Isoform-specific function and distribution of Na/K pumps in the frog lens epithelium

J. Gao, X. Sun, V. Yatsula, R. S. Wymore, R. T. Mathias

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56 Scopus citations

Abstract

Epithelial cells from the anterior and equatorial surfaces of the frog lens were isolated and used the same day for studies of the Na/K ATPase. RNase protection assays showed that all cells express α1- and α2-isoforms of the Na/K pump but not the α3-isoform, however the α2-isoform dominates in anterior cells whereas the α1-isoform dominates in equatorial cells. The whole cell patch-clamp technique was used to record functional properties of the Na/K pump current (I(P)), defined as the current specifically inhibited by dihydro-ouabain (DHO). DHO-I(P) blockade data indicate the α1-isoform has a dissociation constant of 100 μM DHO whereas for the α2-isoform it is 0.75 μM DHO. Both α1- and α2-isoforms are half maximally activated at an intracellular Na+-concentration of 9 mM. The α1-isoform is half maximally activated at an extracellular K+-concentration of 3.9 mM whereas for the α2-isoform, half maximal activation occurs at 0.4 mM. Lastly, transport by the α1-isoform is inhibited by a drop in extracellular pH, which does not affect transport by the α2-isoform. Under normal physiological conditions, I(P) in equatorial cells is approximately 0.23 μA/μF, and in anterior cells it is about 0.14 μA/μF. These current densities refer to the area of cell membrane assuming a capacitance of around 1 μF/cm2. Because cell size and geometry are different at the equatorial vs. anterior surface of the intact lens, we estimate Na/K pump current density per area of lens surface to be around 10 μA/cm2 at the equator vs. 0.5 μA/cm2 at the anterior pole.

Original languageEnglish
Pages (from-to)89-101
Number of pages13
JournalJournal of Membrane Biology
Volume178
Issue number2
DOIs
StatePublished - 15 Nov 2000

Keywords

  • K-dependence
  • Lens
  • Na-dependence
  • Na/K ATPase
  • Whole cell patch clamp
  • pH
  • α-Isoforms

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