Effect of Concanavalin A on Membrane Potential of Polymorphonuclear Leukocyte Monitored by Fluorescent Dye

Utsumi, Kozo; Sugiyama, Katsumi; Miyahara, Masanobu; Naito, Mitsuko; Awai, Michiyasu; Inoue, Masayasu

doi:10.1247/csf.2.203

Cited by 57 publications

(11 citation statements)

References 15 publications

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“…In experiments using one indirect probe, i.e., radiolabeled triphenylmethylphosphonium ion, Korchak and Weissmann (1978) found that changes in the apparent membrane potential of human neutrophils antecede the metabolic responses of these cells to surface stimulation with Con A and immune complexes. Utsumi et al (1977), using the cyanine dye, 3-3'-dipropylthiocarbocyanine iodide, also found evidence that concanavalin A provokes changes in neutrophils compatible with a small membrane hyperpolarization followed by a large depolarization. In studies using yet another membrane potential-sensitive cyanine dye (Le., 3,3'-dipentyloxacarbocyanine), Seligmann et al (1980) described changes in fluorescence after stimulation of neutrophils with chemotactic factors compatible with a rapid depolarization of the plasma membrane followed by a prolonged hyperpolarization.…”

Section: B Changes In Membrane Fluidity and Apparent Membrane Potentialmentioning

confidence: 88%

Neutrophil Degranulation

Goldstein

1984

Regulation of Leukocyte Function

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Section: B Changes In Membrane Fluidity and Apparent Membrane Potentialmentioning

confidence: 88%

Neutrophil Degranulation

Goldstein

1984

Regulation of Leukocyte Function

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“…The use of two different lipophilic probes of membrane potential has provided evidence that neutrophils (24,(26)(27)(28)(29)(30) and monocytes (56) from cultured macrophages using direct intracellular recording techniques (25,56,57). However, results obtained with the indirect probes may be influenced by a number of factors such as changes in membrane lipid composition or interaction (particularly with the fluorescent probes) with reactive molecular species.…”

Section: Discussionmentioning

confidence: 99%

“…kocyte calcium, sodium, and potassium fluxes (2,(18)(19)(20)(21)(22)(23) and alteration of leukocyte membrane electrochemical potential (referred to in this paper as membrane potential) (24)(25)(26)(27)(28)(29)(30). Korchak and Weissmann (24), using the indirect probe of membrane potential triphenylmethylphosphonium ion (TPMP+), have reported that stimulus-induced alteration of the plasma membrane potential precedes superoxide generation in neutrophils.…”

mentioning

confidence: 99%

Use of lipophilic probes of membrane potential to assess human neutrophil activation. Abnormality in chronic granulomatous disease.

Seligmann¹,

Gallin²

1980

J. Clin. Invest.

186

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A B S T R A C T Previous studies using membrane potential sensitive probes have provided evidence that chemotactic factors elicit membrane potential changes in normal human neutrophils (PMN). In addition to stimulation of PMN motility, chemotactic factors also stimulate degranulation and superoxide ion (O°) generation and it has been suggested that alteration of membrane potential activates these events (Korchak, H. M., and G. Weissmann. 1978. Proc. Natl. Acad. Sci. U. S. A. 75: 3818-3822.). To further define the interrelationship of these functions, studies were done with two indirect probes of membrane potential, 3-3'-dipentyloxacarbocyanine and triphenylmethylphosphonium ion (TPMP+) using PMN from normal subjects, from patients with abnormal O2 production (chronic granulomatous disease [CGD]), and from patients with defective degranulation and/or chemotaxis (Chediak-Higashi syndrome and patients with elevated immunoglobulin (Ig)E and recurrent staphylococcal infections). The stimuli used were the chemoattractant N-formyl-methionyl-leucyl-phenylalanine (f-Met-LeuPhe) and the secretagogues ionophore A23187 and phorbol myristate acetate (PMA). The results obtained with 3-3'-dipentyloxacarbocyanine and TPMP were comparable. The apparent membrane potential changes elicited by f-Met-Leu-Phe and PMA in normal PMN were reduced or entirely absent in PMN obtained from patients with CGD but normal in PMN from other patients. PMN from patients with CGD had normal calculated resting membrane potentials and normal responses elicited by the potassium ionophore valinomycin. The responses to calcium ionophore A23187 were only slightly impaired. The abnormality of the elicited response of CGD cells to f-Met-Leu-Phe and PMA could not be attributed to the absence of O2, This work appeared in part in abstract form (1).

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“…Based on studies of altered transmembrane distribution of various lipid-soluble cationic probes, an essential step in the process whereby the NADPH oxidase is activated appears to be a change in the plasma membrane potential. However, there is controversy as to whether the required change is a depolarization, hyperpolarization, or a sequence of potential fluctuations (2)(3)(4)(5)(6)(7)(8)(9)(10)(11). Among the reasons for the uncertainty in these measurements are (i) the time necessary for movement of the cations after perturbation; (ii) accumulation or redistribution (or both) of the cations across the inner mitochondrial membrane, which has a much greater potential difference (negative inside) than that of the plasma membrane (12); and (iii) metabolic alterations of the probes.…”

mentioning

confidence: 99%

Depolarization and increased conductance precede superoxide release by concanavalin A-stimulated rat alveolar macrophages.

Cameron¹,

Nelson²,

Forman³

1983

Proc. Natl. Acad. Sci. U.S.A.

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Rat alveolar macrophages release superoxide into the extracellular medium when stimulated by concanavalin A. This process, the respiratory burst, is characterized by a delay between binding of the stimulus and release of superoxide. It has been proposed that a key event that occurs during this delay period is the alteration of membrane electrical potential. Microelectrode impalement was used to directly measure electrical properties of the plasma membrane. Upon addition of concanavalin A, the membrane potential depolarized 21%, and membrane electrical resistance decreased 16%. Parallel chemical measurement of superoxide release indicated that these changes in electrical properties precede the release of superoxide.Generation of superoxide radical is an integral part of the antimicrobial activity of phagocytic cells (1). The process in which superoxide is released by these cells is characterized by a lag between recognition of a stimulus and activation of a plasma membrane NADPH oxidase, which catalyzes the reduction of oxygen to superoxide. Based on studies of altered transmembrane distribution of various lipid-soluble cationic probes, an essential step in the process whereby the NADPH oxidase is activated appears to be a change in the plasma membrane potential. However, there is controversy as to whether the required change is a depolarization, hyperpolarization, or a sequence of potential fluctuations (2-11). Among the reasons for the uncertainty in these measurements are (i) the time necessary for movement of the cations after perturbation; (ii) accumulation or redistribution (or both) of the cations across the inner mitochondrial membrane, which has a much greater potential difference (negative inside) than that of the plasma membrane (12); and (iii) metabolic alterations of the probes. For example, Whitin et al. (11) In this study of rat alveolar macrophages, we used microelectrodes to obtain a direct measurement of the membrane potential at rest and after addition of concanavalin A, a stimulant of superoxide release, along with measurements of the membrane input resistance under the same conditions. The length of time for the maximal changes in membrane electrical properties was compared with the lag time for superoxide release. Electrophysiologic Measurements. For electrophysiologic measurements, cells were centrifuged onto high-density oil (Fluorinert FC-40; 3M, Minneapolis, MN) and covered with Krebs-Ringer bicarbonate buffer (pH 7.4). Microelectrodes were pulled with final resistances of between 35 MfQ and 55 MfQ and were filled with 3 M KCI, and these were maneuvered with a Leitz micromanipulator. Cells were impaled, and membrane potential was recorded through a high-impedance electrometer. Results were displayed on a dual-beam oscilloscope and recorded on a oscillographic recorder. Input resistance was measured by passing 500-msec square-wave pulses of hyperpolarizing current through the recording electrode. Pulses were generated through the electrometer by using a Grass Instruments type 88...

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Effect of Concanavalin A on Membrane Potential of Polymorphonuclear Leukocyte Monitored by Fluorescent Dye

Cited by 57 publications

References 15 publications

Neutrophil Degranulation

Neutrophil Degranulation

Use of lipophilic probes of membrane potential to assess human neutrophil activation. Abnormality in chronic granulomatous disease.

Depolarization and increased conductance precede superoxide release by concanavalin A-stimulated rat alveolar macrophages.

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