Multimodal convergence in the pedunculopontine tegmental nucleus: Motor, sensory and theta-frequency inputs influence activity of single neurons.

The pedunculopontine tegmental nucleus of the brainstem (PPTg) has extensive interconnections and neuronal-behavioural correlates. It is implicated in movement control and sensorimotor integration. We investigated whether single neuron activity in freely moving rats is correlated with components of skilled forelimb movement, and whether individual neurons respond to both motor and sensory events. We found that individual PPTg neurons showed changes in firing rate at different times during the reach. This type of temporally specific modulation is like activity seen elsewhere in voluntary movement control circuits, such as the motor cortex, and suggests that PPTg neural activity is related to different specific events occurring during the reach. In particular, many neuronal modulations were time-locked to the end of the extension phase of the reach, when fine distal movements related to food grasping occur, indicating strong engagement of PPTg in this phase of skilled individual forelimb movements. In addition, some neurons showed brief periods of apparent oscillatory firing in the theta range at specific phases of the reach-to-grasp movement. When movement-related neurons were tested with tone stimuli, many also responded to this auditory input, allowing for sensorimotor integration at the cellular level. Together, these data extend the concept of the PPTg as an integrative structure in generation of complex movements, by showing that this function extends to the highly coordinated control of the forelimb during skilled reach to grasp movement, and that sensory and motor-related information converges on single neurons, allowing for direct integration at the cellular level.

Hindlimb muscle spindles inform preparatory forelimb coordination prior to landing in toads.

Animals move across a wide range of surface conditions in real-world environments to acquire resources and avoid predation. To effectively navigate a variety of surfaces, animals rely on several mechanisms including intrinsic mechanical responses, spinal-level central pattern generators, and neural commands that require sensory feedback. Muscle spindle Ia afferents play a critical role in providing sensory feedback and informing motor control strategies across legged vertebrate locomotion, which is apparent in cases where this sensory input is compromised. Here, we tested the hypothesis that spindle Ia afferents from hindlimb muscles are important for coordinating forelimb landing behavior in the cane toad. We performed bilateral sciatic nerve reinnervations to ablate the stretch reflex from distal hindlimb muscles while allowing for motor neuron recovery. We found that toads significantly delayed the onset and reduced the activation duration of their elbow extensor muscle following spindle Ia afferent ablation in the hindlimbs. However, reinnervated toads achieved similar elbow extension at touchdown to that of their pre-surgery state. Our results suggest that while toads likely tuned the activation timing of forelimb muscles in response to losing Ia afferent sensation from the hindlimbs they were likely able to employ compensatory strategies that allowed them to continue landing effectively with reduced sensory information during take-off. These findings indicate muscle spindle Ia afferents may contribute to tuning complex movements involving multiple limbs.

Performing a Three-Dimensional Finite Element Analysis to Simulate and Quantify the Contact Pressure in the Canine Elbow Joint: A Pilot Study.

OBJECTIVE: The aim of this study was to measure surface pressures and force distribution on radius and ulna in healthy and dysplastic elbow joints in different positions using the finite element analysis (FEA). STUDY DESIGN: FEA was performed on computed tomographic data of healthy and fragmented coronoid process diseased elbow joints of Labrador Retrievers. It considered the articular cartilage, collateral ligaments, triceps and biceps muscle. The analysis of each joint was performed in four positions (standing position: 145 degrees and three positions of the stance phase of gait: beginning: 115 degrees, middle: 110 degrees, end: 145 degrees joint angle) in consideration of different ground reaction forces (standing: 88.3 N; stance phase of gait: 182.5 N). RESULTS: Mean values of total force of 317.5 N (standing), 590.7 N (beginning), 330.9 N (middle) and 730.9 N (end) were measured. The percentual force distribution resulted in a total of 49.56 ± 26.58% on the ulna with a very inhomogeneous distribution. A significant difference was detected between the positions 'standing' and 'end' (p = 0.0497) regardless of the joint condition. In some FEA results, visual assessment of the surface pressures indicated an increase in pressure in the region of the medial compartment without a uniform pattern. An increase in pressure resulted in an area increase in the pressure marks on the joint surface and measurable pressure was increased at a larger joint angle. CLINICAL SIGNIFICANCE: FEA can provide information about the transmission of force in the joint. Prior to the use of FEA in scientific clinical research for the simulation of force, further model improvements are necessary.

The effect of the interosseous ligament and selected antebrachiocarpal ligaments on rotation of the radius during extension of the carpus.

OBJECTIVE: To evaluate the effect of the interosseous ligament and selected antebrachiocarpal ligaments on the internal rotation of the radius relative to the ulna, during carpal extension. STUDY DESIGN: Cadaveric study. SAMPLE POPULATION: Twenty-four cadaveric canine forelimbs. METHODS: Twenty-four forelimbs were disarticulated at the elbow joint and the antebrachia were prepared for testing. The forelimbs were divided to 6 groups, defined by the order in which ligaments were transected. All specimens were tested intact and after transecting each of the ligaments with the order defined by the group. Rotation of the radius relative to the ulna, caused by extension of the carpus, was measured using a sensor connected to the radius. RESULTS: The mean(±sd) maximum internal rotation of the radius (5.94° ± (1.23°)) with all the ligaments intact was significantly greater (p < 0.012) than the mean(±sd) maximum internal rotation of the radius (3.13° (± 1.13°)) after transecting the interosseus ligament. Transecting the interosseous ligament subsequent to one of the other ligaments caused a decrease in internal radial rotation (p = 0.629), while, transecting the short radial collateral ligament caused an increase in radial rotation (p = 0.629). Transecting the palmar radiocarpal and ulnocarpal ligaments had no effect on radial rotation. CONCLUSION: The interosseous ligament was stretched with internal rotation of the radius due to carpal extension. Carpal flexion resulted in external rotation of the radius. This effect was lost when the interosseus ligament was transected. CLINICAL SIGNIFICANCE: Rotation of the radius is associated with carpal extension, and is likely an intrinsic part of forelimb biomechanics.

Effect of sedation or general anesthesia on elbow goniometry and thoracic limb circumference measurements in dogs with naturally occurring elbow osteoarthritis.

OBJECTIVE: To evaluate the effect of sedation or general anesthesia (GA) on elbow goniometry and thoracic limb circumference (TLC) measurements in dogs with elbow osteoarthritis (OA). STUDY DESIGN: Prospective study. ANIMALS: Twenty-four client-owned dogs with radiographically confirmed elbow OA. METHODS: Elbow goniometry and TLC measurements were made before and after either sedation or GA by using a hand-held goniometer and spring tension measuring tape, respectively. Observers were not allowed to review their pre-sedation or pre-GA measurements at the time of obtaining measurements on dogs under sedation or GA. Mixed analysis of variance models were used to compare elbow goniometry and TLC measurements before and after sedation or GA. RESULTS: Eleven and thirteen dogs were included in the sedation and GA groups, respectively. Mean elbow flexion decreased by 5° and 3° and mean elbow extension increased by 6° and 2° under sedation and GA, respectively. Total range of motion increased by 11° under sedation and by 5° under GA. Each of these changes was statistically significant (P < .05) except elbow extension under GA (P = .129). Sedation and GA did not influence TLC measurements (P > .05). CONCLUSION: Sedation or GA led to slight and similar increase in elbow flexion and extension but did not influence TLC measurements in dogs with elbow OA. CLINICAL SIGNIFICANCE: Sedation or GA can cause slight alterations to goniometric measurements in canine elbows with OA. The protocols used in this study for sedation and GA seem interchangeably acceptable for goniometry and TLC measurements in dogs with elbow OA.

Forelimb movements evoked by optogenetic stimulation of the macaque motor cortex.

Optogenetics has become an indispensable tool for investigating brain functions. Although non-human primates are particularly useful models for understanding the functions and dysfunctions of the human brain, application of optogenetics to non-human primates is still limited. In the present study, we generate an effective adeno-associated viral vector serotype DJ to express channelrhodopsin-2 (ChR2) under the control of a strong ubiquitous CAG promoter and inject into the somatotopically identified forelimb region of the primary motor cortex in macaque monkeys. ChR2 is strongly expressed around the injection sites, and optogenetic intracortical microstimulation (oICMS) through a homemade optrode induces prominent cortical activity: Even single-pulse, short-duration oICMS evokes long-lasting repetitive firings of cortical neurons. In addition, oICMS elicits distinct forelimb movements and muscle activity, which are comparable to those elicited by conventional electrical ICMS. The present study removes obstacles to optogenetic manipulation of neuronal activity and behaviors in non-human primates.

Effect of proximal abducting ulnar osteotomy (PAUL) on frontal plane thoracic limb alignment: An ex vivo canine study.

OBJECTIVE: To determine the effect of proximal abducting ulnar osteotomy (PAUL) on frontal plane thoracic limb alignment in standing and recumbent positions. STUDY DESIGN: Ex vivo cadaveric study. SAMPLE POPULATION: Canine thoracic limbs (n = 15 limb pairs). METHODS: Limbs were acquired from healthy Labrador retrievers that had been euthanized for reasons unrelated to this study. A limb press was used to obtain standing and recumbent caudocranial radiographs before and after PAUL. Foot lateralization and rotation were directly measured in standing position. Mechanical joint angles were determined using full limb radiographic montages and the center of rotation of angulation (CORA) method for pre-PAUL (Pre), 2-mm PAUL (PAUL2), and 3-mm PAUL (PAUL3). Data are reported as mean ± SD and 95% CI. Mixed linear modeling was used to identify differences in limb alignment values and foot position, with significance established at P ≤ .004. RESULTS: There were differences in five of 12 limb alignment values pre-PAUL and post-PAUL in standing and recumbent positions. In the standing position, there was an increase in mechanical medial proximal radioulnar angle (Pre, 80.6° ± 2.5°; PAUL2, 82.6° ± 2.4°; PAUL3, 84° ± 2.4°) and a decrease in elbow compression angle (Pre, 1.4° ± 1.3°; PAUL2, 1° ± 0.9°; PAUL3, 0.8° ± 1°). There was a movement of mechanical humeral radioulnar angle (Pre, -8.9° ± 2.8°; PAUL2, -6.1° ± 2.7°; PAUL3, -5.2 ± 2.7°), mechanical thoracic humeral angle (Pre, 3.9° ± 1.7°; PAUL2, 2.4° ± 1.4°; PAUL3, 2.6° ± 1.5°), and elbow mechanical axis deviation (Pre, 1.9% ± 1.1%; PAUL2, 0.9% ± 1.1%; PAUL3, 0.4% ± 1.4%) toward a value of "0" representing coaxial alignment of the limb. The foot underwent lateralization (Pre, 1.4 ± 0.6 cm; PAUL2, 1.8 ± 0.7 cm; PAUL3, 2.3 ± 0.7 cm) and external rotation (Pre, 10.5° ± 4.7°; PAUL2, 13.7° ± 5.1°; PAUL3, 16° ± 6.6°). CONCLUSION: In the ex vivo setting, PAUL resulted in translation of the mechanical axis of the thoracic limb from a medial to lateral direction through alterations in limb alignment values associated with the elbow, humerus, and proximal radius/ulna. CLINICAL SIGNIFICANCE: Additional studies are required to determine whether PAUL alters thoracic limb alignment in client-owned dogs.

Physiologic effects of long-term immobilization of the equine distal limb.

OBJECTIVE: To describe the effects of distal limb immobilization and remobilization in the equine metacarpophalangeal joint. STUDY DESIGN: Randomized, prospective experimental study. ANIMALS: Eight healthy, skeletally mature horses. METHODS: One forelimb of each horse was immobilized in a fiberglass cast for 8 weeks; this was followed by 12 weeks of a treadmill-based training program after the cast had been removed. Clinical examinations, radiography, computed tomography (CT), nuclear scintigraphy, MRI, and histomorphometry were used to examine the third metacarpal (MC3), proximal phalanx, proximal sesamoid bones, and associated soft tissues in each horse. Serum and synovial fluid were collected for biomarker analyses. RESULTS: Distal limb immobilization resulted in persistent lameness (P < .001), effusion (P = .002), and a decreased range of motion (P = .012) as well as radiographically visible fragments (P = .036) in the cast forelimb. Bone density was decreased (P < .001) in MC3 according to CT, and trabecular bone fluid was increased (P < .001) according to MRI in the cast forelimb. The cast forelimbs had a change (P = .009) in the appearance of the deep digital flexor tendon according to MRI immediately after removal of the cast. Numerous clinical, radiographic, CT, and MR abnormalities were visible at the end of the study period. CONCLUSION: Eights weeks of cast immobilization induced changes in bone, cartilage, and periarticular soft tissues that were not reversed after 12 weeks of remobilization. CLINICAL SIGNIFICANCE: Cast application should be used judiciously in horses with musculoskeletal injuries, balancing appropriate stabilization with potential morbidity secondary to cast placement.

Functional morphology and identity of the thenar pad in the subterranean genus Ctenomys (Rodentia, Caviomorpha).

As in many other fossorial tetrapods, the most obvious adaptations to scratch-digging in the subterranean tuco-tuco (Rodentia, Ctenomyidae, Ctenomys) are found in the hands, which among other adaptations, present the mesaxonic condition; i.e. the central digits are more developed, and also their claws, which are curved and elongated. The thumb is atrophied and aligned with the rest of the digits, showing a small and flat claw. This configuration of digits and claws seems to be in accordance with what it is expected for rodents: rudimentary movements when handling food items. However, on the palmar side of the hand, tuco-tucos have several pads, the thenar (located under the thumb) being the most developed. In this study, we investigated the functional morphology of the thenar pad through different approaches: musculoskeletal anatomy, histology and functionality. The analysis of radiographs and clarified and double-stained hand samples of Ctenomys talarum and C. australis showed that the thenar pad is supported by a paddle-shaped bone that articulates with a protrusion in the scapholunate bone. This bone, flat and long, continues in a flat cartilaginous structure, with a shape similar to a claw. Dissections showed that the thenar pad has several associated muscles: the m. palmaris longus, the m. abductor pollicis longus, and a massive muscular complex located between the thumb and the thenar pad. By topology it might be inferred that this complex is formed by the m. abductor pollicis brevis, the m. flexor pollicis brevis and the m. adductor pollicis brevis. Longitudinal histological sections of the thenar pad stained with hematoxylin-eosin showed a thick layer of keratin at the distal end, external face. The observation of live specimens of C. talarum foraging on two food items of different size and filmed at 300 fps showed that the thenar pad acts as an opposable thumb, with digit-like movements. Tuco-tucos are able to perform more precise movements than expected, and to grasp and manipulate the food with one hand. In previous studies, it was suggested that the thenar pad was supported by a 'palmar ossicle', or 'prepollex' (= radial sesamoid bone). Our results suggest that this sesamoid underwent a radical change on its morphology, making the thenar pad a part of the food handling system in Ctenomys, so the thenar pad might be considered a 'false thumb', rather than a palmar pad. It is suggested to advance on the description and functional analysis of the thenar pad, redefining the structure, since the terms used so far to define it would not be accurate.

Automated Forelimb Tasks for Rodents: Current Advantages and Limitations, and Future Promise.

Rodent tests of function have advanced our understanding of movement, largely through the human training and testing and manual assessment. Tools such as reaching and grasping of a food pellet have been widely adopted because they are effective and simple to use. However, these tools are time-consuming, subjective, and often qualitative. Automation of training, testing, and assessment has the potential to increase efficiency while ensuring tasks are objective and quantitative. We detail new methods for automating rodent forelimb tests, including the use of pellet dispensers, sensors, computer vision, and home cage systems. We argue that limitations in existing forelimb tasks are driving the innovations in automated systems. We further argue that automated tasks partially address these limitations, and we outline necessary precautions and remaining challenges when adopting these types of tasks. Finally, we suggest attributes of future automated rodent assessment tools that can enable widespread adoption and help us better understand forelimb function in health and disease.

Hydrogen gas therapy improves survival rate and neurological deficits in subarachnoid hemorrhage rats: a pilot study.

The high morbidity, high mortality, and significant shortage of effective therapies for subarachnoid hemorrhage (SAH) have created an urgency to discover novel therapies. Human studies in Asia have established the safety of hydrogen gas in the treatment of hepatic, renal, pulmonary, and cardiac diseases. Mechanistically, hydrogen gas has been shown to affect oxidative stress, inflammation, and apoptosis. We hypothesized that hydrogen therapy would improve neurological function and increase survival rate in SAH. High dose hydrogen gas (66% at 3 L/min) was administered for 2 hours at 0.5, 8, and 18 hours after SAH. This treatment increased 72-hour survival rate and provided 24-hour neuroprotection after SAH in rats. To our knowledge, this is the first report demonstrating that high dose hydrogen gas therapy reduces mortality and improves outcome after SAH. Our results correlate well with the proposed mechanisms of hydrogen gas therapy within the literature. We outline four pathways and downstream targets of hydrogen gas potentially responsible for our results. A potentially complex network of pathways responsible for the efficacy of hydrogen gas therapy, along with a limited mechanistic understanding of these pathways, justifies further investigation to provide a basis for clinical trials and the advancement of hydrogen gas therapy in humans. This study was approved by the Institutional Animal Care and Use Committee of Loma Linda University, USA (Approval No. 8160016) in May 2016.

The Effects of Ergosta-7,9(11),22-trien-3ß-ol from Antrodia camphorata on the Biochemical Profile and Exercise Performance of Mice.

Antrodia camphorata (AC) is a rare and unique mushroom that is difficult to cultivate. Previous studies have demonstrated the bioactivity of the compound Ergosta-7,9(11),22-trien-3ß-ol (EK100) from AC in submerged culture. The purpose of this study is to evaluate the potential beneficial effects of EK100 on fatigue and ergogenic functions following physiological challenge. Male ICR (Institute of Cancer Research) mice were randomly divided into three groups (n = 8 per group) and orally administered EK100 for six weeks at 0 (Vehicle), 10 (EK100-1X), and 20 (EK100-2X) mg/kg/day. The six-week Ek100 supplementation significantly increased grip strength (P = 0.0051) in trend analysis. Anti-fatigue activity was evaluated using 15-min. acute exercise testing and measuring the levels of serum lactate, ammonia, glucose, blood urea nitrogen (BUN), and creatine kinase (CK) after a 15-min. swimming exercise. Our results indicate that AC supplementation leads to a dose-dependent decrease in serum lactate, ammonia, BUN, and CK activity after exercise and significantly increases serum glucose and glycogen content in liver tissues. Biochemical and histopathological data demonstrated that long term daily administration of EK100 for over six weeks (subacute toxicity) was safe. EK100's anti-fatigue properties appear to be through the preservation of energy storage, increasing blood glucose and liver glycogen content, and decreasing the serum levels of lactate, ammonia, BUN, and CK. EK100 could potentially be used to improve exercise physiological adaptation, promote health, and as a potential ergogenic aid in combination with different nutrient strategies.

Comparison of ground reaction force measurements in a population of Domestic Shorthair and Maine Coon cats.

Current research on gait analysis mostly involves horses and dogs. Feline kinetics and kinematics are being investigated and receiving more clinical interest at present. Ground reaction forces measured on pressure-sensitive mattresses have been established in healthy Domestic Shorthair cats (DSH). Currently, no further information exists on either breed-specific measured gait reaction forces or comparisons among breeds. Because Maine Coon (MC) cats appear to be over-represented with orthopaedic diseases of the hind limb (hip dysplasia, patellar luxation), we evaluated ground reaction force GRF measurements in MC cats and compared them with those of DSH cats. Pre-evaluation radiological and clinical exams determined that the cats were not lame. The parameters evaluated were peak vertical force (PFz), vertical impulse (IFz), time to PFz (TPFz), step length (SL), paw contact area (PCA), stance phase duration (SPD) and symmetry index (SI) for the fore- and hind limbs. In both breeds, PFz and IFz were greater in forelimbs than in hind limbs. The PFz and IFz in Newtons were higher in the MC cats compared to the DSH cats, but not after normalisation for total force (%TF) and body mass (%BM). Furthermore, due to their body conformation, MC cats have a longer SL, larger PCA, and higher body weight than DSH cats. No other parameters differed significantly, except that the TPFz displayed an earlier value in the MC hind limbs. Measured symmetry indices were similar to those reported in dogs and did not differ between breeds. This is the first study to report GRF values and temporospatial parameters in a healthy MC cat population. However, our results could not confirm differences between normalized PFz and IFz and temporospatial parameters between the breeds. The authors therefore conclude that genetic or other causes may be involved in orthopaedic hind limb pathogenesis seen in MC cats more often than in other breeds.

Biceps Detachment Preserves Joint Function in a Chronic Massive Rotator Cuff Tear Rat Model.

BACKGROUND: Lesions of the long head of the biceps tendon are often associated with massive rotator cuff tears (MRCTs), and biceps tenotomy is frequently performed for pain relief and functional reservation. However, the efficacy and safety of biceps tenotomy regarding the effects on the surrounding tissues in chronic MRCT are unclear. HYPOTHESIS: Biceps tenotomy would result in improved mechanical and histological properties of the intact subscapularis tendon and improved in vivo shoulder function while not compromising glenoid cartilage properties. STUDY DESIGN: Controlled laboratory study. METHODS: Right supraspinatus and infraspinatus tendons were detached in 25 male Sprague-Dawley rats, followed by 4 weeks of cage activity to create a chronic MRCT condition. Animals were randomly divided into 2 groups and received either biceps tenotomy (n = 11) or sham surgery (n = 14) and were sacrificed 4 weeks thereafter. Forelimb gait and ground-reaction forces were recorded 1 day before the tendon detachment (baseline), 1 day before the surgical intervention (biceps tenotomy or sham), and 3, 7, 10, 14, 21, and 28 days after the intervention to assess in vivo shoulder joint function. The subscapularis tendon and glenoid cartilage were randomly allocated for mechanical testing or histologic assessment after the sacrifice. RESULTS: Compared with sham surgery, biceps tenotomy partially restored the in vivo shoulder joint function, with several gait and ground-reaction force parameters returning closer to preinjury baseline values at 4 weeks. With biceps tenotomy, mechanical properties of the subscapularis tendons were improved, while mechanical properties and histological Mankin scores of the glenoid cartilage were not diminished when compared with the sham group. CONCLUSION: Biceps tenotomy in the presence of chronic MRCT partially preserves overall shoulder function and potentially restores subscapularis tendon health without causing detrimental effects to joint cartilage. This laboratory study adds to the growing literature regarding the protective effects of biceps tenotomy on the shoulder joint in a chronic MRCT model. CLINICAL RELEVANCE: This study provides important basic science evidence supporting the use of biceps tenotomy in patients with massive rotator cuff tears.

Optogenetic surface stimulation of the rat cervical spinal cord.

Electrical intraspinal microstimulation (ISMS) at various sites along the cervical spinal cord permits forelimb muscle activation, elicits complex limb movements and may enhance functional recovery after spinal cord injury. Here, we explore optogenetic spinal stimulation (OSS) as a less invasive and cell type-specific alternative to ISMS. To map forelimb muscle activation by OSS in rats, adeno-associated viruses (AAV) carrying the blue-light sensitive ion channels channelrhodopsin-2 (ChR2) and Chronos were injected into the cervical spinal cord at different depths and volumes. Following an AAV incubation period of several weeks, OSS-induced forelimb muscle activation and movements were assessed at 16 sites along the dorsal surface of the cervical spinal cord. Three distinct movement types were observed. We find that AAV injection volume and depth can be titrated to achieve OSS-based activation of several movements. Optical stimulation of the spinal cord is thus a promising method for dissecting the function of spinal circuitry and targeting therapies following injury. NEW & NOTEWORTHY Optogenetics in the spinal cord can be used both for therapeutic treatments and to uncover basic mechanisms of spinal cord physiology. For the first time, we describe the methodology and outcomes of optogenetic surface stimulation of the rat spinal cord. Specifically, we describe the evoked responses of forelimbs and address the effects of different adeno-associated virus injection paradigms. Additionally, we are the first to report on the limitations of light penetration through the rat spinal cord.

A single hydrotherapy session increases range of motion and stride length in Labrador retrievers diagnosed with elbow dysplasia.

Canine elbow dysplasia is a debilitating condition of unknown aetiology and is a common cause of forelimb lameness in dogs. Canine hydrotherapy is a therapeutic approach rapidly increasing in popularity for the treatment of a range of musculoskeletal pathologies. In this study, kinematic analysis was used to assess the effect of a customised hydrotherapy session on the range of motion, stride length and stride frequency of healthy Labrador retrievers (n=6) and Labrador retrievers diagnosed with bilateral elbow dysplasia (n=6). Reflective kinematic markers were attached to bony anatomical landmarks and dogs were recorded walking at their preferred speed on a treadmill before and 10min after a single hydrotherapy session. Range of motion, stride length and stride frequency were calculated for both forelimbs. Data were analysed via a robust mixed ANOVA to assess the effect of hydrotherapy on the kinematic parameters of both groups. Range of motion was greater in the healthy dogs at baseline (P<0.05). Hydrotherapy increased the range of motion of the forelimbs of both groups (P<0.05); dogs with elbow dysplasia demonstrated a greater improvement in range of motion than healthy dogs (P<0.05). Hydrotherapy stride length (P<0.01) of all dogs, but differences were not seen between the two groups. Stride frequency increased after hydrotherapy only in the left limb (P<0.05) in all dogs. These results support the potential of canine hydrotherapy as a therapeutic tool for the rehabilitation and treatment of Labradors with elbow dysplasia. Furthermore, results indicate that hydrotherapy might improve the gait and movement of healthy dogs. However, whether these results are transient or sustained remains undetermined.

Pseudotyped baculovirus is an effective gene expression tool for studying molecular function during axolotl limb regeneration.

Axolotls can regenerate complex structures through recruitment and remodeling of cells within mature tissues. Accessing the underlying mechanisms at a molecular resolution is crucial to understand how injury triggers regeneration and how it proceeds. However, gene transformation in adult tissues can be challenging. Here we characterize the use of pseudotyped baculovirus (BV) as an effective gene transfer method both for cells within mature limb tissue and within the blastema. These cells remain competent to participate in regeneration after transduction. We further characterize the effectiveness of BV for gene overexpression studies by overexpressing Shh in the blastema, which yields a high penetrance of classic polydactyly phenotypes. Overall, our work establishes BV as a powerful tool to access gene function in axolotl limb regeneration.

Cortical Reorganization in Dual Innervation by Single Peripheral Nerve.

BACKGROUND: Functional recovery after peripheral nerve injury and repair is related with cortical reorganization. However, the mechanism of innervating dual targets by 1 donor nerve is largely unknown. OBJECTIVE: To investigate the cortical reorganization when the phrenic nerve simultaneously innervates the diaphragm and biceps. METHODS: Total brachial plexus (C5-T1) injury rats were repaired by phrenic nerve-musculocutaneous nerve transfer with end-to-side (n = 15) or end-to-end (n = 15) neurorrhaphy. Brachial plexus avulsion (n = 5) and sham surgery (n = 5) rats were included for control. Behavioral observation, electromyography, and histologic studies were used for confirming peripheral nerve reinnervation. Cortical representations of the diaphragm and reinnervated biceps were studied by intracortical microstimulation techniques before and at months 0.5, 3, 5, 7, and 10 after surgery. RESULTS: At month 0.5 after complete brachial plexus injury, the motor representation of the injured forelimb disappeared. The diaphragm representation was preserved in the "end-to-side" group but absent in the "end-to-end" group. Rhythmic contraction of biceps appeared in "end-to-end" and "end-to-side" groups, and the biceps representation reappeared in the original biceps and diaphragm areas at months 3 and 5. At month 10, it was completely located in the original biceps area in the "end-to-end" group. Part of the biceps representation remained in the original diaphragm area in the "end-to-side" group. Destroying the contralateral motor cortex did not eliminate respiration-related contraction of biceps. CONCLUSION: The brain tends to resume biceps representation from the original diaphragm area to the original biceps area following phrenic nerve transfer. The original diaphragm area partly preserves reinnervated biceps representation after end-to-side transfer.

Mechanism of Restoration of Forelimb Motor Function after Cervical Spinal Cord Hemisection in Rats: Electrophysiological Verification.

The objective of this study was to electrophysiologically assess the corticospinal tracts of adult rats and the recovery of motor function of their forelimbs after cervical cord hemisection. Of 39 adult rats used, compound muscle action potentials (CMAPs) of the forelimbs of 15 rats were evaluated, before they received left C5 segmental hemisection of the spinal cord, by stimulating the pyramid of the medulla oblongata on one side using an exciting microelectrode. All 15 rats exhibited contralateral electrical activity, but their CMAPs disappeared after hemisection. The remaining 24 rats received hemisection first, and CMAPs of 12 rats were assessed over time to study their recovery time. All of them exhibited electrical activity of the forelimbs in 4 weeks after surgery. The remaining 12 rats received additional right C2 segmental hemisection, and variation of CMAPs between before and after surgery was examined. The right side of the 12 rats that received the additional hemisection exhibited no electrical activity in response to the stimulation of the pyramids on both sides. These results suggest that changes in path between the resected and healthy sides, activation of the ventral corticospinal tracts, and propriospinal neurons were involved in the recovery of motor function after cervical cord injury.

Neurturin overexpression in dopaminergic neurons induces presynaptic and postsynaptic structural changes in rats with chronic 6-hydroxydopamine lesion.

The structural effect of neurturin (NRTN) on the nigrostriatal dopaminergic system in animals remains unknown, although NRTN has been shown to be effective in Parkinson's disease animal models. Herein, we aimed to demonstrate that NRTN overexpression in dopaminergic neurons stimulates both neurite outgrowths in the nigrostriatal pathway and striatal dendritic spines in aging rats with chronic 6-hydroxydopamine (6-OHDA) lesion. At week 12 after lesion, pTracer-mNRTN-His or pGreenLantern-1 plasmids were intranigrally transfected using the NTS-polyplex nanoparticles system. We showed that the transgenic expression in dopaminergic neurons remained until the end of the study (12 weeks). Only animals expressing NRTN-His showed recovery of tyrosine hydroxylase (TH)+ cells (28 ± 2%), their neurites (32 ± 2%) and the neuron-specific cytoskeletal marker ß-III-tubulin in the substantia nigra; striatal TH(+) fibers were also recovered (52 ± 3%), when compared to the healthy condition. Neurotensin receptor type 1 levels were also significantly recovered in the substantia nigra and striatum. Dopamine recovery was 70 ± 4% in the striatum and complete in the substantia nigra. The number of dendritic spines of striatal medium spiny neurons was also significantly increased, but the recovery was not complete. Drug-activated circling behavior decreased by 73 ± 2% (methamphetamine) and 89 ± 1% (apomorphine). Similar decrease was observed in the spontaneous motor behavior. Our results demonstrate that NRTN causes presynaptic and postsynaptic restoration of the nigrostriatal dopaminergic system after a 6-OHDA-induced chronic lesion. However, those improvements did not reach the healthy condition, suggesting that NRTN exerts lesser neurotrophic effects than other neurotrophic approaches.