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Neutrophil Activation Studied Using Two Indirect Probes of Membrane Potential Which Respond by Different Fluorescence Mechanisms

  • Bruce Seligmann
  • John I. Gallin
  • R. E. Basford
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 141)

Abstract

The involvement of ions in the activation of neutrophils (PMN) has been studied by monitoring cell membrane potential.1–11 The relationship of membrane potential to ionic gradients and permeabilities is given by the Goldman Equation and is the basis from which changes in membrane potential can be directly related to ion flux changes:
$$ \begin{matrix}Cellmembrane \\ potential \\ \end{matrix}=\frac{-RT}{F}log\frac{{{P}_{K}}{{[{{K}^{+}}]}_{i}}+{{P}_{Na}}{{[N{{a}^{+}}]}_{i}}+{{P}_{Cl}}{{[C{{l}^{-}}]}_{o}}+\ldots }{{{P}_{K}}{{[{{K}^{+}}]}_{o}}+{{P}_{Na}}{{[N{{a}^{+}}]}_{o}}+{{P}_{Cl}}{{[C{{l}^{-}}]}_{i}}+\ldots } .$$
The factor RT/F is a constant consisting of R, the gas constant; T, temperature; and F, a faraday. The intracellular (i) and extra-cellular (o) concentration of each ion is indicated, weighted by the permeability (P) of each ionic species. Thus, by measuring membrane potential and varying the extracellular concentration of specific ions, one can determine the ionic flux changes giving rise not only to resting membrane potential but also to stimulus-induced changes in membrane potential. Membrane potential of cultured macrophages has been measured by penetrating cells with microelectrodes.12–16 However, PMN are too small to study in this way with the currently available technology. Therefore, it has been necessary to use indirect molecular probes to study PMN membrane potential.1–11 The indirect probes currently in use are charged, lipophilic ions which exhibit some property enabling determination of their intracellular concentration. Two classes of such probes are the fluorescent cyanine dyes and radiolabeled probes such as triphenyl methyl phosphonium ion. A number of reports have appeared using these probes to study PMN membrane potential. However, before conclusive interpretations of results can be drawn, it must be shown that the responses are not affected by cellular artifacts independent of membrane potential.

Keywords

Membrane Potential Xanthine Oxidase Reactive Species Phorbol Myristate Acetate Phorbol Myristate Acetate 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1982

Authors and Affiliations

  • Bruce Seligmann
    • 1
  • John I. Gallin
    • 1
  • R. E. Basford
  1. 1.Bacterial Diseases Section, Laboratory of Clinical Investigation, National Institute of Allergy and Infectious DiseasesNational Institutes of HealthBethesdaUSA

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