Modulation of Cerebral Function by Muscle Afferent Activity, with Reference to Intravenous Succinylcholine.

Cerebral Function and Muscle Afferent Activity Following Intravenous Succinylcholine in Dogs Anesthetized with Halothane: The Effects of Pretreatment with a Defasciculating Dose of Pancuronium. By WL Lanier, PA Iaizzo, and JH Milde. Anesthesiology 1989; 71:87-95. Reprinted with permission. By the mid-1980s, it was widely assumed that if the depolarizing muscle relaxant, succinylcholine, given IV, produced increases in intracranial pressure, it did so because fasciculations produced increases in intrathoracic and central venous pressures that were transferred to the brain; however, there was no direct evidence that this was true. In contrast, we explored the possibility that the succinylcholine effect on the brain was explained by the afferentation theory of cerebral arousal, which predicts that agents or maneuvers that stimulate muscle stretch receptors will tend to stimulate the brain. Our research in tracheally intubated, lightly anesthetized dogs discovered that IV succinylcholine (which does not cross the blood-brain barrier) produced a doubling of cerebral blood flow that lasted for 30 min and corresponded to activation of the electroencephalogram and increases in intracranial pressure. Later, in our Classic Paper, we were able to assess simultaneously cerebral physiology and afferent nerve traffic emanating from muscle stretch receptors (primarily muscle spindles). We affirmed that the cerebral arousal response to succinylcholine was indeed driven by muscle afferent traffic and was independent of fasciculations or increases in intrathoracic or central venous pressures. Later research in complementary models demonstrated that endogenous movement (e.g., coughing, hiccups) produced a cerebral response very similar to IV succinylcholine, apparently as a result of the same muscle afferent mechanisms, independent of intrathoracic and central venous pressures. Thus, the importance of afferentation theory as a driver of the cerebral state of arousal and cerebral physiology during anesthesia was affirmed.

Neuromuscular magnetic stimulation counteracts muscle decline in ALS patients: results of a randomized, double-blind, controlled study.

The aim of the study was to verify whether neuromuscular magnetic stimulation (NMMS) improves muscle function in spinal-onset amyotrophic lateral sclerosis (ALS) patients. Twenty-two ALS patients were randomized in two groups to receive, daily for two weeks, NMMS in right or left arm (referred to as real-NMMS, rNMMS), and sham NMMS (sNMMS) in the opposite arm. All the patients underwent a median nerve conduction (compound muscle action potential, CMAP) study and a clinical examination that included a handgrip strength test and an evaluation of upper limb muscle strength by means of the Medical Research Council Muscle Scale (MRC). Muscle biopsy was then performed bilaterally on the flexor carpi radialis muscle to monitor morpho-functional parameters and molecular changes. Patients and physicians who performed examinations were blinded to the side of real intervention. The primary outcome was the change in the muscle strength in upper arms. The secondary outcomes were the change from baseline in the CMAP amplitudes, in the nicotinic ACh currents, in the expression levels of a selected panel of genes involved in muscle growth and atrophy, and in histomorphometric parameters of ALS muscle fibers. The Repeated Measures (RM) ANOVA with a Greenhouse-Geisser correction (sphericity not assumed) showed a significant effect [F(3, 63) = 5.907, p < 0.01] of rNMMS on MRC scale at the flexor carpi radialis muscle, thus demonstrating that the rNMMS significantly improves muscle strength in flexor muscles in the forearm. Secondary outcomes showed that the improvement observed in rNMMS-treated muscles was associated to counteracting muscle atrophy, down-modulating the proteolysis, and increasing the efficacy of nicotinic ACh receptors (AChRs). We did not observe any significant difference in pre- and post-stimulation CMAP amplitudes, evoked by median nerve stimulation. This suggests that the improvement in muscle strength observed in the stimulated arm is unlikely related to reinnervation. The real and sham treatments were well tolerated without evident side effects. Although promising, this is a proof of concept study, without an immediate clinical translation, that requires further clinical validation.

Neuroanatomical characteristics of deep and superficial needling using LI11 as an example.

OBJECTIVES: To compare the neuroanatomical characteristics of the deep and superficial tissues at acupuncture point LI11 using a neural tracing technique, in order to examine the neural basis of potential differences between deep and superficial needling techniques. METHODS: In order to mimic the situations of the deep and superficial needling, the retrograde neural tracer Alexa Fluor 488 conjugate of cholera toxin subunit B (AF488-CTB) was injected into the muscle or subcutaneous tissue, respectively, at acupuncture point LI11 in eight rats (n=4 each). Three days following injection, the distribution of motor and sensory neurons labelled with AF488-CTB was examined in the spinal cord and dorsal root ganglia (DRG) under a fluorescent microscope. RESULTS: For both types of injection, labelled motor and sensory neurons were distributed on the side ipsilateral to the injection in the spinal cord and DRG between spinal levels C5 and T1. The number of labelled motor neurons following intramuscular injection was significantly higher than subcutaneous injection. By contrast, the number of labelled sensory neurons following subcutaneous injection was significantly higher in number and extended over a greater number of spinal segments compared to intramuscular injection. CONCLUSIONS: These data indicate that the motor and sensory innervation of muscle and subcutaneous tissue beneath LI11 differ, and suggest that acupuncture signals induced by deep and superficial needling stimulation may be transmitted through different neural pathways.

GABA in the nervous system of the cestodes Diphyllobothrium dendriticum (Diphyllobothriidea) and Caryophyllaeus laticeps (Caryophyllidea), with comparative analysis of muscle innervation.

The nervous system (NS) of the cestodes Diphyllobothrium dendriticum (Diphyllobothriidea) and Caryophyllaeus laticeps (Caryophyllidea) was investigated using immunocytochemistry. The GABA neurotransmitter was identified in the NS of both species; GABAergic neurons were detected in the main nerve cords (MC). GABA-like immunoreactive neurons were predominantly unipolar and exhibited more intensive immunoreactivity in the neurite than in the perikaryon. In C. laticeps , GABA-like immunoreactive somas are located in both the MCs and peripheral NS near the longitudinal muscles. The innervation of the body musculature was studied using a combination of antibodies against GABA, serotonin (5-HT), and FMRFamide and with complementary staining of F-actin. In both species, the location of GABAergic neurites is associated with all muscle layers including the subtegumental, longitudinal, transverse, and dorsoventral muscles. The cytomorphology of 5-HTergic motoneurons in the MCs of both species is described and differences in muscle innervation between D. dendriticum and C. laticeps are demonstrated. No evidence for co-localization of GABA with 5-HT or FMRFamide neurotransmitters at any particular neuron was found. Neuropiles in MCs and peripheral NS had separate sets of immunoreactive neurites. A functional role for GABA in muscle innervation is discussed.

Acute electromyostimulation decreases muscle sympathetic nerve activity in patients with advanced chronic heart failure (EMSICA Study).

BACKGROUND: Muscle passive contraction of lower limb by neuromuscular electrostimulation (NMES) is frequently used in chronic heart failure (CHF) patients but no data are available concerning its action on sympathetic activity. However, Transcutaneous Electrical Nerve Stimulation (TENS) is able to improve baroreflex in CHF. The primary aim of the present study was to investigate the acute effect of TENS and NMES compared to Sham stimulation on sympathetic overactivity as assessed by Muscle Sympathetic Nerve Activity (MSNA). METHODS: We performed a serie of two parallel, randomized, double blinded and sham controlled protocols in twenty-two CHF patients in New York Heart Association (NYHA) Class III. Half of them performed stimulation by TENS, and the others tested NMES. RESULTS: Compare to Sham stimulation, both TENS and NMES are able to reduce MSNA (63.5 ± 3.5 vs 69.7 ± 3.1 bursts / min, p < 0.01 after TENS and 51.6 ± 3.3 vs 56.7 ± 3.3 bursts / min, p < 0, 01 after NMES). No variation of blood pressure, heart rate or respiratory parameters was observed after stimulation. CONCLUSION: The results suggest that sensory stimulation of lower limbs by electrical device, either TENS or NMES, could inhibit sympathetic outflow directed to legs in CHF patients. These properties could benefits CHF patients and pave the way for a new non-pharmacological approach of CHF.

Effects of acute and long-term slow breathing exercise on muscle sympathetic nerve activity in untreated male patients with hypertension.

OBJECTIVE: Acute slow breathing (SLOWB) affects sympathetic cardiovascular regulation, but its long-term effects are unknown. Using device-guided breathing we explored short-term and long-term SLOWB effects on blood pressure (BP), heart rate (HR) and muscle sympathetic nerve activity (MSNA) in essential hypertension. METHODS: We measured BP, HR and MSNA in 10 hypertensive individuals at rest, during laboratory stressors, before and after acute SLOWB, and 8 weeks after SLOWB exercise. Twelve matched hypertensive controls underwent a similar protocol excluding SLOWB intervention. Office and 24-h BP were obtained at baseline and at follow-up. RESULTS: Acute SLOWB had no influence on BP, HR, but decreased MSNA (P < 0.01). BP, HR, MSNA responses to handgrip were comparable before and after acute SLOWB. Acute SLOWB tended to reduce SBP (P = 0.09), HR (P = 0.08), but not MSNA (P = 0.20) responses to mental stress. Long-term SLOWB decreased office SBP (P < 0.001), DBP (P < 0.01), HR (P = 0.004), but not 24-h BP. Resting MSNA was unchanged after long-term SLOWB (P = 0.68). Long-term SLOWB did not influence BP, HR or MSNA responses to handgrip and cold pressor, but reduced SBP (P = 0.03), HR (P = 0.03) responses to mental stress without MSNA changes. In controls BP, HR, MSNA responses to laboratory stressors remained unchanged at baseline and at follow-up. CONCLUSION: In essential hypertension, MSNA is reduced during acute SLOWB, but remains unaltered following long-term SLOWB. Long-term SLOWB reduces office, but not ambulatory BP and HR. SLOWB attenuates cardiovascular response to mental stress, but not physical stressors. These findings may be indicative of beneficial SLOWB effects on stress reduction in essential hypertension.

Short-term synaptic plasticity compensates for variability in number of motor neurons at a neuromuscular junction.

We studied how similar postsynaptic responses are maintained in the face of interindividual variability in the number of presynaptic neurons. In the stomatogastric ganglion of the lobster, Homarus americanus, the pyloric (PY) neurons exist in variable numbers across animals. We show that each individual fiber of the stomach muscles innervated by PY neurons received synaptic input from all neurons present. We performed intracellular recordings of excitatory junction potentials (EJPs) in the muscle fibers to determine the consequences of differences in the number of motor neurons. Despite the variability in neuron number, the compound electrical response of muscle fibers to natural bursting input was similar across individuals. The similarity of total synaptic activation was not due to differences in the spiking activity of individual motor neurons across animals with different numbers of PY neurons. The amplitude of a unitary EJP in response to a single spike in a single motor neuron also did not depend on the number of PY neurons present. Consequently, the compound EJP in response to a single stimulus that activated all motor axons present was larger in individuals with more PY neurons. However, when axons were stimulated with trains of pulses mimicking bursting activity, EJPs facilitated more in individuals with fewer PY neurons. After a few stimuli, this resulted in depolarizations similar to the ones in individuals with more PY neurons. We interpret our findings as evidence that compensatory or homeostatic regulatory mechanisms can act on short-term synaptic dynamics instead of absolute synaptic strength.

CGRPα-expressing sensory neurons respond to stimuli that evoke sensations of pain and itch.

Calcitonin gene-related peptide (CGRPα, encoded by Calca) is a classic marker of nociceptive dorsal root ganglia (DRG) neurons. Despite years of research, it is unclear what stimuli these neurons detect in vitro or in vivo. To facilitate functional studies of these neurons, we genetically targeted an axonal tracer (farnesylated enhanced green fluorescent protein; GFP) and a LoxP-stopped cell ablation construct (human diphtheria toxin receptor; DTR) to the Calca locus. In culture, 10-50% (depending on ligand) of all CGRPα-GFP-positive (+) neurons responded to capsaicin, mustard oil, menthol, acidic pH, ATP, and pruritogens (histamine and chloroquine), suggesting a role for peptidergic neurons in detecting noxious stimuli and itch. In contrast, few (2.2±1.3%) CGRPα-GFP(+) neurons responded to the TRPM8-selective cooling agent icilin. In adult mice, CGRPα-GFP(+) cell bodies were located in the DRG, spinal cord (motor neurons and dorsal horn neurons), brain and thyroid-reproducibly marking all cell types known to express Calca. Half of all CGRPα-GFP(+) DRG neurons expressed TRPV1, ∼25% expressed neurofilament-200, <10% contained nonpeptidergic markers (IB4 and Prostatic acid phosphatase) and almost none (<1%) expressed TRPM8. CGRPα-GFP(+) neurons innervated the dorsal spinal cord and innervated cutaneous and visceral tissues. This included nerve endings in the epidermis and on guard hairs. Our study provides direct evidence that CGRPα(+) DRG neurons respond to agonists that evoke pain and itch and constitute a sensory circuit that is largely distinct from nonpeptidergic circuits and TRPM8(+)/cool temperature circuits. In future studies, it should be possible to conditionally ablate CGRPα-expressing neurons to evaluate sensory and non-sensory functions for these neurons.

High dose ascorbic acid does not reverse central sympathetic overactivity in chronic heart failure.

WHAT IS KNOWN AND OBJECTIVE: The increased central sympathetic activity typically associated with chronic heart failure (CHF) is probably mediated by formation of reactive oxygen species (ROS) in the brain. Our objective was to undertake a trial to test our hypothesis that administration of the well-known antioxidant and ROS scavenger ascorbic acid, would reverse or reduce the sympathetic overactivity in CHF patients. METHODS: In a prospective, randomized, placebo-controlled, double-blind, cross-over trial, 11 CHF patients were treated with ascorbic acid 2 g/day or placebo for 3 days. At the end of each treatment period, sympathetic nervous system activity was measured by microneurography for direct muscle sympathetic nerve activity (MSNA) recording, analysis of heart rate variability (HRV) and measurement of plasma norepinephrine concentrations. RESULTS: During ascorbic acid administration, plasma vitamin C levels were higher than during placebo (74·9 ± 6·0 µmol/L vs. 54·8 ± 4·6 µmol/L, P = 0·03). Ascorbic acid had no effect on sympathetic activity: MSNA (ascorbic acid: 66·8 ± 3·3 vs. placebo 66·9 ± 3·2 bursts/100 beats, P = 0·98). In addition, HRV and plasma norepinephrine levels did not differ. WHAT IS NEW AND CONCLUSION: Short-term administration of the antioxidant ascorbic acid in CHF patients does not reverse the increased sympathetic activity as measured by microneurography, HRV and plasma norepinephrine levels. The use of higher oral dosages seems not feasible due to accompanying side effects.

Misdirection of regenerating axons and functional recovery following sciatic nerve injury in rats.

Poor functional recovery found after peripheral nerve injury has been attributed to the misdirection of regenerating axons to reinnervate functionally inappropriate muscles. We applied brief electrical stimulation (ES) to the common fibular (CF) but not the tibial (Tib) nerve just prior to transection and repair of the entire rat sciatic nerve, to attempt to influence the misdirection of its regenerating axons. The specificity with which regenerating axons reinnervated appropriate targets was evaluated physiologically using compound muscle action potentials (M responses) evoked from stimulation of the two nerve branches above the injury site. Functional recovery was assayed using the timing of electromyography (EMG) activity recorded from the tibialis anterior (TA) and soleus (Sol) muscles during treadmill locomotion and kinematic analysis of hindlimb locomotor movements. Selective ES of the CF nerve resulted in restored M-responses at earlier times than in unstimulated controls in both TA and Sol muscles. Stimulated CF axons reinnervated inappropriate targets to a greater extent than unstimulated Tib axons. During locomotion, functional antagonist muscles, TA and Sol, were coactivated both in stimulated rats and in unstimulated but injured rats. Hindlimb kinematics in stimulated rats were comparable to untreated rats, but significantly different from intact controls. Selective ES promotes enhanced axon regeneration but does so with decreased fidelity of muscle reinnervation. Functional recovery is neither improved nor degraded, suggesting that compensatory changes in the outputs of the spinal circuits driving locomotion may occur irrespective of the extent of misdirection of regenerating axons in the periphery.

Brain-computer interface research comes of age: traditional assumptions meet emerging realities.

Brain-computer interfaces (BCIs) could provide important new communication and control options for people with severe motor disabilities. Most BCI research to date has been based on 4 assumptions that: (a) intended actions are fully represented in the cerebral cortex; (b) neuronal action potentials can provide the best picture of an intended action; (c) the best BCI is one that records action potentials and decodes them; and (d) ongoing mutual adaptation by the BCI user and the BCI system is not very important. In reality, none of these assumptions is presently defensible. Intended actions are the products of many areas, from the cortex to the spinal cord, and the contributions of each area change continually as the CNS adapts to optimize performance. BCIs must track and guide these adaptations if they are to achieve and maintain good performance. Furthermore, it is not yet clear which category of brain signals will prove most effective for BCI applications. In human studies to date, low-resolution electroencephalography-based BCIs perform as well as high-resolution cortical neuron-based BCIs. In sum, BCIs allow their users to develop new skills in which the users control brain signals rather than muscles. Thus, the central task of BCI research is to determine which brain signals users can best control, to maximize that control, and to translate it accurately and reliably into actions that accomplish the users' intentions.

Neural correlates of skill acquisition with a cortical brain-machine interface.

Research into the development of brain-machine interfaces (BMIs) has led to demonstrations of rodents, nonhuman primates, and humans controlling prosthetic devices in real time through modulation of neural signals. In particular, cortical BMI studies have shown that improvements in performance require learning and are associated with changes in neuronal tuning properties. These studies have further shown evidence of long-term improvements in performance with practice. The authors conducted experiments to understand long-term skill acquisition with BMIs and to characterize the neural correlates of improvements in task performance. They specifically assessed long-term acquisition of neuroprosthetic skill (i.e., accurate task performance readily recalled across days). In 2 monkeys performing a center-out task using a brain-controlled (BC) computer cursor, they closely monitored daily performance trends and the neural correlates under different conditions. Importantly, they assessed BC performance using a continuous-control multistep task. The authors first conducted experiments that mimicked experimental conditions commonly used. Specifically, a large set of neurons was incorporated with daily recalibration of the transform of neural activity to BC. Under such conditions, they found evidence of variable daily performance. In contrast, when a fixed transform was applied to stable recordings from an ensemble of neurons across days, there was consistent evidence of long-term skill acquisition. Such skill acquisition was associated with the crystallization of a cortical map for prosthetic control. Taken together, the results suggest that the primate motor cortex can achieve skilled control of a neuroprosthetic device through consolidation of a cortical representation.

Modularity for sensorimotor control: evidence and a new prediction.

By combining a few modules, the CNS may learn new control policies quickly and efficiently. Support for a modular organization of the motor system has recently come from the observation of low dimensionality in the motor commands. However, stronger evidence would come from testing the predictions on the effect of an intervention on the mechanisms required to implement a modular controller. Thus, the authors propose to test the predictions of modularity on motor adaptation. They argue that unlike a nonmodular controller, a modular controller must adapt faster to a perturbation that is compatible with the modules (i.e., one that can be compensated by reusing the same modules), than to an incompatible perturbation (i.e., one that requires new modules).

Why professional athletes need a prolonged period of warm-up and other peculiarities of human motor learning.

Professional athletes involved in sports that require the execution of fine motor skills must practice for a considerable length of time before competing in an event. Why is such practice necessary? Is it merely to warm-up the muscles, tendons, and ligaments, or does the athlete's sensorimotor network need to be constantly recalibrated? In this article, the authors present a point of view in which the human sensorimotor system is characterized by: (a) a high noise level and (b) a high learning rate at the synaptic level (which, because of the noise, does not equate to a high learning rate at the behavioral level). They argue that many heuristics of human skill learning, including the need for a prolonged period of warm-up in experts, follow from these assumptions.

Anatomical study of the fibularis longus muscle motor points and electrical stimulation therapy application / Estudio anatómico de los puntos motores del músculo fibularis longus y su aplicación en la terapia de electroestimulación

The fibularis longus muscle (FLM) has an important role in the movement of eversion of the foot and in maintaining the plantar arch. The electrostimulation procedures seek to maintain muscle trophism, increase strength and endurance, and are frequently used in physiotherapy, for which the clinician needs to know the location of the motor points of the FLM. Therefore, the purpose of this study was to determine the number and distribution of motor points of the FLM and relate them to observable parameters in the surface anatomy. Ten formalin-preserved limbs were used, and the lateral regions of the leg were dissected in detail. In all the cases, the muscle presented three fascicular patterns, the superior and anteroinferior fascicles presented two motor points each, while the posteroinferior fascicles were between 2 and 3 motor points. Our results suggest that there is a pattern of distribution of the superficial fibular nerve, whose knowledge is useful for clinical application in the FLM electrostimulation proceedings.

El músculo fibular largo (MFL) tiene una importante función en el movimiento de eversión del pié y en la mantención del arco plantar. Los procedimientos de electroestimulación buscan mantener el trofismo muscular, aumentar la potencia y resistencia y es frecuente su utilización en fisioterapia, para ello el clínico necesita conocer la localización de los puntos motores del MFL, por ello, el propósito de este estudio fue determinar el número y distribución de los puntos motores del MFL y relacionarlos con parámetros observables en la anatomía de superficie. Se utilizaron 10 miembros inferiores conservados y se disecó detalladamente la región lateral de la pierna. El músculo presentó en todos los casos una estructura trifascicular, los fascículos superiores y anteroinferiores presentaron dos puntos motores cada uno, mientras en el fascículo posteroinferior encontramos entre 2 y 3 puntos motores. Nuestros resultados sugieren que existe un patrón de distribución del nervio fibular superficial cuyo conocimiento es de utilidad clínica para los procedimientos de electroestimulación del MFL.

Algesic agents exciting muscle nociceptors.

Morphologically, muscle nociceptors are free nerve endings connected to the CNS by thin myelinated (group III) or unmyelinated (group IV) afferent fibers. Not all of these endings are nociceptive; approximately 40% have a low mechanical threshold and likely fulfill non-nociceptive functions. Two chemical stimuli are particularly relevant as causes of muscle pain. The first is a drop in tissue pH, i.e. an increase in proton (H+) concentration. A large number of painful patho(physio)logical alterations of muscle tissue are associated with an acidic interstitial pH (e.g. tonic contractions, spasm, inflammation). The second important cause of muscle pain is a release of adenosine triphosphate (ATP). ATP is present in all body cells, but in muscle its concentration is particularly high. Any damage of muscle cells (trauma, necrotic myositis) is accompanied by a release of ATP from the cells. Therefore, ATP is considered a general pain stimulus by some. ATP and protons are relatively specific stimuli for muscle pain; in cutaneous pain they play a less important role. The numerous agents that are released in pathologically altered muscle include substances that desensitize mechanosensitive group IV receptors. Capsaicin has a long-lasting desensitizing action, brain-derived neurotrophic factor, and tumor necrosis factor-alpha, a short-lasting one. Most of the agents exciting group IV units (e.g. low pH, ATP, capsaicin) activate not only nociceptive endings but also non-nociceptive ones. The only substance encountered that excites exclusively nociceptive group IV receptors is nerve growth factor (NGF). In rat muscle chronically inflamed with complete Freund's adjuvant, most group IV endings are sensitized to mechanical (and to some) chemical stimuli. However, stimulants such as ATP, NGF, and solutions of low pH were found to be less effective in inflamed muscle. A possible explanation for this surprising finding is that in inflamed muscle the concentrations of ATP and NGF and H+ are increased. Therefore, experimental administration of these agents is a less effective stimulus.

Nerves in the endodermal canals of hydromedusae and their role in swimming inhibition.

N eoturris breviconis (Anthomedusae) has a nerve plexus in the walls of its endodermal canals. The plexus is distinct from the ectodermal nerve plexuses supplying the radial and circular muscles in the ectoderm and no connections have been observed between them. Stimulation of the endodermal plexus evokes electrical events recorded extracellularly as "E" potentials. These propagate through all areas where the plexus has been shown by immunohistology to exist and nowhere else. When Neoturris is ingesting food, trains of "E" potentials propagate down the radial canals to the margin and cause inhibition of swimming. This response is distinct from the inhibition of swimming associated with contractions of the radial muscles but both may play a part in feeding and involve chemoreceptors. Preliminary observations suggest that the "E" system occurs in other medusae including Aglantha digitale (Trachymedusae) where the conduction pathway was previously thought to be an excitable epithelium.

Bone marrow-derived mesenchymal stem cell transplantation does not improve quality of muscle reinnervation or recovery of motor function after facial nerve transection in rats.

Recently, we devised and validated a novel strategy in rats to improve the outcome of facial nerve reconstruction by daily manual stimulation of the target muscles. The treatment resulted in full recovery of facial movements (whisking), which was achieved by reducing the proportion of pathologically polyinnervated motor endplates. Here, we posed whether manual stimulation could also be beneficial after a surgical procedure potentially useful for treatment of large peripheral nerve defects, i.e., entubulation of the transected facial nerve in a conduit filled with suspension of isogeneic bone marrow-derived mesenchymal stem cells (BM-MSCs) in collagen. Compared to control treatment with collagen only, entubulation with BM-MSCs failed to decrease the extent of collateral axonal branching at the lesion site and did not improve functional recovery. Post-operative manual stimulation of vibrissal muscles also failed to promote a better recovery following entubulation with BM-MSCs. We suggest that BM-MSCs promote excessive trophic support for regenerating axons which, in turn, results in excessive collateral branching at the lesion site and extensive polyinnervation of the motor endplates. Furthermore, such deleterious effects cannot be overridden by manual stimulation. We conclude that entubulation with BM-MSCs is not beneficial for facial nerve repair.