A Na+-activated K+ current in cultured brain stem neurones from chicks

Abstract

1. Patch-clamp techniques were used to study the properties of a Na+-activated K+ current (IK(Na) in neurones cultured from embryonic chick brain stem. 2. With whole-cell clamp, a depolarizing voltage command evoked an inward current that was followed by an outward current with two components, the first transient, the second sustained. 3. Tetrodotoxin (TTX, 1 microM) eliminated the inward current and the transient component of the outward current, without affecting the sustained outward current. In addition, the transient outward current was attenuated when all external Na+ was replaced by Li+, suggesting that it was activated specifically by Na+ entry into the cell. 4. The time course of the transient outward current was obtained by subtracting records obtained in Li+ solution from those obtained in Na+ solution. There was significant overlap between the decay of the inward current and the onset of the transient outward current. 5. When just after the peak of the transient outward current, the membrane was stepped to progressively more hyperpolarized levels, the tail currents associated with the current reversed polarity near the calculated K+ equilibrium potential. 6. 4-Aminopyridine (4-AP, 4 mM) abolished the transient outward current and approximately half of the sustained late current. Tetraethylammonium (TEA, 2 mM) had no effect on the transient current, but reduced the sustained current slightly. 7. Inside-out patches, made in LiCl bathing solutions, contained channels that were activated by exposing the cytoplasmic face of the patch to Na+. Channel activity continued as long as Na+ was present. 8. The single-channel currents reversed at the K+ equilibrium potential, and were associated with a main conductance that depended upon K+ concentration (about 50 pS with [K+]o = 15 mM, [K+]i = 5 mM, and 100 pS when [K+]i was increased to 75 mM). 9. The open probability of the channels increased with increasing cytoplasmic Na+ concentration. At [Na+]i = 150 mM (the maximum concentration tested), channels were open almost continuously. Open probability was considerably less at 50 mM, and still measureable at 20 mM. 10. The magnitude of IK(Na) and its overlap with the inward Na+ current indicate that these channels contribute significantly to the repolarizing phase of the action potential. In addition, the relation between channel activity and Na+ concentration suggests that the channels may make a measurable contribution to membrane conductance at resting intracellular Na+ concentrations.