Flow cytometric determination of absolute membrane potential of cells

Krasznai, Zoltán; Márián, Teréz; Balkay, László; Emri, Miklós; Trón, Lajos

doi:10.1016/1011-1344(94)07099-a

Cited by 56 publications

(75 citation statements)

References 19 publications

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“…The protocol developed by Krasznai et al (15) was used that allows the evaluation of the membrane potential in millivolts using fluorescence readings of the cells both in depolarized state and in the state of interest. For the measurements, a modified FACSTAR flow cytometer equipped with an argon ion laser was used (Becton Dickinson, Parsippany, NJ, USA).…”

Section: Flow Cytometric Determination Of the Membrane Potentialmentioning

confidence: 99%

A1 and A2 Adenosine Receptor Activation Inversely Modulates Potassium Currents and Membrane Potential in DDT1 MF-2 Smooth Muscle Cells

Márián

Rubovszky

Szentmiklósi

et al. 2002

Japanese Journal of Pharmacology

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ABSTRACT-Adenosine receptors are widely distributed in mammalian tissues and have been possibly involved through transmembrane potential changes in cell function regulation. The effect of A1 and A2A adenosine receptor ligands on transmembrane potential measured with flow cytometry and potassium conductance measured by the patch-clamp technique was investigated in DDT1 MF-2 smooth muscle cells. The A1 adenosine-receptor agonist CPA (50 nM) and the A2A adenosine-receptor agonist CGS 21680 (50 nM) elicited a rapid and maintained increase and decrease in the potassium conductance, respectively, and a concomitant hyperpolarization and depolarization of the membrane, respectively. These effects were eliminated by subtype-selective adenosine receptor antagonists (DPCPX, CSC, ZM 241385, all 1 mM). The ligand induced membrane potential changes were reversible. Based on these detected membrane potential changes along with the published voltage dependence of the adenylyl cyclase, the regulation of cAMP production by A1-and A2A-receptor activation is suggested to be mediated through the induced early hyperpolarization and depolarization. The interaction between the effects of these receptor subtypes allows for a complex regulation mechanism.

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Section: Flow Cytometric Determination Of the Membrane Potentialmentioning

confidence: 99%

A1 and A2 Adenosine Receptor Activation Inversely Modulates Potassium Currents and Membrane Potential in DDT1 MF-2 Smooth Muscle Cells

Márián

Rubovszky

Szentmiklósi

et al. 2002

Japanese Journal of Pharmacology

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“…Eosinophil transmembrane potential was determined using diBAC4(3), a ratiometric fluorescent membrane potential-sensitive dye (Molecular Probes, Eugene, OR) based on a protocol by Krasznai et al (Krasznai et al, 1995). Cells were resuspended at 0.5×10 6 cells/ml in HybriCare media (no bicarbonate buffer, with 25 mM HEPES) (ATCC, Rockville, MD) in HSA-blocked FACS tubes (Falcon 2052, Fisher Scientific, Lincoln Park, NJ).…”

Section: Transmembrane Potential Measurementsmentioning

confidence: 99%

“…Inhibitors were incubated with cells at least 10 minutes before PMA stimulation. A linear calibration curve was determined from the cells incubated with varying concentrations of diBAC4(3), and this equation was used to calculate the absolute transmembrane potential of the cells as described by Krasznai et al (Krasznai et al, 1995).…”

Section: Transmembrane Potential Measurementsmentioning

confidence: 99%

“…Here, we describe the effects of PMA on the membrane potential of eosinophils using a cell-permeable oxonol membrane potential indicator, bis-barbituric acid oxonol [diBAC4(3)], to determine the extent of depolarization (Krasznai et al, 1995). Like neutrophils, eosinophils depolarize to a membrane potential sufficient to allow activation of the NADPH oxidase-associated proton channel under conditions where superoxide is produced.…”

Section: Introductionmentioning

confidence: 99%

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Regulation of eosinophil membrane depolarization during NADPH oxidase activation

Bankers-Fulbright

Gleich

Kephart

et al. 2003

Journal of Cell Science

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(+44.4±1.4 mV) increased depolarization compared with PMA alone. Additionally, the protein kinase C (PKC) δ-selective blocker, rottlerin, inhibited PMA-stimulated depolarization, indicating that PKCδ was involved in regulating depolarization associated with eosinophil NADPH oxidase activity. Thus, the membrane depolarization that is associated with NADPH oxidase activation in eosinophils is sufficient to produce marked proton channel activation under physiological conditions.

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“…Resting potential against 5 mmol/L KCl was expected to be Ϫ34 mV, 28 owing to the known problems for estimation of absolute membrane potentials with the potentially sensitive fluorescent probes. 29 All data were expressed as changes in millivolts from resting potentials.…”

Section: Membrane Potentialmentioning

confidence: 99%

Effects of Superoxide Anions on Endothelial Ca ²⁺ Signaling Pathways

Graier

Hoebel

Paltauf-Doburzynska

et al. 1998

ATVB

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Abstract-Although the involvement of free radicals in the development of endothelial dysfunction under pathological conditions, like diabetes and hypercholesterolemia, has been proposed frequently, there is limited knowledge as to how superoxide anions (O 2 Ϫ ) might affect endothelial signal transduction. In this study, we investigated the effects of preincubation with the O 2 Ϫ -generating system xanthine oxidase/hypoxanthine (XO/HX) on mechanisms for Ca 2ϩ signaling in cultured porcine aortic endothelial cells. Incubation of cells with XO/HX yielded increased intracellular Ca 2ϩ release and capacitative Ca 2ϩ entry in response to bradykinin and ATP in a time-and concentration-dependent manner. This effect was prevented by superoxide dismutase but not by the tyrosine kinase inhibitor tyrphostin A48. In addition, capacitative Ca 2ϩ entry induced by the receptor-independent stimulus 2,5-di-(tert-butyl)-1,4-benzohydroquinone or thapsigargin was enhanced in O 2 Ϫ -exposed cells (ϩ38% and ϩ32%, respectively). Increased Ca 2ϩ release in response to bradykinin in XO/HX-pretreated cells might be due to enhanced formation of inositol-1,4,5-trisphosphate (ϩ140%). Exposure to XO/HX also affected other signal transduction mechanisms involved in endothelial Ca 2ϩ signaling, such as microsomal cytochrome P450 epoxygenase and membrane hyperpolarization to Ca 2ϩ store depletion with thapsigargin (ϩ103% and ϩ48%, respectively) and tyrosine kinase activity (ϩ97%). A comparison of bradykinin-initiated intracellular Ca 2ϩ release and thapsigargin-induced hyperpolarization with membrane viscosity modulated by XO/HX (decrease in viscosity) or cholesterol (increase in viscosity) reflected a negative correlation between bradykinin-initiated Ca 2ϩ release and membrane viscosity. Because intracellular Ca 2ϩ is a main regulator of endothelial vascular function, our data suggest that O 2 Ϫ anions are involved in regulation of the vascular endothelium.

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Flow cytometric determination of absolute membrane potential of cells

Cited by 56 publications

References 19 publications

A1 and A2 Adenosine Receptor Activation Inversely Modulates Potassium Currents and Membrane Potential in DDT1 MF-2 Smooth Muscle Cells

A1 and A2 Adenosine Receptor Activation Inversely Modulates Potassium Currents and Membrane Potential in DDT1 MF-2 Smooth Muscle Cells

Regulation of eosinophil membrane depolarization during NADPH oxidase activation

Effects of Superoxide Anions on Endothelial Ca ²⁺ Signaling Pathways

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Flow cytometric determination of absolute membrane potential of cells

Cited by 56 publications

References 19 publications

A1 and A2 Adenosine Receptor Activation Inversely Modulates Potassium Currents and Membrane Potential in DDT1 MF-2 Smooth Muscle Cells

A1 and A2 Adenosine Receptor Activation Inversely Modulates Potassium Currents and Membrane Potential in DDT1 MF-2 Smooth Muscle Cells

Regulation of eosinophil membrane depolarization during NADPH oxidase activation

Effects of Superoxide Anions on Endothelial Ca 2+ Signaling Pathways

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Effects of Superoxide Anions on Endothelial Ca ²⁺ Signaling Pathways