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Identification of a voltage- and calcium-dependent non-selective cation channel in cultured adult and fetal human nasal epithelial cells
Authors:Mark Jorissen  Johan Vereecke  Edward Carmeliet  Herman Van den Berghe  Jean -Jacques Cassiman
Affiliation:(1) Center for Human Genetics, University of Leuven, Campus Gasthuisberg, O&N, B-3000 Leuven, Belgium;(2) Laboratory of Physiology, University of Leuven, Campus Gasthuisberg, O&N, B-3000 Leuven, Belgium;(3) ENT Department, University of Leuven, Campus Gasthuisberg, O&N, B-3000 Leuven, Belgium
Abstract:The apical membranes of cultured human nasal epithelial cells from adults and fetuses were investigated with the patch-clamp technique. Amiloride-insensitive, calcium- and voltage-dependent, non-selective cation channels were found in 4% of the cell-attached, and 18% of the inside-out and outside-out patches (n=412). Multiple functional channels were present in more than 90% of these patches, with a mean of 3.9 channels per patch (n=55). The current-voltage relationship can be described by the Goldman equations and the single channel conductance was 20.1±0.3 pS (n=29) in adult and 20.7±0.4 pS (n=44) in fetal cells in symmetrical 150mM NaCl solutions. The channels were highly selective for cations: PNa/PCl was 30 in adult and 45 in fetal experiments. They were equally permeable for K+ and Na+, somewhat less for Cs+, and impermeable for choline+ and tetraethylammonium+. The open probability was voltage dependent: it increased approximately 2-fold with 30mV depolarization in the potential range from −60mV to +60mV. The channels were activated by Ca2+ concentrations of about 10−4M at the cytoplasmic side, but were insensitive to extracellular Ca2+ and amiloride (10−4M). The non-selective cation channels found in apical membranes of cultured fetal nasal epithelial cells were not different from the adult ones.
Keywords:Patch-clamp  Non-selective cation channels  Ca2+ dependence  Voltage dependence  Culture  Fetal nasal epithelium  Adult nasal epithelium
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