The Na+-K+-ATPase α2-subunit isoform modulates contractility in the perinatal mouse diaphragm

Abstract

This study uses genetically altered mice to examine the contribution of the Na+-K+-ATPase α2catalytic subunit to resting potential, excitability, and contractility of the perinatal diaphragm. The α2protein is reduced by 38% in α2-heterozygous and absent in α2-knockout mice, and α1-isoform is upregulated 1.9-fold in α2-knockout. Resting potentials are depolarized by 0.8–4.0 mV in heterozygous and knockout mice. Action potential threshold, overshoot, and duration are normal. Spontaneous firing, a developmental function, is impaired in knockout diaphragm, but this does not compromise its ability to fire evoked action potential trains, the dominant mode of activation near birth. Maximum tetanic force, rate of activation, force-frequency and force-voltage relationships, and onset and magnitude of fatigue are not changed. The major phenotypic consequence of reduced α2content is that relaxation from contraction is 1.7-fold faster. This finding reveals a distinct cellular role of the α2-isoform at a step after membrane excitation, which cannot be restored simply by increasing α1content. Na+/Ca2+exchanger expression decreases in parallel with α2-isoform, suggesting that Ca2+extrusion is affected by the altered α2genotype. There are no major compensatory changes in expression of sarcoplasmic reticulum Ca2+-ATPase, phospholamban, or plasma membrane Ca2+-ATPase. These results demonstrate that the Na+-K+-ATPase α1-isoform alone is able to maintain equilibrium K+and Na+gradients and to substitute for α2-isoform in most cellular functions related to excitability and force. They further indicate that the α2-isoform contributes significantly less at rest than expected from its proportional content but can modulate contractility during muscle contraction.