La technique chirurgicale a été initialement acquis dans le laboratoire du Dr William D. Hill au Medical College of Georgia. Les auteurs tiennent également à remercier le Dr David A. Rempe et Landa Prifti pour l&#39;utilisation de la caméra de dissection. Cette recherche a été financée par le NIH NS041744, NS051279, F31 NS064700 et AHA 30815697D.

Ce protocole a été approuvé par l&#39;Université de Rochester comité consacré à l&#39;utilisation éthique des animaux en recherche (UCAR). Techniques d&#39;asepsie doivent être suivies pendant le protocole. L&#39;utilisation de gants stériles et un masque est nécessaire. Tous les équipements, matériaux, produits chimiques, et des outils qui sont utilisés au cours du protocole sont décrits dans le tableau 1. 1. Préparation préopératoire <ol><li> Injecter des souris par voie sous cutanée à la buprénorphine (0.05mg/kg) 2 h avant la chirurgie, immédiatement après la chirurgie, puis toutes les 3-5 h pendant les 24 premières heures post-opératoire. </li><li> Stériliser tous les instruments chirurgicaux, compresses de gaze, et applicateurs coton-tige par autoclave. Garder les outils chirurgicaux dans l&#39;éthanol à 70% pendant la chirurgie et l&#39;air sec sur la gaze stérile juste avant utilisation. Vaporiser la zone de travail avec de l&#39;éthanol à 70%. </li><li> Anesthésier la souris avec un 3% isofluorane-20% mélange gazeux d&#39;oxygène en utilisant un vaporisateur anesthésique. Ajustez la quantité d&#39;oxygène avec débitmètre. Testez le niveau de l&#39;anesthésie par orteil ou le pincement de la queue (ils doivent être insensible). Maintenir le niveau de l&#39;anesthésie avec 2% (v / v) isofluorane. </li><li> Appliquer des larmes artificielles pour les yeux de la souris et faire preuve de prudence pour éviter d&#39;endommager l&#39;œil pendant l&#39;intervention chirurgicale. Placez la souris sur son côté droit en position latérale. </li><li> Rasez la zone sur le côté gauche, entre l&#39;œil gauche et la base de l&#39;oreille gauche en utilisant la tondeuse pour animaux. Nettoyer la zone en alternant entre une solution bétadine et 70% d&#39;éthanol à l&#39;aide des applicateurs coton-tige et les frottant légèrement la région. Répéter au besoin. </li><li> Placez la souris sur un coussin chauffant relié à une sonde rectale pour maintenir la température du corps à 37 ° C. Insérer la sonde rectale en utilisant une huile minérale. </li><li> Placez la souris sur son côté droit sous le microscope et sécurisé avec du ruban adhésif. Couper une fenêtre dans une compresse de gaze et de couvrir la zone chirurgicale. </li></ol> 2. Intervention chirurgicale et ligature MCA <ol><li> Faire une incision verticale entre l&#39;œil gauche et la base de l&#39;oreille gauche en utilisant beaux ciseaux droits. Utilisez hémostatiques courbes pour garder la zone chirurgicale ouverte. </li><li> Faire une incision horizontale sur le muscle temporal en utilisant des ciseaux à ressort et légèrement séparée du muscle temporal du crâne par tirant lentement avec une pince. </li><li> Faites une petite craniotomie subtemporal à la jonction de l&#39;arcade zygomatique et l&#39;os squamosal en utilisant une aiguille 18G tout en tenant la mâchoire avec une pince courbe. </li><li> Exposer le MCA en enlevant les petits morceaux de crâne avec Os Rongeurs. L&#39;arcade zygomatique et contenu orbitaire ne doit pas être endommagé pendant ce processus. Si une déshydratation excessive du tissu se produit pendant ce processus, appliquer PBS stérile avec des applicateurs de coton-tige. </li><li> Ligaturer la partie distale du MCA en utilisant un petit navire cauterizer. </li><li> Placez le muscle temporal dans sa position originale et fermer le site d&#39;incision chirurgicale avec sutures 5-0 en nylon. </li><li> Cesser d&#39;anesthésie et de retirer la sonde rectale. Injecter de la souris avec une 2e dose de buprénorphine (0.05mg/kg) sous-cutanée. Retour de la souris dans la cage qui a été maintenue à 37 ° C avec un panneau de chauffage. </li><li> Surveiller de près la souris pour les prochaines 24 h pour tout inconfort (diminution de l&#39;appétit / consommation d&#39;eau, une posture voûtée, augmentation de la respiration, pilo-érigé cheveux). Injecter de la souris avec la buprénorphine par voie sous cutanée toutes les deux heures 3-5 post-opératoire jusqu&#39;à 24h. Fournir la souris avec gel de récupération durant cette période. </li></ol> 3. Coloration TTC et détermination du volume d&#39;éjection <ol><li> Vingt-quatre heures après la chirurgie profondément anesthésier la souris, transcardiaque perfuser la souris avec un 4% TTC (p / v) en tampon phosphate salin (PBS) pendant 15 min, puis avec une solution de paraformaldéhyde 4% pendant 10 min en utilisant un mini pompe péristaltique à un débit moyen. Enlever le cerveau et le placer dans la solution de paraformaldéhyde à 4% pendant une nuit. </li><li> Position du cerveau dans le bloc de coupe. Trancher le cerveau en tranches de 1mm d&#39;épaisseur en utilisant des lames de rasoir. </li><li> Placer les tranches à côté d&#39;une règle-échelle millimétrique. Photographier les tranches en utilisant un appareil photo numérique connecté à la loupe binoculaire. </li><li> Calculer la zone de course en soustrayant le secteur non-infarctus du site ipsilatérale de la superficie totale du site en utilisant l&#39;image controlatérale J Software (http://rsbweb.nih.gov/ij/). Calculer le volume d&#39;éjection systolique par l&#39;empilement de la zone de l&#39;accident vasculaire cérébral 22 tranches. </li></ol> 4. Itinéraire d&#39;utiliser un logiciel Image J <ol><li> Ouvrez le fichier image à analyser dans le logiciel Image J en cliquant sur le menu Fichier. </li><li> Cliquez sur l&#39;onglet «ligne droite» dans le programme. Tracez une ligne droite entre les deux marges sur la règle. Cliquez sur &quot;analyser&quot; le menu et sélectionner &#39;échelle définie. Réglez la distance connue comme une unité de longueur en mm. </li><li> Cliquez sur l&#39;onglet «cercle à main levée». Tracez un cercle de décrire l&#39;hémisphère controlatéral. Cliquez sur &quot;analyser&quot; et sélectionnez le menu «mesure». La fenêtre de calcul apparaît montrant la zone du cercle tracé. </li><li> Dessinez un autre cercle autour de l&#39;hémisphère exclusion AVC ipsilatéral (blanc) région. Mesurer la surface, comme indiqué dans l&#39;étape 4.3. La nouvelle valeur sera ajoutée à la fenêtre de calcul. La différence entre la 1ère et 2ème valeurs représente la zone de l&#39;infarctus qui est indiqué comme A de la formule ci-dessous. </li><li> Calculer la zone de course dans chaque tranche en répétant les étapes 4.2 à 4.4. Prenez sommation de la zone de course calculé dans chaque tranche (Ea n). Cela représente le volume d&#39;éjection sachant le fait que l&#39;épaisseur de chaque tranche est de 1mm. Ea n x 1mm (épaisseur de chaque tranche) = volume des maladies </li></ol> 5. Les résultats représentatifs: L&#39;infarctus obtenus par ligature permanente MCA chez la souris sont principalement corticale. Cependant, il est possible d&#39;obtenir des lésions sous-corticales si le MCA est ligaturé en amont de la branche lenticulostriées. Les volumes AVC après ligature MCA pourrait varier d&#39;un 10mm à 35mm ³ ³ 19, 23. Les volumes course calculé par coloration TTC en Moyanova et al. 19 sont entre ³ 10mm à 22mm ³ tandis que les volumes course déterminée à partir d&#39;images IRM dans Filiano et al 23 sont entre ³ 20mm à 35mm ³. Les raisons possibles de ces différences peut-être l&#39;emplacement exact de la ligature et les différentes méthodes utilisées pour mesurer les volumes d&#39;AVC. Dans la figure 1, un teint TTC cérébrales 24 heures après la ligature permanente MCA dans un type C57/BL6 souris sauvage est montré. Le site AVC est blanche en apparence (Fig. 1A, B). Figure 1A et B illustrent le site d&#39;infarctus sous différents angles. La figure 2 montre les tranches de 1mm d&#39;épaisseur de la TTC vasculaires cérébraux teinté d&#39;avant en arrière (Fig. 2A-G). Volume de l&#39;infarctus a été calculée 24 h après la chirurgie. Le volume de l&#39;infarctus est ³ ~ 23mm. <img src="/files/ftp_upload/3039/3039fig1.jpg" alt="Figure 1"/> Figure 1. Représentation du site d&#39;AVC. AB, TTC teinté images du cerveau entier, 24 heures après la ligature du MCA. Zone blanche représente l&#39;infarctus. <img src="/files/ftp_upload/3039/3039fig2.jpg" alt="Figure 2"/> Figure 2. Les résultats représentatifs. (A à G&gt;), TTC sections colorées du cerveau 1mm, 24 h après MCAL. Les tranches sont alignés d&#39;avant en arrière. Zone blanche représente l&#39;infarctus.

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La méthode la ligature permanente MCA donne volumes d&#39;infarctus hautement reproductible et une augmentation des taux de survie post-opératoires par rapport aux autres méthodes disponibles. La facilité et la courte durée (~ 30 min) de la procédure de la rendre encore plus pratique. La méthode est largement utilisée dans les souris et les rats. Cette technique nécessite une chirurgie invasive sous un stéréomicroscope. Par conséquent, expérience dans l&#39;exploitation sous un ...

<table border="1"><tbody><tr><th> NOM </th><th> SOCIETE ET NUMÉRO DE CATALOGUE </th></tr><tr><td> Solutions et produits chimiques A. </td><td></td></tr><tr><td> Solution de polyvidone iodée, lavage chirurgical </td><td> Aplicare, 82-209 </td></tr><tr><td> L&#39;éthanol à 70% </td><td> Koptec, V1001 </td></tr><tr><td> 2,3,5 chlorure de triphényltétrazolium (TTC) </td><td> Sigma, T8877 </td></tr><tr><td> Paraformaldéhyde à 4% </td><td> Sciences Electron Microscopy, 159SP </td></tr><tr><td> Tampon phosphate salin, pH: 7,4 </td><td> Fabriqué et stérilisé en laboratoire. </td></tr><tr><td> Huile minérale </td><td> PML Microbiologicals, R6570 </td></tr><tr><td> Isofluorane </td><td> La société Butler, 029405 </td></tr><tr><td> La buprénorphine (0.3mg/ml) </td><td> Hospira, NDC 0409-2012-32 </td></tr><tr><td> Les larmes artificielles </td><td> La société Butler, 007312 </td></tr><tr><td> Récupération Gel </td><td> Effacer H 2 O, 72-01-5022 </td></tr><tr><td> B. du matériel chirurgical et de l&#39;équipement </td><td></td></tr><tr><td> C57/BL6 la souris, 10-12 semaines </td><td> La colonie a été élevé dans le vivarium universitaires </td></tr><tr><td> 100% d&#39;oxygène du réservoir </td><td> Airgas </td></tr><tr><td> Vaporisateur anesthésie et débitmètre </td><td> Surgivet, Modèle 100 </td></tr><tr><td> Coussin chauffant avec sonde rectale </td><td> Outils Fine Science, TR200 </td></tr><tr><td> Microscope à dissection </td><td> Zeiss, Stemi 2000 </td></tr><tr><td> Panneau de chauffage </td><td> Petco Services Ltd 0307-013 </td></tr><tr><td> Petits Set Cauterizer navire </td><td> Outils Fine Science, 18000-00 </td></tr><tr><td> Mini Pompe péristaltique </td><td> Harvard Apparatus, MPII </td></tr><tr><td> Seringues 1ml </td><td> BD, 329650 </td></tr><tr><td> Aiguille 18G </td><td> Becton Dickinson, 305196 </td></tr><tr><td> Suture chirurgicale Nylon 5.0 </td><td> Ethilon, 698H </td></tr><tr><td> Sectionnement bloc </td><td> Kent scientifique, 1mm </td></tr><tr><td> Simple lames de rasoir bord </td><td> Sciences Electron Microscopy, 71962 </td></tr><tr><td> Ruban d&#39;étiquettes </td><td> Scienceware, Bel-Art Produits F13463-2075 </td></tr><tr><td> Compresses de gaze </td><td> Curity Kendall 2252, Johnson and Johnson 6415 </td></tr><tr><td> Applicateurs coton-tige </td><td> Puritain 806-CMT </td></tr><tr><td> C. Outils chirurgie </td><td></td></tr><tr><td> Ciseaux Spring chirurgicale </td><td> Outils Fine Science 15003-08 </td></tr><tr><td> Pince Microdissecting angle </td><td> Outils Fine Science 5 / 45 </td></tr><tr><td> Pince courbée Microdissecting </td><td> Beaux sciences Outils 5 Ti </td></tr><tr><td> Beaux-ciseaux droits </td><td> Outils Fine Science 14568-12 </td></tr><tr><td> Rongeur d&#39;os incurvé </td><td> Outils Fine Science 16021-14 </td></tr><tr><td> Hémostatique Hartman courbé </td><td> Outils Fine Science 13003-10 </td></tr></tbody></table> Tableau 1. La liste des matériaux, outils et produits chimiques utilisés pour la procédure de ligature MCA. Solutions A. et produits chimiques, matériaux et équipement chirurgical B., C. outils de chirurgie.

the application of permanent middle cerebral artery ligation in the mouse

Ischémie cérébrale focale est parmi les plus fréquentes d&#39;accident vasculaire cérébral chez les patients. En raison de la signification clinique, il ya eu un effort prolongé pour développer des modèles animaux appropriés pour étudier les événements qui se déroulent pendant l&#39;insulte ischémique. Ces techniques comprennent transitoire ou permanent, des modèles d&#39;ischémie focale ou globale à l&#39;aide de nombreux modèles animaux différents, avec les rongeurs les plus communs étant. La méthode la ligature permanente MCA qui est également désigné comme pMCAo dans la littérature est largement utilisé comme un modèle d&#39;ischémie focale chez les rongeurs 1-6. Cette méthode a été initialement décrit pour le rat par Tamura et al. en 1981 7. Dans ce protocole une craniotomie a été utilisé pour accéder à la MCA et les régions proximales ont été occultées par électrocoagulation. L&#39;infarctus impliquer les régions essentiellement corticale et parfois striés selon l&#39;emplacement de l&#39;occlusion. Cette technique est maintenant bien établi et utilisé dans de nombreux laboratoires 8-13. L&#39;utilisation précoce de cette technique ont conduit à la définition et la description du «noyau d&#39;infarctus» et «pénombre» 14-16, et il est souvent utilisé pour évaluer le potentiel des composés neuroprotecteurs 10, 12, 13, 17. Bien que les études initiales ont été effectuées chez le rat, la ligature permanente MCA a été utilisé avec succès chez la souris avec de légères modifications 18-20. Ce modèle donne des infarctus reproductibles et augmenté les taux de survie après. Environ 80% des AVC ischémiques chez l&#39;homme se produisent dans la région de MCA 21 et donc c&#39;est un modèle très pertinent pour les études de course. Actuellement, il ya un manque de traitements efficaces disponibles pour les patients d&#39;AVC, et il existe donc un besoin de bons modèles pour tester le potentiel des composés pharmacologiques et évaluer les résultats physiologiques. Cette méthode peut également être utilisé pour étudier les mécanismes intracellulaires de réponse hypoxie in vivo. Ici, nous présentons la ligature MCA dans une souris C57/BL6. Nous décrivons la préparation pré-chirurgicale, la ligature MCA et 2,3,5 chlorure de triphényltétrazolium (TTC) coloration pour la quantification des volumes d&#39;infarctus.

Watch this Scientific Journal Video about The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse at JoVE.com

The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

L&#39;artère cérébrale moyenne (MCA) ligature est une technique pour étudier l&#39;ischémie cérébrale focale chez des modèles animaux. Dans cette méthode, l&#39;artère cérébrale moyenne est exposée par craniotomie et ligaturée par cautérisation. Cette méthode donne les volumes d&#39;infarctus hautement reproductible et une augmentation des taux de survie post-opératoires par rapport aux autres méthodes disponibles.

L&#39;application de la ligature permanente artère cérébrale moyenne chez la souris

Focal cerebral ischemia is among the most common type of stroke seen in patients. Due to the clinical significance there has been a prolonged effort to ...

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19.5K vues.  University of Rochester. L'artère cérébrale moyenne (MCA) ligature est une technique pour étudier l'ischémie cérébrale focale chez des modèles animaux. Dans cette méthode, l'artère cérébrale moyenne est exposée par craniotomie et ligaturée par cautérisation. Cette méthode donne les volumes d'infarctus hautement reproductible et une augmentation des taux de survie post-opératoires par rapport aux autres méthodes disponibles.

Vidéo: The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

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.

Course de modélisation chez la souris - Moyen-occlusion artérielle cérébrale avec le modèle filamentaire

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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Mouse Model of Middle Cerebral Artery Occlusion

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the brain (ischemic stroke) 2. Since most ischemic strokes (~80%) occur in the territory of middle cerebral artery (MCA) 3, many animal stroke models that have been developed have focused on this artery. The intraluminal monofilament model of middle cerebral artery occlusion (MCAO) involves the insertion of a surgical filament into the external carotid artery and threading it forward into the internal carotid artery (ICA) until the tip occludes the origin of the MCA, resulting in a cessation of blood flow and subsequent brain infarction in the MCA territory 4. The technique can be used to model permanent or transient occlusion 5. If the suture is removed after a certain interval (30 min, 1 h, or 2 h), reperfusion is achieved (transient MCAO); if the filament is left in place (24 h) the procedure is suitable as a model of permanent MCAO. This technique does not require craniectomy, a neurosurgical procedure to remove a portion of skull, which may affect intracranial pressure and temperature 6. It has become the most frequently used method to mimic permanent and transient focal cerebral ischemia in rats and mice 7,8. To evaluate the extent of cerebral infarction, we stain brain slices with 2,3,5-triphenyltetrazolium chloride (TTC) to identify ischemic brain tissue 9. In this video, we demonstrate the MCAO method and the determination of infarct size by TTC staining.

We demonstrate in the video a method for producing a middle cerebral artery occlusion in adult mice using an intraluminal monofilament. We also show how to evaluate the extent of cerebral infarction by 2,3,5-triphenyltetrazolium chloride (TTC) staining.

Modèle murin d&#39;occlusion artère cérébrale moyenne

Stroke is the most common fatal neurological disease in the United States 1. The majority of strokes (88%) result from blockage of blood vessels in the ...

Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be performed in many locations overlying the rat cerebrum, but is most easily carried out by creating a craniectomy over the readily accessible frontal or parietal bones. Most frequently, we have used this technique in combination with the endothelin-1 middle cerebral artery occlusion model of ischemic stroke to quantify the changes in middle cerebral artery vessel diameter that occur with injection of endothelin-1 into the brain parenchyma adjacent to the proximal MCA4, 5. In order to visualize the proximal portion of the MCA during endothelin -1 induced MCAO, we use a technique to create a cranial window through the temporal bone on the lateral aspect of the rat skull (Figure 1). Cerebral arteries can be visualized either with the dura intact or with the dura incised and retracted. Most commonly, we leave the dura intact during visualization since endothelin-1 induced MCAO involves delivery of the vasoconstricting peptide into the brain parenchyma. This bypasses the need to incise the dura directly over the visualized vessels for drug delivery. This protocol will describe how to create a cranial window to visualize cerebral arteries in a step-wise fashion, as well as how to avoid many of the potential pitfalls pertaining to this method.

This article describes a method for visualizing rat cerebral arteries through a cranial window using temporal craniectomy in order to view proximal portions of the middle cerebral artery (Figure 1). This versatile method can be combined with various techniques of drug delivery to measure cerebral artery reactivity in vivo.

L&#39;utilisation d&#39;une fenêtre crânien à la Visualisez l&#39;artère cérébrale moyenne pendant l&#39;endothéline-1 induite par l&#39;occlusion Artère cérébrale moyenne

Creation of a cranial window is a method that allows direct visualization of structures on the cortical surface of the brain1-3. This technique can be ...

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.

Système optimisé pour la surveillance de la perfusion cérébrale chez le rat des maladies du intraluminale occlusion de l&#39;artère cérébrale moyenne

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

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia) 1. Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex 2, is the most common site of human stroke 3, and ischemia within its territory can result in extensive dysfunction or death 1,4,5. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA.
Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

We describe a highly reproducible method for the permanent occlusion of a rodent major cerebral blood vessel. This technique can be accomplished with very little peripheral damage, minimal blood loss, a high rate of long-term survival, and consistent infarct volume commensurate with the human clinical population.

Permanent occlusion des vaisseaux cérébraux<em&gt; Via</em&gt; Double ligature et Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

Modeling Stroke in Mice: Permanent Coagulation of the Distal Middle Cerebral Artery

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical options are available for a small proportion of stroke patients in the acute phase. Current research is intensively searching for novel therapeutic strategies and is increasingly focusing on the sub-acute and chronic phase after stroke because more patients might be eligible for therapeutic interventions in a prolonged time window. These delayed mechanisms include important pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity and regeneration. In order to analyze these mechanisms and to subsequently evaluate novel drug targets, experimental stroke models with clinical relevance, low mortality and high reproducibility are sought after. Moreover, mice are the smallest mammals in which a focal stroke lesion can be induced and for which a broad spectrum of transgenic models are available. Therefore, we describe here the mouse model of transcranial, permanent coagulation of the middle cerebral artery via electrocoagulation distal of the lenticulostriatal arteries, the so-called &#8220;coagulation model&#8221;. The resulting infarct in this model is located mainly in the cortex; the relative infarct volume in relation to brain size corresponds to the majority of human strokes. Moreover, the model fulfills the above-mentioned criteria of reproducibility and low mortality. In this video we demonstrate the surgical methods of stroke induction in the &#8220;coagulation model&#8221; and report histological and functional analysis tools.

Various murine models of middle cerebral artery occlusion (MCAO) are widely used in experimental brain research. Here, we demonstrate the model of transcranial permanent distal MCAO which produces consistent cortical infarction of a size corresponding to damage imposed by the majority of human ischemic strokes.

Modélisation des maladies chez les souris: Coagulation permanente de l&#39;artère cérébrale moyenne distale

Stroke is the third most common cause of death and a main cause of acquired adult disability in developed countries. Only very limited therapeutical ...

Permanent Ligation of the Left Anterior Descending Coronary Artery in Mice: A Model of Post-myocardial Infarction Remodelling and Heart Failure

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It is characterized by fluid retention, shortness of breath, and fatigue, in particular on exertion. Heart failure is a growing public health problem, the leading cause of hospitalization, and a major cause of mortality. Ischemic heart disease is the main cause of heart failure.
Ventricular remodelling refers to changes in structure, size, and shape of the left ventricle. This architectural remodelling of the left ventricle is induced by injury (e.g., myocardial infarction), by pressure overload (e.g., systemic arterial hypertension or aortic stenosis), or by volume overload. Since ventricular remodelling affects wall stress, it has a profound impact on cardiac function and on the development of heart failure. A model of permanent ligation of the left anterior descending coronary artery in mice is used to investigate ventricular remodelling and cardiac function post-myocardial infarction. This model is fundamentally different in terms of objectives and pathophysiological relevance compared to the model of transient ligation of the left anterior descending coronary artery. In this latter model of ischemia/reperfusion injury, the initial extent of the infarct may be modulated by factors that affect myocardial salvage following reperfusion. In contrast, the infarct area at 24 hr after permanent ligation of the left anterior descending coronary artery is fixed. Cardiac function in this model will be affected by 1) the process of infarct expansion, infarct healing, and scar formation; and 2) the concomitant development of left ventricular dilatation, cardiac hypertrophy, and ventricular remodelling.
Besides the model of permanent ligation of the left anterior descending coronary artery, the technique of invasive hemodynamic measurements in mice is presented in detail.

Heart failure is the leading cause of hospitalization and a major cause of mortality. A model of permanent ligation of the left anterior descending coronary artery in mice is applied to investigate ventricular remodelling and cardiac dysfunction post-myocardial infarction. The technique of invasive hemodynamic measurements in mice is presented.

Ligature permanente de l&#39;artère interventriculaire antérieure coronaire chez la souris: Un modèle de post-infarctus du myocarde et de remodelage de l&#39;insuffisance cardiaque

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It ...

Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, obesity and diabetes. Accordingly, when evaluating therapies for stroke in animals, it is important to select a model with excellent face validity. Ischemic stroke accounts for 80% of all strokes, and the majority of these occur in the territory of the middle cerebral artery (MCA), often inducing infarcts that affect the sensorimotor cortex, causing persistent plegia or paresis on the contralateral side of the body. We demonstrate in this video a method for producing ischemic stroke in elderly rats, which causes sustained sensorimotor disability and substantial cortical infarcts. Specifically, we induce permanent distal middle cerebral artery occlusion (MCAO) in elderly female rats by using diathermy forceps to occlude a short segment of this artery. The carotid artery on the ipsilateral side to the lesion was then permanently occluded and the contralateral carotid artery was transiently occluded for 60 min. We measure the infarct size using structural T2-weighted magnetic resonance imaging (MRI) at 24 hr and 8 weeks after stroke. In this study, the mean infarct volume was 4.5% &#177; 2.0% (standard deviation) of the ipsilateral hemisphere at 24 hr (corrected for brain swelling using Gerriet&#8217;s equation, n = 5). This model is feasible and clinically relevant as it permits the induction of sustained sensorimotor deficits, which is important for the elucidation of pathophysiological mechanisms and novel treatments.

Here we present a protocol to produce permanent distal middle cerebral artery occlusion in elderly female rats with simultaneous occlusion of the carotid arteries to generate large cortical infarcts and sustained deficits. We show confirmation of the lesion size using structural MRI at 24 hr and 8 weeks after stroke.

Exécution distale permanente Moyen cérébrale avec l&#39;artère carotide commune occlusion chez les rats âgés pour étudier corticale ischémie avec une incapacité prolongée

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, ...

Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature and mature brain. Stroke occurs commonly in the perinatal period in humans, and transient ischemia-reperfusion is the most common form of stroke in neonates. The reperfusion phase is a critical component of injury progression, which occurs over a period of days to weeks, and of the endogenous response to injury. This postnatal day 10 (p10) rat model of transient middle cerebral artery occlusion (tMCAO) creates a unilateral, non-hemorrhagic focal ischemia-reperfusion injury that can be utilized to study the mechanisms of focal injury and repair in the full-term-equivalent brain. The injury pattern that is produced by tMCAO is consistent and highly reproducible and can be confirmed with MRI or histological analyses. The severity of injury can be manipulated through changes in occlusion time and other methods that will be discussed.

Neonatal stroke is a significant cause of early brain injury requiring a translational model with consistent focal injury patterns and high reproducibility in order to enable study. This study describes the detailed surgical procedure for creating a non-hemorrhagic, unilateral focal ischemia-reperfusion injury in full-term-equivalent rodents.

Transitoire cérébrale moyenne Artère Occlusion Modèle de l&#39;AVC néonatale chez les rats P10

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature ...

Middle Cerebral Artery Occlusion Allowing Reperfusion via Common Carotid Artery Repair in Mice

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue plasminogen activator (tPA) as an approved drug treatment to target ischemic strokes. Current research in the field of ischemic stroke focuses on better understanding the pathophysiology of stroke, to develop and investigate novel pharmaceutical targets. Reliable experimental stroke models are crucial for the progression of potential treatments. The middle cerebral artery occlusion (MCAO) model is clinically relevant and the most frequently used surgical model of ischemic stroke in rodents. However, the outcomes of this model, such as lesion volume, are associated with high levels of variability, particularly in mice. The alternative MCAO model described here allows the reperfusion of the common carotid artery (CCA) and the increased perfusion of the middle cerebral artery (MCA) territory, using a tissue pad with fibrinogen-based sealant to repair the vessel, and the improved welfare of the mice by avoiding external carotid artery (ECA) ligation. This reduces the reliance on the Circle of Willis, which is known to be highly anatomically variable in mice. Representative data show that using this alternative surgical approach decreases the variability in lesion volumes between the traditional MCAO approach and the alternative approach described here.

Intraluminal filament occlusion of the middle cerebral artery is the most frequently used in vivo model of experimental stroke in rodents. An alternative surgical approach to allow common carotid artery repair is performed here, which allows the reperfusion of the common carotid artery and a full reperfusion to the middle cerebral artery territory.

Occlusion de l’artère cérébrale moyenne permettant une Reperfusion par l’intermédiaire de réparation de l’artère carotide commune chez les souris

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue ...