Number Of Motor Endplates Research Articles

Effect of lentiviral vector-mediated overexpression of hypoxia-inducible factor 1 alpha delivered by pluronic F-127 hydrogel on brachial plexus avulsion in rats

Brachial plexus avulsion often results in massive motor neuron death and severe functional deficits of target muscles. However, no satisfactory treatment is currently available. Hypoxia-inducible factor 1α is a critical molecule targeting several genes associated with ischemia-hypoxia damage and angiogenesis. In this study, a rat model of brachial plexus avulsion-reimplantation was established, in which C5-7 ventral nerve roots were avulsed and only the C6 root reimplanted. Different implants were immediately injected using a microsyringe into the avulsion-reimplantation site of the C6 root post-brachial plexus avulsion. Rats were randomly divided into five groups: phosphate-buffered saline, negative control of lentivirus, hypoxia-inducible factor 1α (hypoxia-inducible factor 1α overexpression lentivirus), gel (pluronic F-127 hydrogel), and gel + hypoxia-inducible factor 1α (pluronic F-127 hydrogel + hypoxia-inducible factor 1α overexpression lentivirus). The Terzis grooming test was performed to assess recovery of motor function. Scores were higher in the hypoxia-inducible factor 1α and gel + hypoxia-inducible factor 1α groups (in particular the gel + hypoxia-inducible factor 1α group) compared with the phosphate-buffered saline group. Electrophysiology, fluorogold retrograde tracing, and immunofluorescent staining were further performed to investigate neural pathway reconstruction and changes of neurons, motor endplates, and angiogenesis. Compared with the phosphate-buffered saline group, action potential latency of musculocutaneous nerves was markedly shortened in the hypoxia-inducible factor 1α and gel + hypoxia-inducible factor 1α groups. Meanwhile, the number of fluorogold-positive cells and ChAT-positive neurons, neovascular area (labeled by CD31 around avulsed sites in ipsilateral spinal cord segments), and the number of motor endplates in biceps brachii (identified by α-bungarotoxin) were all visibly increased, as well as the morphology of motor endplate in biceps brachil was clear in the hypoxia-inducible factor 1α and gel + hypoxia-inducible factor 1α groups. Taken together, delivery of hypoxia-inducible factor 1α overexpression lentiviral vectors mediated by pluronic F-127 effectively promotes spinal root regeneration and functional recovery post-brachial plexus avulsion. All animal procedures were approved by the Institutional Animal Care and Use Committee of Guangdong Medical University, China.

The effect of bone marrow mesenchymal stem cells on recovery of skeletal muscle after neurotization surgery in rat.

Objective(s):When the nerve is injured near its entrance to the muscle belly, we cannot perform conventional methods. One useful method in such a situation is neurotization surgery. In this study, Bone marrow mesenchymal stem cells (BMSCs) implanted into the paralyzed muscle after neurotization surgery. These cells can stimulate axon growth and motor endplate formation, also prevent muscle atrophy.Materials and Methods:Thirty-six adult male Sprague-Dawley rats were randomized into six groups: intact group, sham surgery group, control group, DMEM group, cell+DMEM group, denervated group. The motor nerve of the lateral head of gastrocnemius muscle was cut, and the proximal portion of the severed nerve was transplanted to the proximal third of the muscle paralysis. BMSCs with/or DMEM was injected into the site of injury. All animals were evaluated by withdrawal reflex latency (WRL), electromyography, muscle weight, histology and immunohistochemistry.Results:The WRL difference between the control and cell+DMEM groups at weeks 4 and 12 post-operation was statistically significant (P<0.05). The mean number of motor end plates in cell+DMEM group was more than control group (P<0.05). At 12 weeks post-operation, the difference of the mean nerve conduction velocity (NCV) between cell treated group and sham surgery groups were not statistically significant (P>0.05). In weeks 4 and 12 post-operation, the mean fiber diameters in cell+DMEM group were more than control group (P<0.05).Conclusion:The results of this study demonstrate that transplantation of BMSCs after neurotization surgery, prevent muscle atrophy and improve muscle function.

Targeted Muscle Reinnervation of the Brachium: An Anatomic Study of Musculocutaneous and Radial Nerve Motor Points Relative to Proximal Landmarks

Targeted muscle reinnervation (TMR) offers enhanced prosthetic use by harnessing additional neural control from unused nerves in the amputated limb. The purpose of this study was to document the location and number of motor end plates to each muscle commonly used in TMR in the brachium relative to proximally based bony landmarks. We dissected 18 matched upper limbs (9 fresh-frozen cadavers). The locations of each of the nerves' muscular insertions into the medial biceps and brachialis were measured relative to the anterolateral tip of the acromion. The terminal branches to the lateral triceps were measured relative to the posterolateral tip of the acromion. Both the number of branches and the location of the muscular insertions were documented. Common descriptive statistics were used to describe the data. There was a median of 2 branches to the medial biceps located 19.6 cm from the anterolateral tip of the acromion (range, 15-25 cm). There was a median of 3.5 branches to the brachialis located 24.2 cm from the anterolateral tip of the acromion (range, 19-27.5 cm). There was a median of 2.5 branches to the lateral triceps located 21.6 cm from the posterolateral tip of the acromion (range, 11-29 cm). The mean distances to the primary branch muscle and the number of smaller branches were not significantly different when compared by sex or side. Motor points for the medial biceps, brachialis, and lateral triceps can be identified reliably using proximal landmarks in targeted muscle reinnervation. The data obtained from this study may assist the surgeon in localizing the nerve branches and muscular insertions for the commonly used muscles for TMR of the brachium.

A rat model study of atrophy of denervated musculature of the hand being faster than that of denervated muscles of the arm

There are no biological marks to indicate if denervated muscle atrophy after nerve injury is irreversible. Clinically in obstetric brachial plexus palsy (OBPP), atrophy of denervated intrinsic musculature of the hand is much faster to irreversible than that of denervated muscles of the arm. 64 pup rats whose C5C6 had been divided and C7C8T1 avulsed, were divided equally into the reconstruction and control groups. The former had subgroups R1, R5, R10, R15 where the ulnar and musculocutaneous nerves were reconstructed one, five, ten and 15 weeks respectively after injury and efficacy was evaluated 12 weeks later. The latter had C1, C5, C10, C15 subgroups where denervated muscles of the two nerves were assessed one, five, ten and 15 weeks after injury. Results of average cross-sectional area of the muscle fiber for intrinsic musculature of the forepaw showed that the R5, R10, R15 subgroups were not statistically superior to the C5, C10, C15 ones, respectively, though R1 was; those for biceps indicated, however, that the R1, R5, R10 subgroups were better than the C1, C5, C10 ones, respectively, though R15 was not. In the reconstruction subgroups regenerative nerve fibers in each nerve were no less than 53 percent of those on the control side, while number of motor end plates was statistically less in subgroups with irreversible muscle atrophy. We conclude that rat model of OBPP is suitable for simulating clinical appearance of atrophy of denervated intrinsic musculature of the hand being faster than that of denervated muscles of the arm.

Transplanted embryonic spinal tissue promotes severed sciatic nerve regeneration in rats

The effects of transplanted embryonic spinal tissue on host motor nerve regeneration and target muscle reinervation were investigated in severed sciatic nerves of rats. The electromyogram (EMG) responses and number of motor end plates (MEP) in target muscles, number of nerve axons, and retrogradely labeled motor neurons were examined in transplantation-, anastomosis without transplantation-, and naïve groups of the animals. The EMG patterns of the transplantation group returned to nearly normal at the 8th week, but those of the anastomosis group did not. MEP counts in the transplantation group were significantly higher than in the anastomosis group. The myelinated axon counts and myelin sheath thickness in the transplantation group were significantly higher than those in the anastomosis group. The number of retrogradely labeled motor neurons was significantly higher in the transplantation group. We conclude that transplanted embryonic spinal tissue can promote both host motor nerve regeneration and target muscle reinnervation.

Electrical stimulation of paralyzed vibrissal muscles reduces endplate reinnervation and does not promote motor recovery after facial nerve repair in rats

The outcome of peripheral nerve injuries requiring surgical repair is poor. Recent work has suggested that electrical stimulation (ES) of denervated muscles could be beneficial. Here we tested whether ES has a positive influence on functional recovery after injury and surgical repair of the facial nerve. Outcomes at 2 months were compared to animals receiving sham stimulation (SS). Starting on the first day after end-to-end suture (facial-facial anastomosis), electrical stimulation (square 0.1 ms pulses at 5 Hz at an ex tempore established threshold amplitude of between 3.0 and 5.0V) was delivered to the vibrissal muscles for 5 min a day, 3 times a week. Restoration of vibrissal motor performance following ES or SS was evaluated using the video-based motion analysis and correlated with the degree of collateral axonal branching at the lesion site, the number of motor endplates in the target musculature and the quality of their reinnervation, i.e. the degree of mono- versus poly-innervation. Neither protocol reduced collateral branching. ES did not improve functional outcome, but rather reduced the number of innervated motor endplates to approximately one-fifth of normal values and failed to reduce the proportion of poly-innervated motor endplates. We conclude that ES is not beneficial for recovery of whisker function after facial nerve repair in rats.

Morphological and morphometrical characteristics of the esophageal intrinsic nervous system in the golden hamster.

Very little is known about esophageal innervation in the hamster. In the present study, we used protein gene product 9.5 (PGP 9.5) to determine immunohistochemically the architectural features of the enteric nervous system in the hamster esophagus. The myenteric plexus consisted of a loose and irregular network of ganglia and interganglionic nerve bundles. The density of the neurons in the myenteric plexus was relatively low (479 +/- 75/cm(2), n = 5), with a preferentially higher density in the upper cervical portion than other parts of the esophagus. Regional differences in the number of PGP 9.5-positive neurons and ganglia were observed. PGP 9.5-immunoreactive fibers in the ganglia often branched, giving rise to expanding nerve endings of laminar morphology resembling intraganglionic laminar endings described in rats and cats. Fine varicose fibers originating from the secondary plexus were occasionally observed near the motor endplates, suggested a dual innervation of the striated muscle. The submucosal plexus was free from ganglionated plexus. A regional difference in the submucosal nervous network was observed. The number of motor endplates in the inner muscle layer was higher than that in the outer muscle layer.

Fibroblast Growth Factor-Induced Motor End Plate Regeneration in Atrophic Muscle

To determine whether fibroblast growth factor 1 implanted with viable nerve into atrophic muscle will stimulate formation of functional, acetylcholine producing motor end plates. Twelve male Lewis rats underwent predenervation of the hamstring muscle 8 weeks before implantation of the nerve at a site distant from the original motor end plate. Six animals underwent implantation of the tagged nerve ending into atrophic muscle with 50 microgram of fibroblast growth factor 1 in a fibrin adhesive carrier (group 1). Three animals underwent implantation with nerve, fibrin adhesive, and no fibroblast growth factor 1 (group 2); and three animals underwent implantation with fibroblast growth factor 1 and fibrin adhesive with no nerve (group 3). Animals were killed 9 weeks after implantation and nerve and muscle specimens were harvested. Histoenzymologic methods for acetylcholinesterase and silver impregnation of nerve fibers were performed 9 weeks after fibroblast growth factor 1-fibrin adhesive implantation. Variables included the number of motor end plates per highpower field and motor end plate length. Robust axonal sprouting and formation of multiple motor end plates were found arborized in serial fashion equidistant around the implanted nerve ending. Rare extrasynaptic staining occurred. End plate lengths were significantly shorter in the fibroblast growth factor 1-treated muscles (group 1) than in the specimens without fibroblast growth factor 1 (group 2) (31.2 vs 58.5 micron; P>001, paired t test). The arborization of motor end plates, rare extrasynaptic staining, and shorter end plate lengths seen in group 1 were all consistent with mature motor end plates. Controls (group 3) displayed limited motor end plate formation and extensive extrasynaptic staining typical of denervation. This study presents encouraging evidence that fibroblast growth factor 1 with fibrin adhesive carrier can facilitate the reinnervation of atrophied muscle by enhancing the formation or revitalization of motor end plates. Future studies will address muscle function and use of different carrier materials.

Regeneration of hypoglossal nerve axons following blockade of the axotomy-induced microglial cell reaction in the rat.

The aim of the present study was to examine whether inhibition of the microglial cell reaction around axotomized motoneurons affects the subsequent regeneration process of the injured axons. The microglial cell reaction in the hypoglossal nucleus of the rat was blocked by infusion of cytosine-arabinoside (ARA-C) into the ventricular system. Axon regeneration was evaluated by determining the number and size distribution of myelinated axons at a defined level distal to the crush site, the number of neurons which could be retrogradely labelled from the distal stump as well as the number of motor endplates in the tongue at various times following injury. No significant difference was observed for any of these parameters between ARA-C-treated and untreated animals. Therefore, it is concluded that the microglial cell reaction is not necessary for peripheral nerves to regenerate and restore target contact at a normal rate and to a normal extent.

An Organotypic Co-culture of Embryonic Rat Brainstem and Tongue or Skeletal Muscle.

Transverse sections of rat embryo brainstem (embryonic age 18 - 19 days) containing the brainstem motor nuclei were explanted, together with a small piece of tongue or skeletal muscle, in a plasma clot and maintained in culture for 2 - 3 weeks using the roller tube technique. The results show that brainstem motoneurons survived, differentiated and innervated newly formed multinucleated myotubes which displayed large synchronized contractions after 1 week in culture. Muscle fibre contractions could be induced by excitatory amino acid applications and suppressed by curarization. Muscle fibres differentiated normally. During the first week they showed diffuse acetylcholinesterase positivity and multi-innervation. During the second and third weeks the number of motor end-plates was greatly reduced and transverse striation was observed. In the presence of muscle fibres, the brainstem thinned out and spread, becoming one or two cell layers thick, and the motoneurons tended to migrate towards the muscle fibres, becoming clearly observable in the living culture. When the muscle explant was not present, the brainstem explant did not spread and remained several cell layers thick, while acetylcholinesterase-positive cells, presumed to be motoneurons, tended to disappear. The preparation described is well suited for electrophysiological studies of differentiating motoneurons and offers direct access to their dendritic tree, a most desirable feature for patch-clamp or multisite optical recording.

Botulinum toxin treatment of spasmodic dysphonia: Techniques, indications, efficacy

Summary A group of 58 patients with spasmodic dysphonia (SD) was treated perorally or transcutaneously with botulinum toxin (BT), and followed for at least 6 months. Most were treated by peroral injection using indirect laryngoscopy to facilitate precise placement. An effort was made to disperse the toxin over the entire thyroarytenoid muscle in order to affect the greatest number of motor end-plates using the smallest possible effective dose. Severely symptomatic patients with primarily adductor SD had the best results. A potentiation effect is suggested by analysis of a subpopulation of patients, mostly injected unilaterally, for whom the outcome was more favorable for those who had had prior injections. One-month postinjection studies reflect significant improvement in measured readings of a standard passage, and this is consistent with the subjective and perceptual improvements noted. There were no severe complications. Peroral injection seems inadvisable for patients with mixed or abductor SD as well as for those with an uncontrollable gag reflex.

The fine structure of the fin musculature in two teleost species with different swimming modes, the puffer, Tetraodon steindachneri, and the goldfish, Carassius auratus

Puffer fish (Tetraodon steindachneri) can execute precise maneuvers due to their highly specialized mode of propulsion. In the conventional locomotion exemplified by the goldfish (Carassius auratus), the fish thrusts are generated by lateral beating of the caudal fin. In contrast, the puffer generates its propulsive force by very rapid undulating movements of the pectoral, dorsal and anal fins. The fine structure of the fin muscles is identical in the two species of fishes, despite the differences in fin movement; cytologically, the fibers are intermediate between those of red and of white muscle. On the other hand, both the fusion frequency and the number of motor endplates are considerably higher in the fin muscles of the puffer than in those of the goldfish.

The size of motor units in laryngeal muscles of the rat.

The size of motor units in rat laryngeal muscles was determined by correlating the number of neurons labeled by i.m. injections of horseradish peroxidase with the number of motor end plates stained for acetylcholinesterase. The cricothyroid has a motor unit size of 8 muscle fibers per motor neuron and the posterior cricoarytenoid 4-5 muscle fibers per motor neuron.

Effects of chronic paralysis with α-bungarotoxin on development of innervation

Chronic paralysis of the duck embryo with α-bungarotoxin increases the number of trochlear motor neurons that survive by decreasing the magnitude of embryonic cell death. The present study examined how such an increase in the motor neuron projection is accommodated at the peripheral target. The results indicate that the overall size of the duck superior oblique muscle was drastically reduced following chronic paralysis with α-bungarotoxin. The number of motor end-plates was reduced to about 25% of the control. Although the total number of end-plates was significantly reduced, their size was larger and they became innervated by far more nerve terminals than in the control. Thus, increased motor neuron survival occurred while the size of the periphery and the number of synaptic sites (end-plates) were greatly reduced. These observations suggest that the cause of embryonic cell death may not be related to the availability of adequate numbers of synapses with the periphery. They also suggest that the physiologic activity of the embryo may play a significant role in regulation of cell number in the developing nervous system.

Neurophysiological signs of myasthenia in rabbits after receptor antibody development

Antibodies directed against nicotinic cholinergic receptor protein have been obtained in rabbits injected with purified receptor protein. The receptor-immunized rabbits showed a peripheral paralysis after 3–4 weeks. No antibodies were formed in rabbits injected with adjuvant alone and no sign of illness appeared in these animals. The motor performance of the immunized animals improved with neostigmine treatment. Recovery of the evoked muscle action potential on supramaximal stimulation of the nerve to the tibialis anterior showed a low amplitude response to a single stimulus, a decrease in amplitude on repetitive stimulation, particularly pronounced at lower stimulation frequencies and a positive response to neostigmine in the immunized animals. The results resemble those found in patients with myasthenia gravis. No pathological abnormalities were found in the viscera of the sick rabbits Cholinesterase staining showed that the number of motor end-plates was normal. The first sign of neuromuscular disturbance was detected simultaneously with the initial increase in antibody titres, about 1 week after the second injection.