PGE2 activation of apical membrane Cl- channels in A6 epithelia: Impedance analysis

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Abstract

Measurements of transepithelial electrical impedance of continuously short-circuited A6 epithelia were made at audio frequencies (0.244 Hz to 10.45 kHz) to investigate the time course and extent to which prostaglandin E2 (PGE2) modulates Cl- transport and apical membrane capacitance in this cell-cultured model epithelium. Apical and basolateral membrane resistances were determined by nonlinear curve-fitting of the impedance vectors at relatively low frequencies (<50 Hz) to equations (Pǎnescu, T. G., and S. I. Helman. 2001. Biophys. J. 81:838-851) where depressed Nyquist impedance semicircles were characteristic of the membrane impedances under control Na+-transporting and amiloride-inhibited conditions. In all tissues (control, amiloride-blocked, and amilodde-blocked and furosemide-pretreated), PGE2 caused relatively small (<∼3 μA/cm2) and rapid (<60 s) maximal increase of chloride current due to activation of a rather large increase of apical membrane conductance that preceded significant activation of Na+ transport through amiloride-sensitive epithelial Na+ channels (ENaCs). Apical membrane capacitance was frequency-dependent with a Cole-Cole dielectric dispersion whose relaxation frequency was near 150 Hz. Analysis of the time-dependent changes of the complex frequency-dependent equivalent capacitance of the cells at frequencies >1.5 kHz revealed that the mean 9.8% increase of capacitance caused by PGE2 was not correlated in time with activation of chloride conductance, but rather correlated with activation of apical membrane Na+ transport.

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Pǎunescu, T. G., & Helman, S. I. (2001). PGE2 activation of apical membrane Cl- channels in A6 epithelia: Impedance analysis. Biophysical Journal, 81(2), 852–866. https://doi.org/10.1016/S0006-3495(01)75746-3

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