Abnormal acute changes in upper limb muscle cortical representation areas in the patients with writer's cramp during co-activation of distal and proximal muscles.

AIM: We analysed cortical muscle representation areas during single muscle activation and during the co-activation of several upper arm muscles in the patients with writer's cramp to determine the possible occurrence of abnormal dynamic somatotopic changes in M1, in addition to the static map abnormalities already described in this form of dystonia. METHODS: Using transcranial magnetic stimulation, we assessed cortical representations of medial deltoid, extensor carpi radialis and the first dorsal interosseus muscles in eight patients with writer's cramp and in eight healthy control subjects. Cortical maps were obtained during distal muscles' activation either in isolation or in conjunction with voluntary medial deltoid co-activation. RESULTS: This study showed a difference in the organization of cortical representations of these muscles between the patients with dystonia and control subjects. The first dorsal interosseus and the extensor carpi radialis cortical representation areas were larger in the dystonic group. The cortical representations became larger when the medial deltoid was simultaneously co-activated, and this effect was not observed in the control group. In the dystonic group, the three cortical muscle representations largely overlapped and their centres of gravity were closer. CONCLUSION: Patients with dystonia showed not only a different spatial organization of muscle cortical representation areas, but also abnormal acute somatotopic changes during proximal muscle co-activation. Task-specific motor impairment in writer's cramp may result not only from lack of cortical inhibition and the well-known anomalous cortical organization observed in these patients, but also from abnormal patterns of proximo-distal functional muscle coupling.

Writer's cramp: cortical excitability in tasks involving proximo-distal coordination.

AIM: The aim of this work was to analyse how writer's cramp patients coordinate each element of the proximal to distal upper arm muscle chain during voluntary movement. METHODS: Using transcranial magnetic stimulation, we have assessed motor cortex excitability properties in patients by recording motor-evoked potentials and silent periods in both the extensor carpi radialis (ECR) and the first dorsal interosseus muscles (FDI), activated either in isolation, or in conjunction with voluntary medial deltoid (MD) co-activation during performance of precise tasks. Ten dystonic patients and ten healthy controls were tested. RESULTS: In both test groups, the ECR muscle displayed a similar active motor threshold, but the excitability curves reached higher plateau values, when the proximal MD muscle was co-activated. In the dystonic group, the FDI muscle excitability curves reached higher plateau values when the MD was co-activated, whereas co-activation had no effect on the control group. In the control group, silent periods, in both the ECR and the FDI were longer when the MD was co-activated. This effect was not observed in the dystonic group. CONCLUSION: In the dystonic group, facilitation of the FDI was observed during a task involving proximo-distal coordination. No differences in silent periods were observed when the muscle was activated alone. Our results suggest that such abnormal facilitation is not only an impairment of the central inhibitory mechanisms reported for dystonic patients, but, in addition, represents true abnormality in cortical muscle activation strategies.

Plasticity of motor cortex induced by coordination and training.

OBJECTIVE: To study the modifications induced by training of a coordinated movement on the primary motor cortex (M1) maps of one proximal muscle and one distal muscle activated alone and during their co-contraction. METHODS: Six healthy female sport students performed a 6-week training program during which they were trained in darts 3-4 times a week. At the end each subject had made more than 1200 throws. Transcranial magnetic stimulation (TMS) was used to map the proximal medial deltoid (MD) and the distal brachio-radialis (BR) muscle representations on M1. Motor evoked potentials (MEPs) amplitude and excitability curves were used to test corticomotor excitability. RESULTS: The cortical representation areas of each muscle separately increased after training. The cortical representation and the excitability curve of the BR muscle increased during co-activation with the MD. Combining co-contraction and training produced a further enlargement of the M1 representation of the BR muscle. CONCLUSIONS: The enlargement of the BR representation in M1 suggests the development of overlapping zones specifying functional synergies between distal and proximal muscles. SIGNIFICANCE: Our findings support the idea that training of a coordinated movement involving several muscles and joints requires an activity-dependent coupling of cortical networks.

Modification of motoneuron size after partial denervation in neonatal rats.

Our previous studies have shown that partial denervation of extensor digitorum longus muscle (EDL) in the rat at 3 days of age causes an increase in the activity of the intact motoneurons. The originally phasic pattern of activity of EDL became tonic after partial denervation. These modifications of motoneuron activity were associated with the change in the phenotype of the muscle from fast to slow contracting and with a conversion of the muscle fibres from a fast to a slow type. The present study investigates whether the size of the cell body of the active EDL motoneurons change in parallel with the altered muscular activity. The study involved partial denervation of rat EDL muscle by section of the L4 spinal nerve at 3 days of age. Then the remaining motoneurons from L5 spinal nerve supplying the EDL muscle were retrogradly labelled with horseradish peroxidase two months later. The results show a reduction in motoneuron size in parallel with an increase in activity of the motoneurons after partial denervation of EDL muscle.

Motor cortex plasticity induced by extensive training revealed by transcranial magnetic stimulation in human.

This study examines the effect of high-level skilled behaviour on motor cortex representations of upper extremity muscles of ten sportswomen. We used transcranial magnetic stimulation to map proximal medial deltoid and distal extensor carpi radialis muscle representations on both hemispheres during low-level voluntary contraction. We compared cortical representation areas between two groups of subjects and between hemispheres within subjects. The first group comprised five elite volleyball attackers and the second group five runners. Four stimuli were delivered on multiple scalp sites (1.5 cm apart) to induce motor-evoked potentials recorded by surface EMG. Maps were described in terms of excitable scalp positions and of motor-evoked potentials. We observed differences in map areas between the two groups. Volleyball players had larger cortical representations of the proximal medial deltoid muscle than runners. Furthermore, the volleyball players had larger map areas for dominant muscles compared with non-dominant muscles. There was no difference, however, in map area for either muscle between the dominant and non-dominant arm in the runner group. Our results show that heavy training in a specific skill induces an expansion of proximal muscle representation in the contralateral primary motor cortex. This enlarged map area for proximal muscle is accompanied by an increase in the overlapping of proximal and distal muscle representations. This could reflect the fact that motor learning of co-ordinated movement involves a common control of both muscles. This reorganization supports the hypothesis of a cortical plasticity driven by activity.

Re-innervation and recovery of rat soleus muscle and motor unit function after nerve crush.

In this study we have investigated the effects of peripheral nerve crush on the contractile properties of the adult rat soleus muscle. The soleus nerve was crushed close to the muscle and functional re-innervation was assessed by the measurement of the force produced by contraction induced by electrical nerve stimulation. Whole soleus muscle and single motor unit (MU) properties were studied at increasing re-innervation times 7-56 days after crush. Results showed progressive re-innervation as 50% of the axotomized motoneurones had re-innervated their muscle 7 days after crush, 72% at 14 days and re-innervation was complete at 28 days. The force parameters recovered more slowly. Tetanic contractions faded at high frequency stimulation, which did not occur in the control muscle. This disruption in the tetanic response was more pronounced in single MUs. Our results demonstrate for the first time a process of progressive axonal re-innervation by the axotomized motoneurones and provide a functional picture of the effective restoration of the neuromuscular function.

Rescue of motoneurones by MK-801 innervating partially denervated rat muscles.

Partial denervation of the extensor digitorum longus muscle by cutting the L4 spinal nerve in 3-day-old rats causes some of the remaining uninjured motoneurones to die. A previous study has shown that of the 12 motor units usually present in the remaining uninjured L5 spinal nerve, a significantly smaller number of motor units to extensor digitorum longus muscle is found in animals operated on at 3 days. This reduction can be caused by a greater sensitivity of neonatal motoneurones with axons in a partially denervated muscle, to excitotoxic effects of glutamate. Therefore an N-methyl- D-aspartic acid (NMDA) receptor antagonist, MK-801, was injected daily for 12 days after partial denervation at 3 days. Two months after the operation contractile properties, motor unit numbers and sizes were studied. Following MK-801 treatment, the reduction in muscle weight and force output of the partially denervated muscle was less than that in the untreated group. Moreover there were more motor units in MK-801 treated animals. After partial denervation only, 15% of the total number of motor units was present whereas when the same operation was followed by treatment with MK-801, 29% remained. The mean motor unit size in the untreated group was 69% while after treatment with MK-801 it was 152% of the control. Thus treatment with MK-801 after partial denervation of neonatal animals rescued some of the motoneurones destined to die, and allowed expansion of motor unit territory of most of the surviving motoneurones.

The age dependent effect of partial denervation of rat fast muscles on their activity.

In 3 or 18 day old Wistar rats the hindlimb muscles were partially denervated by cutting the L4 spinal nerve. Three months later, the effects of partial denervation of the fast extensor digitorum longus (EDL) muscle on the activity of its remaining motor units were studied using electromyographic (EMG) recordings in freely moving animals. In spite of a reduced number of motor units the amount of aggregate EMG activity was greater in the partially denervated EDL muscle in all experimental conditions. This increase was more obvious at rest than during exploratory behaviour, and was significantly greater in muscles that were partially denervated at 3 days than at 18 days of age. On the other hand, the effect of partial denervation on the EMG activity pattern during locomotion was similar in animals partially denervated at 3 or 18 days of age. Unlike in intact EDL, in the partially denervated EDL muscle the duration of the bursts was influenced by the step cycle duration. Thus, we conclude that although partial denervation of EDL muscle influences the amount and pattern of activity of the remaining undamaged motor units in all animals, some of the alterations of EMG activity were more pronounced in animals denervated at younger age.

Increased activity improves recovery of partially denervated fast rat muscles.

Partial denervation of the neonatal rat extensor digitorum longus muscles by removing the L4 spinal nerve and thus 80% of its innervation [A.L. Connold, T.J. Fisher, S. Maudarbocus, G. Vrbová, Response of developing fast muscles to partial denervation, Neuroscience 46 (1992) 981-988; F. Tyc, G. Vrbová, The effect of partial denervation of developing rat fast muscles on their motor unit properties, J. Physiol. 482 (1995) 651-660] results in its permanent weakness. The possibility that the weakness that follows partial denervation is due to the effects of reducing activity of the muscle during a critical stage of development was studied here. Partial denervation was carried out in 3-day-old pups by removing the L4 spinal nerve. To enhance motor activity two days later some animals had injections of l-Dopa twice a day for 8 days. This treatment induced locomotor activity for at least 2 h/day. The muscles from treated and untreated animals were examined 2 months later. There was a significantly smaller reduction of weight and force production in the muscles from l-Dopa treated animals. Both twitch and tetanic force developed by the EDL muscle from the treated group was twice that of the control untreated group. This effect was due mainly to the larger size of the motor units (MUs) in the l-Dopa treated muscles compared to the controls. The mean motor unit force in the untreated group was 69% of that in the control muscle, whereas this value was 127% in the l-Dopa treated animals. Thus it appears that the activity induced by treatment with l-Dopa could to some extent prevent the loss of weight and force output seen after partial denervation of young fast muscles.

Time course of changes in EMG activity of fast muscles after partial denervation.

After partial denervation, the remaining motor units (MUs) of adult fast extensor digitorum longus muscle (EDL) expand their peripheral field. The time course of this event was studied using tension measurement and recordings of electromyographic (EMG) activity. The results show that after section of the L4 spinal nerve, when only 5.3 +/- 0.63 of the 40 MUs normally supplying EDL muscle remain, the force of individual motor units starts to increase between the 1st and 2nd week after the operation and continues to do so for a further week. The drastic reduction of the number of motoneurones supplying the fast EDL leads to an increase in activity of the remaining MUs. In the 1st week after partial denervation, there was a sharp increase in the EMG activity of remaining motor units. During the next 12 days, this increase became less marked, but EMG activity remained nevertheless significantly higher than that of the unoperated EDL muscle. Many MUs became tonically active during posture. The EMG activity pattern during locomotion was also altered, so that the burst duration was positively correlated with the step cycle duration. Moreover, shortly after partial denervation, the interlimb coordination was disturbed but returned to its original symmetrical use 1-2 weeks later.

A simple method for local delivery of various substances to the rat neuromuscular system.

We report here a simple method for the local delivery of various substances to the neuromuscular system in developing and adult rats. This method permits continuous treatment of tissues with a compound over a period of days. Alternative drug delivery systems are unsuitable in neonates. Osmotic pumps are too large and repetitive injections damage the tissues in neonatal rats. Our delivery system provides an adaptable means by which we can directly apply substances in various concentrations in implants of differing sizes. Substances are incorporated into flexible, non-toxic silicone rubber. Strips are cut from the rubber for implantation alongside the muscle or nerve in the anaesthetised animal. The size of the strip is tailored to the length of the muscle or nerve requiring the treatment. Release of the substance from the implant occurs over a period of days and if a longer period of treatment is required, the initial strip can be replaced with a second and even a third implant. We have tested the effects of the substances applied in this manner both physiologically, by examination of muscle function, and morphologically, by muscle histology and retrograde labelling of motoneurones. We have successfully used this method for the application of various groups of substances, including neurotoxins, channel blockers (K+, Ca2+ and Cl-), calcium-chelating agents, protease inhibitors and ionic salts.

Functional reorganization of the partially denervated hindlimb extensor and flexor muscle in rat.

In view of the neuromuscular system plasticity the functional changes induced by partial denervation are presented. The long-term effects of partial denervation of postural (soleus-SOL) or flexor (extensor digitorum longus-EDL) muscles on their EMG activity were studied in rats. The activity per motor unit was significantly higher both in the partially denervated SOL and EDL muscles. During standing or walking the EMG activity pattern of the partially denervated SOL muscle was similar to normal, while the partially denervated EDL muscle during standing exhibited abnormal tonic activity, and during locomotion its burst duration was strongly correlated to the step cycle duration. Thus, partial denervation led to an overall increase of activity of the remaining motor units in both SOL and EDL muscles, while the temporal pattern of muscle activity during locomotion was drastically altered in EDL muscle only, what indicates that partial denervation influenced the postural activity less than the phasic one.

Stabilisation of neuromuscular junctions by leupeptin increases motor unit size in partially denervated rat muscles.

The effect on inhibiting the calcium activated neutral protease (CANP) by leupeptin on force output and motor unit size of the partially denervated rat EDL muscle was studied. Partial denervation was performed under anaesthesia by section of the L4 ventral ramus in 3- and 18-day-old Wistar rats. Two days after the operation a silicon strip containing the inhibitor of CANP leupeptin was implanted alongside the partially denervated EDL. Two to 3 months later the animals were anaesthetized and the EDL muscles on both sides prepared for tension recording. The results from these recordings show dramatic reduction in force output and muscle weight in animals operated at 3 days and this reduction was less pronounced in muscles treated with leupeptin. The mean force output of individual motor units increases in the leupeptin-treated partially denervated muscle compared to the untreated muscle. The increased fatigue resistance typical of muscles partially denervated at 3 days [37] is less pronounced in the treated muscle. In animals operated at 18 days the individual motor units actually increased in size and the leupeptin treatment had no effect on the partially denervated EDL muscles. The difference between the response to leupeptin of the 3 day and 18 day operated animals could be due to the different patterns of innervation of the muscles at the time of the application of the inhibitor of CANP.

The effect of partial denervation of developing rats fast muscles on their motor unit properties.

1. The effects of partial denervation on motor units of the fast twitch extensor digitorum longus (EDL) muscle of the rat were studied. 2. Partial denervation was performed by surgically removing 2-4 mm of the L4 ventral ramus in 3- and 18-day-old Wistar rats. Two to three months later, EMG activity, contractile properties and muscle fibre types were analysed. 3. After partial denervation the EDL muscle became significantly more active, particularly in the animals operated on at 3 days. The amount of activity during periods of rest was up to 4 times that of the control EDL muscle. 4. The maximum tetanic tension developed by the EDL muscles 2 months after partial denervation at 3 days was only 11 +/- 1.01% (S.E.M., n = 18) of the control. In animals operated on at 18 days this value was larger, i.e. 44 +/- 3.46% (S.E.M., n = 12). The low force output of animals operated on at 3 days was also reflected in the low mean motor unit (MU) force output which was only 69 +/- 5.82% (S.E.M., n = 17) of the contralateral control muscle. In contrast the force generated by MUs of rats operated on at 18 days was larger than that of control muscles, i.e. 151 +/- 13.05% (S.E.M., n = 11). The number of MUs was 6 +/- 0.32 (S.E.M., n = 19) in rats operated on at 3 days and 12 +/- 0.83 (S.E.M., n = 14) in rats operated on at 18 days. 5. The speed of contraction decreased and the resistance to fatigue increased. These changes were greater in animals operated on at 3 days. The proportion of muscle fibres reacting with antibody against slow myosin showed a significant increase, especially in the group of animals operated on at 3 days.