activation trigger was investigated. Stimulation by ADP (10 ¼M) resulted in a significant (p :0.05) depolarization of the membrane potential. Preincubation with 6 mM EGTA failed to inhibit ADP-induced depolarization even though EGTA effectively prevented primary and secondary aggregation but not shape change. Preincubation with PGE1 inhibited shape change, aggregation, and the ADP- induced depolarization. No significant change in membrane potential was observed following stimulation by epinephrine (50 ¼M). These results suggest that the initial interaction of ADP and its receptor may involve an inward positive current which can be determined by thiocyanate distribution.

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The use of a fluorescent probe to monitor alterations in trans-membrane potential in single cell suspensions

Cited by 32 publications

References 13 publications

¹⁹F NMR studies of changes in membrane potential and intracellular volume during dexamethasone‐induced apoptosis in human leukemic cell lines

¹⁹F NMR studies of changes in membrane potential and intracellular volume during dexamethasone‐induced apoptosis in human leukemic cell lines

Optical measurement of membrane potential

ADP Induced Depolarization of Human Platelet Membrane Potential

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The use of a fluorescent probe to monitor alterations in trans-membrane potential in single cell suspensions

Cited by 32 publications

References 13 publications

19F NMR studies of changes in membrane potential and intracellular volume during dexamethasone‐induced apoptosis in human leukemic cell lines

19F NMR studies of changes in membrane potential and intracellular volume during dexamethasone‐induced apoptosis in human leukemic cell lines

Optical measurement of membrane potential

ADP Induced Depolarization of Human Platelet Membrane Potential

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¹⁹F NMR studies of changes in membrane potential and intracellular volume during dexamethasone‐induced apoptosis in human leukemic cell lines

¹⁹F NMR studies of changes in membrane potential and intracellular volume during dexamethasone‐induced apoptosis in human leukemic cell lines