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Neurons of the inferior olive (IO) fire action potentials with large, long-lasting afterdepolarizations (ADPs). Broader ADPs support more spikes in climbing fibre axons and evoke longer bursts of complex spikes in Purkinje cells, which affect the magnitude and sign of cerebellar synaptic plasticity. In the present study, we investigated the ionic mechanisms that regulate IO action potential waveforms by making whole-cell recordings in brainstem slices from C57BL6/J mice. IO spikes evoked from rest had ADPs of ∼30 ms. After 500-ms hyperpolarizations, however, evoked action potentials were brief (1-2 ms), lacking ADPs altogether. Because such preconditioning should maximally recruit depolarizing I h and T-type currents and minimize repolarizing Ca-dependent currents known to shape the ADP, the rapid action potential downstroke suggested additional, dominant recovery of voltage-gated K currents at negative voltages. Under voltage clamp, outward currents evoked from -98 mV included large, voltage-gated, rapidly inactivating 'A-type' K currents. These currents had a steep availability curve with half-inactivation at -85 mV, suitable for recruitment by small hyperpolarizations. The fast decay time constant increased with depolarization, as is typical of K V 4 channels. The K V 4 channel blocker AmmTx3 almost eliminated inactivating currents and broadened action potentials evoked from strongly negative potentials by ∼8-fold. Optogenetic stimulation of inhibitory cerebellar nucleo-olivary terminals hyperpolarized IO cells sufficiently to abolish the ADP. The data support the idea that currents through K V 4 channels control action potential waveforms in IO cells, shortening ADPs during synaptic inhibition or troughs of membrane potential oscillations, thereby controlling the number of climbing fibre action potentials that propagate to the cerebellum. KEY POINTS: Neurons in the mouse inferior olive (IO) express a large, inactivating, voltage-gated A-type K current carried by K V 4 channels. IO action potentials evoked from rest have large, long afterdepolarizations that disappear with pre-spike hyperpolarizations of 5-15 mV. The steep voltage-sensitivity and rapid recovery of K V 4 channels regulates the duration of the afterdepolarization over more than one order of magnitude. Factors such as synaptic inhibition are sufficient to recruit K V 4 channels and eliminate afterdepolarization (ADP). By controlling the ADP, K V 4 channels can set the number of climbing fibre action potentials relayed to the cerebellum and regulate plasticity implicated in motor learning., (© 2024 The Author(s). The Journal of Physiology published by John Wiley & Sons Ltd on behalf of The Physiological Society.)

Many neurons that fire high-frequency action potentials express specialized voltage-gated Na channel complexes that not only conduct transient current upon depolarization, but also pass resurgent current upon repolarization. The resurgent current is associated with recovery of transient current, even at moderately negative potentials where fast inactivation is usually absorbing. The combined results of many experimental studies have led to the hypothesis that resurgent current flows upon repolarization when an endogenous blocking protein that occludes open channels at depolarized potentials is expelled by inwardly permeating Na ions. Additional observations have suggested that the position of the voltage sensor of domain IV regulates the affinity of the channel for the putative blocker. To test the effectiveness of a kinetic scheme incorporating these features, here we develop and justify a Markov model with states grounded in known Na channel conformations. Simulations were designed to investigate whether including a permeation-dependent unblocking rate constant and two open-blocked states, superimposed on conformations and voltage-sensitive movements present in all voltage-gated Na channels, is sufficient to account for the unusual gating of channels with a resurgent component. Optimizing rate constant parameters against a wide range of experimental data from cerebellar Purkinje cells demonstrates that a kinetic scheme for Na channels incorporating the novel aspects of a permeation-dependent unblock, as well as distinct high- and low-affinity blocked states, reproduces all the attributes of experimentally recorded Na currents in a physiologically plausible manner., (© 2023 The Authors. The Journal of Physiology published by John Wiley & Sons Ltd on behalf of The Physiological Society.)

Key Points: The inferior olive sends instructive motor signals to the cerebellum via the climbing fibre projection, which sends collaterals directly to large premotor neurons of the mouse cerebellar nuclei (CbN cells). Optogenetic activation of inferior olivary axons in vitro evokes EPSCs in CbN cells of several hundred pA to more than 1 nA. The inputs are three-fold larger at younger ages, 12 to 14 days old, than at 2 months old, suggesting a strong functional role for this pathway earlier in development. The EPSCs are multipeaked, owing to burst firing in several olivary afferents that fire asynchronously. The convergence of climbing fibre collaterals onto CbN cells decreases from ∼40 to ∼8, which is consistent with the formation of closed-loop circuits in which each CbN neuron receives input from 4-7 collaterals from inferior olivary neurons as well as from all 30-50 Purkinje cells that are innervated by those olivary neurons., Abstract: The inferior olive conveys instructive signals to the cerebellum that drive sensorimotor learning. Inferior olivary neurons transmit their signals via climbing fibres, which powerfully excite Purkinje cells, evoking complex spikes and depressing parallel fibre synapses. Additionally, however, these climbing fibres send collaterals to the cerebellar nuclei (CbN). In vivo and in vitro data suggest that climbing fibre collateral excitation is weak in adult mice, raising the question of whether the primary role of this pathway may be developmental. We therefore examined climbing fibre collateral input to large premotor CbN cells over development by virally expressing channelrhodopsin in the inferior olive. In acute cerebellar slices from postnatal day (P)12-14 mice, light-evoked EPSCs were large (> 1 nA at -70 mV). The amplitude of these EPSCs decreased over development, reaching a plateau of ∼350 pA at P20-60. Trains of EPSCs (5 Hz) depressed strongly throughout development, whereas convergence estimates indicated that the total number of functional afferents decreased with age. EPSC waveforms consisted of multiple peaks, probably resulting from action potential bursts in single collaterals and variable times to spike threshold in converging afferents. Activating climbing fibre collaterals evoked well-timed increases in firing probability in CbN neurons, especially in younger mice. The initially strong input, followed by the decrement in synaptic strength coinciding with the pruning of climbing fibres in the cerebellar cortex, implicates the climbing fibre collateral pathway in early postnatal development. Additionally, the persistence of substantial synaptic input at least to P60 suggests that this pathway may function in cerebellar processing into adulthood., (© 2017 The Authors. The Journal of Physiology © 2017 The Physiological Society.)

Key Points: Large premotor neurons of the cerebellar nuclei (CbN cells) integrate synaptic inhibition from Purkinje neurons and synaptic excitation from mossy fibres to generate cerebellar output. We find that mossy fibre inputs to CbN cells generate unitary AMPA receptor EPSCs of ∼1 nS that decay in ∼1 ms and mildly voltage-dependent NMDA receptor EPSCs of ∼0.6 nS that decay in ∼7 ms. A few hundred mossy fibres active at a few tens of spikes s -1 must converge on CbN cells to generate physiological CbN spike rates (∼60 spikes s -1 ) during convergent inhibition from spontaneously active Purkinje cells. Dynamic clamp studies in cerebellar slices from weanling mice demonstrate that synaptic excitation from mossy fibres becomes more effective at increasing the rate of CbN cell spiking when the coherence (synchrony) of convergent inhibition is increased., Abstract: Large projection neurons of the cerebellar nuclei (CbN cells), whose activity generates movement, are inhibited by Purkinje cells and excited by mossy fibres. The high convergence, firing rates and strength of Purkinje inputs predict powerful suppression of CbN cell spiking, raising the question of what activity patterns favour excitation over inhibition. Recording from CbN cells at near-physiological temperatures in cerebellar slices from weanling mice, we measured the amplitude, kinetics, voltage dependence and short-term plasticity of mossy fibre-mediated EPSCs. Unitary EPSCs were small and brief (AMPA receptor, ∼1 nS, ∼1 ms; NMDA receptor, ∼0.6 nS, ∼7 ms) and depressed moderately. Using these experimentally measured parameters, we applied combinations of excitation and inhibition to CbN cells with dynamic clamp. Because Purkinje cells can fire coincident simple spikes during cerebellar behaviours, we varied the proportion (0-20 of 40) and precision (0-4 ms jitter) of synchrony of inhibitory inputs, along with the rates (0-100 spikes s -1 ) and number (0-800) of excitatory inputs. Even with inhibition constant, when inhibitory synchrony was higher, excitation increased CbN cell firing rates more effectively. Partial inhibitory synchrony also dictated CbN cell spike timing, even with physiological rates of excitation. These effects were present with ≥10 inhibitory inputs active within 2-4 ms of each other. Conversely, spiking was most effectively suppressed when inhibition was maximally asynchronous. Thus, the rate and relative timing of Purkinje-mediated inhibition set the rate and timing of cerebellar output. The results suggest that increased coherence of Purkinje cell activity can facilitate mossy fibre-driven spiking by CbN cells, in turn driving movements., (© 2017 The Authors. The Journal of Physiology © 2017 The Physiological Society.)

Resurgent Na(+) current results from a distinctive form of Na(+) channel gating, originally identified in cerebellar Purkinje neurons. In these neurons, the tetrodotoxin-sensitive voltage-gated Na(+) channels responsible for action potential firing have specialized mechanisms that reduce the likelihood that they accumulate in fast inactivated states, thereby shortening refractory periods and permitting rapid, repetitive, and/or burst firing. Under voltage clamp, step depolarizations evoke transient Na(+) currents that rapidly activate and quickly decay, and step repolarizations elicit slower channel reopening, or a 'resurgent' current. The generation of resurgent current depends on a factor in the Na(+) channel complex, probably a subunit such as NaVβ4 (Scn4b), which blocks open Na(+) channels at positive voltages, competing with the fast inactivation gate, and unblocks at negative voltages, permitting recovery from an open channel block along with a flow of current. Following its initial discovery, resurgent Na(+) current has been found in nearly 20 types of neurons. Emerging research suggests that resurgent current is preferentially increased in a variety of clinical conditions associated with altered cellular excitability. Here we review the biophysical, molecular and structural mechanisms of resurgent current and their relation to the normal functions of excitable cells as well as pathophysiology., (© 2014 The Authors. The Journal of Physiology © 2014 The Physiological Society.)

1. The permeability of AMPA (alpha-amino-3-hydroxy-5-methyl-4- isoxazolepropionate) receptors in the chick cochlear nucleus, the nucleus magnocellularis (nMAG), was examined by measuring the shift in reversal potential (Erev) of current through glutamate or neurotransmitter-gated channels in solutions of different ionic composition. 2. Outwardly rectifying glutamate-activated currents in outside-out membrane patches showed rapid activation and desensitization. The Erev of glutamate-evoked current in zero sodium solutions was dependent on the extracellular Ca2+ concentration. The relation between Erev and Ca2+ ionic activities could be described by the Goldman-Hodgkin-Katz equation with a permeability ratio, PCa/PCs, of 3.3. The PNa/PCs was estimated as 0.66, indicating a PCa/PNa of 5. 3. Evoked excitatory postsynaptic currents (EPSCs) could be recorded during local perfusion of the auditory nerve-nMAG synapse with isotonic Ca2+. The Erev of the EPSC shifted in the positive direction in high-Ca2+ solution as predicted from the preceding analysis. The fraction of current carried by Ca2+ during the AMPA receptor EPSC was estimated as 18%.