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In amyotrophic lateral sclerosis (ALS), an adult onset disease in which there is progressive degeneration of motoneurones, it has been suggested that an intrinsic hyperexcitability of motoneurones (i.e. an increase in their firing rates), contributes to excitotoxicity and to disease onset. Here we show that there is no such intrinsic hyperexcitability in spinal motoneurones. Our studies were carried out in an adult mouse model of ALS with a mutated form of superoxide dismutase 1 around the time of the first muscle fibre denervations. We showed that the recruitment current, the voltage threshold for spiking and the frequency-intensity gain in the primary range are all unchanged in most spinal motoneurones, despite an increased input conductance. On its own, increased input conductance would decrease excitability, but the homeostasis for excitability is maintained due to an upregulation of a depolarizing current that is activated just below the spiking threshold. However, this homeostasis failed in a substantial fraction of motoneurones, which became hypoexcitable and unable to produce sustained firing in response to ramps of current. We found similar results both in lumbar motoneurones recorded in anaesthetized mice, and in sacrocaudal motoneurones recorded in vitro, indicating that the lack of hyperexcitability is not caused by anaesthetics. Our results suggest that, if excitotoxicity is indeed a mechanism leading to degeneration in ALS, it is not caused by the intrinsic electrical properties of motoneurones but by extrinsic factors such as excessive synaptic excitation.

Spinal motoneurons may display a variety of firing patterns including bistability between repetitive firing and quiescence and, more rarely, bistability between two firing states of different frequencies. It was suggested in the past that firing bistability required that the persistent L-type calcium current be segregated in distal dendrites, far away from the spike generating currents. However, this is not supported by more recent data. Using a two compartment model of motoneuron, we show that the different firing patterns may also result from the competition between the more proximal dendritic component of the dendritic L-type conductance and the calcium sensitive potassium conductance responsible for afterhypolarization (AHP). Further emphasizing this point, firing bistability may be also achieved when the L-type current is put in the somatic compartment. However, this requires that the calcium-sensitive potassium conductance be triggered solely by the high threshold calcium currents activated during spikes and not by calcium influx through the L-type current. This prediction was validated by dynamic clamp experiments in vivo in lumbar motoneurons of deeply anesthetized cats in which an artificial L-type current was added at the soma. Altogether, our results suggest that the dynamical interaction between the L-type and afterhyperpolarization currents is as fundamental as the segregation of the calcium L-type current in dendrites for controlling the discharge of motoneurons.

Since their discovery in the late 19th century our conception of motoneurons has steadily evolved. Motoneurons share the same general function: they drive the contraction of muscle fibers and are the final common pathway, i.e., the seat of convergence of all the central and peripheral pathways involved in motricity. However, motoneurons innervate different types of muscular targets. Ordinary muscle fibers are subdivided into three main subtypes according to their structural and mechanical properties. Intrafusal muscle fibers located within spindles can elicit either a dynamic, or a static, action on the spindle sensory endings. No less than seven categories of motoneurons have thereby been identified on the basis of their innervation pattern. This functional diversity has hinted at a similar diversity in the inputs each motoneuron receives, as well as in the electrical, or cellular, properties of the motoneurons that match the properties of their muscle targets. The notion of the diverse properties of motoneurons has been well established by the work of many prominent neuroscientists. But in today's scientific literature, it tends to fade and motoneurons are often thought of as a homogenous group, which develop from a given population of precursor cells, and which express a common set of molecules. We first present here the historical milestones that led to the recognition of the functional diversity of motoneurons. We then review how the intrinsic electrical properties of motoneurons are precisely tuned in each category of motoneurons in order to produce an output that is adapted to the contractile properties of their specific targets.

We explain the mechanism that elicits the mixed mode oscillations (MMOs) and the subprimary firing range that we recently discovered in mouse spinal motoneurons. In this firing regime, high-frequency subthreshold oscillations appear a few millivolts below the spike voltage threshold and precede the firing of a full blown spike. By combining intracellular recordings in vivo (including dynamic clamp experiments) in mouse spinal motoneurons and modeling, we show that the subthreshold oscillations are due to the spike currents and that MMOs appear each time the membrane is in a low excitability state. Slow kinetic processes largely contribute to this low excitability. The clockwise hysteresis in the I-F relationship, frequently observed in mouse motoneurons, is mainly due to a substantial slow inactivation of the sodium current. As a consequence, less sodium current is available for spiking. This explains why a large subprimary range with numerous oscillations is present in motoneurons displaying a clockwise hysteresis. In motoneurons whose I-F curve exhibits a counterclockwise hysteresis, it is likely that the slow inactivation operates on a shorter time scale and is substantially reduced by the de-inactivating effect of the afterhyperpolarization (AHP) current, thus resulting in a more excitable state. This accounts for the short subprimary firing range with only a few MMOs seen in these motoneurons. Our study reveals a new role for the AHP current that sets the membrane excitability level by counteracting the slow inactivation of the sodium current and allows or precludes the appearance of MMOs.

The fast contraction time of mouse motor units creates a unique situation in which motoneurons have to fire at low frequencies to produce small forces but also at very high frequency (much higher than in cat or rat motoneurons) to reach the fusion frequency of their motor units. To understand how this problem is solved, we performed intracellular recordings of adult mouse spinal motoneurons and investigated systematically their subthreshold properties and their discharge pattern. We show that mouse motoneurons have a much wider range of firing frequencies than cat and rat motoneurons because of three salient features. First, they have a short membrane time constant. This results in a higher cutoff frequency and a higher resonance frequency, which allow mouse motoneurons to integrate inputs at higher frequencies. Second, their afterhyperpolarization (AHP) is faster, allowing the motoneurons to discharge at a higher rate. Third, motoneurons display high-frequency (100-150 Hz) subthreshold oscillations during the interspike intervals. The fast membrane kinetics greatly favors the appearance of these oscillations, creating a "subprimary range" of firing. In this range, which has never been reported in cat and in rat spinal motoneurons, the oscillations follow the AHP and trigger spiking after a variable delay, allowing a discharge at low frequency but at the expense of an irregular rate.

Why do motoneurons possess two persistent inward currents (PICs), a fast sodium current and a slow calcium current? To answer this question, we replaced the natural PICs with dynamic clamp-imposed artificial PICs at the soma of spinal motoneurons of anesthetized cats. We investigated how PICs with different kinetics (1-100 ms) amplify proprioceptive inputs. We showed that their action depends on the presence or absence of a resonance created by the I(h) current. In resonant motoneurons, a fast PIC enhances the resonance and amplifies the dynamic component of Ia inputs elicited by ramp-and-hold muscle stretches. This facilitates the recruitment of these motoneurons, which likely innervate fast contracting motor units developing large forces, e.g., to restore balance or produce ballistic movements. In nonresonant motoneurons, in contrast, a fast PIC easily triggers plateau potentials, which leads to a dramatic amplification of the static component of Ia inputs. This likely facilitates the recruitment of these motoneurons, innervating mostly slowly contracting and fatigue-resistant motor units, during postural activities. Finally, a slow PIC may switch a resonant motoneuron to nonresonant by counterbalancing I(h), thus changing the action of the fast PIC. A modeling study shows that I(h) needs to be located on the dendrites to create the resonance, and it predicts that dendritic PICs amplify synaptic input in the same manner as somatic PICs.

Does the afterhyperpolarization current control the gain and discharge variability of motoneurones according to the same law? We investigated this issue in lumbar motoneurones of anaesthetized cats. Using dynamic clamp, we measured the conductance, time constant and driving force of the AHP current in a sample of motoneurones and studied how the gain was correlated to these quantities. To study the action of the AHP on the discharge variability and to compare it to its action on the gain, we injected an artificial AHP-like current in motoneurones. This increased the natural AHP. In three motoneurones, we abolished most of the natural AHP with the calcium chelator BAPTA to investigate the condition where the discharge was essentially controlled by the artificial AHP. Our results demonstrate that both the gain and the coefficient of variation of the firing rate are inversely proportional to the magnitude and to the time constant of the artificial AHP conductance. This indicates that the AHP exerts the same control over the gain and the variability. This mechanism ensures that the variability of the discharge is modulated with the gain. This guarantees a great regularity of the discharge when the motoneurone is in a low excitability state and hence good control of the force produced.

We accurately measured the conductance responsible for the afterhyperpolarization (medium AHP) that follows a single spike in spinal motoneurons of anesthetized cats. This was done by using the dynamic-clamp method. We injected an artificial current in the neurons that increased the AHP amplitude, and we made use of the fact that the intensity of the natural AHP current at the trough of the voltage trajectory was related linearly to the AHP amplitude. We determined at the same time the conductance and the reversal potential of the AHP current. This new method was validated by a simple theoretical model incorporating AHP and hyperpolarization-activated (Ih) currents and could be applied when the decay time constant of the AHP conductance was at least five times shorter than the estimated Ih activation time. This condition was fulfilled in 33 of 44 motoneurons. The AHP conductance varied from 0.3 to 1.4 microS in both slow- and fast-type motoneurons, which was approximately the same range as the input conductance of the entire population. However, AHP and input conductances were not correlated. The larger AHP in slow-type motoneurons was mainly attributable to their smaller input conductance compared with fast motoneurons. The likeness of the AHP conductance in both types of motoneurons is in sharp contrast to differences in AHP decay time and explains why slow- and fast-type motoneurons have similar gain.

Peroneal motoneurons were recorded intracellularly in anesthetized cats during sustained submaximal contractions of peroneus brevis muscle (PB) elicited by repetitive electrical stimulation of motor axons in the distal portion of cut ventral root filaments. Mechanical stimulation of the territory innervated by the superficial peroneal nerve (SP) was applied during contraction to assess the influence of afferents from this territory on the contraction-induced excitation of motoneurons. In 21 peroneal motoneurons in which PB contraction evoked excitatory potentials, a stimulation engaging mechanoreceptors located in the skin around toes was found to either enhance (in 12 motoneurons) or reduce (in 9 motoneurons) the contraction-induced excitatory potentials. Among positive effects, six showed simple summation of the responses to each individual stimulus, suggesting a convergence of afferent pathways on motoneurons. In six other motoneurons, complex interactions were observed, as may result from convergence at a premotoneuronal level. Among negative effects, a single instance was observed of inhibitory facilitation, as may result from convergence of cutaneous and muscular, possibly Ib, afferents on inhibitory interneurons. Several pathways, mediating either facilitory or inhibitory influences, are available for cooperation of muscle and cutaneous input, allowing flexibility of motoneuron activation in different tasks.

The postsynaptic potentials elicited in peroneal motoneurons by either mechanical stimulation of cutaneous areas innervated by the superficial peroneal nerve (SP) or repetitive electrical stimulation of SP were compared in anesthetized cats. After denervation of the foot sparing only the territory of SP terminal branches, reproducible mechanical stimulations were applied by pressure on the plantar surface of the toes via a plastic disk attached to a servo-length device, causing a mild compression of toes. This stimulus evoked small but consistent postsynaptic potentials in every peroneal motoneuron. Weak stimuli elicited only excitatory postsynaptic potentials (EPSPs), whereas increase in stimulation strength allowed distinction of three patterns of response. In about one half of the sample, mechanical stimulation or trains of 20/s electric pulses at strengths up to six times the threshold of the most excitable fibers in the nerve evoked only EPSPs. Responses to electrical stimulation appeared with 3-7 ms central latencies, suggesting oligosynaptic pathways. In another, smaller fraction of the sample, inhibitory postsynaptic potentials (IPSPs) appeared with an increase of stimulation strength, and the last fraction showed a mixed pattern of excitation and inhibition. In 24 of 32 motoneurons where electrical and mechanical effects could be compared, the responses were similar, and in 6 others, they changed from pure excitation on mechanical stimulation to mixed on electrical stimulation. With both kinds of stimulation, stronger stimulations were required to evoke inhibitory postsynaptic potentials (IPSPs), which appeared at longer central latencies than EPSPs, indicating longer interneuronal pathways. The similarity of responses to mechanical and electrical stimulation in a majority of peroneal motoneurons suggests that the effects of commonly used electrical stimulation are good predictors of the responses of peroneal motoneurons to natural skin stimulation. The different types of responses to cutaneous afferents from SP territory reflect a complex connectivity allowing modulations of cutaneous reflex responses in various postures and gaits.

Recent experiments indicate that afferent information is processed in the intraspinal arborisation of mammalian group I fibres. During muscle contraction, Ib inputs arising from tendon organs are filtered out by presynaptic inhibition after their entry in the spinal cord. This paper reviews the mechanisms by which GABAergic axo-axonic synapses, i.e., the morphological substrate of presynaptic inhibition, exert this filtering effect. Using confocal microscopy, axo-axonic synapses were demonstrated on segmental Ib collaterals. Most synapses were located on short preterminal and terminal branches. Using a simple compartmental model of myelinated axon, the primary afferent depolarisation (PAD), generated by such synapses, was predicted to reduce the amplitude of incoming action potentials by inactivating the sodium current, and this prediction was experimentally verified. A further theoretical work, relying on cable theory, suggests that the electrotonic structure of collaterals and the distribution of axo-axonic synapses allow large PADs (about 10 mV) to develop on some distal branches, which is likely to result in a substantial presynaptic inhibition. In addition, the electrotonic structure of group I collaterals is likely to prevent PAD from spreading to the whole arborisation. Such a non-uniform diffusion of the PAD accounts for differential presynaptic inhibition in intraspinal branches of the same fibre. Altogether, our experimental and theoretical works suggest that axo-axonic synapses can control the selective funnelling of sensory information toward relevant targets specified according to the motor task.

Confocal microscopy was used to detect GABA-immunoreactive axo-axonic appositions, indicating possible synaptic contacts, on Ib fiber terminals in the lumbosacral spinal cord. A Ib fiber from posterior biceps-semitendinosus muscles was labeled by intra-axonal ejection of tetramethylrhodamine dextran (red), and serial sections of S1-L7 spinal cord segments were processed for GABA immunocytochemistry revealed by fluorescein isothiocynate (green). Appositions between GABA-immunoreactive structures and the labeled fiber appeared as yellow spots because of the presence of both fluorochromes in small volumes (0.3 * 0.3 * 0.5 micrometer(3)) of tissue. These spots were identified as probable axo-axonic contacts when: (1) they were observed in two to four serial confocal planes, indicating that they did not occur by chance; and (2) their sizes, shapes, and locations were similar to those of axo-axonic contacts found on Ia terminals, known to bear presynaptic boutons, and resembled the axo-axonic synapses described in electron microscope studies of Ib boutons in Clarke's column. A total of 59 presumed axo-axonic contacts was observed on two Ib collaterals, representing an estimated 20% of the total complement. In a three-dimensional reconstruction of one collateral, they were mostly located in terminal positions, and some branches bore more contacts than others. Such differential distribution could not result from chance appositions between GABAergic structures and Ib arborization and further supported the identification of axo-axonic contacts. Segmental Ib collaterals bear axo-axonic synapses that might ensure differential funneling of information toward different targets.

A compartmental model of myelinated nerve fiber was used to show that primary afferent depolarization (PAD), as elicited by axo-axonic synapses, reduces the amplitude of propagating action potentials primarily by interfering with ionic current responsible for the spike regeneration. This reduction adds to the effect of the synaptic shunt, increases with the PAD amplitude, and occurs at significant distances from the synaptic zone. PAD transiently enhances the sodium current activation, which partly accounts for the PAD-induced fiber hyperexcitability, and enhances sodium inactivation on a slower time course, thus reducing the amplitude of action potentials. In vivo, intraaxonal recordings from the intraspinal portion of group I afferent fibers were carried out to verify that depolarizations reduced the amplitude of propagating action potentials as predicted by the model. This article suggests PAD might play a major role in presynaptic inhibition.

In 1- to 72-day-old kittens, motoneurons of the 3 peroneal muscle nuclei were labeled by retrograde axonal transport of horseradish peroxidase from individual muscles. At birth, the locations of peroneal nuclei were similar to those of the adult cat. Counts of motoneurons at different ages indicated that postnatal cell death does not occur in peroneal motor nuclei. Primary dendrites were as numerous in motoneurons of newborn kittens as in adult motoneurons but they were thinner, shorter and poorly ramified. The number of recurrent axon collaterals was higher in the first postnatal week than at later stages. The growth of motoneurons followed similar rates in the 3 peroneal nuclei. Distributions of cell body diameters and volumes were unimodal at birth and became bimodal between 15 and 20 days postnatal. The separation of peroneal motoneurons in two size subgroups, presumably corresponding to alpha and gamma populations, was followed by an increase in growth rate which became faster for alpha than for gamma motoneurons.

Autogenetic inhibition of homonymous and synergist motoneurones can be elicited by very weak partial twitches of gastrocnemius medialis muscle, but during sustained contractions the amplitude of inhibitory post-synaptic potentials decreases quickly. A similar decrease also occurs during stronger contractions. The mechanism responsible for this decrease is still active in low spinal preparations. Pre-synaptic inhibition of Ib afferent fibres might contribute to this reduction of efficiency in the transmission of Ib afferent inputs to motoneurones.