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- W2056519016 abstract "Summary We present a thorough calibration and verification of a combined non‐invasive self‐referencing microelectrode‐based ion‐flux measurement and whole‐cell patch clamp system as a novel and powerful tool for the study of ion transport. The system is shown to be capable of revealing the movement of multiple ions across the plasma membrane of a single protoplast at multiple voltages and in complex physiologically relevant solutions. Wheat root protoplasts are patch clamped in the whole‐cell configuration and current–voltage relations obtained whilst monitoring net K + and Ca 2+ flux adjacent to the membrane with ion‐selective electrodes. At each voltage, net ion flux (nmol m −2 sec −1 ) is converted to an equivalent current density (mA m −2 ) taking into account geometry and electrode efficiency, and compared with the net current density measured with the patch clamp system. Using this technique, it is demonstrated that the K + ‐permeable outwardly rectifying conductance (KORC) is responsible for net outward K + movement across the plasma membrane [1:1 flux‐to‐current ratio (1.21 ± 0.14 SEM, n = 15)]. Variation in the K + flux‐to‐current ratio among single protoplasts suggests a heterogeneous distribution of KORC channels on the membrane surface. As a demonstration of the power of the technique we show that despite a significant Ca 2+ permeability being associated with KORC (analysis of tail current reversal potentials), there is no correlation between Ca 2+ flux and KORC activity. A very significant observation is that large Ca 2+ fluxes are electrically silent and probably tightly coupled to compensatory charge movements. This analysis demonstrates that it is mandatory to measure flux and currents simultaneously to investigate properly Ca 2+ transport mechanisms and selectivity of ion channels in general." @default.
- W2056519016 created "2016-06-24" @default.
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- W2056519016 date "2006-03-08" @default.
- W2056519016 modified "2023-10-11" @default.
- W2056519016 title "Simultaneous flux and current measurement from single plant protoplasts reveals a strong link between K<sup>+</sup>fluxes and current, but no link between Ca<sup>2+</sup>fluxes and current" @default.
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- W2056519016 doi "https://doi.org/10.1111/j.1365-313x.2006.02676.x" @default.
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