Ce travail est soutenu par un financement du programme de l&#39;Union européenne septième cadre (FP7/2007-2013) sous convention de subvention n ° 246556 (projet RBUCE-UP européenne), HandiMedEx attribué par l&#39;Office d&#39;investissement public français. Marcel Bonay a été soutenue par la Chancellerie des Universités de Paris (Legs Poix), le Fonds de Dotation Recherche en Santé Respiratoire, et le Centre d&#39;Assistance Respiratoire à Domicile d&#39;Île de France (CARDIF)

Ce protocole a été approuvé par le comité d&#39;éthique de la chaise RBUCE-UP d&#39;excellence (Université de Paris Sud, accord de subvention n ° 246556) et l&#39;Université de Versailles Saint-Quentin-en-Yvelines. 1. Préparation des instruments chirurgicaux stérilisés <ol><li> Nettoyer les instruments chirurgicaux avec détergent de laboratoire. </li><li> Autoclave les instruments avant la chirurgie. </li><li> Dans une session chirurgicale, stériliser les outils en plaçant les pointes dans un stérilisateur à billes à chaud pendant 10 minutes à 180 ° C entre deux interventions chirurgicales. </li></ol> 2. Préparation des médicaments <ol><li> Préparer 2 x 1 ml seringues pour les médicaments pré-anesthésique cocktail et post-opératoires médicaments. </li><li> Selon le poids du rat, préparer une seringue à médicaments pré-anesthésique: Carprofen (5 mg / kg), la buprénorphine (50 ug / kg), Baytril (5 mg / kg) et de dexmédétomidine (0,5 mg / kg). Compléter le volume à 1 ml avec Ringer lactate. </li><li> Préparez à l&#39;une autre seringue pour l&#39;inversion des médicaments pré-anesthésique: atipamézole (500 ug / kg). </li></ol> 3. Anesthésie du Rat <ol><li> Administrer par voie sous cutanée à l&#39;animal la solution de médicaments pré-anesthésiques décrites à l&#39;étape 2.2. Ensuite, mettez le dos de l&#39;animal en cage et attendre jusqu&#39;à ce que l&#39;effet sédatif apparaît. </li><li> Placez le rat dans une chambre fermée remplie de 5% d&#39;isoflurane dans 100% O 2, et attendre jusqu&#39;à ce que le rythme respiratoire ralentit (environ 30 secondes). Ensuite, retirez le rat de la chambre et le placer sur la table de l&#39;intubation. </li></ol> 4. Orotrachéale intubation <ol><li> Allongez l&#39;animal sur son dos, puis fixer la tête en plaçant une sangle attachée à ses dents de devant à la table. </li><li> Avec une lumière de fibre optique, la lumière l&#39;espace thoracique. Ensuite, placez un laryngoscope (ou une coutume fait un, Jou et al. 20 pour plus de détails) dans la bouche de l&#39;animal. Visualiser les cordes vocales. </li><li> Glissée et placer un guide orotrachéal dans la trachée (entre les cordes vocales). Faites glisser le tube orotrachéal (16 G de taille de cathéter) sur le guide. </li><li> Retirer le guide et vérifier avec un miroir laryngé placé à l&#39;extrémité du tube oro-trachéale pour la présence d&#39;humidité, ce qui confirme la position appropriée du tube dans la trachée et non dans l&#39;œsophage. </li><li> Connectez le tuyau à un ventilateur de rongeur (683 rongeur ventilateur, Harvard Apparatus) et ajuster la concentration d&#39;isoflurane à 2% (100% O 2). </li><li> Fixer le tube orotrachéal avec du ruban adhésif chirurgical. </li></ol> 5. Chirurgie rachidienne <ol><li> Placer l&#39;animal en décubitus ventral sur une plaque chauffée chirurgical, avec le nez pointant à un angle de 90 ° pour le chirurgien. Maintenir la température du corps d&#39;environ 37,5 ° C pendant toute la chirurgie. </li><li> Raser les cheveux avec une tondeuse entre les omoplates et enlever les poils avec de la gaze. </li><li> Nettoyer la peau avec de la bétadine, puis avec 70% d&#39;alcool. Répétez cette étape 3x. </li><li> Une pincée de pointe est effectuée avant début de la chirurgie pour assurer une profondeur d&#39;anesthésie adéquate. &#160; Ensuite, effectuer une incision de la peau rostro-caudale latérale avec des ciseaux entre les omoplates. </li><li> Couper le acromiotrapezius muscle rostro-caudale en suivant le tendon d&#39;éviter tout saignement. Puis dissocier le muscle rhomboïde pour accéder aux muscles de SpinaliS (autour de la vertèbre). </li><li> Rétracter le muscle épineux de C1 à C3 vertèbre. La vertèbre C2 est l&#39;une d&#39;une apophyse proéminent. </li><li> Nettoyez le muscle autour de la partie dorsale de la vertèbre en utilisant des cotons-tiges stériles. </li><li> Commencer à retirer soigneusement l&#39;apophyse de C2 avec un rongeur. Ensuite, continuer méticuleusement jusqu&#39;à la moelle épinière dorsale est exposé. Assurez-vous que la laminectomie est une laminectomie dorsale hémi. Prêtez une attention particulière à la dure qui entoure la moelle épinière, et les artères à proximité de cette zone. </li><li> Avec # 55 pince, disséquer rostro-caudale la dure long C2, à côté de continuer latéralement de chaque côté rostral et caudal. </li><li> Éponger le liquide céphalo-rachidien. </li><li> Faire une coupe latérale sous la dorsale cervicale nombre de racines 2 avec les microciseaux. Vérifier avec un micro scalpel que l&#39;étendue de la lésion est suffisamment proche pour atteindre la ligne médiane de la moelle épinière (voir la figure 2A pour une vue dorsale de la blessure). Si non, alors une autre coupe pourrait être faite pour compléter la blessure. En cas de saignement, utiliser des cotons-tiges stériles. Attention de ne pas aller du côté opposé, sinon, l&#39;animal ne sera pas récupérer de la blessure et aura une insuffisance respiratoire. </li><li> Suturer les muscles comme une couche protectrice et suturer la peau. Nettoyer la plaie avec de la gaze stérile Betadine saturé. </li><li> Tournez-off le vaporisateur isoflurane et injecter les médicaments d&#39;inversion (Atipamézole [500 ug / kg, im]), vérifier la température du corps. </li><li> Quand l&#39;animal commence à respirer contre le ventilateur,débrancher le tube trachéal du ventilateur, puis retirez le tube orotrachéal. Placez l&#39;animal dans une cage chauffée pour la récupération. </li></ol> 6. Soins post-opératoire Après la chirurgie, les animaux sont surveillés en permanence pour assurer le meilleur environnement possible pour la récupération. Antibiotiques (Baytril, 5 mg / kg), anti-inflammatoire (Carprofen, 5 mg / kg) et la buprénorphine (50 ug / kg) médicaments sont donnés toutes les 12 h pour les 2 premiers jours post-opératoires pour prévenir les infections et réduire l&#39;apparition de la douleur post-opératoire. Les rats ont accès ad libitum à la nourriture et de l&#39;eau douce (ou de l&#39;eau gélifiée pour le 1er jour post-opératoire). Fluides sous-cutanés peuvent être utilisés pour prévenir la déshydratation sur les premiers jours post-opératoires. Le poids corporel et la consommation de nourriture sont contrôlés tous les jours. Leur environnement est enrichi tout au long de l&#39;expérience et du temps post-traumatique (double logement, des tubes dans leurs cages).

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Les auteurs déclarent qu&#39;ils n&#39;ont aucun intérêt financier concurrents.

Les difficultés techniques de Faire la blessure modèle C2 Le modèle murin de blessures C2 est un outil intéressant pour étudier la plasticité synaptique respiratoire post-lésionnelle. Cependant, les étapes nécessaires pour produire un modèle reproductible et fiable sont nombreux et chacun pourraient avoir une incidence sur les résultats de l&#39;étude. Par exemple, au cours du processus d&#39;intubation, un soin extrême doit être pris puisque le tube orotraché...

Traumatisme de la moelle épinière est une blessure courante observée dans la population humaine avec des incidences dramatiques, comme une paralysie permanente. Cependant, la gravité de la blessure est fonction du niveau et de l&#39;étendue du traumatisme initial. L&#39;insuffisance respiratoire est la principale cause de mortalité à la suite du col des blessures de la moelle épinière supérieure (SCI) 1. Actuellement, le seul traitement thérapeutique consiste à placer le patient sous assistance ventilatoire. Depuis quelques patients peuvent être sevrés de l&#39;assistance ventilatoire 2, en raison de la récupération spontanée qui se produit avec un retard post-lésionnelle, la nécessité de développer de nouvelles thérapies non invasives novatrices est urgent 3. Avoir un bon modèle pré-clinique normalisée pour étudier l&#39;effet d&#39;une SCI col de l&#39;utérus sur l&#39;insuffisance respiratoire et donc d&#39;étudier l&#39;application de stratégies thérapeutiques putatifs, est essentiel. Dans cet article technique, nous décrivons un modèle pré-clinique de murin spécifique of insuffisance respiratoire induite par une SCI partielle du col au niveau de C2. Ce modèle est actuellement utilisé par plusieurs laboratoires dans le monde (pour avis: 4-13). Cependant, de légères différences dans la procédure chirurgicale peuvent être observées entre les différents enquêteurs pour générer ce modèle murin de lésion cervicale particulier. L&#39;effet d&#39;une SCI C2 sur le débit respiratoire a été décrite pour la première en 1895 par Porter 14. Une hémi col de l&#39;utérus induit une désafférentation des motoneurones phréniques de leur entraînement central (situé dans le rVRG dans le tronc cérébral, la figure 1A) sur le côté ipsilatéral de blessures, conduisant à une activité du nerf phrénique silence et la paralysie du diaphragme ultérieure. Le côté opposé reste intact et permet à l&#39;animal de survivre. Contrairement différent SCI situé dans un segment vertébral inférieur (par exemple une blessure contusive au niveau de C4 15), l&#39;intégrité du noyau du motoneurone phrénique sur les deux côtés est préservée. Après une cervblessures C2 ical, une activité spontanée peut être observée sur le côté ipsilatéral (phrénique et le diaphragme) en raison d&#39;une activation de contralatérales voies synaptiques silencieuses qui ont traversé la ligne médiane de la moelle au niveau segmentaire C3-C6 (Les voies phrénique croisés, RPC, figure 1B) . L&#39;activation de la RPC, qui est, par définition, un hemisection C2 combiné avec un phrenicotomy controlatéral qui induisent une reprise ipsilatérale du nerf phrénique partielle, peut se produire de plusieurs heures à plusieurs semaines après la lésion 16-18. L&#39;effet bénéfique réel de cette voie RPC sur la reprise respiratoire est limitée 19 et une enquête plus approfondie et le traitement doit être développé pour améliorer l&#39;ampleur de la restauration spontanée 3. Ce protocole fournit un type puissant modèle murin pré-clinique pour étudier respiratoire plasticité post-lésionnelle à différents niveaux (physiologie respiratoire de motoneurones pré et phrénique, interneurones, moléculaire et cellular, la locomotion de l&#39;aile avant, par exemple) ainsi que d&#39;un modèle pour tester des stratégies thérapeutiques invasives et non-invasives visant à améliorer les voies respiratoires et la récupération locomotrice après une lésion cervicale C2 partielle de la moelle épinière.

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a murine model of cervical spinal cord injury to study post-lesional respiratory neuroplasticity

Une lésion de la moelle épinière cervicale induit une paralysie permanente, et conduit souvent à une détresse respiratoire. À ce jour, aucun thérapeutiques efficaces ont été développées pour améliorer / améliorer l&#39;insuffisance respiratoire suite à une blessure cervicale haute de la moelle épinière (SCI). Ici, nous proposons un modèle pré-clinique murin de haute SCI au col de l&#39;utérus 2 (C2) niveau métamérique à étudier divers neuroplasticité respiratoire post-lésionnelle. La technique consiste en une lésion chirurgicale partielle au niveau de C2, qui va induire une hémiparalysie de la membrane due à une désafférentation des motoneurones phréniques des centres respiratoires situés dans le tronc cérébral. Le côté opposé de la blessure reste intacte et permet la récupération des animaux. Contrairement à d&#39;autres SIC qui affectent la fonction locomotrice (au niveau thoracique et lombaire), la fonction respiratoire ne nécessite pas la motivation de l&#39;animal et la quantification du déficit / récupération peut être facilement réalisée (diaphragme et l&#39;enregistrement du nerf phréniques, la ventilation du corps entier). Ce modèle C2 SCI pré-clinique est un modèle pré-clinique puissant, utile et fiable pour étudier divers événements de neuroplasticité respiratoires et non respiratoires à différents niveaux (moléculaires à la physiologie) et de tester différentes stratégies thérapeutiques putatifs qui pourrait améliorer la respiration en patients blessés médullaires.

Ampleur de la blessure Le succès et la reproductibilité de ce modèle expérimental particulier dépendent de l&#39;expérience de chaque manipulateur / chirurgien. La quantité suivante de récupération respiratoire (activité du nerf phrénique et le diaphragme d&#39;activité) à la suite d&#39;une blessure de C2 est en corrélation avec le restant ventrolatérale épargné substance blanche 21. Depuis la blessure est &quot;fait main&quot; et nécessite une ...

Watch this Scientific Journal Video about A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity at JoVE.com

A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

L&#39;insuffisance respiratoire est la principale cause de décès suite à une blessure de la moelle épinière cervicale. Avoir un modèle animal pré-clinique reproductible, quantifiable et fiable de l&#39;insuffisance respiratoire provoquée par une blessure cervicale partielle aidera à comprendre la neuroplasticité respiratoire et non respiratoire ultérieure et permettra de tester des stratégies de réparation putatifs.

Un modèle murin de la moelle épinière cervicale blessures à étude post-lésionnelle respiratoire neuroplasticité

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed ...

a-murine-model-cervical-spinal-cord-injury-to-study-post-lesional

Research

JoVE Journal

Medicine

13.4K vues.  Université de Versailles Saint-Quentin-en-Yvelines.  L'insuffisance respiratoire est la principale cause de décès suite à une blessure de la moelle épinière cervicale. Avoir un modèle animal pré-clinique reproductible, quantifiable et fiable de l'insuffisance respiratoire provoquée par une blessure cervicale partielle aidera à comprendre la neuroplasticité respiratoire et non respiratoire ultérieure et permettra de tester des stratégies de réparation putatifs. 

Vidéo: A Murine Model of Cervical Spinal Cord Injury to Study Post-lesional Respiratory Neuroplasticity

Intraspinal Cell Transplantation for Targeting Cervical Ventral Horn in Amyotrophic Lateral Sclerosis and Traumatic Spinal Cord Injury

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) cases 1 and is the ultimate cause of death in patients with the motor neuron disorder, amyotrophic laterals sclerosis (ALS) 2.
ALS is a devastating neurological disorder that is characterized by relatively rapid degeneration of upper and lower motor neurons. Patients ultimately succumb to the disease on average 2-5 years following diagnosis because of respiratory paralysis due to loss of phrenic motor neuron innnervation of the diaphragm 3. The vast majority of cases are sporadic, while 10% are of the familial form. Approximately twenty percent of familial cases are linked to various point mutations in the Cu/Zn superoxide dismutase 1 (SOD1) gene on chromosome 21 4. Transgenic mice 4,5 and rats 6 carrying mutant human SOD1 genes (G93A, G37R, G86R, G85R) have been generated, and, despite the existence of other animal models of motor neuron loss, are currently the most highly used models of the disease.
Spinal cord injury (SCI) is a heterogeneous set of conditions resulting from physical trauma to the spinal cord, with functional outcome varying according to the type, location and severity of the injury 7. Nevertheless, approximately half of human SCI cases affect cervical regions, resulting in debilitating respiratory dysfunction due to phrenic motor neuron loss and injury to descending bulbospinal respiratory axons 1. A number of animal models of SCI have been developed, with the most commonly used and clinically-relevant being the contusion 8.
Transplantation of various classes of neural precursor cells (NPCs) is a promising therapeutic strategy for treatment of traumatic CNS injuries and neurodegeneration, including ALS and SCI, because of the ability to replace lost or dysfunctional CNS cell types, provide neuroprotection, and deliver gene factors of interest 9.
Animal models of both ALS and SCI can model many clinically-relevant aspects of these diseases, including phrenic motor neuron loss and consequent respiratory compromise 10,11. In order to evaluate the efficacy of NPC-based strategies on respiratory function in these animal models of ALS and SCI, cellular interventions must be specifically directed to regions containing therapeutically relevant targets such as phrenic motor neurons. We provide a detailed protocol for multi-segmental, intraspinal transplantation of NPCs into the cervical spinal cord ventral gray matter of neurodegenerative models such as SOD1G93A mice and rats, as well as spinal cord injured rats and mice 11.

Neural precursor transplantation is a promising strategy for protecting and/or replacing lost/dysfunctional cervical phrenic motor neurons in spinal cord injury (SCI) and the motor neuron disorder, amyotrophic laterals sclerosis (ALS). We provide a protocol for cell delivery to cervical spinal cord ventral horn in rodent models of ALS and SCI.

Cell Transplantation intraspinale pour le ciblage du col corne ventrale de la sclérose latérale amyotrophique et les lésions de la moelle épinière

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) ...

Recherche

Médecine

Acute and Chronic Tactile Sensory Testing after Spinal Cord Injury in Rats

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory testing in rat SCI models is needed. However, until recently, no single testing approach had been validated for SCI so that standardized methods have not been implemented across labs. Additionally, available testing methods could not be implemented acutely or when severe motor impairments existed, preventing studies of the development of SCI-induced allodynia3. Here we present two validated sensory testing methods using von Frey Hair (VFH) monofilaments which quantify changes in tactile sensory thresholds after SCI4-5. One test is the well-established Up-Down test which demonstrates high sensitivity and specificity across different SCI severities when tested chronically5. The other test is a newly-developed dorsal VFH test that can be applied acutely after SCI when allodynia develops, prior to motor recovery4-5. Each VFH monofilament applies a calibrated force when touched to the skin of the hind paw until it bends. In the up-down method, alternating VFHs of higher or lower forces are used on the plantar L5 dermatome to delineate flexor withdrawal thresholds. Successively higher forces are applied until withdrawal occurs then lower force VFHs are used until withdrawal ceases. The tactile threshold reflects the force required to elicit withdrawal in 50% of the stimuli. For the new test, each VFH is applied to the dorsal L5 dermatome of the paw while the rat is supported by the examiner. The VFH stimulation occurs in ascending order of force until at least 2 of 3 applications at a given force produces paw withdrawal. Tactile sensory threshold is the lowest force to elicit withdrawal 66% of the time. Acclimation, testing and scoring procedures are described. Aberrant trials that require a retest and typical trials are defined. Animal use was approved by Ohio State University Animal Care and Use Committee.

We describe two tactile sensory testing methods for acute or chronic periods of spinal cord injury in rats. These validated procedures can detect the development and maintenance of allodynia-like sensations.

Aiguë et chronique Test tactile sensorielle après un traumatisme médullaire chez le rat

Spinal cord injury (SCI) impairs sensory systems causing allodynia1-8. To identify cellular and molecular causes of allodynia, sensitive and valid sensory ...

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of spinal cord injury (SCI) in which the thoracic spinal cord is injured. Additionally, because most human cord injuries occur as the result of blunt, non-penetrating trauma (e.g. motor vehicle accident, sporting injury) where the spinal cord is violently struck by displaced bone or soft tissues, the majority of SCI researchers are of the opinion that the most clinically relevant injury models are those in which the spinal cord is rapidly contused.1 Therefore, an important step in the preclinical evaluation of novel treatments on their way to human translation is an assessment of their efficacy in a model of contusion SCI within the cervical spinal cord. Here, we describe the technical aspects and resultant anatomical and behavioral outcomes of an unilateral contusive model of cervical SCI that employs the Infinite Horizon spinal cord injury impactor.
Sprague Dawley rats underwent a left-sided unilateral laminectomy at C5. To optimize the reproducibility of the biomechanical, functional, and histological outcomes of the injury model, we contused the spinal cords using an impact force of 150 kdyn, an impact trajectory of 22.5&deg; (animals rotated at 22.5&deg;), and an impact location off of midline of 1.4 mm. Functional recovery was assessed using the cylinder rearing test, horizontal ladder test, grooming test and modified Montoya's staircase test for up to 6 weeks, after which the spinal cords were evaluated histologically for white and grey matter sparing.
The injury model presented here imparts consistent and reproducible biomechanical forces to the spinal cord, an important feature of any experimental SCI model. This results in discrete histological damage to the lateral half of the spinal cord which is largely contained to the ipsilateral side of injury. The injury is well tolerated by the animals, but does result in functional deficits of the forelimb that are significant and sustained in the weeks following injury. The cervical unilateral injury model presented here may be a resource to researchers who wish to evaluate potentially promising therapies prior to human translation.

A reliable and repeatable way to produce a cervical unilateral spinal cord injury using the Infinite Horizon impactor is described. The method takes advantage of a custom designed frame and clamp to stabilize the spine. The standardized procedure and biomechanical injury parameters result in sufficient and sustained injuries.

Un modèle de contusion unilatérale des blessures moelle épinière cervicale aide de l&#39;impacteur Infinite Horizon

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of ...

Controlled Cervical Laceration Injury in Mice

Use of genetically modified mice enhances our understanding of molecular mechanisms underlying several neurological disorders such as a spinal cord injury (SCI). Freehand manual control used to produce a laceration model of SCI creates inconsistent injuries often associated with a crush or contusion component and, therefore, a novel technique was developed. Our model of cervical laceration SCI has resolved inherent difficulties with the freehand method by incorporating 1) cervical vertebral stabilization by vertebral facet fixation, 2) enhanced spinal cord exposure, and 3) creation of a reproducible laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm in depth without associated contusion. Compared to the standard methods of creating a SCI laceration such as freehand use of a scalpel or scissors, our method has produced a consistent lesion. This method is useful for studies on axonal regeneration of corticospinal, rubrospinal, and dorsal ascending tracts.

A novel technique to create a reproducible in vivo model of cervical spinal cord laceration injury in the mouse is described. This technique is based on spine stabilization by fixation of the cervical facets and laceration of the spinal cord using an oscillating blade with an accuracy of &plusmn;0.01 mm.

Contrôlé blessures de lacération du col chez la souris

Use of genetically modified mice enhances our  understanding of molecular mechanisms underlying several neurological disorders  such as a spinal cord ...

A Contusion Model of Severe Spinal Cord Injury in Rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is described to obtain a consistent model of severe SCI. Use of a stereotactic frame and computer controlled impactor allows for creation of reproducible injury. Hypothermia and urinary tract infection pose significant challenges in the post-operative period. Careful monitoring of animals with daily weight recording and bladder expression allows for early detection of post-operative complications. The functional results of this contusion model are equivalent to transection models. The contusion model can be utilized to evaluate the efficacy of both neuroprotective and neuroregenerative approaches.

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

Un modèle de contusion de graves lésions de la moelle épinière chez les rats

The translational potential of novel treatments should be investigated in severe spinal cord injury (SCI) contusion models. A detailed methodology is ...

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment (RMCA) in Individuals with Chronic Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination is disrupted by spinal cord injury (SCI), control of respiratory muscles innervated below the injury level is compromised1,2 leading to respiratory muscle dysfunction and pulmonary complications. These conditions are among the leading causes of death in patients with SCI3. Standard pulmonary function tests that assess respiratory motor function include spirometrical and maximum airway pressure outcomes: Forced Vital Capacity (FVC), Forced Expiratory Volume in one second (FEV1), Maximal Inspiratory Pressure (PImax) and Maximal Expiratory Pressure (PEmax)4,5. These values provide indirect measurements of respiratory muscle performance6. In clinical practice and research, a surface electromyography (sEMG) recorded from respiratory muscles can be used to assess respiratory motor function and help to diagnose neuromuscular pathology. However, variability in the sEMG amplitude inhibits efforts to develop objective and direct measures of respiratory motor function6. Based on a multi-muscle sEMG approach to characterize motor control of limb muscles7, known as the voluntary response index (VRI)8, we developed an analytical tool to characterize respiratory motor control directly from sEMG data recorded from multiple respiratory muscles during the voluntary respiratory tasks. We have termed this the Respiratory Motor Control Assessment (RMCA)9. This vector analysis method quantifies the amount and distribution of activity across muscles and presents it in the form of an index that relates the degree to which sEMG output within a test-subject resembles that from a group of healthy (non-injured) controls. The resulting index value has been shown to have high face validity, sensitivity and specificity9-11. We showed previously9 that the RMCA outcomes significantly correlate with levels of SCI and pulmonary function measures. We are presenting here the method to quantitatively compare post-spinal cord injury respiratory multi-muscle activation patterns to those of healthy individuals.

Evaluation of Respiratory Muscle Activation Using Respiratory Motor Control Assessment RMCA in Individuals with Chronic Spinal Cord Injury

The purpose of this publication is to present our original work on a multi-muscle surface electromyographic approach to quantitatively characterize respiratory muscle activation patterns in individuals with chronic spinal cord injury using vector-based analysis.

Evaluation de l&#39;activation des muscles respiratoires aide respiratoire évaluation du contrôle moteur (MRAC) chez les personnes atteintes de maladies chroniques Spinal Cord Injury

During breathing, activation of respiratory muscles is coordinated by integrated input from the brain, brainstem, and spinal cord. When this coordination ...

Controlled Cortical Impact Model for Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI). Combined with the incidence of TBIs worldwide, the physical, emotional, social, and economical effects are staggering. Therefore, further research into the effects of TBI and effective treatments is necessary. The controlled cortical impact (CCI) model induces traumatic brain injuries ranging from mild to severe. This method uses a rigid impactor to deliver mechanical energy to an intact dura exposed following a craniectomy. Impact is made under precise parameters at a set velocity to achieve a pre-determined deformation depth. Although other TBI models, such as weight drop and fluid percussion, exist, CCI is more accurate, easier to control, and most importantly, produces traumatic brain injuries similar to those seen in humans. However, no TBI model is currently able to reproduce pathological changes identical to those seen in human patients. The CCI model allows investigation into the short-term and long-term effects of TBI, such as neuronal death, memory deficits, and cerebral edema, as well as potential therapeutic treatments for TBI.

Traumatic brain injuries (TBIs) remain a serious health problem. Using the controlled cortical impact surgery model, research on the effects of TBI and possible treatment methods may be performed.

Contrôlée corticale modèle d&#39;impact pour Traumatic Brain Injury

Every year over a million Americans suffer a traumatic brain injury (TBI).  Combined with the incidence of TBIs worldwide, the physical, emotional, ...

A Radio-telemetric System to Monitor Cardiovascular Function in Rats with Spinal Cord Transection and Embryonic Neural Stem Cell Grafts

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter connected to a radio transmitter into the femoral artery of rats that underwent a T4 spinal cord transection with or without grafting of embryonic brainstem-derived neural stem cells expressing green fluorescent protein. Compared to other methods such as cannula insertion or tail-cuff, telemetry is advantageous to continuously monitor blood pressure and heart rate in freely moving animals. It is also capable of long term multiple data acquisitions. In spinal cord injured rats, basal cardiovascular data under unrestrained condition and autonomic dysreflexia in response to colorectal distension were successfully recorded. In addition, cardiovascular parameters before and after SCI can be compared in the same rat if a transmitter is implanted before a spinal cord transection. One limitation of the described telemetry procedure is that implantation in the femoral artery may influence the blood supply to the ipsilateral hindlimb.

We present a protocol for using a radio-telemetric system to record cardiovascular parameters in T4 spinal cord transected rats eight weeks after embryonic brainstem neural stem cell grafting into the lesion site. Telemetry is an advanced technique to accurately evaluate cardiovascular function in conscious freely moving spinal cord injured rats.

Un système de radio-télémétrique de surveiller la fonction cardiovasculaire chez le rat avec la moelle épinière Transection et embryonnaires souches neurales greffes cellulaires

High thoracic or cervical spinal cord injury (SCI) can lead to cardiovascular dysfunction. To monitor cardiovascular parameters, we implanted a catheter ...

Synergetic Use of Neural Precursor Cells and Self-assembling Peptides in Experimental Cervical Spinal Cord Injury

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. Clinically, more than 50% of SCI affect the cervical spine1. As a consequence of the primary injury, a cascade of secondary mechanisms including inflammation, apoptosis, and demyelination occur finally leading to tissue scarring and development of intramedullary cavities2,3. Both represent physical and chemical barriers to cell transplantation, integration, and regeneration. Therefore, shaping the inhibitory environment and bridging cavities to create a supportive milieu for cell transplantation and regeneration is a promising therapeutic target4. Here, a contusion/compression model of cervical SCI using an aneurysm clip is described. This model is more clinically relevant than other experimental models, since complete transection or ruptures of the cord are rare. Also in comparison to the weight drop model, which in particular damage the dorsum columns, circumferential compression of the spinal cord appears advantageous. Clip closing force and duration can be adjusted to achieve different injury severity. A ring spring facilitates precise calibration and constancy of clip force. Under physiological conditions, synthetic self-assembling peptides (SAP) self-assemble into nanofibers and thus, are appealing for application in SCI5. They can be injected directly into the lesion minimizing damage to the cord. SAPs are biocompatible structures erecting scaffolds to bridge intramedullary cavities and thus, equip the damaged cord for regenerative treatments. K2(QL)6K2 (QL6) is a novel SAP introduced by Dong et al.6 In comparison to other peptides, QL6 self-assembles into &#946;-sheets at neutral pH6.14 days after SCI, after the acute stage, SAPs are injected into the center of the lesion and neural precursor cells (NPC) are injected into adjacent dorsal columns. In order to support cell survival, transplantation is combined with continuous subdural administration of growth factors by osmotic micro pumps for 7 days.

Treating cervical spinal cord injury with both self-assembling peptides (SAP) and neural precursor cells (NPC), together with growth factors, is a promising approach to promote regeneration and recovery. A contusion/compression aneurysm clip rat model of cervical SCI and combined treatment involving SAP injection and NPC transplantation is established.

Utilisation synergique des cellules neurales précurseurs et peptides d&#39;auto-assemblage dans la partie expérimentale moelle épinière cervicale blessures

Spinal cord injuries (SCI) cause serious neurological impairment and psychological, economic, and social consequences for patients and their families. ...

Calibrated Forceps Model of Spinal Cord Compression Injury

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The calibrated forceps model of compression injury is a convenient, low cost, and very reproducible animal model for SCI. We used a pair of modified forceps in accordance with the method published by Plemel et al. (2008) to laterally compress the spinal cord to a distance of 0.35 mm. In this video, we will demonstrate a dorsal laminectomy to expose the spinal cord, followed by compression of the spinal cord with the modified forceps. In the video, we will also address issues related to the care of paraplegic laboratory animals. This injury model produces mice that exhibit impairment in sensation, as well as impaired hindlimb locomotor function. Furthermore, this method of injury produces consistent aberrations in the pathology of the SCI, as determined by immunohistochemical methods. After watching this video, viewers should be able to determine the necessary supplies and methods for producing SCI of various severities in the mouse for studies on SCI and/or treatments designed to mitigate impairment after injury.

Spinal cord injury models should be highly reproducible. We demonstrate that the calibrated forceps compression model of spinal cord injury is an easy to use surgical method for generating reproducible injuries to the murine spinal cord.

Calibré Forceps modèle de compression de la moelle épinière des blessures

Compression injuries of the murine spinal cord are valuable animal models for the study of spinal cord injury (SCI) and spinal regenerative therapy. The ...