Wir möchten die Bemühungen von Dr. Mary-Lou Vallano, Department of Neuroscience und Physiologie, für ihre wertvolle Inputs erkennen im Schreiben bis zu diesem Manuskript.

Ein. Rat Chirurgie für SAH Betreff Mit 0,15 ml Autologe arterielle Blut injiziert <ol><li> Die Ratte ist betäubt mit 0,1 mg / kg Ketamin / Xylazin Nager Cocktail und darf für 5 min zu sitzen. </li><li> Eine adäquate Anästhesie wird durch Reduktion in Hinterbein reflex bestätigt. </li><li> Mit Hilfe eines elektronischen Rasierer a Hals Bereich der Haare rund um die sub-Okzipitalregion Nase ist rasiert. </li><li> Das Tier wird in Rückenlage auf dem OP-Tisch gelegt und der Schwanz ist mit betadine abgetupft, um eine sterile Schnitt zu gewährleisten. </li><li> Ein gerader 1 cm Inzision auf der ventralen Seite des Schwanzes ergriffen </li><li> Die Präparation wird verlängert, bis der Schwanzarterie wird identifiziert und isoliert. </li><li> Verwendung einer sterilen 26-Gauge-Katheters wird der Schwanzarterie kanüliert und 0,15 ml arterielle Blut wird in eine Spritze gezogen. </li><li> Eine sterile Gaze gehüllt um den Einschnitt, um Hämostase vor dem Aufbringen der vetbond den Einschnitt abzudichten gewährleisten. </li><li> TEr rat eingeschaltet ist bäuchlings auf dem Tisch und dem rasierten Bereich über dem Sub-okzipitalen Region mit betadine abgetupft. </li><li> Verwendung einer vertikalen Mittellinieneinschnitt Zugang wird dem cisterna magna gewonnen. </li><li> Sobald sie identifiziert sind eine 25 Gauge-Nadel in die Zisterne magna eingefügt und 0,15 ml CSF wird in eine Spritze zu einer erhöhten intrakraniellen Druck mit Injektion von Eigenblut Volumen vermeiden zurückgezogen. </li><li> Nun wird das 0,15 ml Blut aus der Schwanzarterie entnommen langsam in die Zisterne magna injiziert. </li><li> Die Nadel wird in für 30 sec bleiben, um sicherzustellen Gerinnungszeit im Subarachnoidalraum und dann vorsichtig abgezogen. </li><li> Hämostase gewährleistet ist und der Einschnitt wird geschlossen mit einer Heftvorrichtung. </li><li> Das Tier ist nun platziert neigen mit 20 ° Kopf Position für 20 min auf einer Erwärmung Oberfläche Blut zu ermöglichen, in den Zisternen um die basilaris erstarren. </li><li> Schritte 1.1 bis 1,15 sind während der zweiten Operation 48 hr voneinander wiederholt. </li&gt; </ol> 2. Rat Chirurgie für SAH Betreff mit 0,15 ml Kochsalzlösung injiziert <ol><li> Die Ratte ist betäubt mit 0,1 mg / kg Ketamin / Xylazin Nager Cocktail und darf für 5 min zu sitzen. </li><li> Eine adäquate Anästhesie wird durch Reduktion in Hinterbein reflex bestätigt. </li><li> Mit Hilfe eines elektronischen Rasierer a Hals Bereich der Haare rund um die sub-Okzipitalregion Nase ist rasiert. </li><li> Das Tier wird in Rückenlage auf dem OP-Tisch gelegt und der Schwanz ist mit betadine abgetupft, um eine sterile Schnitt zu gewährleisten. </li><li> Ein gerader 1 cm Inzision auf der ventralen Seite des Schwanzes ergriffen </li><li> Die Präparation wird verlängert, bis der Schwanzarterie wird identifiziert und isoliert. </li><li> Verwendung einer sterilen 26-Gauge-Katheters wird der Schwanzarterie kanüliert und 0,15 ml arterielle Blut wird in eine Spritze gezogen. </li><li> Eine sterile Gaze gehüllt um den Einschnitt, um Hämostase vor dem Aufbringen der vetbond den Einschnitt abzudichten gewährleisten. </li><li> Die rat eingeschaltet ist auf dem Tisch und dem rasierten Gebiet anfällig ist mit betadine übermalt Sub-okzipitalen Region. </li><li> Verwendung einer vertikalen Mittellinieneinschnitt Zugang wird dem cisterna magna gewonnen. </li><li> Sobald sie identifiziert sind eine 25 Gauge-Nadel in die Zisterne magna eingefügt und 0,15 ml CSF wird in eine Spritze aufgezogen und die Probe wird gespeichert. </li><li> Nun wird das 0,15 ml physiologischer Kochsalzlösung (37 ° C) langsam in die Zisterne magna injiziert. </li><li> Die Nadel wird in für 30 sec links und vorsichtig abgezogen. </li><li> Hämostase gewährleistet ist und der Einschnitt wird geschlossen mit einer Heftvorrichtung. </li><li> Das Tier ist nun platziert neigen mit 20 ° Kopf Position für 20 min auf einer Erwärmung Oberfläche. </li><li> Schritte 2,1-2,15 werden während der zweiten Operation 48 hr voneinander wiederholt. </li></ol> 3. Rat Sacrifice <ol><li> Am Tag 5 nach der zweiten Operation werden die Ratten durch kardiale Perfusion geopfert. </li><li> Die Ratte wird eine tödliche Dosis (0 gegeben0,2 ml / kg) von Fatal Plus (Vortech PHARMACEUTICALS LTD., Dearborn, MI) </li><li> Mit einer vertikalen Inzision wird die Bauchhöhle näherte und das Peritoneum eröffnet. </li><li> Eine anteriore Thorakotomie durchgeführt wird und das Herz freigelegt ist. </li><li> Verwendung einer 26-Gauge-Katheter mit einer Phosphatpufferlösung (PBS, pH 7,4 und bei 37 °) wird das Tier von Blut entleert und danach mit 4% Paraformaldehyd perfundiert. </li><li> Nach Sicherstellung einer angemessenen Perfusion wird die Perfusion gestoppt und die Ratte wird über die Enthauptung Tisch gebracht. </li><li> Nach Enthauptung, ist ein Knochen Rongeur verwendet werden, um den Schädel für Gehirn Entfernen entfernen. </li><li> Das Gehirn und Hirnstamm werden sorgfältig aus der Schädelhöhle extrahiert und in eine 4% Paraformaldehyd-Lösung und bei 4 ° C für 48 Stunden. </li></ol> 4. Erstellen von Abschnitten, um Vasospasmus beurteilen <ol><li> Die Rattengehirn, die nun in 30% Saccharose wurde für 4 Tage eingetaucht ist, um th gebrachte Kryostat zum Schneiden. </li><li> Sobald cryoprotected, 12 uM Abschnitte werden mit dem Kryostaten mit dem Anterior Inferior Kleinhirnarterie (AICA) als Ausgangspunkt, um inter-subject Konsistenz zu gewährleisten. </li><li> 20 Abschnitten werden für jedes Tier geschaffen, endend bei dem Superior Kleinhirnarterie (SCA). </li><li> Die Schnitte werden auf einen Objektträger platziert und auf eine mögliche Verwendung von histologischen Vasospasmus Methodik </li></ol>

<ol>
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M., Guarino, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+blood+flow+and+cerebral+perfusion+pressure+in+a+new+noncraniotomy+model+of+subarachnoid+hemorrhage+in+the+rat.">Cortical blood flow and cerebral perfusion pressure in a new noncraniotomy model of subarachnoid hemorrhage in the rat.</a> Stroke. 26, 1086-1091 (1995).</li><li>Cheng, G., Wei, L., Zhi-Dan, S., Shi-Guang, Z., Xiang-Zhen, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Atorvastatin+ameliorates+cerebral+vasospasm+and+early+brain+injury+after+subarachnoid+hemorrhage+and+inhibits+caspase-dependent+apoptosis+pathway.">Atorvastatin ameliorates cerebral vasospasm and early brain injury after subarachnoid hemorrhage and inhibits caspase-dependent apoptosis pathway.</a> BMC Neurosci. 10, 7-17 (2009).</li><li>Jackowski, A., Crockard, A., Burnstock, G., Russell, R. 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Methods. 50, 301-307 (1993).</li><li>Karaoglan, A., Akdemir, O., Barut, S., Kokturk, S., Uzun, H., Tasyurekli, M., Colak, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+resveratrol+on+vasospasm+after+experimental+subarachnoid+hemorrhage+in+rats.">The effects of resveratrol on vasospasm after experimental subarachnoid hemorrhage in rats.</a> Surg. Neurol. 70, 337-343 (2008).</li><li>Lee, J. Y., Huang, D. L., Keep, R., Sagher, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+an+improved+double+hemorrhage+rat+model+for+the+study+of+delayed+cerebral+vasospasm.">Characterization of an improved double hemorrhage rat model for the study of delayed cerebral vasospasm.</a> J. Neurosci. Methods. 168, 358-366 (2008).</li><li>Lee, J. Y., Sagher, O., Keep, R., Hua, Y., Xi, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+experimental+rat+models+of+early+brain+injury+after+subarachnoid+hemorrhage.">Comparison of experimental rat models of early brain injury after subarachnoid hemorrhage.</a> Neurosurgery. 65 (2), 331-343 (2009).</li><li>Megyesi, J. F., Vollrath, B., Cook, D. A., Findlay, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+animal+models+of+cerebral+vasospasm:+a+review.">In vivo animal models of cerebral vasospasm: a review.</a> Neurosurgery. 46, 448-460 (2000).</li><li>Prunell, G. F., Mathiesen, T., Diemer, N. H., Svendgaard, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+subarachnoid+hemorrhage:+Subarachnoid+blood+volume,+mortality+rate,+neuronal+death,+cerebral+blood+flow,+and+perfusion+pressure+in+three+different+rat+models.">Experimental subarachnoid hemorrhage: Subarachnoid blood volume, mortality rate, neuronal death, cerebral blood flow, and perfusion pressure in three different rat models.</a> Neurosurgery. 52, 165-176 (2003).</li><li>Prunell, G. F., Mathiesen, T., Svendgaard, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+subarachnoid+hemorrhage:+Cerebral+blood+flow+and+brain+metabolism+during+the+acute+phase+in+three+different+models+in+the+rat.">Experimental subarachnoid hemorrhage: Cerebral blood flow and brain metabolism during the acute phase in three different models in the rat.</a> Neurosurgery. 54, 426-436 (2004).</li><li>Ryba, M. S., Gordon-Krajcer, W., Walski, M., Chalimoniuk, M., Chrapusta, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hydroxylamine+attenuates+the+effects+of+simulated+subarachnoid+hemorrhage:+implication+for+the+role+of+oxidative+stress+in+cerebral+vasospasm.">Hydroxylamine attenuates the effects of simulated subarachnoid hemorrhage: implication for the role of oxidative stress in cerebral vasospasm.</a> Neurol. Res. 31, 195-199 (1999).</li><li>Satoh, M., Parent, A. D., Zhang, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhibitory+effect+with+antisense+mitogen-activated+protein+kinase+oligodeoxynucleotide+against+cerebral+vasospasm+in+rats.">Inhibitory effect with antisense mitogen-activated protein kinase oligodeoxynucleotide against cerebral vasospasm in rats.</a> Stroke. 33, 775-781 (2002).</li><li>Suzuki, H., Kanamaru, K., Tsunoda, H., Inada, H., Kuroki, M., Sun, H., Waga, S., Tanaka, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heme+oxygenase-1+gene+induction+as+an+intrinsic+regulation+against+delayed+cerebral+vasospasm+in+rats.">Heme oxygenase-1 gene induction as an intrinsic regulation against delayed cerebral vasospasm in rats.</a> J. Clin. Invest. 104, 59-66 (1999).</li><li>Swift, D. M., Solomon, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subarachnoid+hemorrhage+fails+to+produce+vasculopathy+or+chronic+blood+flow+changes+in+rats.">Subarachnoid hemorrhage fails to produce vasculopathy or chronic blood flow changes in rats.</a> Stroke. 19, 878-882 (1988).</li><li>Vatter, H., Weidauer, S., Konczalla, J., Dettmann, E., Zimmermann, M., Raabe, A., Preibisch, C., Zanella, F., Seifert, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Time+course+in+the+development+of+cerebral+vasospasm+after+experimental+subarachnoid+hemorrhage:+clinical+and+neuroradiological+assessment+of+the+rat+double+hemorrhage+model.">Time course in the development of cerebral vasospasm after experimental subarachnoid hemorrhage: clinical and neuroradiological assessment of the rat double hemorrhage model.</a> Neurosurgery. 58, 1190-1197 (2006).</li><li>Veelken, J. A., Laing, R. J., Jakubowski, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Sheffield+model+of+subarachnoid+hemorrhage+in+rats.">The Sheffield model of subarachnoid hemorrhage in rats.</a> Stroke. 26, 1279-1283 (1995).</li><li>Zubkov, A. Y., Nanda, A., Zhang, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Signal+transduction+pathways+in+cerebral+vasospasm.">Signal transduction pathways in cerebral vasospasm.</a> Pathophysiology. 9, 47-61 (2003).</li></ol>

Wir haben nichts zu im Zusammenhang mit dieser Studie offen zu legen.

Primaten, eine weitere ähnliche genetische Zusammensetzung und anatomischen Merkmalen der Mensch, genauer nachahmen die Ereignisse des verzögerten zerebralen Vasospasmus und kann leichter zu unterziehen nicht-invasive Bildgebung (MRI und Angiographie), um arterielle Veränderungen, als Nagetiere 8 überwachen. Allerdings sind Primatenmodelle kostspielig und mit komplexer Pflege und ethische Fragen, als kleine Tiermodellen. Kleintier-SAH Modelle, die entwickelt wurden vorher für drei Methoden zur Induktion ...

<table border="1"><tbody><tr><td> Name der Ausrüstung / Reagenz </td><td> Firma </td><td> Katalog-Nummer </td></tr><tr><td> Männliche SD-Ratten (250-300 g) </td><td> Taconic </td><td> SD-M </td></tr><tr><td> 26 G Katheter </td><td> Webster </td><td> 8416683 </td></tr><tr><td> 25 G Nadeln </td><td> Büffel </td><td> 305122 </td></tr><tr><td> 1 cc Spritzen </td><td> Central speichert </td><td> 54245 </td></tr><tr><td> Ketamin / Xylazin-Cocktail </td><td> Animal Care (SUNY) * </td><td> - </td></tr><tr><td> Betadine </td><td> Central speichert </td><td> 51458 </td></tr><tr><td> Saccharose </td><td> Sigma </td><td> S9378-1 kg </td></tr><tr><td> Paraformaldehyd </td><td> Sigma </td><td> P6148-500G </td></tr><tr><td> Phosphatpufferlösung </td><td> Fischer </td><td> BP-399-4 </td></tr><tr><td> OP-Tisch </td><td> Harvard </td><td> PY2 72-2590 </td></tr><tr><td> Oktober Compound (Kryokonservierung) </td><td> VWR </td><td> 25608-930 </td></tr><tr><td> Superfrost Slides </td><td> Fischer </td><td> 12-550-15 </td></tr></tbody></table> * Bei Department of Laboratory Animal Care, SUNY Upstate Medical University synthetisiert. Fügen Sie 1 cc [100 mg / ml] von Xylazin bis 10 ml [100 mg / ml] von Ketamin.

a low mortality rat model to assess delayed cerebral vasospasm after experimental subarachnoid hemorrhage

Zielsetzung: charakterisieren und festzustellen, die ein reproduzierbares Modell verzögert zerebralen Vasospasmus nach aneurysmatischer veranschaulicht Subarachnoidalblutung (SAB) bei Ratten, um die auslösenden Ereignissen, pathophysiologische Veränderungen und potentielle Ziele für die Behandlung zu identifizieren. Methoden: Achtundzwanzig männliche Sprague-Dawley-Ratten (250 - 300 g) - SAH oder Kochsalzlösung Kontrolle wurden willkürlich einer von zwei Gruppen zugeordnet. Rat Subarachnoidalblutung in der SAH-Gruppe (n = 15) wurde durch doppelte Injektion von Eigenblut induziert, 48 hr auseinander, in die Cisterna magna. Ähnlich normaler Kochsalzlösung (n = 13) wurde in die Zisterne magna der Salzlösungs-Kontrollgruppe injiziert. Die Ratten wurden am Tag fünf nach dem zweiten Blut-Injektion getötet und die Gehirne wurden für die histologische Analyse aufbewahrt. Der Grad der Vasospasmus wurde unter Verwendung Abschnitten der Arteria basilaris, durch Messung der internen luminale Querschnittsfläche mit NIH Image Software-J. Die Bedeutung wargetestet Tukey / Kramer &#39;s statistische Analyse. Ergebnisse: Nach der Analyse von histologischen Schnitten, waren basilaris luminale Querschnittsfläche kleiner in der SAH als in der Kochsalz-Gruppe, die mit zerebralen Vasospasmus in der ersten Gruppe. Im SAH Gruppe waren basilaris Innenraum (0,056 um ± 3) deutlich kleiner ab Vasospasmus fünf Tage nach der zweiten Injektion Blut (sieben Tage nach der ersten Injektion Blut), verglichen mit der Kontrollgruppe, die mit Kochsalzlösung Innenraum (0,069 ± 3, p = 0,004). Es gab keine Todesfälle von zerebralen Vasospasmus. Fazit: Die Ratte Doppel SAH-Modell führt zu einem milden, überlebensfähige, Basilararterie Vasospasmus, die verwendet werden, um die pathophysiologischen Mechanismen der zerebralen Vasospasmus in einem kleinen Tiermodell studieren können. Eine niedrige und akzeptablen Mortalitätsrate ist ein wesentliches Kriterium für eine ideale SAH Tiermodell befriedigt werden, so dass die Mechanismen des Vasospasmus kann elucidated 7, 8. Weitere Modifikationen des Modells können, um für erhöhte Schwere Vasospasmus und neurologischen Untersuchungen einzustellen.

Innerhalb der Protokolle oben beschrieben, gibt es mehrere Schritte, die wir glauben erfordern eine bessere Charakterisierung des Modells als das, was zuvor in der Literatur beschrieben. Hier haben wir auf den Stufen, die notwendig sind, um eine reproduzierbare niedrige Sterblichkeit zerebralen Vasospasmus kleinen Tiermodell zu erreichen und vermeiden potenzielle Fallstricke bei diesem Modell verbunden, wenn nicht richtig gemacht zu konzentrieren. Ein. Eigenblut Draw aus dem Schwanzarterie: </p...

Watch this Scientific Journal Video about A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage at JoVE.com

A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage

Aneurysmal Subarachnoidalblutung (SAB) blutet, das auftritt, in den Subarachnoidalraum, wenn ein Aneurysma platzt. Während die Morbidität und Mortalität von dieser Veranstaltung wurde auf einen Rückgang aufgrund der verbesserten Behandlungsansätze, die Gefahr des Vasospasmus wurde nach Subarachnoidalblutung weiterhin die gleichen wie noch vor einigen Jahren war. Die Bedeutung der Schaffung eines umfassenden und reproduzierbare Tiermodell, um auslösende Ereignisse des zerebralen Vasospasmus zu identifizieren hat sich der Schwerpunkt der Forschung seit dem ersten Einsatz von Ratten in einem experimentellen Vasospasmus-Modell 1979 von Barry<em&gt; Et al.</em&gt; Frühe Arbeiten bei Ratten gezeigt, dass eine einzige Injektion von Eigenblut in die Cisterna magna zu akuten geführt (in Minuten), aber nicht verzögert zerebralen Vasospasmus<sup&gt; 3, 6, 14</sup&gt;. Hier charakterisieren wir eine niedrige Sterblichkeit SAH Rattenmodell, dass die Ergebnisse reproduzierbar verzögerte Vasospasmus.

Eine niedrige Sterblichkeit Rattenmodell Verzögerte zerebralen Vasospasmus nach experimenteller Subarachnoidalblutung beurteilen

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) ...

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14.2K Aufrufe.  SUNY Upstate Medical University. Aneurysmal Subarachnoidalblutung (SAB) blutet, das auftritt, in den Subarachnoidalraum, wenn ein Aneurysma platzt. Während die Morbidität und Mortalität von dieser Veranstaltung wurde auf einen Rückgang aufgrund der verbesserten Behandlungsansätze, die Gefahr des Vasospasmus wurde nach Subarachnoidalblutung weiterhin die gleichen wie noch vor einigen Jahren war. Die Bedeutung der Schaffung eines umfassenden und reproduzierbare Tiermodell, um auslösende Ereignisse de...

Serie: A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage

Modeling Stroke in Mice - Middle Cerebral Artery Occlusion with the Filament Model

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are very limited. To meet the need for novel therapeutic approaches, experimental stroke research frequently employs rodent models of focal cerebral ischaemia. Most researchers use permanent or transient occlusion of the middle cerebral artery (MCA) in mice or rats.
Proximal occlusion of the middle cerebral artery (MCA) via the intraluminal suture technique (so called filament or suture model) is probably the most frequently used model in experimental stroke research. The intraluminal MCAO model offers the advantage of inducing reproducible transient or permanent ischaemia of the MCA territory in a relatively non-invasive manner. Intraluminal approaches interrupt the blood flow of the entire territory of this artery. Filament occlusion thus arrests flow proximal to the lenticulo-striate arteries, which supply the basal ganglia. Filament occlusion of the MCA results in reproducible lesions in the cortex and striatum and can be either permanent or transient. In contrast, models inducing distal (to the branching of the lenticulo-striate arteries) MCA occlusion typically spare the striatum and primarily involve the neocortex. In addition these models do require craniectomy. In the model demonstrated in this article, a silicon coated filament is introduced into the common carotid artery and advanced along the internal carotid artery into the Circle of Willis, where it blocks the origin of the middle cerebral artery. In patients, occlusions of the middle cerebral artery are among the most common causes of ischaemic stroke. Since varying ischemic intervals can be chosen freely in this model depending on the time point of reperfusion, ischaemic lesions with varying degrees of severity can be produced. Reperfusion by removal of the occluding filament at least partially models the restoration of blood flow after spontaneous or therapeutic (tPA) lysis of a thromboembolic clot in humans.
In this video we will present the basic technique as well as the major pitfalls and confounders which may limit the predictive value of this model.

Filamentous occlusion of the Middle cerebral artery is a common model for studying ischemic stroke in mice.

Modeling Stroke in Mice - Middle Zerebralarterie mit dem Filament-Modell

Stroke is among the most frequent causes of death and adult disability, especially in highly developed countries. However, treatment options to date are ...

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Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

The number of patients with end-stage renal disease, and the number of kidney allograft recipients continuously increases. Episodes of acute cellular allograft rejection (AR) are a negative prognostic factor for long-term allograft survival, and its timely diagnosis is crucial for allograft function 1. At present, AR can only be definitely diagnosed by core-needle biopsy, which, as an invasive method, bares significant risk of graft injury or even loss. Moreover, biopsies are not feasible in patients taking anticoagulant drugs and the limited sampling site of this technique may result in false negative results if the AR is focal or patchy. As a consequence, this gave rise to an ongoing search for new AR detection methods, which often has to be done in animals including the use of various transplantation models.
Since the early 60s rat renal transplantation is a well-established experimental method for the examination and analysis of AR 2. We herein present in addition small animal positron emission tomography (PET) using 18F-fluorodeoxyglucose (FDG) to assess AR in an allogeneic uninephrectomized rat renal transplantation model and propose graft FDG-PET imaging as a new option for a non-invasive, specific and early diagnosis of AR also for the human situation 3. Further, this method can be applied for follow-up to improve monitoring of transplant rejection 4.

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.

Nicht-invasive Bildgebung des akuten Abstoßung nach Nierentransplantation mit Rat<sup&gt; 18</sup&gt; F-FDG PET

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

Surgical Procedures for a Rat Model of Partial Orthotopic Liver Transplantation with Hepatic Arterial Reconstruction

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as studies on graft preservation and ischemia-reperfusion injury 1,2, immunological responses 3,4, hemodynamics 5,6, and small-for-size syndrome 7. The rat OLT is among the most difficult animal models in experimental surgery and demands advanced microsurgical skills that take a long time to learn. Consequently, the use of this model has been limited. Since the reliability and reproducibility of results are key components of the experiments in which such complex animal models are used, it is essential for surgeons who are involved in rat OLT to be trained in well-standardized and sophisticated procedures for this model.
While various techniques and modifications of OLT in rats have been reported 8 since the first model was described by Lee et al. 9 in 1973, the elimination of the hepatic arterial reconstruction 10 and the introduction of the cuff anastomosis technique by Kamada et al. 11 were a major advancement in this model, because they simplified the reconstruction procedures to a great degree. In the model by Kamada et al., the hepatic rearterialization was also eliminated. Since rats could survive without hepatic arterial flow after liver transplantation, there was considerable controversy over the value of hepatic arterialization. However, the physiological superiority of the arterialized model has been increasingly acknowledged, especially in terms of preserving the bile duct system 8,12 and the liver integrity 8,13,14.
In this article, we present detailed surgical procedures for a rat model of OLT with hepatic arterial reconstruction using a 50% partial graft after ex vivo liver resection. The reconstruction procedures for each vessel and the bile duct are performed by the following methods: a 7-0 polypropylene continuous suture for the supra- and infrahepatic vena cava; a cuff technique for the portal vein; and a stent technique for the hepatic artery and the bile duct.

Orthotopic liver transplantation in rats is an indispensable experimental model for biomedical research. Here we present our surgical procedures for orthotopic rat liver transplantation with hepatic arterial reconstruction using a 50% partial graft.

Chirurgische Verfahren für eine Ratte Modell Teilweise Lebertransplantation mit arteriellen Rekonstruktion

Orthotopic liver transplantation (OLT) in rats using a whole or partial graft is an indispensable experimental model for transplantation research, such as ...

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Optimierte System for Cerebral Perfusion Überwachung im Rat Stroke Modell Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

Einem Mausmodell der Subarachnoidalblutung

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and controllable animal models that simulate both conditions are presently uncommon. Therefore, new models are needed in order to mimic human pathophysiological conditions resulting from SAH.
This report describes the technical nuances of a rabbit blood-shunt SAH model that enables control of intracerebral pressure (ICP). An extracorporeal shunt is placed between the arterial system and the subarachnoid space, which enables examiner-independent SAH in a closed cranium. Step-by-step procedural instructions and necessary equipment are described, as well as technical considerations to produce the model with minimal mortality and morbidity. Important details required for successful surgical creation of this robust, simple and consistent ICP-controlled SAH rabbit model are described.

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

Das Kaninchen Blut-Shunt-Modell für die Untersuchung von akuten und Spätfolgen Subarachnoidalblutung: Technische Aspekte

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

Leprdb Mouse Model of Type 2 Diabetes: Pancreatic Islet Isolation and Live-cell 2-Photon Imaging Of Intact Islets

Type 2 diabetes is a chronic disease affecting 382 million people in 2013, and is expected to rise to 592 million by 2035 1. During the past 2 decades, the role of beta-cell dysfunction in type 2 diabetes has been clearly established 2. Research progress has required methods for the isolation of pancreatic islets. The protocol of the islet isolation presented here shares many common steps with protocols from other groups, with some modifications to improve the yield and quality of isolated islets from both the wild type and diabetic Leprdb (db/db) mice. A live-cell 2-photon imaging method is then presented that can be used to investigate the control of insulin secretion within islets.

Leprdb Mouse Model of Type 2 Diabetes: Pancreatic Islet Isolation and Live-cell 2-Photon Imaging Of Intact Islets

We present here a protocol for the isolation of islets from the mouse model of type 2 diabetes, Leprdb and details of a live-cell assay for measurement of insulin secretion from intact islets that utilizes 2 photon microscopy.

Lepr<sup&gt; Db</sup&gt; Mausmodell des Typ-2-Diabetes: Pancreatic Islet Isolation und Live-cell 2-Photonen-Imaging von intakten Inseln

Type 2 diabetes is a chronic disease affecting 382 million people in 2013, and is expected to rise to 592 million by 2035 1. During the past 2 decades, ...

Integrated Compensatory Responses in a Human Model of Hemorrhage

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of hemorrhaging patients is difficult because current clinical tools provide measures of vital signs that remain stable during the early stages of bleeding due to compensatory mechanisms. Consequently, there is a need to understand and measure the total integration of mechanisms that compensate for reduced circulating blood volume and how they change during ongoing progressive hemorrhage. The body&#39;s reserve to compensate for reduced circulating blood volume is called the &#39;compensatory reserve&#39;. The compensatory reserve can be accurately evaluated with real-time measurements of changes in the features of the arterial waveform measured with the use of a high-powered computer. Lower Body Negative Pressure (LBNP) has been shown to simulate many of the physiological responses in humans associated with hemorrhage, and is used to study the compensatory response to hemorrhage. The purpose of this study is to demonstrate how compensatory reserve is assessed during progressive reductions in central blood volume with LBNP as a simulation of hemorrhage.

The purpose of this protocol is to demonstrate the techniques for measuring compensatory responses to reduced central blood volume using lower body negative pressure as a noninvasive experimental model of human hemorrhage which can be used to quantify the total integration of compensatory mechanisms to blood volume deficit in humans.

Integrierte kompensatorische Antworten in einem menschlichen Modell von Einblutung

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of ...

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of clinical efficacy, they are until now imperfect solutions. Stem cell therapy using ASC sheets could provide a solution to this problem. ASCs are considered as promising candidates for promoting tissue healing because of their trophic and immunomodulatory properties. Here, we provide methods to produce high-density ASC sheets, that are transplanted onto a colorectal anastomosis in a rat model to reduce the leakage. ASCs formed cell sheets in thermo-responsive culture dishes that could be easily detached. On the day of the transplantation, a partial colectomy with a 5-suture colorectal anastomosis was performed. Animals were immediately transplanted with 1 ASC sheet per rat. ASC sheets adhered spontaneously to the anastomosis without any glue, suture, or any biomaterial. Animal groups were sacrificed 3 and 7 days postoperatively. Compared to transplanted animals, the incidence of anastomotic abscesses and leakage was higher in control animals. In our model, the transplantation of ASC sheets after colorectal anastomosis was successful and associated with a lower leakage rate.

The preparation and transplantation of adipose tissue derived stem cell (ASC) sheets onto insufficiently sutured colorectal anastomoses in a rat model is presented. This novel application shows that ASC sheets can reduce colorectal anastomosis leakage.

Transplantation von Fettgewebe-Derived Stem Cell Blatt, Leckage nach teilweiser Colectomy in einem Rattenmodell zu reduzieren

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

Implantation of hiPSC-derived Cardiac-muscle Patches after Myocardial Injury in a Guinea Pig Model

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss of relevant amounts of heart muscle mass can lead to the heart failure. Besides heart transplantation, there is no curative treatment option for the end-stage heart failure. In times of organ donor shortage, organ independent treatment modalities are needed. Left-ventricular assist devices are a promising therapy option, however, especially as destination therapy, limited by its side-effects like stroke, infections and bleedings. In recent years, several cardiac repair strategies including stem cell injection, cardiac progenitors or myocardial tissue engineering have been investigated. Recent improvements in cell biology allow for the differentiation of large amounts of cardiomyocytes derived from human induced pluripotent stem cells (iPSC). One of the cardiac repair strategies currently under evaluation is to transplant artificial heart tissue. Engineered heart tissue (EHT) is a three-dimensional in vitro created cardiomyocyte network, with functional properties of native heart tissue. We have created EHT-patches from hiPSC derived cardiomyocytes. Here we present a protocol for the induction of left ventricular myocardial cryoinjury in a guinea pig, followed by implantation of hiPSC derived EHT on the left ventricular wall.

Here we present a protocol for the induction of left ventricular cryoinjury followed by the implantation of a cardiac muscle patch, derived from human iPS-cell cardiomyocytes in a guinea pig model.

Implantation von HiPSC abgeleitete Herzmuskel Patches nach Myokard-Schädigung in einem Meerschweinchen-Modell

Due to the limited regeneration capacity of the heart in adult mammals, myocardial infarction results in an irreversible loss of cardiomyocytes. This loss ...