Kv 2.1 K+ channels underlie major voltage-gated K+ outward current in H9c2 myoblasts

Abstract

The H9c2 clonal cell line derived from embryonic rat ventricle is an in vitro model system for cardiac and skeletal myocytes. We used the whole-cell patch clamp technique to characterize the electrophysiological and pharmacological properties of an outward K+ current (IKv) and determined its molecular correlate in H9c2 myoblasts. IKv was activated by threshold depolarization to -30 mV, and its current amplitude and rate of activation increased with further depolarizations. IKv inactivated slowly with a time constant of 1-2 s, and the V0.5 for steady-state inactivation was -37.9±4.6 mV (n= 10). Tetraethylammonium and quinidine suppressed IKv with IC50's of 3.7 mM and 11.6 μM, respectively. Using RT-PCR analysis we found that the Kv2.1 gene is the most abundantly expressed among genes for Kv 1.2, 1.4, 1.5, 2.1, 4.2, and 4.3, and by Western blotting we confirmed the synthesis of the Kv2.1 α-subunit protein. We conclude that IKv, the predominant voltage-gated outward current in H9c2 myoblasts, flows through the channels comprised of the Kv 2.1-subunit gene product.