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A highly-selective chloride microelectrode based on a mercuracarborand anion carrier

Marino DiFranco, Marbella Quinonez, Rafal M. Dziedzic, Alexander M. Spokoyny, Stephen C. Cannon
doi: https://doi.org/10.1101/629691
Marino DiFranco
1Department of Physiology, David Geffen School of Medicine at UCLA
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Marbella Quinonez
1Department of Physiology, David Geffen School of Medicine at UCLA
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Rafal M. Dziedzic
2Department of Chemistry and Biochemistry, UCLA
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Alexander M. Spokoyny
2Department of Chemistry and Biochemistry, UCLA
3California NanoSystems Institute (CNSI), UCLA
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Stephen C. Cannon
1Department of Physiology, David Geffen School of Medicine at UCLA
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  • For correspondence: sccannon@mednet.ucla.edu
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ABSTRACT

The chloride gradient plays an important role in regulating cell volume, membrane potential, pH, secretion, and the reversal potential of inhibitory GABAA receptors. Measurement of intracellular chloride activity, Embedded Image, using liquid membrane ion-selective microelectrodes (ISM), however, has been limited by the physiochemical properties of Cl ionophores which have caused poor stability, drift, sluggish response times, and interference from other biologically relevant anions. Most importantly, intracellular HCO3− may be up to 4 times more abundant than Cl− (e.g. skeletal muscle) which places severe constraints on the required selectivity of a Cl – sensing ISM.

Previously, a sensitive and highly-selective Cl sensor was developed in a polymeric membrane electrode (Badr et al. 1999) using a trinuclear Hg(II) complex containing carborane-based ligands, [9]-mercuracarborand-3, or MC3 for short. Here, we have adapted the use of the MC3 anion carrier in a liquid membrane ion-selective microelectrode and show the MC3-ISM has a linear Nernstian response over a wide range of aCl (0.1 mM to 100 mM), is highly selective for Cl over other biological anions or inhibitors of Cl transport, and has a response time of less than 5 sec. Importantly, over the physiological range of aCl (1 mM to 100 mM) the potentiometric response of the MC3-ISM is insensitive to HCO3− or changes in pH. Finally, we demonstrate the biological application of an MC3-ISM by measuring intracellular aCl, and the response to an external Cl-free challenge, for an isolated skeletal muscle fiber.

  • Abbreviations

    9-ACA
    9-anthracenecarboxylic acid
    BMT
    bumetanide
    ISM
    ion selective microelectrode
    MC3
    [9]mercuracarborand-3
    NPOE
    2-nitrophenyl octyl ether
    TDMAC
    tridodecylmethyl ammonium chloride
  • Copyright 
    The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license.
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    Posted May 07, 2019.
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    A highly-selective chloride microelectrode based on a mercuracarborand anion carrier
    Marino DiFranco, Marbella Quinonez, Rafal M. Dziedzic, Alexander M. Spokoyny, Stephen C. Cannon
    bioRxiv 629691; doi: https://doi.org/10.1101/629691
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    A highly-selective chloride microelectrode based on a mercuracarborand anion carrier
    Marino DiFranco, Marbella Quinonez, Rafal M. Dziedzic, Alexander M. Spokoyny, Stephen C. Cannon
    bioRxiv 629691; doi: https://doi.org/10.1101/629691

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