Forelimb Muscles Research Articles

G.P.211: Combination therapy with Glatiramer acetate and FTS improves strength and fibrosis in the dy2J/dy2Jmouse model of MDC1A

Merosin deficient congenital muscular dystrophy type 1A (MDC1A) is a devastating incurable disorder of childhood. We previously studied the effect of Glatiramer acetate (GA), an anti-inflammatory agent and S- farnesylthiosalicilate acid (FTS), an anti-fibrotic agent, as a single agent in the dy2J/dy2J mouse model of MDC1A. Each of the two medications showed a different pattern of disease on three primary outcome measures: strength, fibrosis and animal mobility. GA monotheraphy resulted in significant improvement in hind limb muscle strength and movement parameters, but without a decrease in fibrosis measurement. FTS monotheraphy was associated with increased hind limb muscle strength, decreased fibrosis, but with no improvement in movement parameters. In the present study, the combination of GA and FTS was found to increase both fore (32%) and hind limb (75.8%) muscle strength, more than treatment with either drug alone (fore limb: no significant change for GA and for FTS; hind limb: GA 52.7% vs. FTS 42%). In addition, GA-FTS treatment resulted in significant reduction in muscle fibrosis (35.4%) compared to untreated dy2J/dy2J mice. However, no improvement in animal mobility was shown. According to the results, combined GA and FTS increased both fore and hind limb muscle strength more than either drug by itself and significantly ameliorated muscle fibrosis. Thus, GA-FTS treatment might be considered as a potential candidate for future clinical trials in children with CMD.

Specialize or risk disappearance - empirical evidence of anisomerism based on comparative and developmental studies of gnathostome head and limb musculature.

William K. Gregory was one of the most influential authors defending the existence of an evolutionary trend in vertebrates from a higher degree of polyisomerism (more polyisomeric or 'serial' anatomical structures arranged along any body axis) to cases of anisomerism (specialization or loss of at least some original polyisomeric structures). Anisomerism was the subject of much interest during the 19th and the beginning of the 20th centuries, particularly due to the influence of the Romantic German School and the notion of 'primitive archetype' and because it was conceptually linked to other crucial biological issues (e.g. complexity, scala naturae, progress, modularity or phenotypic integration). However, discussions on anisomerism and related issues (e.g. Williston's law) have been almost exclusively based on hard tissues. Here we provide the first detailed empirical test, and discussion, of anisomerism based on quantitative data obtained from phylogenetic and comparative analyses of the head and forelimb muscles of gnathostomes. Our results strongly support the existence of such a trend in both forelimb and head musculature. For instance, the last common ancestor (LCA) of extant tetrapods likely had 38 polyisomeric muscles (PMs) out of a total of 70 forelimb muscles (i.e. 54%), whereas in the LCAs of extant amniotes and of mammals these numbers were 38/73 (52%) and 21/67 (31%), and in humans are 11/59 (19%). Interestingly, the number of PMs that became specialized during the forelimb evolutionary transition from the LCA of extant tetrapods to humans (13) is very similar to the number of PMs that became lost (14), indicating that both specialization and loss contributed equally to the trend towards anisomerism. By contrast, during the evolution of the head musculature from the LCA of gnathostomes to humans a total of 27 PMs were lost whereas only one muscle became specialized. Importantly, the evolutionary trend towards anisomerism is not related to a general trend leading to the presence of fewer muscles in derived taxa, because for instance humans have more head muscles in total, but many less head polyisomeric muscles than early gnathostomes and extant fish such as sharks, and than early tetrapods and amphibians such as salamanders. This is because new muscles have also been acquired during gnathostome evolution (e.g. facial muscles of mammals). Interestingly, many new PMs have also been acquired during head evolution (but subsequently lost during the transitions towards humans), whereas only a few new PMs were acquired during forelimb evolution. Our comparisons and review of the literature indicate that there is also a trend towards anisomerism during development, thus providing a further example of a parallel between ontogeny and phylogeny, e.g. some forelimb PMs (e.g. contrahentes, intermetacarpales) become specialized or lost (re-absorbed) during human ontogeny and some head PMs (e.g. constrictores branchiales) become lost during salamander ontogeny. This review will inform future discussions on modularity, complexity, body plans, phenotypic integration and macroevolution, which should ideally include soft tissues and the use of new tools (e.g. anatomical networks) in order to provide a broader and more integrative understanding of these relevant subjects.

Generalizability of EMG decoding using local field potentials.

Motor cortical local field potentials (LFPs) have been successfully used to decode both kinematics and kinetics of arm movement. For future clinically viable prostheses, however, brain activity decoders will have to generalize well under a wide spectrum of behavioral conditions. This property has not yet been demonstrated clearly. Here, we provide evidence for the first time, that an LFP-based electromyogram (EMG) decoder can generalize reasonably well across two different types of behavior. We implanted intracortical microelectrode arrays in the primary motor (M1) and ventral pre-motor (PMv) cortices of a rhesus macaque, and recorded LFP and EMG activity from arm and hand muscles of the contralateral forelimb during a two-dimensional (2-D) centre-out isometric wrist torque task (TT), and during free reach and grasp behavior (FB). Selected temporal and spectral features of the LFP signals were used to train EMG decoders using data from both types of behavior separately. We assessed the decoding performance for both within- and across-task cases. The average achieved generalization score was 65 ± 20%, while in many cases individual scores reached 100%.

Injection of WGA-Alexa 488 into the ipsilateral hemidiaphragm of acutely and chronically C2 hemisected rats reveals activity-dependent synaptic plasticity in the respiratory motor pathways

WGA-Alexa 488 is a fluorescent neuronal tracer that demonstrates transsynaptic transport in the central nervous system. The transsynaptic transport occurs over physiologically active synaptic connections rather than less active or silent connections. Immediately following C2 spinal cord hemisection (C2Hx), when WGA-Alexa 488 is injected into the ipsilateral hemidiaphragm, the tracer diffuses across the midline of the diaphragm and retrogradely labels the phrenic nuclei (PN) bilaterally in the spinal cord. Subsequently, the tracer is transsynaptically transported bilaterally to the rostral Ventral Respiratory Groups (rVRGs) in the medulla over physiologically active connections. No other neurons are labeled in the acute C2Hx model at the level of the phrenic nuclei or in the medulla. However, with a recovery period of at least 7weeks (chronic C2Hx), the pattern of WGA-Alexa 488 labeling is notably changed. In addition to the bilateral PN and rVRG labeling, the chronic C2Hx model reveals fluorescence in the ipsilateral ventral and dorsal spinocerebellar tracts, and the ipsilateral reticulospinal tract. Furthermore, interneurons are labeled bilaterally in laminae VII and VIII of the spinal cord as well as neurons in the motor nuclei bilaterally of the intercostal and forelimb muscles. Moreover, in the chronic C2Hx model, there is bilateral labeling of additional medullary centers including raphe, hypoglossal, spinal trigeminal, parvicellular reticular, gigantocellular reticular, and intermediate reticular nuclei. The selective WGA-Alexa 488 labeling of additional locations in the chronic C2Hx model is presumably due to a hyperactive state of the synaptic pathways and nuclei previously shown to connect with the respiratory centers in a non-injured model. The present study suggests that hyperactivity not only occurs in neuronal centers and pathways caudal to spinal cord injury, but in supraspinal centers as well. The significance of such injury-induced plasticity is that hyperactivity may be a mechanism to re-establish lost function by compensatory routes which were initially physiologically inactive.

Complete forelimb myology of the basal theropod dinosaur Tawa hallae based on a novel robust muscle reconstruction method.

The forelimbs of nonavian theropod dinosaurs have been the subject of considerable study and speculation due to their varied morphology and role in the evolution of flight. Although many studies on the functional morphology of a limb require an understanding of its musculature, comparatively little is known about the forelimb myology of theropods and other bipedal dinosaurs. Previous phylogenetically based myological reconstructions have been limited to the shoulder, restricting their utility in analyses of whole-limb function. The antebrachial and manual musculature in particular have remained largely unstudied due to uncertain muscular homologies in archosaurs. Through analysis of the musculature of extant taxa in a robust statistical framework, this study presents new hypotheses of homology for the distal limb musculature of archosaurs and provides the first complete reconstruction of dinosaurian forelimb musculature, including the antebrachial and intrinsic manual muscles. Data on the forelimb myology of a broad sample of extant birds, crocodylians, lizards, and turtles were analyzed using maximum likelihood ancestral state reconstruction and examined together with the osteology of the early theropod Tawa hallae from the Late Triassic of New Mexico to formulate a complete plesiomorphic myology for the theropod forelimb. Comparisons with previous reconstructions show that the shoulder musculature of basal theropods is more similar to that of basal ornithischians and sauropodomorphs than to that of dromaeosaurids. Greater development of the supracoracoideus and deltoideus musculature in theropods over other bipedal dinosaurs correlates with stronger movements of the forelimb at the shoulder and an emphasis on apprehension of relatively large prey. This emphasis is further supported by the morphology of the antebrachium and the intrinsic manual musculature, which exhibit a high degree of excursion and a robust morphology well-suited for powerful digital flexion. The forelimb myology of Tawa established here helps infer the ancestral conformation of the forelimb musculature and the osteological correlates of major muscle groups in early theropods. These data are critical for investigations addressing questions relating to the evolution of specialized forelimb function across Theropoda.

Retrograde migration of pectoral girdle muscle precursors depends on CXCR4/SDF-1 signaling.

In vertebrates, muscles of the pectoral girdle connect the forelimbs with the thorax. During development, the myogenic precursor cells migrate from the somites into the limb buds. Whereas most of the myogenic precursors remain in the limb bud to form the forelimb muscles, several cells migrate back toward the trunk to give rise to the superficial pectoral girdle muscles, such as the large pectoral muscle, the latissimus dorsi and the deltoid. Recently, this developing mode has been referred to as the "In-Out" mechanism. The present study focuses on the mechanisms of the "In-Out" migration during formation of the pectoral girdle muscles. Combining in ovo electroporation, tissue slice-cultures and confocal laser scanning microscopy, we visualize live in detail the retrograde migration of myogenic precursors from the forelimb bud into the trunk region by live imaging. Furthermore, we present for the first time evidence for the involvement of the chemokine receptor CXCR4 and its ligand SDF-1 during these processes. After microsurgical implantations of CXCR4 inhibitor beads in the proximal forelimb region of chicken embryos, we demonstrate with the aid of in situ hybridization and live-cell imaging that CXCR4/SDF-1 signaling is crucial for the retrograde migration of pectoral girdle muscle precursors. Moreover, we analyzed the MyoD expression in CXCR4-mutant mouse embryos and observed a considerable decrease in pectoral girdle musculature. We thus demonstrate the importance of the CXCR4/SDF-1 axis for the pectoral girdle muscle formation in avians and mammals.

Functional osteology of the forelimb digging apparatus of badgers

Badgers (Carnivora, Mustelidae) constitute a nonmonophyletic assemblage of mammals that are characterized by a general similarity of appearance and a broad geographic distribution. A comparative analysis of the forelimb osteology of badgers was performed to assess how observed differences in their scratch-digging behavior are related to variation of forelimb structure. We measured forelimb bone proportions of 87 skeletal specimens and calculated 8 functional indexes that were analyzed using analysis of variance and principal component analysis to identify the osteological correlates of fossorial ability in this clade. The indexes that best characterize forelimb specialization for scratch-digging in badgers are related to a robust humerus, large humeral epicondyles, and a long olecranon process, and these features increase from digger to semifossorial forms. A large scapular area for muscle attachment and a robust ulna also are correlated with fossorial ability. Semifossorial forms are indicated to have greater forelimb muscle mass, increased mechanical advantage of the elbow extensors, and the ability of the elbow extensor and carpal and digital flexor muscles to apply high out-force to the substrate. Digger forms are indicated to have relatively less robust and longer distal limb bones, and lower limb out-force capability. This study reveals that for badgers there is a distinct relationship between the proportions of the forelimb bones and fossorial ability. The indexes tested that relate to mechanical advantage of the elbow extensors and mass and force production capacity of the musculature have the discriminatory ability to distinguish between degrees of forelimb specialization and fossorial ability of badgers.

Voltage-sensitive dye imaging of primary motor cortex activity produced by ventral tegmental area stimulation.

The primary motor cortex (M1) receives dopaminergic projections from the ventral tegmental area (VTA) through the mesocortical dopamine pathway. However, few studies have focused on changes in M1 neuronal activity caused by VTA activation. To address this issue, we used voltage-sensitive dye imaging (VSD) to reveal the spatiotemporal dynamics of M1 activity induced by single-pulse stimulation of VTA in anesthetized rats. VSD imaging showed that brief electrical stimulation of unilateral VTA elicited a short-latency excitatory-inhibitory sequence of neuronal activity not only in the ipsilateral but also in the contralateral M1. The contralateral M1 response was not affected by pharmacological blockade of ipsilateral M1 activity, but it was completely abolished by corpus callosum transection. Although the VTA-evoked neuronal activity extended throughout the entire M1, we found the most prominent activity in the forelimb area of M1. The 6-OHDA-lesioned VTA failed to evoke M1 activity. Furthermore, both excitatory and inhibitory intact VTA-induced activity was entirely extinguished by blocking glutamate receptors in the target M1. When intracortical microstimulation of M1 was paired with VTA stimulation, the evoked forelimb muscle activity was facilitated or inhibited, depending on the interval between the two stimuli. These findings suggest that VTA neurons directly modulate the excitability of M1 neurons via fast glutamate signaling and, consequently, may control the last cortical stage of motor command processing.

Biomechanics and control of landing in toads.

Anything that jumps must land, but unlike during jumping when muscles produce energy to accelerate the body into the air, controlled landing requires muscles to dissipate energy and decelerate the body. Among anurans, toads (genus Bufo) exhibit highly coordinated landing behaviors, using their forelimbs to stabilize the body after touch-down as they lower their hindlimbs to the ground. Moreover, toads land frequently, as they cover distances by stringing together long series of relatively short hops. We have been using toads as a model to understand the biomechanics and motor control strategies of coordinated landing. Our results show that toads prepare for landing differently depending on how far they hop. For example, the forelimbs are extended farther prior to impact after long hops than after short ones. Such kinematic alterations are mirrored by predictable modulation of the recruitment intensity of forelimb muscles before impact, such that longer hops lead to higher levels of pre-landing recruitment of muscles. These differences in kinematics and muscular activity help to control the most flexed configuration of the elbow that is achieved after impact, which in turn constrains the extent to which muscles involved in dissipating energy are stretched. Indeed, a combination of in vivo and in vitro experiments has shown that the elbow-extending anconeus muscle, which is stretched during landing as the elbow flexes, rarely reaches lengths longer than those on the plateau of the muscle's length-tension curve (where damage becomes more likely). We have also been studying how movements of the hindlimbs after take-off help to stabilize animals during landing. In particular, the immediate and rapid flexion of a toad's knees after take-off leads to a repositioning of the animal's center of mass (COM) that better aligns it with ground-reaction forces (GRFs) at impact and reduces torques that would destabilize the animal. Finally, recent work on sensory feedback involved in preparation for landing demonstrates that vision is not required for coordinated landing. Toads can effectively utilize proprioceptive and/or vestibular information during take-off to help inform themselves about landing conditions, but may also use other sensory modalities after take-off to modulate landing behavior.

Locomotor function shapes the passive mechanical properties and operating lengths of muscle

Locomotor muscles often perform diverse roles, functioning as motors that produce mechanical energy, struts that produce force and brakes that dissipate mechanical energy. In many vertebrate muscles, these functions are not mutually exclusive and a single muscle often performs a range of mechanically diverse tasks. This functional diversity has obscured the relationship between a muscle's locomotor function and its mechanical properties. I use hopping in toads as a model system for comparing muscles that primarily produce mechanical energy with muscles that primarily dissipate mechanical energy. During hopping, hindlimb muscles undergo active shortening to produce mechanical energy and propel the animal into the air, whereas the forelimb muscles undergo active lengthening to dissipate mechanical energy during landing. Muscles performing distinct mechanical functions operate on different regions of the force-length curve. These findings suggest that a muscle's operating length may be shaped by potential trade-offs between force production and sarcomere stability. In addition, the passive force-length properties of hindlimb and forelimb muscles vary, suggesting that passive stiffness functions to restrict the muscle's operating length in vivo. These results inform our understanding of vertebrate muscle variation by providing a clear link between a muscle's locomotor function and its mechanical properties.

A pre-clinical evaluation of silver, iodine and Manuka honey based dressings in a model of traumatic extremity wounds contaminated with Staphylococcus aureus

Prevention of extremity war wound infection remains a clinical challenge. Staphylococcus aureus is the most common pathogen in delayed infection. We hypothesised that choice of wound dressings may affect bacterial burden over 7 days reflecting the current practice of delayed primary closure of wounds within this timeframe.A randomised controlled trial of 3 commercially available dressings (Inadine® (Johnson & Johnson, NJ, USA), Acticoat® (Smith & Nephew, Hull, UK), Activon Tulle (Advancis Medical, Nottingham, UK)) was conducted in a rabbit model of contaminated forelimb muscle injury. A positive control group treated with antibiotics was included. Groups were compared to a saline soaked gauze control. The primary outcome was a statistically significant reduction (p<0.05) in tissue S. aureus at 7 days post-injury. Secondary outcome measurements included bacteraemias, observational data, whole blood determination, ELISA for plasma biomarkers, PCR array analysis of wound healing gene expression and muscle/lymph node histopathology.Antibiotic, Inadine and Acticoat groups had statistically significant lower bacterial counts (mean 7.13 [95% CI 0.00–96.31]×102; 1.66 [0.94–2.58]×105; 8.86 [0.00–53.35]×104cfu/g, respectively) and Activon Tulle group had significantly higher counts (2.82 [0.98–5.61]×106cfu/g) than saline soaked gauze control (7.58 [1.65–17.83]×105cfu/g). There were no bacteraemias or significant differences in observational data or whole blood determination. There were no significant differences in muscle/loss or pathology and lymph node cross-sectional area or morphology. There were some significant differences between treatment groups in the plasma cytokines IL-4, TNFα and MCP-1 in comparison to the control. PCR array data demonstrated more general changes in gene expression in the muscle tissue from the Activon Tulle group than the Inadine or Acticoat dressings with a limited number of genes showing significantly altered expression compared to control.This study has demonstrated that both Acticoat® and Inadine® dressings can reduce the bacteria burden in a heavily contaminated soft tissue wound and so they may offer utility in the clinical setting particularly where surgical treatment is delayed.

Comparative anatomy, evolution, and homologies of tetrapod hindlimb muscles, comparison with forelimb muscles, and deconstruction of the forelimb-hindlimb serial homology hypothesis.

For more than two centuries, the idea that the forelimb and hindlimb are serially homologous structures has been accepted without serious question. This study presents the first detailed analysis of the evolution and homologies of all hindlimb muscles in representatives of each major tetrapod group and proposes a unifying nomenclature for these muscles. These data are compared with information obtained previously about the forelimb muscles of tetrapods and the muscles of other gnathostomes in order to address one of the most central and enigmatic questions in evolutionary and comparative anatomy: why are the pelvic and pectoral appendages of gnathostomes generally so similar to each other? An integrative analysis of the new myological data, combined with a review of recent paleontological, developmental, and genetic works and of older studies, does not support serial homology between the structures of these appendages. For instance, many of the strikingly similar forelimb and hindlimb muscles found in each major extant tetrapod taxon were acquired at different geological times and/or have different embryonic origins. These similar muscles are not serial homologues, but the result of evolutionary parallelism/convergence due to a complex interplay of ontogenetic, functional, topological, and phylogenetic constraints/factors.

Is Salamander Limb Regeneration Really Perfect? Anatomical and Morphogenetic Analysis of Forelimb Muscle Regeneration in GFP‐Transgenic Axolotls as a Basis for Regenerative, Developmental, and Evolutionary Studies

The axolotl Ambystoma mexicanum is one of the most commonly used model organisms in developmental and regenerative studies because it can reconstitute what is believed to be a completely normal anatomical and functional forelimb/hindlimb after amputation. However, to date it has not been confirmed whether each regenerated forelimb muscle is really a "perfect" copy of the original muscle. This study describes the regeneration of the arm, forearm, hand, and some pectoral muscles (e.g., coracoradialis) in transgenic axolotls that express green fluorescent protein (GFP) in muscle fibers. The observations found that: (1) there were muscle anomalies in 43% of the regenerated forelimbs; (2) however, on average in each regenerated forelimb there are anomalies in only 2.5% of the total number of muscles examined, and there were no significant differences observed in the specific insertion and origin of the other muscles analyzed; (3) one of the most notable and common anomalies (seen in 35% of the regenerated forelimbs) was the presence of a fleshy coracoradialis at the level of the arm; this is a particularly outstanding configuration because in axolotls and in urodeles in general this muscle only has a thin tendon at the level of the arm, and the additional fleshy belly in the regenerated arms is strikingly similar to the fleshy biceps brachii of amniotes, suggesting a remarkable parallel between a regeneration defect and a major phenotypic change that occurred during tetrapod limb evolution; (4) during forelimb muscle regeneration there was a clear proximo-distal and radio-ulnar morphogenetic gradient, as seen in normal development, but also a ventro-dorsal gradient in the order of regeneration, which was not previously described in the literature. These results have broader implications for regenerative, evolutionary, developmental and morphogenetic studies.

Muscular Anatomy of the Forelimbs of the Lesser Grison (Galictis cuja), and a Functional and Phylogenetic Overview of Mustelidae and Other Caniformia

The extrinsic and intrinsic forelimb musculature of the lesser grison (Galictis cuja), a short-legged mustelid of southern South America, is studied for the first time. We present descriptions, muscular maps, and weight data. Muscular anatomy description of the lesser grison provides the framework for discussing the myological diversity of mustelids and other caniforms, for addressing nomenclatural problems (such as synonymy and homonymy), and for highlighting some functional and phylogenetically informative traits. We recognize in the lesser grison features shared by all mustelid species, and some other caniforms, especially mephitids, such as the presence of rhomboideus profundus, an angular head of triceps brachii, and, apparently, the absence of a flexor digitorum brevis manus. An unexpected record of articularis humeri, a proximal origin of the brachioradialis, and the absence of the tensor fascia antebrachii are recorded for this species. As other ictonychines and mustelines, Galictis cuja possesses stronger and subdivided protractors and sagittal rotators of the forelimbs, as well as shoulder and elbow extensors. These features allow for resistance in landing during bounds and increase the stride length during epigean and subterranean crouched locomotion. Powerful neck musculature assists during hunting and carrying of prey. Weakness of some retractors and intrinsic flexors is related to a relatively minor importance of the forelimbs as propellers during bounding and the lack of other specializations. The configuration of the rhomboideus and the absence of coracobrachialis seem to be informative at the subfamiliar level within Mustelidae. The comprehensive and comparative review of available information leads us to propose possible solutions to old nomenclatural problems and of identification. This allows us to reassess of some myological features as diagnostic of caniform clades.

Stimulation Selectivity of the “Thin-Film Longitudinal Intrafascicular Electrode” (tfLIFE) and the “Transverse Intrafascicular Multi-Channel Electrode” (TIME) in the Large Nerve Animal Model

Neural prostheses are limited by the availability of peripheral neural electrodes to record the user's intention or provide sensory feedback through functional electrical stimulation. Our objective was to compare the ability of the novel “transverse intrafascicular multi-channel electrode” (TIME) and an earlier generation “thin-film longitudinal intrafascicular electrode” (tfLIFE) to selectively stimulate nerve fascicles and activate forelimb muscles in pigs. TIME was designed to access a larger subpopulation of fascicles than tfLIFE and should therefore be able to selectively activate a larger number of muscles. Electrodes were implanted in the median nerve, and sequential electric stimulation was applied to individual contacts. The compound muscle action potentials of seven muscles were recorded to quantify muscle recruitment. As expected, TIME was able to recruit more muscles with higher selectivity than tfLIFE (significant difference when comparing the performance of an entire electrode); a similar activation current was used (no significant difference). Histological analysis revealed that electrodes were located between fascicles, which influenced the selectivity and activation current level. In conclusion, TIME is a viable neural interface for selective activation of multiple fascicles in human-sized nerves that may assist to pave the way for future neuroprosthesis applications.

Distribution patterns and predilection muscles of Trichinella zimbabwensis larvae in experimentally infected Nile crocodiles (Crocodylus niloticus Laurenti).

No controlled studies have been conducted to determine the predilection muscles of Trichinella zimbabwensis larvae in Nile crocodiles (Crocodylus niloticus) or the influence of infection intensity on the distribution of the larvae in crocodiles. The distribution of larvae in muscles of naturally infected Nile crocodiles and experimentally infected caimans (Caiman crocodilus) and varans (Varanus exanthematicus) have been reported in literature. To determine the distribution patterns of T. zimbabwensis larvae and predilection muscles, 15 crocodiles were randomly divided into three cohorts of five animals each, representing high infection (642 larvae/kg of bodyweight average), medium infection (414 larvae/kg of bodyweight average) and low infection (134 larvae/kg of bodyweight average) cohorts. In the high infection cohort, high percentages of larvae were observed in the triceps muscles (26%) and hind limb muscles (13%). In the medium infection cohort, high percentages of larvae were found in the triceps muscles (50%), sternomastoid (18%) and hind limb muscles (13%). In the low infection cohort, larvae were mainly found in the intercostal muscles (36%), longissimus complex (27%), forelimb muscles (20%) and hind limb muscles (10%). Predilection muscles in the high and medium infection cohorts were similar to those reported in naturally infected crocodiles despite changes in infection intensity. The high infection cohort had significantly higher numbers of larvae in the sternomastoid, triceps, intercostal, longissimus complex, external tibial flexor, longissimus caudalis and caudal femoral muscles (p &lt; 0.05) compared with the medium infection cohort. In comparison with the low infection cohort, the high infection cohort harboured significantly higher numbers of larvae in all muscles (p &lt; 0.05) except for the tongue. The high infection cohort harboured significantly higher numbers of larvae (p &lt; 0.05) in the sternomastoid, triceps, intercostal, longissimus complex, external tibial flexor, longissimus caudalis and caudal femoral muscles compared with naturally infected crocodiles. Results from this study show that, in Nile crocodiles, larvae of T. zimbabwensis appear first to invade predilection muscles closest to their release site in the small intestine before occupying those muscles situated further away. The recommendation for the use of masseter, pterygoid and intercostal muscles as sampling sites for the detection of T. zimbabwensis in crocodiles is in contrast to the results from this study, where the fore- and hind limb muscles had the highest number of larvae. This study also supports the use of biopsy sampling from the dorso-lateral regions of the tail for surveillance purposes in both wild and commercial crocodile populations.

Chronic cortical and electromyographic recordings from a fully implantable device: preclinical experience in a nonhuman primate

Objective. Analysis of intra- and perioperatively recorded cortical and basal ganglia local field potentials in human movement disorders has provided great insight into the pathophysiology of diseases such as Parkinson's, dystonia, and essential tremor. However, in order to better understand the network abnormalities and effects of chronic therapeutic stimulation in these disorders, long-term recording from a fully implantable data collection system is needed. Approach. A fully implantable investigational data collection system, the Activa® PC + S neurostimulator (Medtronic, Inc., Minneapolis, MN), has been developed for human use. Here, we tested its utility for extended intracranial recording in the motor system of a nonhuman primate. The system was attached to two quadripolar paddle arrays: one covering sensorimotor cortex, and one covering a proximal forelimb muscle, to study simultaneous cortical field potentials and electromyography during spontaneous transitions from rest to movement. Main results. Over 24 months of recording, movement-related changes in physiologically relevant frequency bands were readily detected, including beta and gamma signals at approximately 2.5 μV/ and 0.7 μV/, respectively. The system architecture allowed for flexible recording configurations and algorithm triggered data recording. In the course of physiological analyses, sensing artifacts were observed (∼1 μVrms stationary tones at fixed frequency), which were mitigated either with post-processing or algorithm design and did not impact the scientific conclusions. Histological examination revealed no underlying tissue damage; however, a fibrous capsule had developed around the paddles, demonstrating a potential mechanism for the observed signal amplitude reduction. Significance. This study establishes the usefulness of this system in measuring chronic brain and muscle signals. Use of this system may potentially be valuable in human trials of chronic brain recording in movement disorders, a next step in the design of closed-loop neurostimulation paradigms.

Patterns of cortical input to the primary motor area in the marmoset monkey

In primates the primary motor cortex (M1) forms a topographic map of the body, whereby neurons in the medial part of this area control movements involving trunk and hindlimb muscles, those in the intermediate part control movements involving forelimb muscles, and those in the lateral part control movements of facial and other head muscles. This topography is accompanied by changes in cytoarchitectural characteristics, raising the question of whether the anatomical connections also vary between different parts of M1. To address this issue, we compared the patterns of cortical afferents revealed by retrograde tracer injections in different locations within M1 of marmoset monkeys. We found that the entire extent of this area is unified by projections from the dorsocaudal and medial subdivisions of premotor cortex (areas 6DC and 6M), from somatosensory areas 3a, 3b, 1/2, and S2, and from posterior parietal area PE. While cingulate areas projected to all subdivisions, they preferentially targeted the medial part of M1. Conversely, the ventral premotor areas were preferentially connected with the lateral part of M1. Smaller but consistent inputs originated in frontal area 6DR, ventral posterior parietal cortex, the retroinsular cortex, and area TPt. Connections with intraparietal, prefrontal, and temporal areas were very sparse, and variable. Our results demonstrate that M1 is unified by a consistent pattern of major connections, but also shows regional variations in terms of minor inputs. These differences likely reflect requirements for control of voluntary movement involving different body parts.

The Effects of Inclination (Up and Down) of the Treadmill on the Electromyogram Activities of the Forelimb and Hind limb Muscles at a Walk and a Trot in Thoroughbred Horses.

ABSTRACTIt is important to know the effects of the inclination of a slope on the activity of each muscle, because training by running on a sloped track is commonly used for Thoroughbred racehorses. The effects of incline (from −6 to +6%) on the forelimbs and hind limbs during walking and trotting on a treadmill were evaluated by an integrated electromyogram (iEMG). The muscle activities in the forelimbs (5 horses) and hind limbs (4 horses) were measured separately. Two stainless steel wires were inserted into each of the brachiocephalicus (Bc), biceps brachii (BB), splenius (Sp), and pectoralis descendens (PD) in the forelimb experiment and into the longissimus dorsi (LD), vastus lateralis (VL), gluteus medius (GM), and biceps femoris (BF) in the hind limb experiment. The EMG recordings were taken at a sampling rate of 1,000 Hz. At a walk, the iEMG values for the forelimb were not significantly different under any of the inclinations. In the hind limb, the iEMG values for the GM and BF significantly decreased as the inclination decreased. At a trot, the iEMG values for the Bc in the forelimb significantly decreased as the inclination of the treadmill decreased. In the hind limb, the iEMG values for the LD, GM, and BF significantly decreased as the inclination decreased. Uphill exercise increased the iEMG values for the Bc, LD, GM, and BF, while downhill exercise resulted in little increase in the iEMG values. It was concluded that the effects of inclination on the muscle activities were larger for the uphill exercises, and for the hind limb muscles compared with the forelimb muscles.

Forelimb muscle architecture and myosin isoform composition in the groundhog (Marmota monax).

Scratch-digging mammals are commonly described as having large, powerful forelimb muscles for applying high force to excavate earth, yet studies quantifying the architectural properties of the musculature are largely unavailable. To further test hypotheses about traits that represent specializations for scratch-digging, we quantified muscle architectural properties and myosin expression in the forelimb of the groundhog (Marmota monax), a digger that constructs semi-complex burrows. Architectural properties measured were muscle moment arm, muscle mass (MM), belly length (ML), fascicle length (l(F)), pennation angle and physiological cross-sectional area (PCSA), and these metrics were used to estimate maximum isometric force, joint torque and power. Myosin heavy chain (MHC) isoform composition was determined in selected forelimb muscles by SDS-PAGE and densitometry analysis. Groundhogs have large limb retractors and elbow extensors that are capable of applying moderately high torque at the shoulder and elbow joints, respectively. Most of these muscles (e.g. latissimus dorsi and pectoralis superficialis) have high l(F)/ML ratios, indicating substantial shortening ability and moderate power. The unipennate triceps brachii long head has the largest PCSA and is capable of the highest joint torque at both the shoulder and elbow joints. The carpal and digital flexors show greater pennation and shorter fascicle lengths than the limb retractors and elbow extensors, resulting in higher PCSA/MM ratios and force production capacity. Moreover, the digital flexors have the capacity for both appreciable fascicle shortening and force production, indicating high muscle work potential. Overall, the forelimb musculature of the groundhog is capable of relatively low sustained force and power, and these properties are consistent with the findings of a predominant expression of the MHC-2A isoform. Aside from the apparent modifications to the digital flexors, the collective muscle properties observed are consistent with its behavioral classification as a less-specialized burrower and these may be more representative of traits common to numerous rodents with burrowing habits or mammals with some fossorial ability.