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Am J Physiol Renal Physiol 288: F493-F504, 2005. First published November 16, 2004; doi:10.1152/ajprenal.00301.2004
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Basolateral K+ conductance in principal cells of rat CCD

Daniel A. Gray, Gustavo Frindt, Yu-Yang Zhang, and Lawrence G. Palmer

Department of Physiology and Biophysics, Weill Medical College of Cornell University, New York, New York

Submitted 11 August 2004 ; accepted in final form 18 October 2004

Whole cell K+ current was measured by forming seals on the luminal membrane of principal cells in split-open rat cortical collecting ducts. The mean inward, Ba2+-sensitive conductance, with 40 mM extracellular K+, was 76 ± 12 and 141 ± 22 nS/cell for animals on control and high-K+ diets, respectively. The apical contribution to this was estimated to be 3 and 16 nS/cell on control and high-K+ diets, respectively. To isolate the basolateral component of whole cell current, we blocked ROMK channels with either tertiapin-Q or intracellular acidification to pH 6.6. The current was weakly inward rectifying when bath K+ was ≥40 mM but became more strongly rectified when bath K+ was lowered into the physiological range. Including 1 mM spermine in the pipette moderately increased rectification, but most of the outward current remained. The K+ current did not require intracellular Ca2+ and was not inhibited by 3 mM ATP in the pipette. The negative log of the acidic dissociation constant (pKa) was ~6.5. Block by extracellular Ba2+ was voltage dependent with apparent Ki at –40 and –80 mV of ~160 and ~80 µM, respectively. The conductance was TEA insensitive. Substitution of Rb+ or NH4+ for K+ led to permeability ratios of 0.65 ± 0.07 and 0.15 ± 0.02 and inward conductance ratios of 0.17 ± 0.03 and 0.57 ± 0.09, respectively. Analysis of Ba2+-induced noise, with 40 mM extracellular K+, yielded single-channel currents of 0.39 ± 0.04 and –0.28 ± 0.04 pA at voltages of 0 and –40 mV, respectively, and a single-channel conductance of 17 ± 1 pS.

renal K+ channels; renal K+ transport; noise analysis; tertiapin; K+ adaptation


Address for reprint requests and other correspondence: D. A. Gray, Dept. of Physiology and Biophysics, Weill Medical College of Cornell Univ., 1300 York Ave., New York, NY 10021 (E-mail: dag2007{at}med.cornell.edu)




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