The transient outward current in mice lacking the potassium channel gene Kv1.4

1. The transient outward current (I-to) plays a prominent role in the repolarization phase of the cardiac action potential. Several K+ channel genes, including Kv1.4, are expressed in the heart, produce rapidly inactivating currents when heterologously expressed, and may be the molecular basis of I-to. 2. We engineered mice homozygous for a targeted disruption of the K+ channel gene Kv1.4 and compared I-to in wild-type (Kv1.4(+/+)), heterozygous (Kv1.4(+/-)) and homozygous 'knockout' (Kv1.4(-/-)) mice. Kv1.4 RNA was truncated in Kv1.4(-/-) mice and protein expression was absent. 3. Adult myocytes isolated from Kv1.4(+/+), Kv1.4(+/-) and Kv1.4(-/-) mice had large rapidly inactivating outward currents. The peak current densities at 60 mV (normalized by cellular capacitance, in pA pF(-1); means +/- S.E.M.) were 53.8 +/- 5.3, 45.3 +/- 2.2 and 44.4 +/- 2.8 in cells from Kv1.4(+/+), Kv1.4(+/-) and Kv1.4(-/-) mice, respectively (P < 0.02 for Kv1.4(+/+) vs. Kv1.4(-/-)). The steady-state values (800 ms after the voltage clamp step) were 30.9 +/- 2.9, 26.9 +/- 3.8 and 23.5 +/- 2.2, respectively (P < 0.02 for Kv1.4(+/+) vs. Kv1.4(-/-)). The inactivating portion of the current was unchanged in the targeted mice. 4. The voltage dependence and time course of inactivation were not changed by targeted disruption of Kv1.4. The mean best-fitting V-1/2 (membrane potential at 50% inactivation) values for myocytes from Kv1.4 (+/+), Kv1.4(+/-) and Kv1.4(-/-) mice were -53.5 +/- 3.7, -51.1 +/- 2.6 and -54.2 +/- 2.4 mV, respectively The slope factors (k) were -10.1 +/- 1.4, -8.8 +/- 1.4 and -9.5 +/- 1.2 mV, respectively. The fast time constants for development of inactivation at -30 mV were 27.8 +/- 2.2, 28.2 +/- 5.1 and 19.6 +/- 2.1 ms in Kv1.4(+/+) Kv1.4(+/-) and Kv1.4(-/-) myocytes, respectively. At +30 mV, they were 35.5 +/- 2.8, 30.0 +/- 2.1 and 28.7 +/- 1.6 ms, respectively. The time constants for the rapid phase of recovery from inactivation at -80 mV were 32.5 +/- 8.2, 23.3 +/- 1.8 and 39.0 +/- 3.7 ms, respectively. 5. Nearly the entire inactivating component as well as more than 60% of the steady-state outward current was eliminated by 1 mM 4-aminopyridine in Kv1.4(+/+), Kv1.4(+/-) and Kv1.4(-/-) myocytes. 6. Western blot analysis of heart membrane extracts showed no significant upregulation of the Kv4 subfamily of channels in the targeted mice. 7. Thus, Kv1.4 is not the molecular basis of I-to in adult murine ventricular myocytes.