Diese Arbeit wird mit Mitteln aus der Europäischen Union Siebten Rahmenprogramms (FP7/2007-2013) unter Finanzhilfevereinbarung Nr. 246556 (europäische Projekt RBUCE-UP) unterstützt, HandiMedEx zugeteilt von der Französisch Öffentliche Investment Board. Marcel Bonay wurde von der Chancellerie des Universités de Paris (Beine Poix), dem Fonds de Recherche en Santé Dotation Respiratoire, und dem Centre d&#39;Assistance Respiratoire à Domicile d&#39;Île de France (CARDIF) unterstützt

Dieses Protokoll wurde von der Ethik-Ausschuss des RBUCE-UP Stuhl of Excellence (Universität Paris Sud, Finanzhilfevereinbarung Nr. 246556) und der Université de Versailles Saint-Quentin-en-Yvelines genehmigt. 1. Vorbereitung der sterilisierte chirurgische Instrumente <ol><li> Reinigen Sie die chirurgischen Instrumente mit Laborreinigungsmittel. </li><li> Autoklaven werden die Instrumente vor der Operation. </li><li> In einer chirurgischen Sitzung, die Werkzeuge zu sterilisieren, indem Sie die Tipps in eine heiße Perle Sterilisator für 10 min bei 180 ° C zwischen zwei Operationen. </li></ol> 2. Herstellung von Drogen <ol><li> Bereiten Sie 2 x 1 ml-Spritzen für Pre-Anästhetika Cocktail und nach der Operation Medikamente. </li><li> Entsprechend dem Gewicht der Ratte Herstellung einer Spritze mit vorge Anästhetika: Carprofen (5 mg / kg), Buprenorphin (50 ug / kg), Baytril (5 mg / kg) und Dexmedetomidin (0,5 mg / kg). Füllen Sie das Volumen auf 1 ml mit Ringer-Laktat. </li><li> Bereiten Sie imeine andere Spritze die Auflösung für die Pre-Anästhetika: Atipamezol (500 ug / kg). </li></ol> 3. Anästhesie der Ratte <ol><li> Subkutan verabreichen, um dem Tier die Lösung der in Schritt 2.2 beschrieben vorge Anästhetika. Dann, setzen Sie das Tier im Käfig zurück und warten, bis die sedierende Wirkung erscheint. </li><li> Legen Sie die Ratte in eine geschlossene Kammer mit 5% Isofluran in 100% O 2 gefüllt und warten, bis der Atemrhythmus verlangsamt (ca. 30 sec). Entfernen Sie dann die Ratte aus der Kammer und legen Sie sie auf den Tisch Intubation. </li></ol> 4. Orotracheale Intubation <ol><li> Legen Sie sich das Tier auf dem Rücken, dann sichern Sie den Kopf, indem sie einen Gurt um seine Vorderzähne an den Tisch befestigt. </li><li> Mit einem Glasfaser-Licht, leuchten den Brustraum. Legen Sie dann einen Kehlkopfspiegel (oder eine maßgeschneiderte ein, Jou et al. 20 für Details) in das Maul des Tieres. Visualisieren Sie die Stimmbänder. </li><li> Geschobene und platzieren Sie eine orotracheale Führung in der Luftröhre (zwischen den Stimmbändern). Schieben Sie die Orotrachealtubus (16 G Kathetergröße) auf der Führungs. </li><li> Entfernen der Führung und Überprüfung mit einer Kehlkopfspiegel am Ende der Orotrachealtubus für die Anwesenheit von Feuchtigkeit platziert, Bestätigung der entsprechenden Position der Röhre in die Luftröhre und in die Speiseröhre. </li><li> Schließen Sie den Schlauch an einem Nagetier Ventilator (683 Nagetier Ventilator, Harvard Apparatus) und stellen die Konzentration von Isofluran bis 2% (in 100% O 2). </li><li> Sichern Sie die Orotrachealtubus mit chirurgischen Band. </li></ol> 5. Wirbelsäulenchirurgie <ol><li> Legen Sie das Tier in Bauch Dekubitus-Position auf einem beheizten Operationsplatte, mit der Nase, die auf 90 °-Winkel an den Chirurgen. Pflegen Sie die Körpertemperatur um 37,5 ° C in der gesamten Chirurgie. </li><li> Rasieren Sie die Haare mit Klipper zwischen den Schulterblättern und entfernen Sie die Haare mit Gaze. </li><li> Reinigen Sie die Haut mit betadine, dann mit 70% Alkohol. Wiederholen Sie diesen Schritt 3x. </li><li> Eine Zehe Prise vor Beginn der Operation durchgeführt, um die ordnungsgemäße Narkosetiefe zu gewährleisten. &#160; Führen Sie dann einen seitlichen Hautschnitt rostro-kaudal mit der Schere zwischen den Schulterblättern. </li><li> Schneiden Sie die acromiotrapezius Muskel rostro-kaudal durch folgende Sehne, um Blutungen zu verhindern. Dann distanzieren den Rautenmuskel, um die Muskeln spinalis (rund um die Wirbel) zugreifen. </li><li> Fahren Sie den spinalis Muskel von C1 bis C3 Wirbel. Die C2-Wirbels ist das mit einem prominenten apophysis. </li><li> Reinigen Sie die Muskulatur rund um den dorsalen Teil der Wirbel mit sterilen Wattestäbchen. </li><li> Beginnen Sie vorsichtig mit einer Zange entfernen Sie den Fortsatz des C2. Dann weiter akribisch, bis der dorsalen Rückenmark freiliegt. Stellen Sie sicher, dass Laminektomie ist ein Rücken Hemi Laminektomie. Achten Sie besonders auf die Dauer, die das Rückenmark, und die Arterien in der Nähe von diesem Bereich umschließt. </li><li> Mit einem # 55 Pinzetten, sezieren rostro-kaudal die Dura entlang C2, nächste seitlich weiterhin auf jedem rostral und kaudal. </li><li> Schwamm den Liquor. </li><li> Machen Sie einen seitlichen Schnitt unter der zervikalen Hinterwurzel Nummer 2 mit der Mikroschere. Verifizierung mit einem Mikroskalpell, dass das Ausmaß der Läsion nahe genug ist, um die Mittellinie des Rückenmarks (siehe Fig. 2A eine dorsale Ansicht der Verletzung) zu erreichen. Wenn nicht, dann könnte ein weiterer Schnitt vorgenommen werden, um die Schädigung zu vervollständigen. Im Falle von Blutungen, sterile Wattestäbchen. Achten Sie darauf, nicht auf die Gegenseite zu gehen, sonst wird das Tier nicht von der Verletzung erholen und wird ein Atemversagen haben. </li><li> Nähen Sie die Muskeln als Schutzschicht und vernähen wieder die Haut. Reinigen Sie die Wunde mit steriler Gaze Betadine gesättigt. </li><li> Schalten Sie die Isofluran-Verdampfer und injizieren die Umkehrung Medikamente (Atipamezol [500 ug / kg, im]), überprüfen Sie die Körpertemperatur. </li><li> Wenn das Tier beginnt, gegen den Ventilator atmen,trennen Sie den Trachealtubus vom Beatmungsgerät, und nehmen Sie die Orotrachealtubus. Legen Sie das Tier in einem beheizten Käfig für die Verwertung. </li></ol> 6. Post-chirurgische Versorgung Nach der Operation werden die Tiere kontinuierlich überwacht, um die beste Umgebung für die Wiederherstellung möglich zu gewährleisten. Antibiotika (Baytril, 5 mg / kg), entzündungshemmende (Carprofen, 5 mg / kg) und Buprenorphin (50 ug / kg) Arzneimittel alle 12 h für die ersten 2 Tage nach der Operation, um Infektionen zu verhindern, und reduziert das Auftreten gegeben von postoperativen Schmerzen. Ratten haben Zugriff ad libitum zu weiche Nahrung und Wasser (oder geliert Wasser für die ersten postoperativen Tag). Subkutanen Flüssigkeiten können verwendet werden, um Austrocknung zu der ersten Tage nach der Operation zu verhindern. Das Körpergewicht und die Nahrungsaufnahme werden täglich überwacht. Ihre Umgebung wird während des Experiments und die Zeit nach der Verletzung (Dual-Gehäuse, Schläuche in ihren Käfigen) bereichert.

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Technische Schwierigkeiten bei der Herstellung der C2 Verletzungsmodell Die C2-Verletzung Mausmodell ist ein interessantes Tool, um Atem Post-läsionaler Neuroplastizität zu studieren. Allerdings sind die notwendigen Schritte, um eine reproduzierbare und zuverlässige Modell produzieren zahlreiche und jeder über die Ergebnisse der Studie beeinflussen können. Zum Beispiel während der Intubation Verfahren ist extrem darauf geachtet werden, da die Orotrachealtubus können e...

Rückenmarksverletzung ist eine häufige Verletzung in der menschlichen Bevölkerung mit dramatischen Vorfälle, wie Dauer Paralyse beobachtet. Die Schwere der Verletzung, hängt jedoch von der Höhe und dem Umfang des anfänglichen Traumas. Atemversagen ist die häufigste Todesursache folgenden oberen zervikalen Rückenmarksverletzung (SCI) ein. Derzeit ist die einzige therapeutische Behandlung des Patienten unter Atemunterstützung zu platzieren. Da nur wenige Patienten von der Atemunterstützung entwöhnt werden 2, durch spontane Erholung, die mit Post-läsionaler Verzögerung auftritt, ist die Notwendigkeit, neue innovative nicht-invasive Therapien entwickeln dringenden 3. Eine gute standardisierte präklinischen Modell, um die Wirkung einer zervikalen SCI auf respiratorische Insuffizienz zu untersuchen und somit die Anwendung der vermeintlichen therapeutischen Strategien zu untersuchen, ist unerlässlich. In diesem technischen Artikel beschreiben wir einen spezifischen präklinischen Mausmodell of Beeinträchtigung der Atemwege durch eine teilweise Hals-SCI an der C2-Ebene induziert. Dieses Modell wird derzeit von mehreren Labors auf der ganzen Welt verwendet werden (Bewertungen: 4-13). Allerdings können geringe Unterschiede in den chirurgischen Eingriff unter den verschiedenen Forschern beobachtet, dass diese besondere zervikalen Verletzungen Mausmodell zu erzeugen. Die Wirkung einer C2 SCI auf die Atemleistung wurde zuerst 1895 von Porter 14 beschrieben. Ein Hals-Hemisektion induziert eine Deafferentation der Zwerchfellmotoneuronen aus ihrer zentralen Antrieb (in der RVRG im Hirnstamm, 1A befindet) auf der gleichen Seite der Verletzung, die zu einer stillen Zwerchfellnervenaktivität und der anschließenden Membran Lähmung. Die Gegenseite erhalten bleibt und ermöglicht es dem Tier, um zu überleben. Im Gegensatz zu verschiedenen SCI in einem unteren Wirbelsäulensegment (zum Beispiel ein kontusive Verletzungen an C4 Ebene 15) befindet, wird die Integrität des Zwerchfellmotoneuron Kern auf beiden Seiten erhalten. Nach einer cervschen C2 Verletzungen können einige spontane Aktivität auf der ipsilateralen Seite (Zwerchfell und Zwerchfell) aufgrund einer Aktivierung des kontralateralen synaptischen stille Wege, die das Rückenmittellinie auf der Segmentebene C3-C6 (Gekreuzte Wege phrenic, CPP, 1B) gekreuzt beobachtet werden . Die Aktivierung der CPP, das ist per Definition ein C2 Hemisektion, kombiniert mit einer kontralateralen phrenicotomy die eine ipsilaterale Teil Phrenikus Erholung zu induzieren, kann von Stunden bis Wochen nach der Verletzung auftreten, 16-18. Die eigentliche positive Wirkung dieser CPP Weg auf die Atem Erholung ist begrenzt 19 und eine weitere Untersuchung und Behandlung zu entwickeln, um das Ausmaß der spontanen Wiederherstellung 3 zu verbessern. Dieses Protokoll bietet eine leistungsstarke Art der präklinischen Mausmodell die Atmungs Post-läsionaler Plastizität auf verschiedenen Ebenen zu studieren (Atemphysiologie von der Vor-und Zwerchfellmotoneuronen, Inter, Molekular-und cellular, Fortbewegung der Vordergliedmaße zum Beispiel) sowie ein Modell für invasive und nicht-invasive Therapiestrategien das Ziel, die Atemwege und des Bewegungsapparates Erholung nach C2 Teil zervikalen Rückenmarksverletzungen verbessern zu testen.

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			<td>Fine Science Tool</td>
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			<td height="25" style="height:25px;">O2</td>
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			<td height="21" style="height:21px;">Stand-Alone Vaporizer</td>
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			<td height="21" style="height:21px;">Thin line heated bed</td>
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			<td height="21" style="height:21px;">Air canister</td>
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a murine model of cervical spinal cord injury to study post-lesional respiratory neuroplasticity

Ein zervikalen Rückenmarksverletzung induziert dauerhafte Lähmung und führt zu Atemnot oft. Bisher wurden keine wirksamen Therapeutika entwickelt worden, um zu verbessern / verbessern die Atemversagen nach hohen zervikalen Rückenmarksverletzung (SCI). Hier schlagen wir eine Maus präklinischen Modell der Hoch SCI am zervikalen 2 (C2) metamere Ebene diverse Post-läsionaler Atemneuroplastizität zu studieren. Die Technik besteht aus einem Operationsteilschäden an der C2-Ebene, die eine Halbseitenlähmung der Membran durch eine Deafferentierung der Zwerchfellmotoneuronen aus den Atemzentren im Hirnstamm induziert wird. Die Gegenseite der Verletzung bleibt erhalten und ermöglicht dem Tier Erholung. Anders als andere Gebiete von gemeinschaftlicher Bedeutung, die die Bewegungsfunktion (bei der Brust-und Lendenbereich) betreffen, wird die Atemfunktion nicht erforderlich Tier Motivation und die Quantifizierung des Defizits / Recovery leicht durchgeführt werden (Membran-und Zwerchfellnerv Aufnahmes, Ganzkörper Lüftung). Diese präklinischen C2 SCI Modell ist ein leistungsfähiges, nützliche und zuverlässige präklinischen Modell, um verschiedene Atem-und nicht-respiratorischen Ereignisse Neuroplastizität auf verschiedenen Ebenen (molekulare Physiologie) zu studieren und diverse vermeintliche therapeutische Strategien, die die Atmung verbessern könnten, testen SCI-Patienten.

Ausmaß der Verletzung Der Erfolg und die Reproduzierbarkeit dieser besonderen experimentellen Modell sind abhängig von der Erfahrung jedes Manipulator / Chirurgen. Die anschließende Höhe von Atemwiederherstellung (Zwerchfellnervenaktivität und Membranaktivität) nach einer Verletzung C2 korreliert mit der Rest ventrolateralen verschont weißen Substanz 21. Da die Verletzung ist &quot;handmade&quot; und erfordert einige Übung von dem Chirurgen, der Umfang der...

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

Atemversagen ist die häufigste Todesursache nach einer zervikalen Rückenmarksverletzungen. Nachdem eine reproduzierbare, messbare und verlässliche präklinischen Tiermodell der Atemversagen durch eine teilweise zervikalen Verletzungen induziert wird helfen, die Atemwege und des anschließenden nicht-respiratorischen Neuroplastizität zu verstehen und ermöglichen die Prüfung vermeintlichen Reparaturstrategien.

Einem Mausmodell der zervikalen Rückenmarksverletzung zu Post-läsionaler Atemneuroplastizität Studieren

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 Aufrufe.  Université de Versailles Saint-Quentin-en-Yvelines.  Atemversagen ist die häufigste Todesursache nach einer zervikalen Rückenmarksverletzungen. Nachdem eine reproduzierbare, messbare und verlässliche präklinischen Tiermodell der Atemversagen durch eine teilweise zervikalen Verletzungen induziert wird helfen, die Atemwege und des anschließenden nicht-respiratorischen Neuroplastizität zu verstehen und ermöglichen die Prüfung vermeintlichen Reparaturstrategien. 

Serie: 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.

Intraspinale Cell Transplantation für Targeting Cervical Vorderhorn in Amyotrophe Lateralsklerose und traumatische Rückenmarksverletzungen

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

Forschung

Medizin

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.

Akute und chronische taktile Sensorische Testung nach Verletzungen des Rückenmarks bei der Ratte

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.

Ein Modell von gequetscht Einseitige Cervical Spinal Cord Injury Mit dem 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 ...

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.

Controlled Cervical Laceration Verletzungen in Mäuse

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.

Eine Prellung 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 ...

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.

Auswertung der Respiratory Muscle Activation Mit Respiratory Motor Control Assessment (RMCA) bei Personen mit chronischen 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.

Controlled Cortical Impact-Modell für traumatische Hirnverletzung

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.

Ein Radio-Telemetrie-System zur Herz-Kreislauf-Funktion bei Ratten mit Rückenmarksdurchtrennung und embryonalen Neural Stem Cell Grafts überwachen

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.

Synergetische Nutzung von neuralen Vorläuferzellen und Self-Montage Peptide 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. ...

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.

Calibrated Zange Modell Spinal Cord Injury Compression

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