AN ANALYSIS OF THE LONG-LASTING AFTER-HYPERPOLARIZATION OF GUINEA-PIG VAGAL MOTONEURONS

1. The long-lasting after-hyperpolarization which characterizes the neurones of the dorsal motor nucleus of the vagus in the guinea-pig was studied in vitro. 2. Following a train of action potentials, vagal motoneurones develop a long-lasting after-hyperpolarization. Two different shapes of long-lasting after-hyperpolarization were encountered: an after-hyperpolarization which slowly (0.6-1.2 s) and monotonically developed to peak value; and a second type of long-lasting after-hyperpolarization where the onset of the slow component appears to be masked by an early, relatively fast component. Both shapes of long-lasting after-hyperpolarization depend on Ca2+ influx and increase as a function of the number of action potentials in the train. 3. A novel procedure was used to analyse the ionic processes which underlie the long-lasting after-hyperpolarization. The neuronal responses to a series of long (7 s) hyperpolarizing current pulses during the long-lasting after-hyperpolarization were recorded and the voltage-current curves at 600 different time points along the long-lasting after-hyperpolarization were plotted. The conductance and the reversal potential at each time point were calculated from the slope and the intersection of these curves, respectively. 4. Using this procedure it was found that the long-lasting after-hyperpolarization consists of two conductances that differ in kinetic properties and reversal potential: an early conductance which peaks shortly after the end of the train and decays in a few tenths of seconds (EAHP), and a late conductance which develops slowly (time to peak about 1 s) and decays in 3-8 s (LAHP). The reversal potential for the early conductance is 10 mV more positive than the reversal potential for the late conductance (-84 mV); the latter reversal potential is in agreement with the K+ equilibrium potential. The different shapes of long-lasting after-hyperpolarization can be explained by different ratios of these two conductances. 5. Noradrenaline (10 muM) selectively blocks the late conductance, without an observable effect on the Ca2+ action potential. 6. The behaviour of the noradrenaline-sensitive late conductance was analysed. The amplitude of the conductance change increased sigmoidally as a function of the number of spikes in the train. A log-log plot suggests that at least two Ca2+ ions participate in the opening of a K+ channel. 7. A model that accounts for the slow kinetics of the late conductance was constructed. The model assumes that two Ca2+ ions are needed to open a potassium channel and that the intracellular Ca2+ concentration reaches its maximum value at the end of the train of action potentials. It was found that under these assumptions, the onset of the LAHP is governed by the channel's rate constants and the initial increase in Ca2+ concentration, whereas its duration is determined by the rate of removal of Ca2+ from the cell. The processes which might underlie the slow rate constants are discussed.