Voltage-Gated Potassium Channels Activated During Action Potentials in Layer V Neocortical Pyramidal Neurons

Abstract

To investigate voltage-gated potassium channels underlying action potentials (APs), we simultaneously recorded neuronal APs and single K+ channel activities, using dual patch-clamp recordings (1 whole cell and 1 cell-attached patch) in single-layer V neocortical pyramidal neurons of rat brain slices. A fast voltage-gated K+ channel with a conductance of 37 pS (Kf) opened briefly during AP repolarization. Activation of Kf channels also was triggered by patch depolarization and did not require Ca2+influx. Activation threshold was about −20 mV and inactivation was voltage dependent. Mean duration of channel activities after single APs was 6.1 ± 0.6 ms (mean ± SD) at resting membrane potential (−64 mV), 6.7 ± 0.7 ms at −54 mV, and 62 ± 15 ms at −24 mV. The activation and inactivation properties suggest that Kf channels function mainly in AP repolarization but not in regulation of firing. Kf channels were sensitive to a low concentration of tetraethylammonium (TEA, 1 mM) but not to charybdotoxin (ChTX, 100 nM). Activities of A-type channels (KA) also were observed during AP repolarization. KA channels were activated by depolarization with a threshold near −45 mV, suggesting that KA channels function in both repolarization and timing of APs. Inactivation was voltage dependent with decay time constants of 32 ± 6 ms at −64 mV (rest), 112 ± 28 ms at −54 mV, and 367 ± 34 ms at −24 mV. KA channels were localized in clusters and were characterized by steady-state inactivation, multiple subconductance states (36 and 19 pS), and inhibition by 5 mM 4-aminopyridine (4-AP) but not by 1 mM TEA. A delayed rectifier K+ channel (Kdr) with a unique conductance of 17 pS was recorded from cell-attached patches with TEA/4-AP-filled pipettes. Kdrchannels were activated by depolarization with a threshold near −25 mV and showed delayed long-lasting activation. Kdr channels were not activated by single action potentials. Large conductance Ca2+-activated K+ (BK) channels were not triggered by neuronal action potentials in normal slices and only opened as neuronal responses deteriorated (e.g., smaller or absent spikes) and in a spike-independent manner. This study provides direct evidence for different roles of various K+ channels during action potentials in layer V neocortical pyramidal neurons. Kf and KA channels contribute to AP repolarization, while KA channels also regulate repetitive firing. Kdr channels also may function in regulating repetitive firing, whereas BK channels appear to be activated only in pathological conditions.