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The spinal motoneuron has long been a good model system for studying neural function because it is a neuron of the central nervous system with the unique properties of (1) having readily identifiable targets (the muscle fibers) and therefore having a very well-known function (to control muscle contraction); (2) being the convergent target of many spinal and descending networks, hence the name of "final common pathway"; and (3) having a large soma which makes it possible to penetrate them with sharp intracellular electrodes. Furthermore, when studied in vivo, it is possible to record simultaneously the electrical activity of the motoneurons and the force developed by their muscle targets. Performing intracellular recordings of motoneurons in vivo therefore put the experimentalist in the unique position of being able to study, at the same time, all the compartments of the "motor unit" (the name given to the motoneuron, its axon, and the muscle fibers it innervates(1)): the inputs impinging on the motoneuron, the electrophysiological properties of the motoneuron, and the impact of these properties on the physiological function of the motoneurons, i.e. the force produced by its motor unit. However, this approach is very challenging because the preparation cannot be paralyzed and thus the mechanical stability for the intracellular recording is reduced. Thus, this kind of experiments has only been achieved in cats and in rats. However, the study of spinal motor systems could make a formidable leap if it was possible to perform similar experiments in normal and genetically modified mice. For technical reasons, the study of the spinal networks in mice has mostly been limited to neonatal in vitro preparations, where the motoneurons and the spinal networks are immature, the motoneurons are separated from their targets, and when studied in slices, the motoneurons are separated from most of their inputs. Until recently, only a few groups had managed to perform intracellular recordings of motoneurons in vivo(2-4 ), including our team who published a new preparation which allowed us to obtain very stable recordings of motoneurons in vivo in adult mice(5,6). However, these recordings were obtained in paralyzed animals, i.e. without the possibility to record the force output of these motoneurons. Here we present an extension of this original preparation in which we were able to obtain simultaneous recordings of the electrophysiological properties of the motoneurons and of the force developed by their motor unit. This is an important achievement, as it allows us to identify the different types of motoneurons based on their force profile, and thereby revealing their function. Coupled with genetic models disturbing spinal segmental circuitry(7-9), or reproducting human disease(10,11), we expect this technique to be an essential tool for the study of spinal motor system.

Movement is accomplished by the controlled activation of motor unit populations. Our understanding of motor unit physiology has been derived from experimental work on the properties of single motor units and from computational studies that have integrated the experimental observations into the function of motor unit populations. The article provides brief descriptions of motor unit anatomy and muscle unit properties, with more substantial reviews of motoneuron properties, motor unit recruitment and rate modulation when humans perform voluntary contractions, and the function of an entire motor unit pool. The article emphasizes the advances in knowledge on the cellular and molecular mechanisms underlying the neuromodulation of motoneuron activity and attempts to explain the discharge characteristics of human motor units in terms of these principles. A major finding from this work has been the critical role of descending pathways from the brainstem in modulating the properties and activity of spinal motoneurons. Progress has been substantial, but significant gaps in knowledge remain., (© 2012 American Physiological Society)

Spasticity is a condition that can include increased muscle tone, clonus, spasms, and hyperreflexia. In this study, we report the effect of manually stimulating the dorsal lumbosacral skin on spontaneous locomotor-like activity and on a variety of reflex responses in 5 decerebrate chronic spinal cats treated with clonidine. Cats were spinalized 1 month before the terminal experiment. Stretch reflexes were evoked by stretching the left triceps surae muscles. Crossed reflexes were elicited by electrically stimulating the right tibial or superficial peroneal nerves. Wind-up of reflex responses was evoked by electrically stimulating the left tibial or superficial peroneal nerves. We found that pinching the skin of the back abolished spontaneous locomotor-like activity. We also found that back pinch abolished the rhythmic activity observed during reflex testing without eliminating the reflex responses. Some of the rhythmic episodes of activity observed during reflex testing were consistent with clonus with an oscillation frequency greater than 3 Hz. Pinching the skin of the back effectively abolished rhythmic activity occurring spontaneously or evoked during reflex testing, irrespective of oscillation frequency. The results are consistent with the hypothesis that locomotion and clonus are produced by common central pattern-generators. Stimulating the skin of the back could prove helpful in managing undesired rhythmic activity in spinal cord-injured humans., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Clonidine, an α-noradrenergic agonist, facilitates hindlimb locomotor recovery after complete spinal transection (i.e. spinalization) in adult cats. However, the mechanisms involved in clonidine-induced functional recovery are poorly understood. Sensory feedback from the legs is critical for hindlimb locomotor recovery in spinalized mammals and clonidine could alter how spinal neurons respond to peripheral inputs in adult spinalized cats. To test this hypothesis we evaluated the effect of clonidine on the responses of hindlimb muscles, primarily in the left hindlimb, evoked by stretching the left triceps surae muscles and by stimulating the right tibial and superficial peroneal nerves in eight adult decerebrate cats that were spinalized 1 month before the terminal experiment. Cats were not trained following spinalization. Clonidine had no consistent effect on responses of ipsilateral muscles evoked by triceps surae muscle stretch. However, clonidine consistently potentiated the amplitude and duration of crossed extensor responses. Moreover, following clonidine injection, stretch and tibial nerve stimulation triggered episodes of locomotor-like activity in approximately one-third of trials. Differential effects of clonidine on crossed reflexes and on ipsilateral responses to muscle stretch indicate an action at a pre-motoneuronal site. We conclude that clonidine facilitates hindlimb locomotor recovery following spinalization in untrained cats by enhancing the excitability of central pattern generating spinal neurons that also participate in crossed extensor reflex transmission.

Earlier studies in multifunctional muscles such as the first dorsal interosseous (FDI) have demonstrated that the selection and control of motor units (MUs) can vary as a function of generated force direction. While directionally dependent motor unit recruitment and rate properties imply that there may also be differential mechanical action, this has yet to be directly demonstrated. Our objective was to determine whether there exists a range of force vectors from different motor units in the FDI muscle within individual subjects. We utilized the spike-triggered averaging (STA) method to derive force twitch estimates from single motor units. We derived MU twitch direction from the ratio of individual twitch estimates recorded concurrently from the load cell. Fifteen units from 2 subjects were used to determine MU force vectors. We were able to estimate force twitch vectors from 7-8 different MUs in each subject. The results of our study suggest that there is varied mechanical action of motor units in the FDI. It is thus possible that differential activation of individual MUs in the FDI is a function of varied mechanical action.

Spinal motoneurons are highly vulnerable in amyotrophic lateral sclerosis (ALS).Previous research using a standard animal model, the mutant superoxide dismutase-1 (SOD1)mouse, has revealed deficits in many cellular properties throughout its lifespan. The electrical properties underlying motoneuron excitability are some of the earliest to change; starting at 1 week postnatal, persistent inward currents (PICs) mediated by Na+ are upregulated and electrical conductance, a measure of cell size, increases. However, during this period these properties and many others undergo large developmental changes which have not been fully analysed.Therefore, we undertook a systematic analysis of electrical properties in more than 100 normal and mutant SOD1 motoneurons from 0 to 12 days postnatal, the neonatal to juvenile period.We compared normal mice with the most severe SOD1 model, the G93A high-expressor line. We found that the Na+ PIC and the conductance increased during development. However, mutant SOD1 motoneurons showed much greater increases than normal motoneurons; the mean Na+PIC in SOD1 motoneurons was double that of wild-type motoneurons. Additionally, in mutant SOD1 motoneurons the PIC mediated by Ca2+ increased, spike width decreased and the time course of the after-spike after-hyperpolarization shortened. These changes were advances of the normal effects of maturation. Thus, our results show that the development of normal and mutant SOD1 motoneurons follows generally similar patterns, but that the rate of development is accelerated in the mutant SOD1 motoneurons. Statistical analysis of all measured properties indicates that approximately 55% of changes attributed to the G93A SOD1 mutation can be attributed to an increased rate of maturation.

The objective of our study was to assess the role of persistent inward currents, or PICs, on the excitability of motoneurons innervating spastic muscle in hemi-spastic stroke individuals. This was accomplished by examining the effects of tonic vibration applied to the tendon of the biceps brachii muscle. The elicited TVR (tonic vibration reflex) provides a useful way to assess the degree of excitability of spinal neurons in spastic syndromes, and it has additional features that may signify the presence of PICs in spastic motoneurons. We applied sinusoidal stretches of varied duration to the biceps tendon of two hemi-spastic stroke individuals and one neurologically intact individual. We recorded the resulting TVR response from electromyographic(EMG) signals obtained from the biceps as well as force recorded at the wrist. The results of our preliminary study show that the initial rise of the TVR force response as well as the force magnitude are generally greater in spastic muscle, perhaps a marker of motoneuron excitability. Additionally, a shorter vibration duration was sufficient to evoke a response on the spastic side of our tested stroke subjects. However, the key marker of PICs--the decay of the force response as well as sustained after-discharge did not exhibit clear differences. Our present data suggests that motoneurons innervating spastic muscle are more readily activated, and thus exhibit increased excitability, which could possibly be a function of greater depolarization, without a change in PIC magnitude. Our data does not rule out the possibility of subthreshold activation of the PIC resulting in enhanced motoneuron depolarization.

Meaningful body movements depend on the interplay between synaptic inputs to motoneurons and their intrinsic properties. Injury and disease often alter either or both of these factors and cause motoneuron and movement dysfunction. The ability of the motoneuronal membrane to generate persistent inward currents (PICs) is especially potent in setting the intrinsic excitability of motoneurons and can drastically change the motoneuron output to a given input. In this article, we review the role of PICs in modulating the excitability of spinal motoneurons during health, and their contribution to motoneuron excitability after spinal cord injury (SCI) and in amyotrophic lateral sclerosis (ALS) leading to exaggerated long-lasting reflexes and muscle spasms, and contributing to neuronal degeneration, respectively., (Copyright 2010 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.)

Spinal motoneurons (MNs) amplify synaptic inputs by producing strong dendritic persistent inward currents (PICs), which allow the MN to generate the firing rates and forces necessary for normal behaviors. However, PICs prolong MN depolarization after the initial excitation is removed, tend to "wind-up" with repeated activation and are regulated by a diffuse neuromodulatory system that affects all motor pools. We have shown that PICs are very sensitive to reciprocal inhibition from Ia afferents of antagonist muscles and as a result PIC amplification is related to limb configuration. Because reciprocal inhibition is tightly focused, shared only between strict anatomical antagonists, this system opposes the diffuse effects of the descending neuromodulation that facilitates PICs. Because inhibition appears necessary for PIC control, we hypothesize that Ia inhibition interacts with Ia excitation in a "push-pull" fashion, in which a baseline of simultaneous excitation and inhibition allows depolarization to occur via both excitation and disinhibition (and vice versa for hyperpolarization). Push-pull control appears to mitigate the undesirable affects associated with the PIC while still taking full advantage of PIC amplification.

The excitability of spinal motoneurons is both fundamental for motor behavior and essential in diagnosis of neural disorders. There are two mechanisms for altering this excitability. The classic mechanism is mediated by synaptic inputs that depolarize or hyperpolarize motoneurons by generating postsynaptic potentials. This "ionotropic" mechanism works via neurotransmitters that open ion channels in the cell membrane. In the second mechanism, neurotransmitters bind to receptors that activate intracellular signaling pathways. These pathways modulate the properties of the voltage-sensitive channels that determine the intrinsic input-output properties of motoneurons. This "neuromodulatory" mechanism usually does not directly activate motoneurons but instead dramatically alters the neuron's response to ionotropic inputs. We present extensive evidence that neuromodulatory inputs exert a much more powerful effect on motoneuron excitability than ionotropic inputs. The most potent neuromodulators are probably serotonin and norepinephrine, which are released by axons originating in the brainstem and can increase motoneuron excitability fivefold or more. Thus, the standard tests of motoneuron excitability (H-reflexes, tendon taps, tendon vibration and stretch reflexes) are strongly influenced by the level of neuromodulatory input to motoneurons. This insight is likely to be profoundly important for clinical diagnosis and treatment.

The primary control of spinal motoneurone excitability is mediated by descending monoaminergic systems, which have diffuse effects on multiple motor pools. Much of the sensory input evoked by movement is also distributed broadly to multiple joints. The muscle spindle Ia afferent system, however, is sharply focused, with Ia excitation restricted to close synergists and Ia reciprocal inhibition only shared between antagonists acting at a single joint. We studied the interaction of neuromodulatory and sensory inputs in determining the movement-related receptive field (MRRF) of motoneurones during passive joint movements of the cat hindlimb. In a decerebrate preparation with tonic monoaminergic input to the cord, the MRRFs tended to be focused for the ankle and knee extensor motor pools studied. Ankle rotation produced larger synaptic currents in ankle extensors than knee or hip rotations and knee rotation dominated input to the knee extensors. The persistent inward current (PIC) in motoneurone dendrites, which is facilitated by monoaminergic input, amplified the MRRF about 2-fold, consistent with its effects on other inputs. Acute spinal transaction markedly broadened MRRFs, with hip rotation generating large currents in both ankle and knee extensors. Spinalization also eliminated amplification of MRRFs, as expected from elimination of descending monoaminergic input. Ia reciprocal inhibition is very effective in suppressing dendritic PICs and thus provides a local and specific PIC control system to oppose the diffuse PIC facilitation from descending monoaminergic systems. The focused MRRF seen in the intact cord state would allow reciprocal inhibition to fulfil this role without undue interference from multijoint input from other afferent systems.

The dendrites of spinal motoneurones are highly active, generating a strong persistent inward current (PIC) that has an enormous impact on processing of synaptic input. The PIC is subject to regulation by descending neuromodulatory systems releasing the monoamines serotonin and noradrenaline. At high monoaminergic drive levels, the PIC dominates synaptic integration, generating an intrinsic dendritic current that is as much as 5-fold larger than the current entering via synapses. Without the PIC, motoneurone excitability is very low. Presumably, this descending control of the synaptic integration via the PIC is used to adjust the excitability (gain) of motoneurones for different motor tasks. A problem with this gain control is that monoaminergic input to the cord is very diffuse, affecting many motor pools simultaneously, probably including both agonists and antagonists. The PIC is, however, exquisitely sensitive to the reciprocal inhibition mediated by length sensitive muscle spindle Ia afferents and Ia interneurones. Reciprocal inhibition is tightly focused, shared only between strict mechanical antagonists, and thus can act to 'sculpt' specific movement patterns out of a background of diffuse neuromodulation. Thus it is likely that motoneurone gain is set by the interaction between diffuse descending neuromodulation and specific and focused local synaptic inhibitory circuits.

Throughout the mammalian spinal cord, interneurones have been shown to exhibit distinct firing patterns in response to a step of injected current. In this study of ventral horn interneurones in a thick slice preparation of the lumbar cord of 11-19-day-old-rats, four distinct firing patterns were observed and classified as repetitive-firing, repetitive/burst, initial-burst or single-spiking. The hypothesis that a persistent sodium current was the predominant determinant of cell firing behaviour was investigated. A slow voltage ramp was used to assess persistent inward currents (PICs). Cells with repetitive-firing patterns had significantly larger PICs than cells displaying repetitive/burst, initial-burst or single-spiking patterns. Repetitive-firing, repetitive/burst and initial-burst-firing cells were reduced to a single-spiking pattern with the application of riluzole, which also markedly reduced the persistent sodium current. Persistent sodium current was found to account for most of the PIC with only a small contribution from L-type calcium current. These results suggest that the persistent sodium current plays a major role in determining firing patterns in these cells.

In the presence of the monoamines serotonin and norepinephrine, motoneurons readily generate large persistent inward currents (PICs). The resulting plateau potentials amplify and sustain motor output. Monoaminergic input to the cord originates in the brainstem and the sharp reduction in monoamine levels that occurs following acute spinal cord injury greatly decreases motoneuron excitability. However, recent studies in the adult sacral cord of the rat have shown that motoneurons reacquire the ability to generate PICs and plateau potentials within 1-2 months following spinal transection. Ca(v)1.3 L-type calcium channels are involved in generating PICs in both healthy and injured animals. Additionally, expression of Ca(v)1.2 and Ca(v)1.3 L-type calcium channels is altered in several pathological conditions. Therefore, in this paper we analyzed the expression of L-type calcium channel alpha(1) subunits within the motoneuron pool following a complete transection of the spinal cord at the level of the sacral vertebra (S)2 segment. The analysis was done both caudally (S4 segment) and rostrally [thoracic vertebra (T)6 segment] from the injury site. The S4 segment was significantly reduced in diameter when compared with control animals, and this reduction was more evident in the white matter. Ca(v)1.2 alpha(1) subunit expression significantly increased (26%) in the motoneuron pool located caudally but not rostrally from the injury site. In contrast, the expression of Ca(v)1.3 alpha(1) subunit remained unchanged in both S4 and T6 segments. The differential expression of the two alpha(1) subunits in spinal injury suggests that Ca(v)1.2 and Ca(v)1.3 channels have different functions in neuronal adaptation following spinal cord injury.

We report the development of an intracellular recording technique for adult mouse motoneurons in sacral spinal cord. Based on a similar preparation for adult rat, we modified the cord preparation solution and filled the sharp electrode with a solution that has physiological osmolarity and pH. The viability of the preparation was examined by recording root reflexes. Short-latency reflexes mediated through monosynaptic transmission between S1 and S3 ventral root were reliably produced by dorsal root electrical stimuli and were stably recorded for more than eight hours. Long-lasting potentiation of the root reflex was observed by bath application of methoxamine, a noradrenergic alpha1 receptor agonist. Bath application of strychnine and picrotoxin, antagonists for glycine and GABA(A) receptors respectively, unmasked long-lasting reflexes that may contain polysynaptic components. In addition, on the background of strychnine and picrotoxin, adding methoxamine induced spontaneous ventral root activity. For intracellular recording, the motoneurons could be reliably penetrated and held for up to 30 min. In all 16 motoneurons recorded, resting membrane potential, input resistance, action potentials and repetitive firing were comparable to those of rat motoneurons. Thus, this preparation is viable and provides a new method for combined electrophysiological and genetic studies of the adult mouse spinal cord.

Neurons perform much of their integrative work in the dendritic tree, and spinal motoneurons have the largest tree of any cell. Electrical excitability is strongly influenced by dendrite membrane properties, which are difficult to measure directly. We describe a method to measure the distribution of ion channel membrane densities along dendritic trajectories. The method combines standard immunohistochemistry with reconstruction procedures for both large-scale and small-scale optical microscopy. Software written for Matlab then extracts the colocalization of the target ion channel with the target dye injected cell, and calculates the relative channel density per square micron of cell surface area, as a function of distance from the cell body. The technique can be used to quantify the localization and distribution of any immunoreactive moiety, and the software provides a flexible vehicle for sensitivity analysis, to validate heuristics for selecting thresholds.

Calcium binding proteins (CBPs) regulate intracellular levels of calcium (Ca(2+)) ions. CBPs are particularly interesting from a morphological standpoint, because they are differentially expressed in certain sub-populations of cells in the nervous system of various species of vertebrate animals. However, knowledge on the cellular regulation governing such cell-specific CBP expression is still incomplete. In this work on the L7 segment of the cat spinal cord, we analyzed the localization and morphology of neurons expressing the CBPs calbindin-28 KD (CB), parvalbumin (PV), and calretinin (CR), and co-expressing CB and PV, CB and CR, and PV and CR. Single CBP-positive ((+)) neurons showed specific distributions: (1) CB was present in small neurons localized in laminae I, II, III and X, in small to medium size neurons in laminae III-VI, and in medium to large neurons in laminae VI-VIII; (2) PV was present in small size neurons in laminae III and IV and in medial portions of laminae V and VI, medium neurons and in lamina X at the border with lamina VII, in medium to large neurons in laminae VII and VIII; (3) CR labeling was detected in small size neurons in laminae I, II, III and VIII, in medium to large size neurons in laminae I and III-VII, and in small to medium size neurons in lamina X. Double labeled neurons were a small minority of the CBP(+) cells. Co-expression of CB and PV was seen in 1 to 2% of the CBP(+) cells, and they were detected in the ventral and intermediate portions of lamina VII and in lamina X. Co-localization of CB and CR was present in 0.3% of the cells and these cells were localized in lamina II. Double labeling for PV and CR occurred in 6% of the cells, and the cells were localized in ventral part of lamina VII and in lamina VIII. Overall, these results revealed distinct and reproducible patterns of localization of the neurons expressing single CBPs and co-expressing two of them. Distinct differences of CBP expression between cat and other species are discussed. Possible relations between the cat L7 neurons expressing different CBPs with the neurons previously analyzed in cat and other animals are suggested.

Mutations in the enzyme superoxide dismutase 1 (SOD1) initiate a progressive motoneurone degeneration in amyotrophic lateral sclerosis (ALS). Transgenic mice overexpressing this mutation develop a similar progressive motoneurone degeneration. In spinal motoneurones cultured from presymptomatic mice expressing the glycine to alanine mutation at base pair 93 (G93A) SOD1 mutation, a marked increase in the persistent component of the Na(+) current was observed, without changes in passive properties. This increase only enhanced neuronal excitability in high input conductance cells, as low input conductance cells exhibited a compensatory outward shift in the current remaining after Na(+) blockade. High input conductance motoneurones tend to be large, so these results may explain the tendency of large motoneurones to degenerate first in ALS. Riluzole, at the therapeutic concentration used to treat ALS, decreased neuronal excitability and persistent Na(+) current in G93A motoneurones to levels observed in the control motoneurones. Aberrations in the intrinsic electrical properties may be among the first symptoms to emerge in SOD1-linked ALS.

Spinal interneurons are essential integrators of descending and peripheral input that receive profuse monoaminergic influence from brainstem nuclei. In this study, the effects of the monoamines serotonin and norepinephrine on the intrinsic properties of ventral horn interneurons were investigated in a slice preparation of the lumbar cord of 7-19 day old rats. Three cell groups with distinct firing patterns in response to steps of injected current were observed and classified as repetitive-firing, initial-burst or single-spiking. Input conductance tended to be largest in single-spiking cells whereas repetitive-firing cells showed the greatest tendency for spontaneous firing and had the fastest rate of rise for the action potential. Rhythmic firing behaviors were defined by the frequency-current relation evoked by linearly increasing current ramps. The monoaminergic modulation of firing patterns and frequency-current relations was primarily studied in repetitive-firing cells. The frequency-current threshold current was decreased in cells with high pre-drug values and increased in cells with low pre-drug values. Therefore, monoamine administration decreased the input-output heterogeneity of the repetitive-firing cells by compressing the range of frequency-current threshold currents. This action of monoamines may have a key role in the suppression of sensory-evoked reflexes and the production of coordinated movement.

Motoneurons have extensive dendritic trees that receive the numerous inputs required to produce movement. These dendrites are highly active, containing voltage-sensitive channels that generate persistent inward currents (PICs) that can enhance synaptic input 5-fold or more. However, this enhancement is proportional to the level of activity of monoaminergic inputs from the brainstem that release serotonin and noradrenalin. The higher this activity, the larger the dendritic PIC and the higher the firing rate evoked by a given amount of excitatory synaptic input. This brainstem control of motoneuron input-output gain translates directly into control of system gain of a motor pool and its muscle. Because large dendritic PICs are probably necessary for motoneurons to have sufficient gain to generate large forces, it is possible that descending monoaminergic inputs scale in proportion to voluntary force. Inhibition from sensory inputs has a strong suppressive effect on dendritic PICs: the stronger the inhibition, the smaller the PIC. Thus, local inhibitory inputs within the cord may oppose the descending monoaminergic control of PICs. Most motor behaviors evoke a mixture of excitation and inhibition (e.g., the reciprocal inhibition between antagonists). Therefore, normal joint movements may involve constant adjustment of PIC amplitude.

Synaptic integration in vivo often involves activation of many afferent inputs whose firing patterns modulate over time. In spinal motoneurons, sustained excitatory inputs undergo enormous enhancement due to persistent inward currents (PICs) that are generated primarily in the dendrites and are dependent on monoaminergic neuromodulatory input from the brain stem to the spinal cord. We measured the interaction between dendritic PICs and inhibition generated by tonic electrical stimulation of nerves to antagonist muscles during voltage clamp in motoneurons in the lumbar spinal cord of the cat. Separate samples of cells were obtained for two different states of monoaminergic input: standard (provided by the decerebrate preparation, which has tonic activity in monoaminergic axons) and minimal (the chloralose anesthetized preparation, which lacks tonic monoaminergic input). In the standard state, steady inhibition that increased the input conductance of the motoneurons by an average of 38% reduced the PIC by 69%. The range of this reduction, from <10% to >100%, was proportional to the magnitude of the applied inhibition. Thus nearly linear integration of synaptic inhibition may occur in these highly active dendrites. In the minimal state, PICs were much smaller, being approximately equal to inhibition-suppressed PICs in the standard state. Inhibition did not further reduce these already small PICs. Overall, these results demonstrate that inhibition from local spinal circuits can oppose the facilitation of dendritic PICs by descending monoaminergic inputs. As a result, local inhibition may also suppress active dendritic integration of excitatory inputs.

Dendrites contain voltage-sensitive conductances that, in vivo, can be influenced by neuromodulatory inputs. In spinal motoneurons, dendrites have voltage-activated persistent inward currents that are facilitated by neuromodulatory input from monoaminergic axons originating in the brainstem. The highest levels of monoamine input are likely to occur during 'fight or flight' behavioral situations. At these high levels, the persistent currents are so large that the dendrites of motoneurons become hyperexcitable, enhancing ionotropic inputs by fivefold or more and allowing the firing rates required for maximal activation of muscle fibers to be generated by surprisingly small inputs. Moderate dendritic excitability (twofold to threefold enhancement) is likely to be a standard component of many normal motor behaviors, such as locomotion. Thus, during normal motor behavior, synaptic integration might be dominated by active currents intrinsic to the dendritic tree rather than by the synaptic current entering via ionotropic channels.

Understanding how synaptic inputs from segmental and descending systems shape motor output from the spinal cord requires information on the relative magnitudes of the synaptic currents produced by the different systems and their patterns of distribution within a motoneurone pool. Equally important are quantitative descriptions of how different synaptic inputs are integrated when they are concurrently active and of how voltage- and ligand-gated conductances on the dendrites of motoneurones affect the transfer of synaptic currents to the soma. We have carried out a number of experimental studies of these inter-related problems on motoneurones in the cat spinal cord and have explored the implications of our findings with computer simulations utilizing a synthetic model of the cat medial gastrocnemius motoneurone pool. This paper provides a brief review of the principal results of our studies.

Enhancement of bistability in spinal motoneurons in vivo by the noradrenergic alpha1 agonist methoxamine. Like many types of motoneurons, spinal motoneurons in the adult mammal can exhibit bistable behavior. This means that short periods of excitatory input can initiate long periods of self-sustained firing and that equally short periods of inhibition can return the cell to the quiescent state. Usually, the presence of one of the monoamines (either serotonin or norepinephrine) is required for spinal motoneurons to express bistable behaviors. Because the decerebrate cat preparation has tonic activity in monoaminergic fibers that originate in the brain stem and project to spinal motoneurons, these cells sometimes exhibit bistable behavior. However, exogenous application of the noradrenergic alpha1 agonist methoxamine greatly enhances bistable behavior in the decerebrate. The goal of this study was to identify the mechanisms of this action of methoxamine. The total persistent inward current (IPIC) in spinal motoneurons in the decerebrate cat was measured from I-V functions generated by triangular voltage commands applied using discontinuous single electrode voltage clamp. The effect of methoxamine on IPIC was assessed by comparing its properties in a control cell sample without methoxamine to its properties in a sample of cells obtained after application of methoxamine. In most experiments, at least one cell was obtained from each sample. Our results showed that methoxamine approximately doubled the amplitude of IPIC without changing its onset voltage, its offset voltage, or its persistence. The reduced amplitude was a consistent finding within experiments and so was unlikely to be caused by interanimal variability. In addition, methoxamine depolarized motoneurons without altering their input conductances, so that a smaller amount of current was required to reach the onset voltage of IPIC. These effects of methoxamine were approximately equal in all cells. As a result of these changes, methoxamine greatly enhanced the tendency for motoneurons to become bistable. It is proposed that the methoxamine-induced increase in the amplitude of IPIC is effective in enhancing the duration of bistable firing because this increase makes IPIC more resistant to the deactivating effects of the afterhyperpolarizations between spikes.

Motoneurons receive a dense innervation from fibers that descend from the brainstem and release the monoamines serotonin and noradrenalin. When monoamines are present, motoneurons can produce plateau potentials, which are sustained depolarizations that outlast a brief excitatory input. During voltage clamp, steady monosynaptic input from Ia afferents produced a current that persisted after the Ia input ceased. The likely origin of this current was in dendritic regions, where plateau potential channels were probably activated due to the lack of voltage clamp control during the synaptic input. The functional significance of these data is discussed.

An important function of the stretch reflex in the soleus muscle in the decerebrate cat preparation is to compensate for the tendency of muscle suddenly to yield during ramp increases in length. As the level of background (i.e. pre-stretch) force increases, there is a systematic change in the curvature of the force trajectory during this reflex compensation, from concave to convex with respect to increasing force. The hypothesis that this change in curvature was due to background force-dependent changes in the recruitment pattern of motor units was investigated with a combined computer simulation/experimental technique. The simulation consisted of 20 model motor units for the soleus muscle, each based on a distributed moment muscle model. The timing of recruitment of the motor units was optimized to allow the simulation outputs to fit a set of experimental data records on the reflex response to stretch initiated at five different levels of pre-stretch force. The resulting recruitment patterns showed that a tendency for recruitment to be concentrated progressively in the early portion of the stretch as pre-stretch force increased could account for the changes in reflex force curvature. These results are consistent with the skewed distribution of intrinsic electrical thresholds of motoneurons, in which low-threshold units are much more frequent than high-threshold ones. Therefore the changes in recruitment pattern and reflex force curvature may be due primarily to the intrinsic properties of motoneurons.

The effects of bilateral focal cooling of dorsolateral thoracic spinal cord on segmental reflex pathways to the triceps surae muscles were assessed in decerebrate cats from the reflex forces produced by single shocks or trains of electrical stimuli applied to the ipsilateral caudal cutaneous sural and the contralateral tibial nerves. The validity of the dorsal cold block technique as a substitute for acute surgical dorsal hemisection was established by showing that focal cooling reliably reproduced the stretch-induced "clasp knife" inhibition of triceps surae reflexive force seen following dorsal hemisection. Under control (warm) conditions, the inhibitory components of electrically evoked ipsilateral sural and contralateral tibial reflexes faded rapidly during sustained trains, with a resultant production of large-amplitude reflex force as measured from either the entire triceps surae or from the medial gastrocnemius muscle alone. Dorsal cold block greatly reduced the amplitude of reflexive force evoked by sustained electrical stimulation of either nerve. Indeed, the cold block completely reversed the sign of train-evoked reflexes to a net inhibition of reflex force output in one-half of the sural and one-half of the contralateral tibial stimulation experiments. Peak transient forces evoked by single shocks to the sural or contralateral tibial nerves were also sometimes reduced, but this result was more variable than for prolonged nerve stimulation. The persistence of activity in segmental inhibitory pathways during dorsal cold block, as indicated by instances of reflex sign reversal, suggests that descending bulbospinal pathways traversing the dorsolateral funiculi may be responsible for "fading" of segmental inhibitory reflex components in decerebrate cats with intact spinal cords during sustained afferent input. The possibility that the enhanced magnitude and duration of segmental inhibition during cold block will increase the likelihood of disruption of the size principle for motoneuron recruitment is also discussed.

In theory, there are at least two distinct mechanisms by which afferent inputs could alter motoneuron discharge and shape the output of a motoneuron pool: either by delivering synaptic current to the motoneurons' somata ('classic' synaptic transduction); or by altering the motoneurons' voltage-sensitive conductances (neuromodulation). Recent work has confirmed the operation of both of these mechanisms. It has been shown that the effect of a 'classic' synaptic input on motoneuron firing rate is predicted by the product of the effective synaptic current and the slope of the motoneuron's frequency-current relation. It has also been shown that neuromodulators can alter both the slope of a motoneuron's frequency-current relation and its threshold for repetitive firing. It is argued here, however, that when two or more sources of synaptic input are activated concurrently, the distinction between these two mechanisms is blurred. Computer simulations of motoneuron and motor pool behavior have proved extremely useful in understanding these processes.

In cat medial gastrocnemius motoneurons, single stimuli to the cutaneous sural nerve evoke a post-synaptic potential with a mixture of depolarization and hyperpolarization, depolarization being dominant in type F cells and hyperpolarization in type S cells. This pattern is consistent with previous reports showing that activation of the sural nerve can sometimes reverse the normal order of motor unit recruitment by inhibiting S motor units while simultaneously exciting F motor units. However, during repetitive stimulation for 1-2 s, we found that the hyperpolarizing component of the sural input to medial gastrocnemius motoneurons was not persistent, but instead gave way to depolarization after the first 30 ms. The net steady-state response after 0.5-1.0 s of stimulation was depolarization in all cells, regardless of motor unit type. This suggests that tonic sural input may be incapable of producing prolonged recruitment reversals.

A tendon-vibration technique, used to raise the electrical threshold of muscle spindle Ia afferent fibers above that of Golgi tendon organ Ib afferent fibers in animals, was tested on human subjects. After prolonged tendon vibration, electrical stimulation of the posterior tibial nerve was ineffective or markedly less effective in eliciting Hoffmann (H-) reflexes in the soleus muscle at previbration threshold intensities. With stimulus intensity held constant at values between 1.0 to 1.4 X threshold, postvibration H-reflex myoelectric amplitudes returned to previbration values usually within 60 min. However, at higher electrical stimulus intensities (1.8 X threshold), postvibration H-reflex amplitudes were produced at or near previbration values irrespective of postvibration recovery time; in contrast, initial postvibration tendon tap reflexes were potentiated. Findings suggest that it is indeed possible to selectively activate Ib afferent fibers in humans by electrical stimuli.

Pulmonary blastoma is a rare lung tumor composed of epithelial and mesenchymal elements; the latter element may show various patterns of differentiation toward mature tissue, such as cartilage, smooth muscle, and bone. Rhabdomyoblastic differentiation in pulmonary blastoma is quite rare; only five such cases have been reported. We report two cases of pulmonary blastoma with rhabdomyoblastic differentiation documented for the first time by electron microscopy and immunohistochemistry including documentation for myoglobin, actin, vimentin and desmin. The diffuse and prominent rhabdomyoblastic differentiation in one case is most unusual.

In humans, prolonged vibration of the Achilles tendon produced transient depression or abolition of the soleus H-reflex. Recovery of the electrical reflex threshold to previbration values at a constant lower stimulus intensity usually occurred between 10 to 55 min. Electrical stimulation at higher multiples of the reflex threshold produced reflex EMG amplitudes more immediately comparable to previbration controls. When postvibration H-reflexes were completely abolished, poststimulus averaging of voluntarily maintained tonic EMG activity showed evidence of inhibition at a 46-ms latency in contrast to a 32-ms previbration H-reflex latency. In cat, observation of H-reflexes were rare, but stimulus-evoked changes in EMG activity mimicked the postvibration depression seen in humans. Ventral root postvibration reflexes from triceps surae varied in magnitude but were usually depressed or abolished at 1.0 to 1.2 times the electrical reflex threshold. These responses returned to previbration control amplitudes within 20 to 35 min. Magnitude of depression and time to recovery were dependent on the intensity of the electrical stimulus. In five experiments, depression of postvibration reflex activity and recovery were accompanied by gradual recovery in amplitude of the group I volley to previbration amplitudes. Elevated group Ia axonal electrical thresholds, monitored from seven isolated units, were observed to recover to previbration values in parallel with postvibration reflex recovery to control amplitudes. At electrical stimulus intensities greater than 1.4 times the reflex threshold, postvibration reflex responses were often potentiated, probably reflecting posttetanic potentiation of group Ia pathways activated at their higher axonal thresholds. In two observations, postvibration Ib axonal electrical thresholds did not change. Overall, the findings supported the proposal that postvibration depression of soleus H-reflexes in humans or cats is caused by both disfacilitation and autogenetic inhibition due to withdrawal of Ia afferent activation and increased selectivity of Ib afferent fiber stimulation, respectively.