Ce projet est partiellement financé par le NIH T32 GM008440 à Espen Walker, R01 NS27713 à William L.Young, P01 NS44155 à William L.Young et Hua Su, R21 NS070153 Hua SU et par l&#39;American Heart Association AHA 10GRNT3130004 Hua Su. Dr Ana Rodríguez Hernández-est soutenu par une subvention de &quot;Obra Social La Caixa&quot;

1. Préparation de la souris <ol><li> Induire une anesthésie générale chez la souris avec le gaz isoflurane. Injecter 0,15 ml de brupenorphine intrapéritonéale pour la gestion de la douleur. Avant de commencer, vérifiez si le niveau d&#39;anesthésie est satisfaisante en piquant les pattes de la souris. </li><li> Mettez la souris en décubitus dorsal avec les quatre membres fixées par un ruban adhésif. Enlever les poils du cou et la poitrine avec des ciseaux. Par injection sous-cutanée, administrer 0,2 à 0,4 ml de solution saline à 0,9% pour maintenir la souris hydraté au cours de la procédure chirurgicale. </li><li> Préparer le champ opératoire stérile suivant une méthode rigoureuse. La zone de l&#39;incision de la peau doit être nettoyée avec 90% d&#39;alcool. </li></ol> 2. Dissection l&#39;artère carotide commune et la veine jugulaire externe <ol><li> Faire une incision médiane cervicale horizontale à travers la région du cou inférieure de la souris. Après l&#39;approfondissement de la plaie, élever les glandes salivaires et les tissus mous col de l&#39;utérus en utilisant auraction suture (figure 1). Exposer la veine jugulaire externe droite (CPA) latérale au muscle sterno (SMC). Cette étape doit être effectuée sous le microscope, depuis traction excessive sur la veine pourrait l&#39;endommager et induire sa thrombose. </li><li> Disséquer soigneusement le droit EJV long de son cours de la clavicule à la base du crâne. Habituellement pince bipolaire électriques, coagulent et se divisent toutes les branches de préparer une longueur suffisante pour le placement du clip temporaire plus tard et anastomose. </li><li> Latéral à la trachée et en dedans de l&#39;Accord SMC, d&#39;explorer l&#39;artère carotide commune (CCA). Il doit être soigneusement exposée de la clavicule juste au-delà de sa bifurcation dans les artères carotides externe et interne. Lors de cette étape, une attention devrait être accordée à nouveau pour éviter une traction excessive à la CPA qui pourraient plus tard compromettre la perméabilité de l&#39;anastomose. </li></ol> 3. Préparation de l&#39;anastomose <ol><li> Ligaturer le CCA avec 10-0 Nylon juste proximale par rapport à sa bifurcation. Ensuite, appliquez un clip temporaire sur la proximale CCA aussi près de la clavicule que possible. </li><li> Une fois que le flux a été interrompue, sectionner l&#39;artère un peu moins de la ligature de bifurcation et irriguer avec une solution saline pour laver du sang restant à l&#39;intérieur de la lumière. Éviter la coagulation bipolaire dans cette étape, depuis la blessure thermique à la paroi de l&#39;artère pourrait mettre l&#39;anastomose avenir en péril. </li><li> Afin d&#39;améliorer la visibilité des bords de la CPA, la paroi médiale est marqué avec un marqueur bleu le long du parcours de la veinotomie prévu. Une fois que la CPA est marqué, en utilisant un fil de suture 10-0 pour ligaturer l&#39;extrémité distale comme caudale que possible et appliquer un clip vasculaire temporaire sur l&#39;extrémité proximale du crâne comme possible. </li><li> Avec une amende de 30 aiguille G et 0,5 ml seringue, faire une ouverture initiale sur la zone marquée de la CPA et immédiatement irriguer la lumière avec une solution saline pour éviter la formation de thrombus. Ensuite, étendre la veinotomie avec les microciseauxjusqu&#39;à ce que la longueur est d&#39;environ 2 à 3 fois le diamètre de l&#39;ACC. Évitez violente étirement et faire attention à conserver un avantage de tranchant et soigné. </li><li> Rapprocher la fin de la CCA à la CPA. Faire une incision latérale en coupe de l&#39;extrémité de donneur de l&#39;ACC pour régler le diamètre de la longueur à la taille de Phlebotomie (figure 2). </li></ol> 4. End-to-anastomose <ol><li> Utilisez un suture en nylon 11-0 de monofilament pour l&#39;anastomose-CCA à EJV fin à côte. Suturer la paroi médiale de l&#39;anastomose dans une direction d&#39;crânien-caudal est la première étape. Chaque point doit être placé de l&#39;extérieur dans la paroi veineuse première (figure 3) et intérieur à l&#39;extérieur de la paroi artérielle (Figure 4) suivant. Cela permet de garder le nœud sur la surface extérieure des vaisseaux anastomotiques en tout temps (Figure 5). Soit interrompus ou sutures continues peuvent être utilisés, mais tous les sutures continues doivent être serrées que l&#39;étape finale. </li><li> Once la paroi médiale a été suturée, répéter la procédure à partir caudale à crânienne avec la paroi latérale. Maintenant, chaque point doit être placé de l&#39;extérieur dans la paroi artérielle et de première intérieur vers l&#39;extérieur la paroi veineuse suivant. irrigation Saline aidera à garder la lumière de l&#39;anastomose visible en tout temps pendant la procédure. </li><li> Après la fin de toutes les étapes de l&#39;anastomose, retirer le clip temporaire de la première veine et de l&#39;artère suivant. Le sang artériel se écoule dans la CPA avec peu ou pas de suintement de l&#39;anastomose (voir vidéos modèle VH et le modèle VH 2). Le saignement minime devrait cesser sans utiliser la compression de coton sur l&#39;anastomose. Cela doit être évité afin de prévenir la thrombose de la veine délicate. </li><li> Une fois pulsatile circuler à travers l&#39;anastomose est confirmé et aucun saignement apparent est observé, irriguer le champ opératoire avec une solution saline et fermer l&#39;incision cervicale avec une suture 6-0 en nylon. Enfin, administrer 0,15 ml plus intrapbrupenorphine eritoneal pour la gestion de la douleur postopératoire et 0,2 à 0,4 ml de solution saline sous-cutanée de 0,9% pour reconstituer la perte de sang pendant la chirurgie. </li></ol>

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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonischemic+cerebral+venous+hypertension+promotes+a+pro-angiogenic+stage+through+HIF-1+downstream+genes+and+leukocyte-derived+MMP-9.">Nonischemic cerebral venous hypertension promotes a pro-angiogenic stage through HIF-1 downstream genes and leukocyte-derived MMP-9.</a> J. Cereb. Blood Flow Metab. 29, 1482-1490 (2009).</li><li>Kim, H., Su, H., Weinsheimer, S., Pawlikowska, L., Brain Young, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=arteriovenous+malformation+pathogenesis:+a+response-to-injury+paradigm.">arteriovenous malformation pathogenesis: a response-to-injury paradigm.</a> Acta Neurochir. Suppl. 111, 83-92 (2011).</li><li>Lawton, M. T., Arnold, C. M., Kim, Y. J., Bogarin, E. A., Stewart, C. L., Wulfstat, A. A., Derugin, N., Deen, D., Young, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiation+arteriopathy+in+the+transgenic+arteriovenous+fistula+model.">Radiation arteriopathy in the transgenic arteriovenous fistula model.</a> Neurosurgery. 62, 1129-1138 (2008).</li><li>Lawton, M. T., Stewart, C. L., Wulfstat, A. A., Derugin, N., Hashimoto, T., Young, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+transgenic+arteriovenous+fistula+in+the+rat:+an+experimental+model+of+gene+therapy+for+brain+arteriovenous+malformations.">The transgenic arteriovenous fistula in the rat: an experimental model of gene therapy for brain arteriovenous malformations.</a> Neurosurgery. 54, 1463-1471 (2004).</li><li>Schaller, B., Graf, R., Sanada, Y., Tolnay, M., Rosner, G., Wienhard, K., Heiss, 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=Hemodynamic+changes+after+occlusion+of+the+posterior+superior+sagittal+sinus:+an+experimental+PET+study+in+cats.">Hemodynamic changes after occlusion of the posterior superior sagittal sinus: an experimental PET study in cats.</a> AJNR Am J Neuroradiol. 24, 1876-1880 (2003).</li><li>Zhu, Y., Lawton, M. T., Du, R., Shwe, Y., Chen, Y., Shen, F., Young, W. L., Yang, G. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+hypoxia-inducible+factor-1+and+vascular+endothelial+growth+factor+in+response+to+venous+hypertension.">Expression of hypoxia-inducible factor-1 and vascular endothelial growth factor in response to venous hypertension.</a> Neurosurgery. 59, 687-696 (2006).</li><li>Herman, J. M., Spetzler, R. F., Bederson, J. B., Kurbat, J. M., Zabramski, 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=Genesis+of+a+dural+arteriovenous+malformation+in+a+rat+model.">Genesis of a dural arteriovenous malformation in a rat model.</a> J. Neurosurg. 83, 539-545 (1995).</li><li>Terada, T., Higashida, R. T., Halbach, V. V., Dowd, C. F., Tsuura, M., Komai, N., Wilson, C. B., Hieshima, G. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+acquired+arteriovenous+fistulas+in+rats+due+to+venous+hypertension.">Development of acquired arteriovenous fistulas in rats due to venous hypertension.</a> J. Neurosurg. 80, 884-889 (1994).</li><li>Yassari, R., Sayama, T., Jahromi, B. S., Aihara, Y., Stoodley, M., Macdonald, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Angiographic,+hemodynamic+and+histological+characterization+of+an+arteriovenous+fistula+in+rats.">Angiographic, hemodynamic and histological characterization of an arteriovenous fistula in rats.</a> Acta Neurochir (Wien). 146, 495-504 (2004).</li><li>Solheim, O., Skeidsvoll, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transient+global+amnesia+may+be+caused+by+cerebral+vein+thrombosis.">Transient global amnesia may be caused by cerebral vein thrombosis.</a> Med. Hypotheses. 65, 1142-1149 (2005).</li><li>Yang, B., Shergill, U., Fu, A. A., Knudsen, B., Misra, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mouse+arteriovenous+fistula+model.">The mouse arteriovenous fistula model.</a> J Vasc Interv Radiol. 20, 946-950 (2009).</li><li>Choi, E. J., Choi, E. J., Walker, E. J., Shen, F., Oh, S. P., Arthur, H. M., Young, W. L., Su, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimal+Homozygous+Endothelial+Deletion+of+Eng+with+VEGF+Stimulation+Is+Sufficient+to+Cause+Cerebrovascular+Dysplasia+in+the+Adult+Mouse.">Minimal Homozygous Endothelial Deletion of Eng with VEGF Stimulation Is Sufficient to Cause Cerebrovascular Dysplasia in the Adult Mouse.</a> Cerebrovascular diseases. 33, 540-547 (2012).</li><li>Hao, Q., Su, H., Marchuk, D. A., Rola, R., Wang, Y., Liu, W., Young, W. L., Yang, G. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+tissue+perfusion+promotes+capillary+dysplasia+in+the+ALK1-deficient+mouse+brain+following+VEGF+stimulation.">Increased tissue perfusion promotes capillary dysplasia in the ALK1-deficient mouse brain following VEGF stimulation.</a> Am. J. Physiol. Heart Circ. Physiol. 295, H2250-H2256 (2008).</li><li>Su, H., Hao, Q., Shen, F., Zhu, Y., Lee, C. Z., Young, W. L., Yang, G. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+cerebral+microvascular+dysplasia+model+in+rodents.">Development of a cerebral microvascular dysplasia model in rodents.</a> Acta Neurochir. Suppl. 105, 185-189 (2008).</li><li>Walker, E. J., Su, H., Shen, F., Degos, V., Jun, K., Young, W. 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Les procédures expérimentales des animaux de laboratoire ont été approuvés par le soin et l&#39;utilisation des animaux Commission institutionnelle de l&#39;Université de Californie, San Francisco (UCSF). Les auteurs ne ont aucun conflit d&#39;intérêts potentiels liés aux médicaments et les matériaux utilisés dans cette procédure.

Hypertension veineuse cérébrale soutenue a été étroitement liée à des manifestations cliniques plus graves et mauvais pronostic chez les patients atteints FAV durale et MAV cérébrales 3. Ces effets de CVH ont été largement étudiés dans des modèles de rats 1,2,8. Un modèle équivalent chez la souris permettrait l&#39;utilisation d&#39;animaux génétiquement modifiés qui finirait par permettre l&#39;analyse des voies moléculaires impliquées sur la pathogenèse de l&#39;hypertension...

Les modèles animaux d&#39;hypertension veineuse cérébrale se sont révélés être un outil essentiel dans la compréhension de la physiopathologie de malformations artério-veineuses cérébrales et fistules artério-veineuses 1-7. Le plus utilisé est le modèle de rat créée par une fistule artificielle entre l&#39;artère carotide commune (CCA) et la veine jugulaire externe (CPA), qui provoque une hypertension veineuse cérébrale cohérente (CVH) chez le rat 1,8-10. Modèles de souris équivalent, en ouvrant la possibilité d&#39;utiliser différentes souches de souris transgéniques, permettrait une étude plus approfondie non seulement sur la physiopathologie, mais aussi des thérapies génétiques potentiels pour ces maladies cérébrovasculaires. En outre, le produit d&#39;assemblage expérimentale pourrait également être adapté à l&#39;étude d&#39;autres maladies cérébro-vasculaires liées à l&#39;hypertension veineuse telles que la migraine, amnésie globale transitoire, la cécité monoculaire transitoire, etc. 11 Cependant, les tentatives antérieures pour construire ces modèles de souris have démontré les difficultés avec la perméabilité de la fistule en raison de l&#39;anatomie diminutif 5,12. Ici, nous décrivons notre protocole étape par étape pour une anastomose succès de la CCA et EJV murin qui se traduit à long terme de la fistule de brevet et un hypertension veineuse durable chez la souris.

<table border="1" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>10-0 Sterile Microsuture</td>
			<td>Arosurgical Ic.</td>
			<td>VT5A010Q10</td>
		</tr>
		<tr>
			<td>11-0 Sterile Microsuture</td>
			<td>Arosurgical Ic</td>
			<td>VT4A00N07</td>
		</tr>
		<tr>
			<td>DUROTIP Scissors</td>
			<td>Aesculap</td>
			<td>BC210R</td>
		</tr>
		<tr>
			<td>Micro-Adson Tissue Forceps</td>
			<td>Aesculap</td>
			<td>BD510R</td>
		</tr>
		<tr>
			<td>Microscissors</td>
			<td>Aesculap</td>
			<td>OC496R</td>
		</tr>
		<tr>
			<td>Micro Forceps #5 Jewelers</td>
			<td>Aesculap</td>
			<td>BD331R</td>
		</tr>
		<tr>
			<td>Angled Jewelers Forceps</td>
			<td>Aesculap</td>
			<td>BD329R</td>
		</tr>
		<tr>
			<td>Micro Suture Forceps</td>
			<td>Aesculap</td>
			<td>BD338R</td>
		</tr>
		<tr>
			<td>DUROGRIP Needle Holder</td>
			<td>Aesculap</td>
			<td>BM009R</td>
		</tr>
	</tbody>
</table>

adult mouse venous hypertension model: common carotid artery to external jugular vein anastomosis.

La compréhension de la physiopathologie de malformations artério-veineuses cérébrales et fistules artério-veineuses se est améliorée grâce à des modèles animaux. Un modèle de rat créant une fistule artificielle entre l&#39;artère carotide commune (CCA) et la veine jugulaire externe (CPA) a été largement décrit et prouvé techniquement possible. Cette construction provoque une hypertension veineuse cérébrale cohérente (CVH), et a donc contribué à l&#39;étude de la contribution de l&#39;hypertension veineuse à la formation, les symptômes cliniques, et le pronostic de MAV cérébrales et FAV durale. Modèles de souris ont été équivalentes ne décrit guère et ont montré des problèmes avec une sténose de la fistule. Un modèle murin établie permettrait l&#39;étude de la physiopathologie non seulement mais aussi des thérapies génétiques potentiels pour ces maladies cérébrovasculaires. Nous présentons un modèle de fistule artério-veineuse qui produit une hypertension veineuse intracrânienne durable chez la souris. Microsurgical anastomose of murin CCA et EJV peut être difficile en raison de l&#39;anatomie diminutif et entraîner fréquemment dans une fistule non-brevet. Dans ce protocole, étape par étape, nous adressons à tous les défis importants rencontrés au cours de cette procédure. Éviter la rétraction excessive de la veine lors de l&#39;exposition, en utilisant 11-0 au lieu de 10-0 sutures, et de faire une anastomose soigneusement planifiée fin à côte sont quelques-unes des étapes critiques. Bien que cette méthode nécessite des compétences de microchirurgie de pointe et une courbe d&#39;apprentissage plus que l&#39;équivalent chez le rat, il peut être constamment développé. Ce modèle de roman a été conçu pour intégrer les techniques de souris transgéniques avec un système expérimental déjà bien établie qui se est avéré utile d&#39;étudier les MAV cérébrales et FAV durale. En ouvrant la possibilité d&#39;utiliser des souris transgéniques, un plus large éventail de modèles valides peut être réalisée et les traitements génétiques peut également être testée. Construction expérimentale pourrait également être adapté à l&#39;étude des otses maladies cérébro-vasculaires liés à l&#39;hypertension veineuse comme la migraine, l&#39;amnésie globale transitoire, la cécité monoculaire transitoire, etc.

Le succès du modèle est une fistule artério-veineuse de brevet qui induit l&#39;hypertension veineuse dans le cerveau de souris. Pour valider le modèle nous avons d&#39;abord mesuré la pression veineuse intracrânienne dans le sinus sagittal de la souris à 2, 3 et 4 semaines après la chirurgie. 6 souris ont été affectés à différents chaque groupe de temps. La pression des sinus était de 8,8 ± 1,2 mmHg dans le groupe mesurés deux semaines après la chirurgie. Dans les 6 souris mesurées 3 semaines après l...

Watch this Scientific Journal Video about Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis. at JoVE.com

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

Nous décrivons un procédé pour créer un modèle fiable de l&#39;hypertension veineuse cérébrale chez la souris adulte. Ce modèle a été largement décrits et testés chez le rat. Ce nouvel homologue de la souris ouvre la possibilité d&#39;utiliser des animaux génétiques modifiés et élargit ainsi les applications du modèle.

Adulte souris veineuse Modèle d&#39;hypertension: l&#39;artère carotide commune à externe Veine jugulaire anastomose.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

adult-mouse-venous-hypertension-model-common-carotid-artery-to

Research

JoVE Journal

Medicine

14.0K vues.  University of California, San Francisco.  Nous décrivons un procédé pour créer un modèle fiable de l'hypertension veineuse cérébrale chez la souris adulte. Ce modèle a été largement décrits et testés chez le rat. Ce nouvel homologue de la souris ouvre la possibilité d'utiliser des animaux génétiques modifiés et élargit ainsi les applications du modèle. 

Vidéo: Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

Training a Sophisticated Microsurgical Technique: Interposition of External Jugular Vein Graft in the Common Carotid Artery in Rats

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in peripheral arterial bypass surgery as well as in arteriovenous fistulas.1-5 The experimental procedure of vein graft interposition in the common carotid artery by using the cuff-technique has been applied in several research projects to examine the aetiology of neointimal hyperplasia and therapeutic options to address it. 6-8 The cuff prevents vessel anastomotic remodeling and induces turbulence within the graft and thereby the development of neointimal hyperplasia.
Using the superior caval vein graft is an established small-animal model for venous arterialization experiment.9-11 This current protocol refers to an established jugular vein graft interposition technique first described by Zou et al., 9 as well as others.12-14 Nevertheless, these cited small animal protocols are complicated.
To simplify the procedure and to minimize the number of experimental animals needed, a detailed operation protocol by video training is presented. This video should help the novice surgeon to learn both the cuff-technique and the vein graft interposition. Hereby, the right external jugular vein was grafted in cuff-technique in the common carotid artery of 21 female Sprague Dawley rats categorized in three equal groups that were sacrificed on day 21, 42 and 84, respectively. Notably, no donor animals were needed, because auto-transplantations were performed. The survival rate was 100 % at the time point of sacrifice. In addition, the graft patency rate was 60 % for the first 10 operated animals and 82 % for the remaining 11 animals. The blood flow at the time of sacrifice was 8&plusmn;3 ml/min. In conclusion, this surgical protocol considerably simplifies, optimizes and standardizes this complicated procedure. It gives novice surgeons easy, step-by-step instruction, explaining possible pitfalls, thereby helping them to gain expertise fast and avoid useless sacrifice of experimental animals.

Neointimal hyperplasia is the primary cause of stenosis in arterialized veins. We propose a new protocol whereby the right external jugular vein is grafted using the cuff technique in the common carotid artery of Sprague Dawley rats. The survival rate was 100 % at the time point of sacrifice.

Former une technique microchirurgicale sophistiquée: Interposition de greffe veine jugulaire externe dans l&#39;artère carotide commune chez les rats

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in ...

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Médecine

Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We have established bilateral common carotid artery occlusion as a preconditioning stimulus to induce early ischemic tolerance to transient focal cerebral ischemia in C57Bl6/J mice. In this video, we will demonstrate the methodology used for this study.

There is accumulating evidence, that ischemic preconditioning (PC) &#x2013; a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We established bilateral common carotid artery occlusion (BCCAO) as a preconditioning stimulus to induce early ischemic tolerance (IT) to transient focal cerebral ischemia (induced by middle cerebral artery occlusion, MCAO) in C57Bl6/J mice.

Occlusion bilatérale de l&#39;artère carotide commune comme un stimulus suffisant pour provoquer le préconditionnement précoce Tolérance ischémique à une ischémie cérébrale focale

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a ...

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. Intra-tracheal inoculation of a known amount of bacteria has been successfully used to reproduce the pathogenesis of pneumonia which then develops into sepsis. Monitoring hemodynamic parameters and providing standard clinical treatment including infusion of antibiotics, fluids and drugs to maintain blood pressure is critical to simulate routine supportive care in this model but to do so requires both arterial and venous vascular access. The video details the surgical technique for implanting carotid artery and common jugular vein catheters in an anesthetized rat. Following a 72 hr recovery period, the animals will be re-anesthetized and connected to a tether and swivel setup attached to the rodent housing which connects the implanted catheters to the hemodynamic monitoring system. This setup allows free movement of the rat during the study while continuously monitoring pressures, infusing fluids and drugs (antibiotics, vasopressors) and performing blood sampling.

Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

Cathétérisme de l&#39;artère carotide et Veine jugulaire faire pour effectuer des mesures hémodynamiques, Infusions et prélèvement sanguin dans un modèle de rat Conscients

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Chlorure ferrique induite souris thrombose type sur l&#39;artère carotide et Mesentery navire

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

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, ...

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves focal endothelial-selective transgenesis, thereby allowing an investigator to determine the biological roles of endothelial-expressed transgenes and to quantify the in vivo transcriptional activity of DNA sequences in large artery endothelial cells. The method uses surgical isolation of rabbit common carotid arteries and an arteriotomy to deliver a transgene-expressing viral vector into the arterial lumen. A short incubation period of the vector in the lumen, with subsequent aspiration of the lumen contents, is sufficient to achieve efficient and durable expression of the transgene in the endothelium, with no detectable transduction or expression outside of the isolated arterial segment. The method allows assessment of the biological activities of transgene products both in normal arteries and in models of human vascular disease, while avoiding systemic effects that could be caused either by targeting gene delivery to other sites (e.g. the liver) or by the alternative approach of delivering genetic constructs to the endothelium by germ line transgenesis. Application of the method is limited by the need for a skilled surgeon and anesthetist, a well-equipped operating room, the costs of purchasing and housing rabbits, and the need for expertise in gene-transfer vector construction and use. Results obtained with this method include: transgene-related alterations in arterial structure, cellularity, extracellular matrix, or vasomotor function; increases or reductions in arterial inflammation; alterations in vascular cell apoptosis; and progression, retardation, or regression of diseases such as intimal hyperplasia or atherosclerosis. The method also allows measurement of the ability of native and synthetic DNA regulatory sequences to alter transgene expression in endothelial cells, providing results that include: levels of transgene mRNA, levels of transgene protein, and levels of transgene enzymatic activity.

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

This method is to introduce a transgene into the endothelium of rabbit carotid arteries. Introduction of the transgene allows the assessment of the biological role of the transgene product either in normal arteries or disease models. The method is also useful for measuring activity of DNA regulatory sequences.

In Vivo Transfert de gènes à l’endothélium de l’artère carotide commune lapin

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves ...

A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, intimal hyperplasia, and atherosclerosis. The goal of this article is to demonstrate a method for placing bilateral venous interposition grafts in a rabbit, then transducing the grafts with a gene transfer vector that achieves durable transgene expression. The method allows the investigation of the biological roles of genes and their protein products in normal vein graft homeostasis. It also allows the testing of transgenes for the activities that could prevent vein graft failure, e.g., whether the expression of a transgene prevents the neointimal growth, reduces the vascular inflammation, or reduces atherosclerosis in rabbits fed with a high-fat diet. During an initial survival surgery, the segments of right and left external jugular vein are excised and placed bilaterally as reversed end-to-side common carotid artery interposition grafts. During a second survival surgery, performed 28 days later, each of the grafts is isolated from the circulation with vascular clips and the lumens are filled (via an arteriotomy) with a solution containing a helper-dependent adenoviral (HDAd) vector. After a 20-min incubation, the vector solution is aspirated, the arteriotomy is repaired, and flow is restored. The veins are harvested at time points dictated by individual experimental protocols. The 28-day delay between the graft placement and the transduction is necessary to ensure the adaptation of the vein graft to the arterial circulation. This adaptation avoids rapid loss of transgene expression that occurs in vein grafts transduced before or immediately after grafting. The method is unique in its ability to achieve durable, stable transgene expression in grafted veins. Compared to other large animal vein graft models, rabbits have advantages of low cost and easy handling. Compared to rodent vein graft models, rabbits have larger and easier-to-manipulate blood vessels that provide abundant tissue for analysis.

This method describes the placement of interposition vein grafts in rabbits, the transduction of the grafts, and the achievement of durable transgene expression. This allows the investigation of physiological and pathological roles of transgenes and their protein products in grafted veins, and testing of gene therapies for vein graft disease.

Un modèle de lapin d’Expression du transgène durables dans la veine jugulaire à des greffes d’Interposition artère carotide commune

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

Phase contraste d’imagerie par résonance magnétique dans l’artère carotide commune de Rat

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Mesure de la contractilité isométrique de l’artère mésentérique de souris à l’aide de fil myographie

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

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 ...