Activity dependence and functional role of the apamin-sensitive K+ current in rat supraoptic neurones in vitro

1. Intracellular recordings were obtained from seventy-two magnocellular neurosecretory cells (MNCs) in superfused explants of rat hypothalamus. The current underlying the after-hyperpolarization (I-AHP) following spike-evoked trains of action potentials was characterized using the hybrid-clamp technique. The activity-dependent requirements for the genesis of the AHP were determined. The functional role of the conductance was investigated using saturating concentrations (50-300 nM) of apamin, a selective blocker of the AHP in MNCs. 2. I-AHP was reversibly abolished by the removal of extracellular Ca2+. The amplitude of I-AHP varied linearly as a function of voltage and reversed at -100 +/- 3 mV in 3 mM external K+. Changes in the concentration of extracellular K+ resulted in shifts of the reversal potential consistent with Nernst equation predictions for a K+-selective conductance. 3. Action potentials triggered by brief depolarizing pulses elicited an AHP during trains evoked at frequencies > 1 Hz. Onset of the AHP progressed exponentially, reaching a maximum after the first fifteen to twenty impulses. The steady-state amplitude of the AHP increased logarithmically between 1 and 20 Hz. 4. Switching to voltage clamp during periods of continuous cell activity (firing rate > 4 Hz) confirmed the presence of an apamin-sensitive Ca2+-dependent K+ current. 5. Application of apamin produced a threefold increase in the mean firing rate of spontaneously active cells, but was without effect when applied to silent cells (firing rate < 0.5 Hz). 6. Apamin did not affect the ability of MNCs to fire in a phasic manner but caused a dramatic increase in the mean intraburst firing rate. Moreover, inhibition of I-AHP by apamin strongly attenuated spike accommodation normally seen at the onset of phasic bursts. 7. While apamin did not enhance the amplitude of depolarizing after-potentials following single spikes, post-train plateau potentials and associated after-discharges were enhanced. 8. The possible consequences of I-AHP modulation are discussed in the context of the regulation of firing rate and pattern in MNCs.