Anatomical analysis of the motor endplate zones of the suprascapular nerve to the infraspinatus muscle and its clinical significance in managing pain disorder.

Myofascial pain syndrome caused by myofascial trigger points is a musculoskeletal disorder commonly encountered in clinical practice. The infraspinatus muscle is the region most frequently involved in the myofascial pain syndrome in the scapular region. The characteristics of the myofascial trigger points are that they can be found constantly in the motor endplate zone. However, localizing myofascial trigger points within the motor endplate zone and establishing an accurate injection site of the infraspinatus muscle has been challenging because the anatomical position of the motor endplate zone of the infraspinatus muscle is yet to be described. Therefore, this cadaveric study aimed to scrutinize the motor endplate zone of the infraspinatus muscle, propose potential myofascial trigger points within the muscle, and recommend therapeutic injection sites. Twenty specimens of the infraspinatus muscle for nerve staining and 10 fresh frozen cadavers for evaluation of the injection were used in this study. The number of nerve branches penetrating the infraspinatus muscle and their entry locations were analyzed and photographed. Modified Sihler's staining was performed to examine the motor endplate regions of the infraspinatus muscle. The nerve entry points were mostly observed in the center of the muscle belly. The motor endplate was distributed equally throughout the infraspinatus muscle, but the motor endplate zone was primarily identified in the B area, which is approximately 20-40% proximal to the infraspinatus muscle. The second-most common occurrence of the motor endplate zone was observed in the center of the muscle. These detailed anatomical data would be very helpful in predicting potential pain sites and establishing safe and effective injection treatment using botulinum neurotoxin, steroids, or lidocaine to alleviate the pain disorder of the infraspinatus muscle.

Ultra-Low Frequency Transcutaneous Electrical Nerve Stimulation on Pain Modulation in a Rat Model with Myogenous Temporomandibular Dysfunction.

Masticatory myofascial pain (MMP) is one of the most common causes of chronic orofacial pain in patients with temporomandibular disorders. To explore the antinociceptive effects of ultra-low frequency transcutaneous electrical nerve stimulation (ULF-TENS) on alterations of pain-related biochemicals, electrophysiology and jaw-opening movement in an animal model with MMP, a total of 40 rats were randomly and equally assigned to four groups; i.e., animals with MMP receiving either ULF-TENS or sham treatment, as well as those with sham-MMP receiving either ULF-TENS or sham treatment. MMP was induced by electrically stimulated repetitive tetanic contraction of masticatory muscle for 14 days. ULF-TENS was then performed at myofascial trigger points of masticatory muscles for seven days. Measurable outcomes included maximum jaw-opening distance, prevalence of endplate noise (EPN), and immunohistochemistry for substance P (SP) and µ-opiate receptors (MOR) in parabrachial nucleus and c-Fos in rostral ventromedial medulla. There were significant improvements in maximum jaw-opening distance and EPN prevalence after ULF-TENS in animals with MMP. ULF-TENS also significantly reduced SP overexpression, increased MOR expression in parabrachial nucleus, and increased c-Fos expression in rostral ventromedial medulla. ULF-TENS may represent a novel and applicable therapeutic approach for improvement of orofacial pain induced by MMP.

Herbal complex 'Buyang Huanwu Tang' improves motor endplate function of denervated-dependent skeletal muscle atrophy in rat.

Denervated-dependent skeletal muscle atrophy is a disease induced by skeletal muscle associated peripheral neuro-disconnection. Its specific molecular mechanisms remain unknown. The treating for denervated-dependent skeletal muscle atrophy is applied with an herbal complex Buyang Huanwu Tang used in traditional Chinese medicine and subjected to the established denervated-dependent skeletal muscle atrophy in rat models, and the therapeutic effects and associated mechanisms were evaluated in the pathogenesis of denervated-dependent skeletal muscle atrophy. Denervated-dependent skeletal muscle atrophy in rats was established and randomly divided into eight groups, including Normal control, Model, Positive control, Model + Buyang Huanwu Tang, Model + astragalus extracts, Model + Buyang Huanwu Tang-astragalus, Buyang Huanwu Tang + LY294002, and astragalus extract + LY294002 group. Hematoxylin-eosin staining and quantitative RT-PCR (qRT-PCR) assay were used to examine the inflammatory response of muscle tissues. Quantitative RT-PCR and Western blotting assay were utilized to analyze mRNA and protein expression. Immunohistochemistry assay was used to detect molecule expression in anterior cervical muscle tissues. Motor endplate activity was examined using the wholemount acetylcholinesterase staining method. The wet mass ratio of anterior cervical muscle was measured. The results indicated that Buyang Huanwu Tang treatment significantly alleviated inflammatory response, enhanced acetylcholinesterase activity, and motor endplate functions, and promoted wet mass of anterior cervical muscle compared to denervated-dependent skeletal muscle atrophy rat models (P < 0.05). Buyang Huanwu Tang regulated molecules of PI3K/PKB/GSK3ß/FOXO1 signaling pathway. Buyang Huanwu Tang significantly reduced muscle atrophy F-box protein, MuFR-1, Bax and caspase 9 expression, significantly enhanced Bcl-2 expression, and remarkably increased element-binding protein and vascular endothelial growth factor levels, compared to Model group (P < 0.05). Buyang Huanwu Tang suppressed caspase 9 and caspase 3 activity and associated apoptosis. Moreover, PI3K specific blocker, LY294002, significantly inhibited the effects of Buyang Huanwu Tang on the above molecule expression (P < 0.05). In conclusion, Buyang Huanwu Tang improved motor endplate functions of denervated-dependent skeletal muscle atrophy rat model through suppressing mitochondria-mediated apoptosis and activating PI3K/PKB/FOXO1 signaling pathway.

Remote dose-dependent effects of dry needling at distant myofascial trigger spots of rabbit skeletal muscles on reduction of substance P levels of proximal muscle and spinal cords.

BACKGROUND: Dry needling at distant myofascial trigger points is an effective pain management in patients with myofascial pain. However, the biochemical effects of remote dry needling are not well understood. This study evaluates the remote effects of dry needling with different dosages on the expressions of substance P (SP) in the proximal muscle, spinal dorsal horns of rabbits. METHODS: Male New Zealand rabbits (2.5-3.0 kg) received dry needling at myofascial trigger spots of a gastrocnemius (distant muscle) in one (1D) or five sessions (5D). Bilateral biceps femoris (proximal muscles) and superficial laminaes of L5-S2, T2-T5, and C2-C5 were sampled immediately and 5 days after dry needling to determine the levels of SP using immunohistochemistry and western blot. RESULTS: Immediately after dry needling for 1D and 5D, the expressions of SP were significantly decreased in ipsilateral biceps femoris and bilateral spinal superficial laminaes (P < .05). Five days after dry needling, these reduced immunoactivities of SP were found only in animals receiving 5D dry needling (P < .05). CONCLUSIONS: This remote effect of dry needling involves the reduction of SP levels in proximal muscle and spinal superficial laminaes, which may be closely associated with the control of myofascial pain.

Electrical stimulation modulates Wnt signaling and regulates genes for the motor endplate and calcium binding in muscle of rats with spinal cord transection.

BACKGROUND: Spinal cord injury (SCI) results in muscle atrophy and a shift of slow oxidative to fast glycolytic fibers. Electrical stimulation (ES) at least partially restores muscle mass and fiber type distribution. The objective of this study was to was to characterize the early molecular adaptations that occur in rat soleus muscle after initiating isometric resistance exercise by ES for one hour per day for 1, 3 or 7 days when ES was begun 16 weeks after SCI. Additionally, changes in mRNA levels after ES were compared with those induced in soleus at the same time points after gastrocnemius tenotomy (GA). RESULTS: ES increased expression of Hey1 and Pitx2 suggesting increased Notch and Wnt signaling, respectively, but did not normalize RCAN1.4, a measure of calcineurin/NFAT signaling, or PGC-1ß mRNA levels. ES increased PGC-1α expression but not that of slow myofibrillar genes. Microarray analysis showed that after ES, genes coding for calcium binding proteins and nicotinic acetylcholine receptors were increased, and the expression of genes involved in blood vessel formation and morphogenesis was altered. Of the 165 genes altered by ES only 16 were also differentially expressed after GA, of which 12 were altered in the same direction by ES and GA. In contrast to ES, GA induced expression of genes related to oxidative phosphorylation. CONCLUSIONS: Notch and Wnt signaling may be involved in ES-induced increases in the mass of paralyzed muscle. Molecular adaptations of paralyzed soleus to resistance exercise are delayed or defective compared to normally innervated muscle.

The effect of lateral electrical surface stimulation (LESS) on motor end-plates in an animal model of experimental scoliosis.

The treatment of idiopathic scoliosis is challenging because of its diverse etiology, age of onset, and long duration of intensive treatment. We examined the effect of lateral electrical surface stimulation (LESS) in an animal model of experimental scoliosis (ES) assessing the number of motor end-plates (MEPs) as a study end-point. The control group (n=5) was adapted to the experimental apparatus without stimulation, whereas ES was induced in rabbits by one-sided LESS of the longissimus dorsi muscle (LDM) for a duration of 2 months. The ES group (n=5) were subjected to a short-term corrective electrostimulation applied at the contralateral side of the spine compared to the previous LESS stimulation for 2 h daily for 3 (n=5) or 6 months (n=5). Another group of ES rabbits was subjected to a long-term corrective electrostimulation applied for 9 h daily for 3 (n=5) or 6 months (n=5). LESS applied for 2 months (ES), significantly increased the number of MEPs in LDM. The short-term corrective electrostimulation for 3 months resulted in an increased number of MEPs. However, a decrease was observed in the animals treated for 6 months. The long-term corrective electrostimulation for 3 months did not change the density of MEPs in the LDM, but for 6 months the number of MEPs in the LMD significantly decreased by ES and control groups. Thus, the results of the present study clearly show that the short-term LESS is able to influence both the number of MEPs and the effectiveness of muscle correctional adaptation in a more efficient and harmless manner than the long-term procedure.

Spinal cord mechanism involving the remote effects of dry needling on the irritability of myofascial trigger spots in rabbit skeletal muscle.

OBJECTIVE: To elucidate the neural mechanisms underlying the remote effects produced by dry needling rabbit skeletal muscle myofascial trigger spots (MTrSs) via analyses of their endplate noise (EPN) recordings. DESIGN: Experimental animal controlled trial. SETTING: An animal laboratory of a university. ANIMALS: Male New Zealand rabbits (N=96) (body weight, 2.5-3.0kg; age, 16-20wk). INTERVENTION: Animals received no intervention for neural interruption in group I, transection of the tibial nerve in group II, transection of L5 and L6 spinal cord in group III, and transection of the T1 and T2 spinal cord in group IV. Each group was further divided into 4 subgroups: animals received ipsilateral dry needling, contralateral dry needling, ipsilateral sham needling, or contralateral sham needling of gastrocnemius MTrSs. MAIN OUTCOME MEASURES: EPN amplitudes of biceps femoris (BF) MTrSs. RESULTS: BF MTrS mean EPN amplitudes significantly increased (P<.05) initially after gastrocnemius verum needling but reduced to a level significantly lower (P<.05) than the preneedling level in groups I and IV with ipsilateral dry needling or contralateral dry needling, and in group II with contralateral dry needling (but not ipsilateral dry needling). No significant EPN amplitude changes were observed in BF MTrS in group III or in the control animals receiving superficial needling (sham). CONCLUSION: This remote effect of dry needling depends on an intact afferent pathway from the stimulating site to the spinal cord and a normal spinal cord function at the levels corresponding to the innervation of the proximally affected muscle.

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.

The influence of lidocaine and racemic bupivacaine on neuromuscular blockade produced by rocuronium. A study in rat phrenic nerve-diaphragm preparation.

PURPOSE: To evaluate in vitro lidocaine and racemic bupivacaine effects in neuromuscular transmission and in neuromuscular blockade produced by rocuronium. METHODS: Rats were distributed in 5 groups (n = 5) in agreement with the studied drugs: lidocaine, racemic bupivacaine, rocuronium, separately (Groups I, II, III); rocuronium in preparations exposed to local anesthetics (Groups IV, V). The concentrations used were: 20 microg/mL, 5 microg/mL and 4 microg/mL, for lidocaine, bupivacaine and rocuronium, respectively. It was evaluated: 1) amplitude of diaphragm muscle response to indirect stimulation, before and 60 minutes after separately addition of lidocaine, racemic bupivacaine and rocuronium and the association of local anesthetics - rocuronium; 2) membrane potentials (MP) and miniature end-plate potentials (MEPP). RESULTS: Lidocaine and bupivacaine separately didn't alter the amplitude of muscle response and MP. In preparations previously exposed to lidocaine and racemic bupivacaine, the rocuronium blockade was significantly larger (90.10 +/- 9.15% and 100%, respectively), in relation to the produced by rocuronium separately (73.12 +/- 9.89%). Lidocaine caused an increase in the frequency of MEPP, being followed by blockade; racemic bupivacaine produced decrease being followed by blockade. CONCLUSIONS: Local anesthetics potentiated the blockade caused by rocuronium. The alterations of MEPP identify presynaptic action.

Remote influences of acupuncture on the pain intensity and the amplitude changes of endplate noise in the myofascial trigger point of the upper trapezius muscle.

OBJECTIVE: To investigate the remote effect of acupuncture on the pain intensity and the endplate noise (EPN) recorded from a myofascial trigger point (MTrP) of the upper trapezius muscle. DESIGN: Randomized controlled trial. SETTING: University hospital. PARTICIPANTS: Patients (N=20) with active MTrPs in upper trapezius muscles and no experience in acupuncture therapy. INTERVENTIONS: Patients were divided into 2 groups. Those in the control group received sham acupuncture, and those in the acupuncture group received modified acupuncture therapy with needle insertion into multiple loci to elicit local twitch responses. The acupuncture points of Wai-guan and Qu-chi were treated. MAIN OUTCOME MEASURES: Subjective pain intensity (numerical pain rating scale) and mean EPN amplitude in the MTrP of the upper trapezius muscle. RESULTS: The pain intensity in the MTrP was significantly reduced after remote acupuncture (from 7.4+/-0.8 to 3.3+/-1.1; P<.001), but not after sham acupuncture (from 7.4+/-0.8 to 7.1+/-0.9; P>.05). The mean EPN amplitude was significantly lower than the pretreatment level after acupuncture treatment (from 21.3+/-9.5 microV to 9.5+/-3.5 microV; P<.01), but not after sham acupuncture treatment (from 19.6+/-7.6 microV to 19.3+/-7.8 microV; P>.05). The change in the pain intensity was significantly correlated with the change of EPN amplitude (r=0.685). CONCLUSIONS: Both subjective changes in the pain intensity and objective changes of the EPN amplitude in the MTrP region of the upper trapezius muscle were found during and after acupuncture treatment at the remote ipsilateral acupuncture points. This study may further clarify the physiological basis of the remote effectiveness of acupuncture therapy for pain control.

The influence of lidocaine and racemic bupivacaine on neuromuscular blockade produced by rocuronium: a study in rat phrenic nerve-diaphragm preparation / Influência da lidocaína e da bupivacaína racêmica no bloqueio neuromuscular produzido pelo rocurônio: estudo em preparação nervo frênico-diafragma de rato

PURPOSE: To evaluate in vitro lidocaine and racemic bupivacaine effects in neuromuscular transmission and in neuromuscular blockade produced by rocuronium. METHODS: Rats were distributed in 5 groups (n = 5) in agreement with the studied drugs: lidocaine, racemic bupivacaine, rocuronium, separately (Groups I, II, III); rocuronium in preparations exposed to local anesthetics (Groups IV, V). The concentrations used were: 20 µg/mL, 5 µg/mL and 4 µg/mL, for lidocaine, bupivacaine and rocuronium, respectively. It was evaluated: 1) amplitude of diaphragm muscle response to indirect stimulation, before and 60 minutes after separately addition of lidocaine, racemic bupivacaine and rocuronium and the association of local anesthetics - rocuronium; 2) membrane potentials (MP) and miniature end-plate potentials (MEPP). RESULTS: Lidocaine and bupivacaine separately didn't alter the amplitude of muscle response and MP. In preparations previously exposed to lidocaine and racemic bupivacaine, the rocuronium blockade was significantly larger (90.10 ± 9.15 percent and 100 percent, respectively), in relation to the produced by rocuronium separately (73.12 ± 9.89 percent). Lidocaine caused an increase in the frequency of MEPP, being followed by blockade; racemic bupivacaine produced decrease being followed by blockade. CONCLUSIONS: Local anesthetics potentiated the blockade caused by rocuronium. The alterations of MEPP identify presynaptic action.

OBJETIVO: Avaliar in vitro os efeitos da lidocaína e bupivacaína racêmica na transmissão neuromuscular e no bloqueio neuromuscular produzido pelo rocurônio. MÉTODOS: Ratos foram distribuídos em 5 grupos (n = 5) de acordo com a droga estudada: lidocaina, bupivacaína racêmica, rocurônio, isoladamente (Grupos I, II, III); rocurõnio em preparações expostas aos anestésicos locais (Grupos IV, V). As concentrações utilizadas foram: 20 µg/mL, 5 µg/mL e 4 µg/mL, para lidocaína, bupivacaína e rocurônio, respectivamente. Avaliou-se: 1) amplitude das respostas do músculo diafragma à estimulação indireta, antes e 60 minutos após a adição da lidocaína, bupivacaína racêmica e rocurônio isoladamente e da associação anestésicos locais - rocurônio; 2) potenciais de membrana (PM) e potenciais de placa terminal em miniatura (PPTM). RESULTADOS: A lidocaína e a bupivacaína isoladamente não alteraram a amplitude das respostas musculares e os PM. Nas preparações previamente expostas a lidocaína e a bupivacaína racêmica, o bloqueio com o rocurônio foi significativamente maior (90,10 ± 9,15 por cento e 100 por cento, respectivamente), em relação ao produzido pelo rocurônio isoladamente (73,12 ± 9,89 por cento). A lidocaína causou aumento na freqüência dos PPTM, seguido de bloqueio; a bupivacaína racêmica produziu diminuição seguida de bloqueio. CONCLUSÕES: Os anestésicos locais potencializaram o bloqueio causado pelo rocurônio. As alterações do PPTM identificam ação pré-sináptica.

The History of Myasthenia Gravis

Since Willis described 'fatigable weakness' in 1672, most physicians consider it as a kind of hysteria due to the inconsistent fluctuation of symptoms. Erb presented three cases of 'bulbal palsy' in the 1870s, and Oppenheim and Hopper considered myasthenia gravis as a disease similar to curare poisoning and as a disease induced by attack of the motor centers by intrinsic toxins, respectively. In 1903, Elliot suggested that a 'chemical substance' mediates the nerve impulses at synapse. However, it was not until 1921 that this was demonstrated by Loewi, who provided evidence from the famous two-frog-hearts experiment. Dale later revealed the substance to be acetylcholine, and he also suggested that myasthenia gravis is due to a problem with the motor end plate. In 1934, Walker was prompted by the resemblance between myasthenia gravis and curare poisoning to apply physostigmine, a curare-poisoning antidote, to a patient, which produced a dramatic result. Since then the use of anticholinesterase inhibitors has been adopted for standard therapeutic modality. Some prominent surgeons have also applied thymectomy as a surgical modality. The most recent focus of myasthenia gravis has been immunological. In 1960, Simpson proposed the autoimmune hypothesis, and Chang et al. showed that snake venom contained a selective antagonist of the nicotinic acetylcholine receptor, alpha-bungarotoxin. The immunization of rabbits with acetylcholine receptor purified from the electrical organs of electric eels by Patrick et al. induced myasthenic symptoms and signs, and these were reversed by acetylcholinesterase inhibitors. The role of the autoimmune system has led to the introduction of an immunosuppressive modality and plasma exchange to the field of clinical neurology.

Function of neuromuscular synapses in the zebrafish choline-acetyltransferase mutant bajan.

We have identified a zebrafish mutant line, bajan, in which compromised motility and fatigue result from a point mutation in the gene coding choline acetyltransferase (ChAT), the enzyme responsible for acetylcholine (ACh) synthesis. Although the mutation predicts loss of ChAT function, bajan inexplicably retains low levels of neuromuscular transmission. We exploited this residual activity and determined the consequences for synaptic function. The attenuated synaptic responses were a direct consequence of a decrease in both resting mean quantal size and quantal content. To replicate behavioral fatigue in swimming, motorneurons were stimulated at high frequencies. A prominent reduction in quantal content, reflecting vesicle depletion, was coincident with a small additional reduction in quantal size. In humans, defective ChAT leads to episodic apnea, a form of congenital myasthenic syndrome characterized by use-dependent fatigue. In contrast to bajan, however, afflicted individuals exhibit a normal resting quantal size and quantal content. The fatigue in humans results from a pronounced long-lasting drop in quantal size with little or no change in quantal content. These differences have important implications for interpreting fatigue as well as on understanding the impact of ACh availability on vesicle filling and recycling.

Electrophysiologic effects of a therapeutic laser on myofascial trigger spots of rabbit skeletal muscles.

OBJECTIVE: To better understand the mechanisms of therapeutic lasers for treating human myofascial trigger points, we designed a blinded controlled study of the effects of a therapeutic laser on the prevalence of endplate noise (EPN) recorded from the myofascial trigger spot (MTrS) of rabbit skeletal muscle. DESIGN: In eight rabbits, one MTrS in each biceps femoris muscle was irradiated with a 660-nm, continuous-wave, gallium-aluminum-arsenate (GaAlAs) laser, at 9 J/cm2. The contralateral side of muscle was treated with a sham laser. Each rabbit received six treatments. The immediate and cumulative effects were assessed by the prevalence of EPN with electromyographic (EMG) recordings after the first and last treatments. RESULTS: Compared with pretreatment values, the percentages of EPN prevalence in the experimental side after the first and last treatments were significantly reduced (P < 0.01 for both). The change in EPN prevalence in the experimental side was significantly greater than in the control side immediately after the first and last treatments (P < 0.05). However, no significant differences were noted between the first and last treatments (P > 0.05). CONCLUSIONS: In our study, immediate and cumulative effects of a GaAlAs laser applied on MTrS were demonstrated on the basis of the assessment of EPN prevalence. It seems that laser irradiation may inhibit the irritability of an MTrS in rabbit skeletal muscle. This effect may be a possible mechanism for myofascial pain relief with laser therapy.

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.

Muscle-specific calpain is localized in regions near motor endplates in differentiating lobster claw muscles.

Calpains are Ca2+-dependent proteinases that mediate protein turnover in crustacean skeletal muscles. We used an antibody directed against lobster muscle-specific calpain (Ha-CalpM) to examine its distribution in differentiating juvenile lobster claw muscles. These muscles are comprised of both fast and slow fibers early in development, but become specialized into predominantly fast or exclusively slow muscles in adults. The transition into adult muscle types requires that myofibrillar proteins specific for fast or slow muscles to be selectively removed and replaced by the appropriate proteins. Using immunohistochemistry, we observed a distinct staining pattern where staining was preferentially localized in the fiber periphery along one side of the fiber. Immunolabeling with an antibody directed against synaptotagmin revealed that the calpain staining was greatest in the cytoplasm adjacent to synaptic terminals. In complementary analyses, we used sequence-specific primers with real-time PCR to quantify the levels of Ha-CalpM in whole juvenile claw muscles. These expression levels were not significantly different between cutter and crusher claws, but were positively correlated with the expression of fast myosin heavy chain. The anatomical localization of Ha-CalpM near motor endplates, coupled with the correlation with fast myofibrillar gene expression, suggests a role for this intracellular proteinase in fiber type switching.

Estudios neurofisiológicos de la unión neuromuscular / Neurophysiological studies of the neuromuscular junction

Introducción. El papel de los estudios neurofisiológicos (ENF) en el diagnóstico de las enfermedades de la transmisión neuromuscular (TNM) se fundamenta en el estudio del fallo de la fibra muscular para alcanzar una despolarización suficiente para que el potencial de placa alcance el umbral adecuado y conseguir un potencial de acción muscular. Este impulso bloqueado total o parcialmente originará los distintos tipos de respuestas en los tests neurofisiológicos. Objetivo. Analizar los distintos ENF aplicados como método diagnóstico en las enfermedades que alteran la TNM. Desarrollo. Se revisa el concepto de margen de seguridad de la placa motora y se describen las técnicas neurofisiológicas más utilizadas en la actualidad –estimulación repetitiva, electromiografía (EMG) convencional, de fibra aislada con activación voluntaria y con activación eléctrica axonal– y los hallazgos más frecuentes en las enfermedades de la TNM. Conclusiones. Los ENF servirán para confirmar o no el diagnóstico clínico, excluir otras enfermedades neuromusculares concomitantes, determinar si el proceso es pre o postsináptico, monitorizar el curso clínico de la enfermedad, tanto si es natural o en respuesta al tratamiento médico o quirúrgico, y permiten, además, determinar el estado de la TNM en los casos de remisión clínica, así como detectar trastornos subclínicos. Los estudios de EMG de fibra aislada son el diagnóstico neurofisiológico que muestra una mayor sensibilidad en el diagnóstico de estas enfermedades (AU)

Introduction. The role played by neurophysiological studies (NPS) in the diagnosis of diseases affecting neuromuscular transmission (NMT) is based on the study of the failure of muscle fibres to achieve a sufficient degree of depolarisation for the junction potential to reach the appropriate threshold and attain a muscular action potential. This totally or partially blocked impulse will give rise to different types of responses in neurophysiological tests. Aims. To analyse the different NPS as diagnostic methods in diseases that affect NMT. Development. The article offers a review of the concept of the safety margin at the neuromuscular junction and a description of the most common neurophysiological techniques currently in use –repetitive stimulation, as well as conventional or single fibre electromyography (EMG) with voluntary activation or axonal electrical activation–. The most frequent findings in diseases affecting NMT are also discussed. Conclusions. NPS will be useful to confirm or reject the clinical diagnosis, to exclude other concomitant neuromuscular diseases, to establish whether the process is pre- or post-synaptic, to monitor the clinical course of the disease (when it is both natural or in response to the medical or surgical treatment) and also to enable the physician to determine the status of NMT in cases of clinical remission, as well as to detect subclinical disorders. Single fibre EMG studies are the most sensitive method of neurophysiological diagnosis when dealing with these diseases (AU)

Early postdenervation depolarization develops faster at endplates of hibernating golden hamsters where spontaneous quantal and non-quantal acetylcholine release is very small.

The hyperpolarization produced by the application of curare to the postsynaptic membrane of the diaphragm neuromuscular synapse (H-effect) is a measure of non-quantal release (NQR) of acetylcholine (ACh) from the motor nerve ending. In mouse diaphragm, H-effect was 9.3 mV, significantly lower in awake hamsters (7.1 mV) and very small (1.1 mV) in hibernating hamsters. Also, the initial resting membrane potential (RMP) after dissection was highest in mouse (81.5 mV, inside negative), significantly smaller in awake hamsters (77.9 mV) and lowest in hibernating hamsters (75.1 mV). The early postdenervation depolarization of muscle fiber RMP to about 66-68 mV developed with half-decay time (T1/2) of 120 min in mouse, more rapidly in active hamsters (T1/2=60 min) and even faster in hibernating hamsters (T1/2=25 min) muscles. This reciprocal correlation between the H-effect and the rate of early depolarization indicates that non-quantal release is important for maintaining the resting membrane potential [Vyskocil, F. 2003. Early postdenervation depolarization is controlled by acetylcholine and glutamate via nitric oxide regulation of the chloride transporter. Neurochem. Res. 28, 575-585]. The amplitude of H-effect in mouse and hamster was proportional to the spontaneous quantal release. The frequency of miniature endplate potentials was highest in mouse (1.6 s-1), much smaller in awake hamsters (0.51 s-1) and very small in hibernating hamsters (0.08 s-1). This is in accordance with the idea that non-quantal release depends on the number of vesicles fused with the presynaptic membrane during quantal release [Edwards et al., 1985; Ferguson, S.M., Savchenko, V., Apparsundaram, S., Zwick, M., Wright J., Heilman, C.J., Yi, H., Levey, A.I., Blakely R.D. Vesicular localization and activity-dependent trafficking of presynaptic choline transporters. J. Neurosci. 23 (2003) 9697-9709].

Travell trigger points--molecular and osteopathic perspectives.

The proposed etiology of Travell trigger points (TrPs) has undergone a fundamental revision in recent years. New research results suggest that TrPs are evoked by the abnormal depolarization of motor end plates. This article expands the proposed etiology to include presynaptic, synaptic, and postsynaptic mechanisms of abnormal depolarization (ie, excessive release of acetycholine [ACh], defects of acetylcholinesterase, and upregulation of nicotinic ACh-receptor activity, respectively). This working hypothesis regarding the etiology of TrPs has changed the approach to treating TrPs. As an example, Travell and Simons abandoned the application of ischemic compression to TrPs; instead the authors adopted several techniques associated with osteopathic medicine (ie, muscle-energy, myofascial, counterstrain; high-velocity, low-amplitude). Scientists are now proposing and reporting the results of new approaches using capsaicin, a vanilloid-receptor agonist, and ACh antagonists (eg, dimethisoquin hydrochloride, botulinum toxin, quinidine, linalool). The purpose of this article is to review these new concepts and describe new resulting approaches to the treatment of TrPs.

Effect of low power 655 nm diode laser irradiation on the neuromuscular junctions of the mouse diaphragm.

BACKGROUND AND OBJECTIVES: Low level laser therapy (LLLT) in specific wavelengths and fluence maintains the electrophysiological activity of injured peripheral nerve in rats, preventing scar formation (at injury site) as well as degenerative changes in the corresponding motor neurons of the spinal cord, thus accelerating regeneration of the injured nerve. We studied the effect of LLLT on the neurotransmitter release in neuromuscular junctions of the mouse diaphragm. STUDY DESIGN/MATERIALS AND METHODS: Thirty-nine diaphragm muscles were studied. LLLT with GaAlAs 655 nm (1-12 J/cm(2)) was used. Neurotransmitter release was studied by conventional intracellular recording techniques on curarised or high magnesium media. Quantal content, amplitude, latency and rise time were analysed for end-plate potentials (EPPs). Frequency and amplitude were evaluated for the miniature end-plate potentials (MEPPs). Short-term plasticity of the neurotransmitter release (fast facilitation) was also evaluated by paired pulse stimulation. RESULTS AND CONCLUSIONS: This study showed that LLLT (655 nm) in these doses has no detectable physiological effect on the motor end-plate neurotransmitter release in mice.