Name: reproducable paraplegia by thoracic aortic occlusion in a murine model of spinal cord ischemia-reperfusion
Uploaded: 2014-03-03T21:55:00.000+00:00
Duration: 5 min 59 s
Description: Watch this Scientific Journal Video about Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion at JoVE.com

Wir möchten die Thoraxchirurgie-Stiftung für Bildung und Forschung für die finanzielle Unterstützung dieses Projekts danken.

1. Die präoperative Vorbereitung und Anästhesie <ol><li> Achten Sie darauf, steriler Technik während des gesamten Verfahrens zu beobachten. Legen Sie alle Instrumente. </li><li> Einschalten des Temperatur-Steuerbett vor Narkoseeinleitung, so daß sie auf die geeignete Temperatur (36,5 ° C) zu erwärmen. Schalten Sie den Laser-Doppler-Perfusion-Monitor, so dass es während der Induktion kann booten. </li><li> Legen Sie die Maus in die Induktionskammer. <ol><li> Die Atemfrequenz der Maus während der Induktion sorgfältig überwachen. </li><li> Sobald die Atemfrequenz ist visuell verlangsamt, entfernen Sie die Maus aus der Induktionskammer. </li><li> Führen Zehe Prise zu Angemessenheit der Narkose zu beurteilen. </li></ol></li><li> Mit der Maus richtig betäubt, statt der Maus in Rückenlage. </li><li> Legen Gesicht in Nasenkegel und sichern Sie alle Extremitäten an die Heizungs Tisch. <ol><li> Achten Sie besonders auf, um sicherzustellen, dass die Extremitäten sind in anatomischen Position gesichert, ohne deviatiauf der einen Seite. Wenn die Maus nicht richtig positioniert ist, ist es schwierig, Brustwandarterie Trennung während Sternotomie zu vermeiden. </li></ol></li><li> Mit Knipser oder handelsübliche Haarentfernungscreme, entfernen Haare von der Mittellinie Thorax und linken unteren Extremität Bauchseite. <ol><li> Bei der Verwendung von Haarentfernungscreme, Sahne vermeiden, dass an Ort und Stelle für mehr als 30 Sekunden, kann als Verätzungen auftreten. </li></ol></li><li> Titriert flüchtigen Narkosezerstäubers Konzentration, um eine ausreichende Anästhesie erhalten. <ol><li> Erwartete Verdampfer Fraktionen sind zwischen 1-5% mit Isofluran mit hohen Durchfluss O 2. </li><li> Flüchtige Narkosezerstäubers Konzentration sollte titriert werden, um Anästhesie bei chirurgischen Stimulierung während spontane Atmung. </li></ol></li></ol> 2. Rektale Sonde Laser-Doppler-Placement <ol><li> Legen geschmiert rektale Sonde in der Maus die Rektum. Sichern Sie in Ort, um opertriebs Bett. </li><li> Stellen Sie die Heizung Bett für eine Ziel rektale Temperatur von 36,5 ° C </li><li> Machen Sie kleine Inzision über Oberschenkelarterie Maus und sezieren Haut vom Unterhautgewebe. </li><li> Legen Laser-Doppler-Sonde über Oberschenkelarterie. </li><li> Stellen Sondenpositionen bis die Perfusion Monitor registriert mehr als 800 Einheiten Perfusion. <ol><li> Fest sichere Sonde an Ort und Stelle. Nicht ordnungsgemäß gesichert Sonden können falsch niedrige Perfusionsmessungen haben. </li></ol></li></ol> 3. Dissektion der Aorta Arch / subclavia <ol><li> Machen Sie eine 2 cm Hautschnitt über dem Brustbeinende und sanft sezieren Haut vom Unterhautgewebe. </li><li> Präparieren Sie die Unterkieferspeicheldrüse kostenlos. <ol><li> Wenn die Blutung auftritt, kann sanften Druck mit einem Wattestäbchen aufgetragen werden. </li><li> Teilen Sie die Unterkieferspeicheldrüse durch Mittellinie in der avaskulären-Ebene. </li></ol></li><li> Heben Sie mit einer Pinzette und Brustbein mit der scissors machen 1 cm Sternotomie Mittellinie durch die Mittellinie des Brustbeins. Jede Abweichung von der Mittellinie in der inneren Brustarterie Blutung, die schwer zu kontrollieren sein wird, führen. </li><li> Platzieren 5-0 Rückzugs Nähte auf jeder Seite an der Kante des Brustbein-und Einfahren Brustbein seitlich Befestigungsnähte an den Betriebs Bett. Vermeiden Rückzugs Nähte seitlich Pneumothorax zu verhindern. </li><li> Mit stumpf freie Band Muskeln entlang der Luftröhre. Die linke Gurt Muskel kann mit einer Schere geteilt werden, um die Exposition zu verbessern. </li><li> Präparieren Sie befreien den Thymus vom umgebenden Gewebe. Weiter stumpf, bis die großen Gefäße sichtbar gemacht werden. Seien Sie äußerst vorsichtig, um Eingang in den Brustraum zu verhindern. </li><li> Zeigen Gefäßklemmen auf Aortenbogen und linken Schlüsselbeinarterie. </li><li> Stellen Sie sicher, distalen Fluss hat entsprechend gestört. Dies wird als eine Reduktion von&gt; 90% in Perfusion Einheiten gesehen werden. <ol><li> Weiter Okklusion für desirot für 4-8 min. </li></ol></li><li> Gefäßklemme entfernen und überprüfen, Blutstillung vor Schließung der Brust. </li></ol> 4. Schließung von Sternotomy und Haut <ol><li> Entfernen der Rückzugs Naht auf der linken Seite der Maus. </li><li> Schließen Sie die Sternotomie mit der rechten Rückzug Naht. <ol><li> Eine einzelne Brustbeinnaht (mit der zuvor platziert Rückzug Stich) ist ausreichend für die Brustbeinverschluss. Platzieren anderen Stich ist unnötig und erhöht die Risiken der Pneumothorax und Blutungen. </li></ol></li><li> Schließen Haut mit fließendem 5-0 Stich. </li></ol> 5. Recovery-und postoperative Beurteilung <ol><li> Übertragen Sie die Maus, um Recovery-Käfig. Cage sollte auf einem Heizkissen, um die Umgebungstemperatur der Rückgewinnungskammer zu erhöhen und den Wärmeverlust an die Umwelt zu reduzieren platziert werden. </li><li> Die Maus auf Anzeichen Atemnot oder Krampfaktivität genau beobachten. Verwalten Analgesie pro Einrichtung Richtlinien. Euthanize Mäuse sofort, wenn Beschlagnahme oder Atemnot beobachtet. <ol><li> CO 2-Kammer-Euthanasie ist unsere bevorzugte Methode. Genickbruch ist eine weitere Option, wenn CO 2 ist nicht verfügbar. </li><li> Vollständige Rückgewinnung kann in 1-2 Stunden erwartet werden, abhängig von der Länge der flüchtigen Anästhetikums und der verwendeten Konzentration. </li></ol></li><li> Bringen Sie die Maus, um normalen Käfig. Zeigen Nahrung und Wasser statt auf dem Boden des Käfigs. </li><li> Bewerten neurologischen Status bei 12 h Abständen mit Basso Maus Skala für Locomotion 8. </li></ol>

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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protecting+the+ischemic+spinal+cord+during+aortic+clamping.+The+influence+of+anesthetics+and+hypothermia.">Protecting the ischemic spinal cord during aortic clamping. The influence of anesthetics and hypothermia.</a> Ann. Surg. , 409-515 (1992).</li><li>Qayumi, A. K., Janusz, M. T., Lyster, D. M., Gillespie, K. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+model+for+investigation+of+spinal+cord+injury+caused+by+aortic+cross-clamping.">Animal model for investigation of spinal cord injury caused by aortic cross-clamping.</a> J. Invest. Surg. 10, 47-52 (1997).</li><li>Lang-Lazdunski, L., Matsushita, K., Hirt, L., Waeber, C., Vonsattel, J. P., Moskowitz, M. A., Dietrich, W. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+Cord+Ischemia:+Development+of+a+model+in+the+mouse.">Spinal Cord Ischemia: Development of a model in the mouse.</a> Stroke. 31, 208-213 (2000).</li><li>Basso, D. M., Fisher, L. C., Anderson, A. J., Jakeman, L. B., McTigue, D. M., Popovich, P. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Basso+Mouse+Scale+for+locomotion+detects+differences+in+recovery+after+spinal+cord+injury+in+five+common+mouse+strains.">Basso Mouse Scale for locomotion detects differences in recovery after spinal cord injury in five common mouse strains.</a> J. Neurotrauma. 23, 635-659 (2006).</li><li>Kwon, B. K., Tetzlaff, W., Grauer, J. N., Beiner, J., Vaccaro, A. 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F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+cord+infarction:+etiology+and+outcome.">Spinal cord infarction: etiology and outcome.</a> Neurology. 47, 321-330 (1996).</li><li>Kakinohana, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delayed+paraplegia+after+spinal+cord+ischemic+injury+requires+caspase-3+activation+in+mice.">Delayed paraplegia after spinal cord ischemic injury requires caspase-3 activation in mice.</a> Stroke. 42 (8), 2302-2307 (2011).</li><li>Wang, Z., Yang, W., Britz, G. W., Lombard, F. W., Warner, D. 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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Post-stroke+hypothermia+provides+neuroprotection+through+inhibition+of+AMP-activated+protein+kinase.">Post-stroke hypothermia provides neuroprotection through inhibition of AMP-activated protein kinase.</a> J. Neurotrauma. 28 (7), 1281-1288 (2011).</li></ol>

Die Autoren erklären, keine finanziellen Interessen konkurrieren.

Paraplegie sekundäre Rückenmark-Ischämie Reperfusion ist das Ergebnis eines komplexen schlecht verstanden Pathologien 9. Während dieser wird am häufigsten nach thorakoabdominellen Aorten-Operation gesehen hat, kann eine Vielzahl von anderen Beleidigungen wie Aortendissektion, Trauma, Embolie, Vaskulitis, Hypotonie und systemischen 10 in Querschnittslähmung führen. Zum weiteren Verständnis dieser Verletzung zu gewinnen und Ziele für die Zukunft stellen, um diese Verletzung zu beseitigen, ha...

Untere Extremität Lähmung weiterhin thorakoabdominellen Interventionen erschweren. Die Verletzung, wie Rückenmark Ischämie-Reperfusion Verletzung (SCIR) bekannt ist, führt zu Lähmungen in bis zu 20% der Patienten mit hohem Risiko ein. Chirurgische Hilfsstoffe wie links Herz-Bypass-, Lenden-cerbrospinal Flüssigkeit Kanalisation, hypothermen Kreislaufstillstand und Zwischenrippenarterie Reimplantation haben die Inzidenz dieser Komplikation 2 reduziert, aber viel zu viele Patienten weiterhin betroffen sein. Klinisch, Rückenmark Ischämie und Reperfusion wird entweder sofort oder verzögert Lähmung nach der Intervention 3 zu sehen. Allerdings hat unser Verständnis von dieser Verletzung durch einen Mangel an mechanistischen Detail erstickt worden. Als Ergebnis sind einige Optionen zur Verfügung, um die Schädigung zu dämpfen, wenn es aufgetreten ist. Wir haben so ein kleines Tier angeworben, murinen Modell des Rückenmarks Ischämie und Reperfusion zubesser zu charakterisieren seine Pathogenese. Die Mehrzahl der Studien haben bisher größere Tiermodelle verwendet werden, um diese Verletzung, nämlich Ratte 4, 5 Kaninchen, Schwein und 6 Modelle charakterisieren. Diese sind aber durch ihre Kosten, Komplexität, variable Reproduzierbarkeit, und, am wichtigsten, der Mangel an verfügbaren Techniken zur genetischen Manipulation beschränkt. Die zuverlässigste dieser veröffentlicht Tiermodellen beinhaltet infra Abklemmen der Bauchaorta bei Kaninchen. Allerdings menschlichen vorderen Rückenmarkneuronen am häufigsten leiten ihre Gefäßversorgung von mehr proximalen Äste 7. Variable vaskulären Anatomie der Wirbelsäule in diesen Modellen addiert zu Schwierigkeiten beim Übergang ihre Ergebnisse in die klinische Verwendung. Dieses Manuskript präsentiert ein Modell für die sofortige oder spätere Querschnittslähmung nach Thorax Aortenokklusionsvorrichtung, die klinisch relevant und einfach zu verwenden ist. Die Belichtung der Aortenbogen über Mini sternotomeine ist weniger invasiv und hoch reproduzierbare Ergebnisse mit minimaler Morbidität und Mortalität zu entlocken. Während dieses Modell in nicht ohne Herausforderungen und technischen Nuancen, können diese mit einer sorgfältigen Präparation und Gewebebehandlung überwunden werden, um ein Modell der Hinterbeinlähmung, die leicht implementiert werden kann produzieren.

<table><tbody><tr><td>VMS Anesthesia Machine</td><td>MDS Matrx</td><td></td><td></td></tr><tr><td>Isoflurane</td><td>Vet One</td><td>13985-528-60</td><td>2.0% through nose cone&amp;nbsp;</td></tr><tr><td>Induction Chamber</td><td>Vet Equip</td><td>941444</td><td></td></tr><tr><td>Heating Bed</td><td>Vestavia Scientific</td><td></td><td></td></tr><tr><td>Lazer Doppler Monitor</td><td>Moor Instruments</td><td>VMS-LDF1</td><td></td></tr><tr><td>5-0 Suture, Polyester</td><td>Surgidac</td><td>VD-551</td><td>Taper Needle</td></tr><tr><td>Microdissecting Clips</td><td>Biomedical Research Instruments</td><td>14-1030, 14-1060</td><td></td></tr><tr><td>Surgical Instruments</td><td>Fine Surgical Instruments</td><td></td><td>Forceps, needle holder</td></tr></tbody></table>

reproducable paraplegia by thoracic aortic occlusion in a murine model of spinal cord ischemia-reperfusion

Hintergrund Untere Extremität Lähmung weiterhin Aorten-Interventionen erschweren. Der Mangel an Verständnis für die zugrunde liegende Pathologie hat Fortschritte behindert, um das Auftreten dieser Verletzung zu verringern. Das aktuelle Modell zeigt reproduzierbare Lähmung der unteren Extremitäten nach thorakalen Aorta Okklusion. Methoden Erwachsene männliche C57BL6 Mäuse wurden mit Isofluran betäubt. Durch eine cervicosternal Einschnitt wurde die Aorta freigelegt. Die absteigenden Aorta und linken Schlüsselbeinarterien wurden ohne Eintritt in Pleuraraum identifiziert. Skelettierung dieser Arterien wurde durch sofortige Schließung (Sham) oder Verschluss für 4 min (moderaten Ischämie) oder 8 min (verlängerte Ischämie) gefolgt. Die Sternotomie und die Haut wurden geschlossen und die Maus wurde für die Wiederherstellung, um Wärmebett übertragen. Nach Wiederherstellung der Funktionsanalyse wurde bei 12 h Intervallen bis zu 48 h erhalten. Ergebnisse Mäuse, die Schein-Operation unterzogen zeigte keine beobachtbare Hinterbein Defizit. Mäuse zu moderaten Ischämie für 4 min unterzogen hatten minimale Funktionsdefizit auf 12 Stunden, gefolgt von Progression zu einer Lähmung bei 48 Stunden zu beenden. Mäuse verlängert Ischämie hatte eine sofortige Lähmung ohne beobachtbare Hintergliedbewegung an einem beliebigen Punkt in der postoperativen Phase. Es gab keine beobachtet intraoperative oder postoperative Mortalität. Abschluss Reproduzierbare Lähmung der unteren Extremitäten, ob sofort oder verzögert kann in einem Mausmodell erreicht werden. Zusätzlich wird durch Verwendung eines Längssternotomie und sorgfältige Präparation, hohe Überlebensraten und Reproduzierbarkeit erreicht werden kann.

Mäuse Scheinoperation unterzogen (n = 3) oder Aortenokklusionsvorrichtung 4 (n = 3) bis 8 min (n = 3). Postoperativ wurden die Mäuse durch das Basso Maus Punktzahl (1) bewertet. Mäuse, die Scheinoperation unterzogen hatte keine feststellbaren Funktionsdefizite an jedem Punkt postoperativ. Mäuse zu moderaten Ischämie (4 min) unterzogen hatten in der Nähe von normalen Hintergliedmaßenfunktion bei 12 h mit progressiven Funktionsabnahme zu Lähmungen von 48 Stunden zu beenden. Mäuse in längerer Isc...

Watch this Scientific Journal Video about Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion at JoVE.com

Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion

Der Mangel an Verständnis der Mechanismen des Rückenmarks Ischämie-Reperfusion Verletzungen hat weitere Zusätze behindert eine Querschnittslähmung nach Aorten-Operationen mit hohem Risiko zu verhindern. Somit ist die Entwicklung von Tiermodellen zwingend. Dieses Manuskript zeigt, reproduzierbare Lähmung der unteren Extremitäten nach Thorax Aortenokklusionsvorrichtung in einem Mausmodell.

Reproduzierbare Paraplegie von thorakalen Aorta Occlusion in einem Mausmodell der Spinal Cord Ischämie-Reperfusion

Background
Lower extremity paralysis continues to complicate aortic interventions. The lack of understanding of the underlying pathology has hindered ...

reproducable-paraplegia-thoracic-aortic-occlusion-murine-model-spinal

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

12.0K Aufrufe.  University of Colorado.  Der Mangel an Verständnis der Mechanismen des Rückenmarks Ischämie-Reperfusion Verletzungen hat weitere Zusätze behindert eine Querschnittslähmung nach Aorten-Operationen mit hohem Risiko zu verhindern. Somit ist die Entwicklung von Tiermodellen zwingend. Dieses Manuskript zeigt, reproduzierbare Lähmung der unteren Extremitäten nach Thorax Aortenokklusionsvorrichtung in einem Mausmodell. 

Serie: Reproducable Paraplegia by Thoracic Aortic Occlusion in a Murine Model of Spinal Cord Ischemia-reperfusion

Murine Model of Hindlimb Ischemia 

In the United States, peripheral arterial disease (PAD) affects about 10 million individuals, and is also prevalent worldwide. Medical therapies for symptomatic relief are limited. Surgical or endovascular interventions are useful for some individuals, but long-term results are often disappointing. As a result, there is a need for developing new therapies to treat PAD. The murine hindlimb ischemia preparation is a model of PAD, and is useful for testing new therapies. When compared to other models of tissue ischemia such as coronary or cerebral artery ligation, femoral artery ligation provides for a simpler model of ischemic tissue. Other advantages of this model are the ease of access to the femoral artery and low mortality rate. 
In this video, we demonstrate the methodology for the murine model of unilateral hindimb ischemia. The specific materials and procedures for creating and evaluating the model will be described, including the assessment of limb perfusion by laser Doppler imaging. This protocol can also be utilized for the transplantation and non-invasive tracking of cells, which is demonstrated by Huang et al.1.

Murine Model of Hindlimb Ischemia

The surgical procedure for induction of unilateral hindlimb ischemia is demonstrated, with confirmation of ischemia by laser Doppler perfusion imaging.

Mausmodell der hinteren Gliedmaßen Ischämie

In the United States, peripheral arterial disease (PAD) affects about 10 million individuals, and is also prevalent worldwide.  Medical therapies for ...

Forschung

Biologie

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

A cervical spinal cord injury induces permanent paralysis, and often leads to respiratory distress. To date, no efficient therapeutics have been developed to improve/ameliorate the respiratory failure following high cervical spinal cord injury (SCI). Here we propose a murine pre-clinical model of high SCI at the cervical 2 (C2) metameric level to study diverse post-lesional respiratory neuroplasticity. The technique consists of a surgical partial injury at the C2 level, which will induce a hemiparalysis of the diaphragm due to a deafferentation of the phrenic motoneurons from the respiratory centers located in the brainstem. The contralateral side of the injury remains intact and allows the animal recovery. Unlike other SCIs which affect the locomotor function (at the thoracic and lumbar level), the respiratory function does not require animal motivation and the quantification of the deficit/recovery can be easily performed (diaphragm and phrenic nerve recordings, whole body ventilation). This pre-clinical C2 SCI model is a powerful, useful, and reliable pre-clinical model to study various respiratory and non-respiratory neuroplasticity events at different levels (molecular to physiology) and to test diverse putative therapeutic strategies which might improve the respiration in SCI patients.

Respiratory failure is the leading cause of death following a cervical spinal cord injury. Having a reproducible, quantifiable, and reliable pre-clinical animal model of respiratory failure induced by a partial cervical injury will help to understand the subsequent respiratory and non-respiratory neuroplasticity and allow testing putative repair strategies.

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 ...

Medizin

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms at work during MI and developing effective therapeutic approaches requires methodology to reproducibly simulate the clinical incidence and reflect the pathophysiological changes associated with MI. Here, we describe a surgical method to induce MI in mouse models that can be used for short-term ischemia-reperfusion (I/R) injury as well as permanent ligation. The major advantage of this method is to facilitate location of the left anterior descending artery (LAD) to allow for accurate ligation of this artery to induce ischemia in the left ventricle of the mouse heart. Accurate positioning of the ligature on the LAD increases reproducibility of infarct size and thus produces more reliable results. Greater precision in placement of the ligature will improve the standard surgical approaches to simulate MI in mice, thus reducing the number of experimental animals necessary for statistically relevant studies and improving our understanding of the mechanisms producing cardiac dysfunction following MI. This mouse model of MI is also useful for the preclinical testing of treatments targeting myocardial damage following MI.

We introduce a surgical method to induce experimental ischemia/reperfusion (I/R) injury to simulate myocardial infarction (MI) in mouse models that allows for more clarity in positioning of the ligation on the left anterior descending artery (LAD) to increase the reproducibility of MI experiments in mice.

Einem Mausmodell der myokardialen Ischämie-Reperfusion Injury durch Ligation des linken vorderen absteigenden Arterie

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) below the site of injury. The secondary ischemia that develops following the initial mechanical insult is a serious complication of the SCI and severely impairs the function and viability of surviving neuronal and non-neuronal cells in the SC. In addition, ischemia is also responsible for the growth of lesion during chronic phase of injury and interferes with the cellular repair and healing processes. Thus there is a need to develop a spinal cord ischemia model for studying the mechanisms of ischemia-induced pathology. Focal ischemia induced by photothrombosis (PT) is a minimally invasive and very well established procedure used to investigate the pathology of ischemia-induced cell death in the brain. Here, we describe the use of PT to induce an ischemic lesion in the spinal cord of mice. Following retro-orbital sinus injection of Rose Bengal, the posterior spinal vein and other capillaries on the dorsal surface of SC were irradiated with a green light resulting in the formation of a thrombus and thus ischemia in the affected region. Results from histology and immunochemistry studies show that PT-induced ischemia caused spinal cord infarction, loss of neurons and reactive gliosis. Using this technique a highly reproducible and relatively easy model of SCI in mice can be achieved that would serve the purpose of scientific investigations into the mechanisms of ischemia induced cell death as well as the efficacy of neuroprotective drugs. This model will also allow exploration of the pathological changes that occur following SCI in live mice like axonal degeneration and regeneration, neuronal and astrocytic Ca2+ signaling using two-photon microscopy.

Photothrombosis is a minimally invasive and highly reproducible procedure to induce focal ischemia in the spinal cord and serves as a model of spinal cord injury in mice.

Photothrombosis induzierte fokale Ischämie als ein Modell der Rückenmarksverletzungen in Mäusen

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal cord. A major current area of research is focused on the mechanisms of adaptive plasticity that underlie spontaneous or induced functional recovery following SCI. Spontaneous functional recovery is reported to be greater early in life, raising interesting questions about how adaptive plasticity changes as the spinal cord develops. To facilitate investigation of this dynamic, we have developed a SCI model in the neonatal mouse. The model has relevance for pediatric SCI, which is too little studied. Because neural plasticity in the adult involves some of the same mechanisms as neural plasticity in early life1, this model may potentially have some relevance also for adult SCI. Here we describe the entire procedure for generating a reproducible spinal cord compression (SCC) injury in the neonatal mouse as early as postnatal (P) day 1. SCC is achieved by performing a laminectomy at a given spinal level (here described at thoracic levels 9-11) and then using a modified Yasargil aneurysm mini-clip to rapidly compress and decompress the spinal cord. As previously described, the injured neonatal mice can be tested for behavioral deficits or sacrificed for ex vivo physiological analysis of synaptic connectivity using electrophysiological and high-throughput optical recording techniques1. Earlier and ongoing studies using behavioral and physiological assessment have demonstrated a dramatic, acute impairment of hindlimb motility followed by a complete functional recovery within 2 weeks, and the first evidence of changes in functional circuitry at the level of identified descending synaptic connections1.

This article describes a method for generating a reproducible spinal cord compression injury (SCI) in the neonatal mouse. The model provides an advantageous platform for studying mechanisms of adaptive plasticity that underlie spontaneous functional recovery.

Ein Neonatal Maus Spinal Cord Injury Kompression Modell

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Murine Modell der intestinalen Ischämie-Reperfusionsschaden

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These complications can be attributed to impairments in re-endothelialization within the wound margins. Yet, the cellular and molecular mechanisms of re-endothelialization remain to be defined. While several animal models to study re-endothelialization after arterial denudation are available, few are performed in the mouse because of surgical limitations. This undermines the opportunity to exploit transgenic mouse lines and investigate the contribution of specific genes to the process of re-endothelialization. Here, we present a step-by-step protocol for creating a highly reproducible murine model of arterial denudation injury in the infrarenal abdominal aorta using external vascular clamping. Immunocytochemical staining of injured aortas for fibrinogen and &#946;-catenin demonstrate the exposure of a pro-thrombotic surface and the border of intact endothelium, respectively. The method presented here has the advantages of speed, excellent overall survival rate, and relative technical ease, creating a uniquely practical tool for imposing arterial denudation injury in transgenic mouse models. Using this method, investigators may elucidate the mechanisms of re-endothelialization under normal or pathological conditions.

Understanding the cellular and molecular mechanisms of re-endothelialization following arterial denudation injury is of paramount importance in preventing thrombosis and restenosis of arteries. Here we describe a protocol for reproducible arterial denudation injury of the infrarenal abdominal aorta. The procedure was developed to investigate the underlying mechanisms that regulate endothelial regeneration using mouse models.

Reproduzierbare Arterielle Denudation Verletzung von Infra Abdominal Aortic Spannen in einem Mausmodell

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

Reproducible Motor Deficit Following Aortic Occlusion in a Rat Model Of Spinal Cord Ischemia

Spinal cord ischemia is a fatal complication following thoracoabdominal aortic aneurysm surgery. Researchers can investigate the strategies for preventing and treating this complication using experimental models of spinal cord ischemia. The model described here demonstrates varying degrees of paraplegia that relate to the length of occlusion following thoracic aortic occlusion in a rat spinal cord ischemia model.
A 2-Fr. balloon-tipped catheter was advanced through the femoral artery into the descending thoracic aorta until the catheter tip was placed at the left subclavian artery in anesthetized male Sprague-Dawley rats. Spinal cord ischemia was induced by inflating the catheter balloon. After a set period of occlusion (9, 10, or 11 min), the balloon was deflated. Neurologic assessment was performed using the motor deficit index at 24 h after surgery, and the spinal cord was harvested for histopathological examination.
Rats that underwent 9 min of aortic occlusion showed mild and reversible motor impairment in the hind limb. Rats subjected to 10 min of aortic occlusion presented with moderate but reversible motor impairment. Rats subjected to 11 min of aortic occlusion displayed complete and persistent paralysis. The motor neurons in the spinal cord sections were more preserved in rats subjected to shorter duration of aortic occlusion.
Researchers can achieve a reproducible hind limb motor deficit following thoracic aortic occlusion using this spinal cord ischemia model.

This study demonstrates the technique to make a minimally invasive and easily reproducible model of spinal cord ischemia in rats. Various degrees of hind limb motor deficit can be produced by controlling the aortic occlusion time.

Reproduzierbare motorische Defizit nach Aortengelenk in einem Rattenmodell der Rückenmark Ischämie

Spinal cord ischemia is a fatal complication following thoracoabdominal aortic aneurysm surgery. Researchers can investigate the strategies for preventing ...

Two-vessel Occlusion Mouse Model of Cerebral Ischemia-reperfusion

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse model is useful for investigating the pathophysiology of cerebral ischemia-reperfusion and determining potential therapeutic strategies for patients with stroke. Variations in the anatomy of the circle of Willis of C57BL/6 mice affects their infarct volume after cerebral-ischemia-induced injury. Studies have indicated that distal MCA occlusion (MCAO) can overcome this problem and result in a stable infarct size. In this study, we establish a two-vessel occlusion mouse model of cerebral ischemia-reperfusion through the interruption of the blood flow to the right MCA. We distally ligate the right MCA and right common carotid artery (CCA) and restore blood flow after a certain period of ischemia. This ischemia-reperfusion injury induces an infarct of stable size and a behavioral deficit. Peripheral immune cells infiltrate the ischemic brain within the 24 h infiltration period. Additionally, the neuronal loss in the cortical area is less for a longer reperfusion duration. Therefore, this two-vessel occlusion model is suitable for investigating the immune response and neuronal recovery during the reperfusion period after cerebral ischemia.

A mouse model of cerebral ischemia-reperfusion is established to investigate the pathophysiology of stroke. We distally ligate the right middle cerebral artery and right common carotid artery and restore blood flow after 10 or 40 min of ischemia.

Zwei-Schiff Okklusion Mausmodell der zerebralen Ischämie-reperfusion

In this study, a middle cerebral artery (MCA) occlusion mouse model is employed to study cerebral ischemia-reperfusion. A reproducible and reliable mouse ...

Induction of Complete Transection-Type Spinal Cord Injury in Mice

Spinal cord injury (SCI) largely leads to irreversible and permanent loss of function, most commonly as a result of trauma. Several treatment options, such as cell transplantation methods, are being researched to overcome the debilitating disabilities arising from SCI. Most pre-clinical animal trials are conducted in rodent models of SCI. While rat models of SCI have been widely used, mouse models have received less attention, even though mouse models can have significant advantages over rat models. The small size of mice equates to lower animal maintenance costs than for rats, and the availability of numerous transgenic mouse models is advantageous for many types of studies. Inducing repeatable and precise injury in the animals is the primary challenge for SCI research, which in small rodents requires high-precision surgery. The transection-type injury model has been a commonly used injury model over the last decade for transplantation-based therapeutic research, however a standardized method for inducing a complete transection-type injury in mice does not exist. We have developed a surgical protocol for inducing a complete transection type injury in C57BL/6 mice at thoracic vertebral level 10 (T10). The procedure uses a small tip drill instead of rongeurs to precisely remove the lamina, after which a thin blade with rounded cutting edge is used to induce the spinal cord transection. This method leads to reproducible transection-type injury in small rodents with minimal collateral muscle and bone damage and therefore minimizes confounding factors, specifically where behavioral functional outcomes are analyzed.

This protocol describes how to create a precise laminectomy for induction of stable transection-type spinal cord injury in the mouse model, with minimal collateral damage for spinal cord injury research.

Induktion von vollständiger Transektion-Typ Rückenmarksverletzung bei Mäusen

Spinal cord injury (SCI) largely leads to irreversible and permanent loss of function, most commonly as a result of trauma. Several treatment options, ...