Calmodulin antagonists suppress gap junction coupling in isolated Hensen cells of the guinea pig cochlea
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The effect of calmodulin (CaM) antagonists W7, trifluoperazine (TFP) and a calmodulin inhibitory peptide on gap junction coupling in isolated Hensen cells of the organ of Corti was analysed by the double whole-cell patch-clamp technique. Addition of the conventional antagonists W7 and TFP in the micromolar range caused a rapid decrease of gap junction conductance after a delay of a few minutes in a dose-dependent manner. Fluorescence spectroscopy of cytoplasmic free calcium concentration ([Ca2+]i) by Fura-2 showed no significant change of [Ca2+]i by W7. Chelation of [Ca2+]i by 10 mM BAPTA or use of nominally Ca2+-free external bath did not suppress the W7-induced gap junction uncoupling. The results suggest that W7 and TFP induce gap junction uncoupling at unchanged global [Ca2+]i in Hensen cells. To obtain additional evidence for an involvement of CaM in regulating gap junction conductance a calmodulin inhibitory peptide, the MLCK peptide (250 nM), was added to the standard pipette solution. Again gap junction uncoupling was observed, but on a significantly slower time scale. This is the first study of an effect of calmodulin antagonists on gap junction coupling in isolated Hensen cells. The question whether the effect of calmodulin inhibitors is specific and involves CaM-dependent gating of gap junction coupling in Hensen cells is discussed.
KeywordsCalmodulin antagonists Cochlea Double whole-cell patch-clamp Gap junctions Hensen cells
The authors would like to thank Drs. C. Peracchia, H. Evans, and K. Török for helpful discussions and Dr. C. Zeilinger for calmodulin alignment to connexins. This work was partially supported by a grant from the Fritz-Thyssen-Stiftung.
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