Analysis of whole-cell currents by patch clamp of guinea-pig myenteric neurones in intact ganglia

Authors

  • François Rugiero,

    1. Laboratoire ‘Intégration des Informations Sensorielles’ (ITIS), CNRS, Bâtiment LNB, No. 31, Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France
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  • Maurice Gola,

    Corresponding author
    1. Laboratoire ‘Intégration des Informations Sensorielles’ (ITIS), CNRS, Bâtiment LNB, No. 31, Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France
    • Corresponding author M. Gola: Laboratoire Intégration des Informations Sensorielles (ITIS), CNRS, Bâtiment LNB, 31 Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France. Email: gola@lnb.cnrs-mrs.fr

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  • Wolf A. A. Kunze,

    1. Department of Anatomy and Cell Biology, University of Melbourne, Parkville, VIC 3010, Australia
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  • Jean-Claude Reynaud,

    1. Laboratoire ‘Intégration des Informations Sensorielles’ (ITIS), CNRS, Bâtiment LNB, No. 31, Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France
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  • John B. Furness,

    1. Department of Anatomy and Cell Biology, University of Melbourne, Parkville, VIC 3010, Australia
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  • Nadine Clerc

    1. Laboratoire ‘Intégration des Informations Sensorielles’ (ITIS), CNRS, Bâtiment LNB, No. 31, Chemin Joseph Aiguier, 13402 Marseille Cedex 20, France
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Abstract

Whole-cell patch-clamp recordings taken from guinea-pig duodenal myenteric neurones within intact ganglia were used to determine the properties of S and AH neurones. Major currents that determine the states of AH neurones were identified and quantified. S neurones had resting potentials of −47 ± 6 mV and input resistances (Rin) of 713 ± 49 MΩ at voltages ranging from −90 to −40 mV. At more negative levels, activation of a time-independent, caesium-sensitive, inward-rectifier current (IKir) decreased Rin to 103 ± 10 MΩ. AH neurones had resting potentials of −57 ± 4 mV and Rin was 502 ± 27 MΩ. Rin fell to 194 ± 16 MΩ upon hyperpolarization. This decrease was attributable mainly to the activation of a cationic h current, Ih, and to IKir. Resting potential and Rin exhibited a low sensitivity to changes in [K+]o in both AH and S neurones. This indicates that both cells have a low background K+ permeability. The cationic current, Ih, contributed about 20 % to the resting conductance of AH neurones. It had a half-activation voltage of −72 ± 2 mV, and a voltage sensitivity of 8.2 ± 0.7 mV per e-fold change. Ih has relatively fast, voltage-dependent kinetics, with on and off time constants in the range of 50–350 ms. AH neurones had a previously undescribed, low threshold, slowly inactivating, sodium-dependent current that was poorly sensitive to TTX. In AH neurones, the post-action-potential slow hyperpolarizing current, IAHP, displayed large variation from cell to cell. IAHP appeared to be highly Ca2+ sensitive, since its activation with either membrane depolarization or caffeine (1 mm) was not prevented by perfusing the cell with 10 mm BAPTA. We determined the identity of the Ca2+ channels linked to IAHP. Action potentials of AH neurones that were elongated by TEA (10 mm) were similarly shortened and IAHP was suppressed with each of the three Ω-conotoxins GVIA, MVIIA and MVIIC (0.3–0.5 μm), but not with Ω-agatoxin IVA (0.2 μm). There was no additivity between the effects of the three conotoxins, which indicates the presence of N- but not of P/Q-type Ca2+ channels. A residual Ca2+ current, resistant to all toxins, but blocked by 0.5 mm Cd2+, could not generate IAHP. This patch-clamp study, performed on intact ganglia, demonstrates that the AH neurones of the guinea-pig duodenum are under the control of four major currents, IAHP, Ih, an N-type Ca2+ current and a slowly inactivating Na+ current.

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