Slowly inactivating potassium channels induced in Xenopus oocytes by messenger ribonucleic acid from Torpedo brain.

Abstract

Poly(A+) messenger RNA was extracted from the electric lobe and medulla of Torpedo and injected into oocytes of Xenopus laevis. The synthesis and processing of proteins coded by the injected messenger RNA led to the incorporation of voltage-activated channels in the oocyte membrane. A large, well maintained outward current was recorded from injected oocytes in response to depolarization. Non-injected oocytes did not show this current. The reversal potential of the current varied according to the Nernst equation with external potassium concentration, indicating that it was largely carried by potassium ions. The maintained potassium current was not reduced by manganese (5 mM) or lanthanum ions (0.1 mM). Tetraethylammonium and aminopyridines blocked the potassium current. The block produced by 3,4-diaminopyridine was enhanced by previous activation, but diminished by strong depolarization. The amplitude of the potassium current increased over the approximate voltage range -30 to +50 mV, but reduced at more positive potentials. The decline of the current during maintained depolarization became slower as the membrane potential was made more positive, and the rate of onset of the current became faster. Estimates from noise analysis indicated that the slow potassium current passes through channels with a mean lifetime of about 14 ms and conductance of 14 pS (at -10 mV and room temperature). Injection of the messenger RNA also induced the formation of fast sodium and potassium channels activated by voltage, and channels activated by kainate.