Cytochemical and patch-clamp studies of calcium influx through voltage-dependent Ca2+ channels in vestibular supporting cells of guinea pigs

European Archives of Oto-rhino-laryngology : Official Journal of the European Federation of Oto-Rhino-Laryngological Societies (EUFOS) : Affiliated with the German Society for Oto-Rhino-Laryngology - Head and Neck Surgery
Y MoriK Yajin

Abstract

To clarify whether or not vestibular supporting cells have voltage-dependent Ca2+ channels, cytochemical and patch-clamp studies were performed using cells isolated from the ampullae of the semicircular canal of the guinea pig. Image analysis used fura-2 as a Ca(2+)-sensitive fluorescence dye and showed that the intracellular Ca2+ concentration ([Ca2+]i) increased with bath application of high (150 mM)K+, but was unaffected by 80 mM K+. The increase in [Ca2+]i induced by high K+ was completely blocked by 1 microM nifedipine as an L-type Ca2+ channel antagonist. In the patch-clamp whole-cell recording of the isolated supporting cells, the voltage-dependent inward current was induced by a depolarizing pulse lasting 2 s in a high (50 mM) Ca2+ and tetraethylammonium-containing external solution replaced by choline chloride and a Cs(+)-containing internal solution. The inward current was obtained when the membrane was depolarized to -50 mV and maximum current was observed at -10 to +10 mV. This inward current was completely blocked by 1 microM nifedipine. These findings strongly suggest that voltage-dependent Ca2+ channels exist in the vestibular supporting cells and regulate Ca2+ concentration in the vestibular endolymph.

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