Les auteurs remercient Laurie von Melchner, Hôpital universitaire de Berne, Département de Neurochirurgie, Berne, Suisse, pour la relecture et l&#39;édition du manuscrit et Paskus Jérémie, Hôpital pour enfants de Boston, Boston, MA pour la relecture du projet initial. Nous apprécions la gestion habile des soins aux animaux, l&#39;anesthésie et l&#39;assistance opérationnelle de Daniel Mettler, DVM, Max Müller, DVM, Daniel Zalokar, et Olgica Beslac, Expérimental Surgical Institute, Département de Recherche Clinique, Université de Berne, Berne, Suisse. Nous remercions Michael Lensch, chef Infirmière de recherche, Département de médecine de soins intensifs de l&#39;hôpital universitaire de Berne et de l&#39;Université de Berne, Berne, Suisse, pour la surveillance des données en temps réel et post-traitement des paramètres physiologiques. Nous remercions Edin Nevzati, Carl Muroi, et Salomé Erhardt, pour leur excellente assistance technique et opérationnelle laboratoire. Ce travail a été soutenu par le ministère de Intensive Care Medicine, University Hospital de Berne et de l&#39;Université de Berne, Berne, Suisse, le Département de Recherche Clinique, Université de Berne, Berne, Suisse, et le Fonds de recherche de l&#39;Hôpital cantonal d&#39;Aarau, Aarau, Suisse. Nous remercions Elsevier, la permission de réimpression pour les figures 1 et 2.

Trois mois vieilles femelles lapins néo-zélandais pesant 2.5-3.5 kg ont été utilisés pour cette procédure. L&#39;étude a été réalisée en conformité avec les Instituts nationaux de la santé des lignes directrices pour les soins et l&#39;utilisation des animaux de laboratoire et avec l&#39;approbation du Comité du canton de Berne, Suisse (approbation n ° 105/13) de protection des animaux. Toutes les interventions chirurgicales ont été réalisées dans des conditions stériles à l&#39;Institut de chirurgie expérimentale du Département de recherche clinique à l&#39;hôpital universitaire de Berne à Berne, Suisse. Un anesthésiste vétérinaire a suivi les animaux pendant la chirurgie et tout au long de la récupération. 1 Préparation des animaux, de positionnement et artère sous-clavière Canulation <ol><li> Induire une anesthésie générale chez le lapin avec une injection intramusculaire de kétamine (30 mg / kg; Ketalar, 50 mg / ml) et de xylazine (6 mg / kg; Xylapan 20 mg / ml) et de la profondeur de l&#39;anesthésie de commande en vérifiant la réponse du lapin à nocive stimulation (par exemple, pincement de l&#39;orteil). Voir 1.7 en cas de réponse positive. </li><li> Déroulez les paupières inférieures des deux yeux et appliquez une petite quantité de pommade sur les paupières pour prévenir la sécheresse et l&#39;irritation. </li><li> Sonder la veine latérale de l&#39;oreille avec un Gbutterfly Venflon® 20 (20 G de cathéter vasculaire), fixer avec du ruban adhésif, et se connecter à un sac de gravité contenant du chlorure de sodium à 0,9% (500 ml / 24 h) et la kétamine (40 mg / kg / h) / xylazine (4 mg / kg / h) pour une administration intraveineuse continue (iv) l&#39;anesthésie. Administrer des analgésiques supplémentaires toutes les 15 min iv (fentanyl, 1 mcg / kg). Remarque: Évitez les anesthésiques de gaz volatils, qui est associée à une diminution du RPC, l&#39;augmentation de la CBF, et la baisse des taux métabolique cérébrale pour l&#39;oxygène 11 anesthésiques intraveineux offrent des caractéristiques plus idéales pour neuroanesthésie en préservant CBF et la vasoconstriction cérébrale, 12, qui est d&#39;une importance supérieure lors de l&#39;étude cérébrale. vasospasme. En outre, bien que la mortalité est augmentée dans spontrespiration aneously animaux, il peut mieux imiter la situation humaine de SEP aiguë. </li><li> Fournir de l&#39;oxygène (1 - 2 L / min) par l&#39;intermédiaire d&#39;un masque respiratoire qui permet à du dioxyde de carbone en fin d&#39;expiration (EtCO 2) la surveillance. </li><li> Installez un électrocardiogramme à 3 canaux (ECG) .Positionner trois électrodes sous-cutanées dans un arrangement triangulaire sur la face ventrale du lapin; spécifiquement, placer une électrode sur la région midthoracic droite (avec la distance sur le terrain rasé stérile pour le cathétérisme de l&#39;artère sous-clavière) et deux électrodes dans le bas-ventre répartis sur les deux branches. </li><li> Surveiller la profondeur d&#39;anesthésie toutes les 15 min pendant la chirurgie par suite de la fréquence respiratoire, la fréquence cardiaque (HR) contrôlée à partir du signal ECG, et réaction à une stimulation nociceptive. </li><li> En cas de réponse positive à une stimulation nociceptive (orteil de pincement), adapter la profondeur de l&#39;anesthésie par kétamine bolus (6 mg / kg) iv et xylazine bolus (0,05 mg / kg) iv et / ou un bolus analgésie supplémentaire avec Fentanyl (1 mcg / kg) iv </li><li> Fixer le lapin dans une position couchée sur une plaque de réchauffement du corps, en inclinant la tête 20 ° vers le bas et en la tournant légèrement controlatérale au côté sur lequel l&#39;artère sous-clavière sera exposé. </li><li> appliquer une pommade ophtalmique et préparer le terrain pour la chirurgie par le rasage des poils sur le pectoral droit dans le tiers médian de la clavicule, et sur le crâne frontal-, parietal- et occipital, le cou, et sur l&#39;artère fémorale commune droite. </li><li> Désinfectez la peau pendant 3 min avec un large spectre antiseptique, par exemple., Povidone iodée. </li><li> Couvrir le lapin avec des draps stériles. Effectuer toutes autres procédures dans des conditions stériles et fréquemment l&#39;application 4% papavérine HCl et une solution antibiotique (sulfate de néomycine 5 mg / ml) par voie topique pour prévenir le vasospasme artériel par la manipulation de la cuve et des infections locales. </li><li> Infiltrer le muscle pectoral avec des anesthésiques locaux (lidocaïne 1% maximum de 6 mg / kg). Faire une incision parasternale peau et la préparer lamuscle pectoral. Utilisation du microscope, disséquer l&#39;artère sous-clavière et le fixer avec une extrémité proximale et distale ligature (4-0 sutures polyfilament) autour de l&#39;extrémité exposée. Gardez une ligature à proximité de la commande proximale en place pour fixer le cathéter et ligaturer le vaisseau distal. </li><li> Effectuer une artériotomie dans la paroi de l&#39;artère sous-clavière par incision de l&#39;artère avec un microscissor courbe et cathétériser l&#39;artère sous-clavière rétrograde avec un petit intravasculaire robinet à 3 voies. Fixer le cathéter en double ligature vers le noeud de ligature distale afin d&#39;empêcher la torsion de l&#39;artère ou de la flexion de la partie proximale de l&#39;artère et à éviter un glissement ou un saignement massif. </li></ol> 2. pression artérielle et de la surveillance Gaz du sang artériel <ol><li> Connectez le robinet d&#39;arrêt à 3 voies à i) la canule intravasculaire pour gaz du sang artériel (ABG) analyse, le pH, la PaCO 2, PaO 2, le bicarbonate, l&#39;excès de base, et de SO 2, ii) le sang artériel invasifpression de l&#39;appareil de mesure, et iii) le dispositif de dérivation. </li><li> Prélever des échantillons de sang pour le statut ABG (PaCO 2, PaO 2) et de surveiller en permanence les paramètres standards cardiovasculaires et respiratoires (pression artérielle, RH, ECG, fréquence respiratoire et de fin-de-marée de CO 2) et de transfert de données via l&#39;interface de sortie analogique à un Analog- convertisseur / enregistreur de données numérique et magasin. REMARQUE: Les pressions seront remis à zéro au niveau du cœur avant et après chaque session, et étalonnage de la pression du convertisseur analogique / numérique convertisseur et système d&#39;enregistrement des données se fera une fois avant la série commence. </li></ol> 3. de base par soustraction numérique angiographie <ol><li> Passer une dispositif de calibrage externe (petite sphère) sur les deux angles de la mandibule afin de calibrer l&#39;angiographie. NOTE: Ceci permet la comparaison exacte des mesures a posteriori de la ligne de base et le suivi diamètre du vaisseau. </li><li> Effectuez l&#39;angiographie numérique (DSA) par rétrograde intra-uninjection d&#39;un bolus de rterial Iopamidol non ionique (0,6 ml / kg, 5 ml / s pendant 2 secondes) à travers l&#39;artère canulée et rincer immédiatement la canule après injection d&#39;un bolus de solution saline afin d&#39;empêcher l&#39;occlusion de celui-ci. </li><li> Obtenir des images (7 images à 14 sec) du système vertébro utilisant un enregistrement rapide d&#39;angiographie séquentielle dans une position oblique 5 ° à gauche antérieure. </li><li> Infiltrez la zone autour de l&#39;artère fémorale commune droite avec des anesthésiques locaux (lidocaïne 1%, maximum 6 mg / kg). Faire une petite incision inguinale de la peau. Utilisation du microscope pour la visualisation, disséquer l&#39;artère fémorale commune et le fixer avec une extrémité proximale et distale ligature (4-0 sutures de polyfilament) autour de l&#39;extrémité exposée. </li><li> Après artériotomie, cathétériser l&#39;artère fémorale d&#39;une gaine 5-F. Rincer l&#39;orifice latéral de la gaine avec une solution saline. </li><li> Progresser un cathéter 5-F dans l&#39;artère brachiocéphalique à travers la gaine sous fluoroscopie. Créer une feuille de route, puis avancez un fil de guidage à the système vertébro. Injecter un bolus de Iopamidol non-ionique (0,6 ml / kg, 5 ml / sec en 2 sec) pour DSA comme décrit dans l&#39;étape 3.2. </li></ol> 4 Rotation à position couchée <ol><li> Après base DSA, repositionner le lapin de la position couchée à la position couchée. Veillez à ne pas manipuler ou déplacer la position des cathéters intra-artériels. </li><li> Placez la tête dans un porte-tête à un angle de 30 °, orienté tête vers le bas. </li></ol> 5. Cisterna Magna Puncture <ol><li> Désinfectez la peau sur la tête et le cou avec Povidone iodée 3 fois pendant 1 min chacun, et couvrir la zone chirurgicale avec des draps stériles. </li><li> Insérez un x 40 mm aiguille d&#39;accès moelle pédiatrique de 22 G transcutanée dans la grande citerne sans incision de la peau avant ou déplacement de muscle. </li><li> Assurez-vous que l&#39;animal est complètement anesthésié en assurant une absence de réponse orteil-pincement avant de glisser l&#39;aiguille le long de la osseux protubérance occipitale externejusqu&#39;à ce qu&#39;un écart a été détecté; ne pas pousser l&#39;aiguille plus loin. </li><li> Confirmer le bon positionnement de l&#39;aiguille en observant des gouttes spontanée de liquide céphalo-rachidien avec la tête du lapin incliné à un angle de 20 ° pendant quelques minutes. </li></ol> 6 Installation de la pression intracrânienne et cérébrale surveillance du flux sanguin <ol><li> Après cutanée médiane et Galea incision, insérer un petit écarteur chirurgical. </li><li> Faire trois ostéotomies rond (diamètre de 2 mm) à l&#39;aide d&#39;un micro-foret à haute vitesse dans la partie frontale du crâne selon les points de repère du crâne extérieures (Figure 1) 9, c&#39;est à dire, sur le bulbe olfactif et frontal bilatéral pour le placement d&#39;un dispositif de neuromonitorage si nécessaire. Utilisez une règle à l&#39;échelle du millimètre à déterminer les coordonnées de bavure emplacement des trous comme suit: pression intracrânienne (PIC) suivi dans la ligne midpupillary, une à deux mm de la ligne sagittale médiane; intraparenchymateuses sondes laser Dopplerquatre à cinq mm antérieure et latérale par rapport au bregma (figure 1). </li><li> Visualisez la dure-mère, et effectuer l&#39;hémostase méticuleuse: utiliser de la cire d&#39;os pour l&#39;hémostase osseuse en raison de son action de tamponnement et effectuer l&#39;hémostase locale à l&#39;aide de la coagulation bipolaire de la dure-mère. </li><li> Placez la pression intracrânienne (PIC) pointe du moniteur intraparenchymateuse dans le bulbe olfactif droit à une profondeur de 2 mm, puis étalonner. </li><li> Placez les sondes à l&#39;aiguille fine de deux laser-Doppler en utilisant un système de serrage externe et de les insérer dans les trous de bavures correspondants dans les deux hémisphères droit et frontal latéral gauche du système ventriculaire, c&#39;est à dire, sur la ligne médiane pour éviter toute interférence avec le fluide céphalo-rachidien. Placer les sondes d&#39;aiguilles jusqu&#39;à une profondeur de 2,5 mm. </li><li> Après mise en place des sondes de neuromonitoring, sceller tous les trous de bavures avec un bouchon de cire d&#39;épaisseur de l&#39;os afin de maintenir le crâne étanche. </li><li> Mesurer les paramètres de base de la moyenne de la pression artérielle (MAP), ICP et la circulation sanguine cérébrale (CBF) à l&#39;aide d&#39;un moniteur à paramètres multiples et quatre canaux tissu laser-Doppler moniteur de perfusion sanguine. </li></ol> 7. Shunt induction <ol><li> Connectez l&#39;aiguille d&#39;accès moelle dans la grande citerne de l&#39;artère sous-clavière auparavant un cathéter par une tubulure de surveillance de la pression artérielle remplie. Utiliser le robinet à 3 voies pour la mesure de la pression artérielle et que le port de prélèvement de sang. NOTE: La gravité de SEP dépend de la quantité de sang, et peut être grossièrement estimé par le ampleur de caillots subarachnoïdiens au moment de la récolte du cerveau 5,11. </li><li> Surveiller en permanence MAP, RH, ECG, la fréquence respiratoire et la fin-de-marée de CO 2 à un taux d&#39;échantillonnage de 1 Hz à partir de 6 min avant au moins jusqu&#39;à 20 min après HSA. </li><li> Assurez-vous que l&#39;animal est complètement anesthésié en assurant une absence de réponse orteil de pincement avant d&#39;ouvrir la connexion de dérivation entre l&#39;artère sous-clavière et la grande citerne pour induire SAH par la Gradie de pressionnt. REMARQUE: Un SAH contrôlée peut être obtenue par la fermeture de la dérivation à n&#39;importe quel moment (par exemple, à un niveau souhaité de ICP.). </li><li> Enregistrer des valeurs de régime permanent pendant une période de temps d&#39;environ 15 min. </li><li> Après ICP atteint son apogée, maintenir l&#39;aiguille d&#39;accès vertébrale en place jusqu&#39;à ce que l&#39;ICP retourne à un état d&#39;équilibre proche des valeurs de référence. Si le plateau du PCI est maintenue pendant plus de 10 secondes ou si ICP diminue spontanément, fermer le shunt. </li><li> Retirer CBF sondes à l&#39;aiguille fine et la sonde ICP, boucher les trous de bavures avec de la cire d&#39;os, retirez tous les cathéters (y compris cathéter sous-clavière, depuis la manipulation du cathéter avec saignement consécutif est associée à une forte morbidité et la mortalité, et augmente le taux d&#39;infection), effectuer rigoureuse irrigation de la plaie avec sulfate de néomycine et suturer la peau. </li></ol> 8. gestion postopératoire <ol><li> La procédure dure environ 2 heures. En raison de la demi-vie de la kétamine et de la xylazine, le temps de récupération de l&#39;animal est assez courte - environ 1 h. Les animaux sont maintenus sous une lampe chauffante pendant la récupération. D&#39;autres liquides ne sont pas fournis. Au cours de cette phase initiale de récupération post-opératoire, appliquer la buprénorphine 0,02 mg / kg sc toutes les 8 heures pendant 24 heures. </li><li> Appliquer transdermique de fentanyl matrice patchs libérant 12,5 pg / h dans la région du cou rasé des animaux pour l&#39;analgésie efficace au cours de la prochaine 72 heures. </li><li> Ne pas laisser un animal sans surveillance tant qu&#39;il a repris conscience suffisante pour maintenir décubitus sternal. </li><li> Ne retournez pas un animal qui a subi une intervention chirurgicale dans la compagnie d&#39;autres animaux jusqu&#39;à guérison complète. </li></ol> 9 Suivi angiographie par soustraction numérique pour évaluer DCVS au Jour 3 <ol><li> Effectuez les étapes 1.1 à 3.6 comme décrit ci-dessus. </li><li> Euthanasier les animaux par injection intra-artérielle de bolus de thiopental de sodium (40 mg / kg) (thiopental, Ospedalia AG, Hünenberg, Suisse). Dans les cas où l&#39;histologie et immunohistochemistry est nécessaire, effectuer une perfusion intracardiaque de fixation à la température ambiante sous une pression de perfusion de 100 cm H 2 O. </li></ol>

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R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+targeting+in+mice:+functional+analysis+of+the+mammalian+genome+for+the+twenty-first+century.">Gene targeting in mice: functional analysis of the mammalian genome for the twenty-first century.</a> Nature reviews. Genetics. 6, 507-512 (2005).</li><li>Flisikowska, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+immunoglobulin+gene+disruption+and+targeted+replacement+in+rabbit+using+zinc+finger+nucleases.">Efficient immunoglobulin gene disruption and targeted replacement in rabbit using zinc finger nucleases.</a> PloS one. 6, e21045 (2011).</li><li>Nakajima, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+aging+on+cerebral+vasospasm+after+subarachnoid+hemorrhage+in+rabbits.">Effects of aging on cerebral vasospasm after subarachnoid hemorrhage in rabbits.</a> Stroke. 32, 620-628 (2001).</li></ol>

None. The authors have no financial or commercial interest in any of the drugs, materials, or equipment used. No specific funding was received for this work. The authors are solely responsible for the design and conduct of the presented study and report no conflict of interest concerning the materials and methods used in this study or the findings specified in the paper. They confirm the adherence of ethical standards. The study was performed in accordance with the National Institutes of Health guidelines for the care and use of experimental animals and with the approval of the Animal Care Committee of the Canton of Bern, Switzerland (approval #109/07 and #107/09).

Le modèle de dérivation produit pathologie similaire à celle observée chez l&#39;homme après aiguë SAH 3,8,10. Il a été suggéré que EBI peut exacerber, maintenir et même déclencher DCVS 12, et que ce tel modèle peut aider à enquêter sur les phases précoces et tardives DCVS, y compris les interactions EBI et DCVS suivantes SAH. En particulier, reproductible in vivo DCVS techniques de surveillance, y compris DSA 13, calculé angiographie par tomographie 14,</s...

Hémorragie hémorragie méningée (HSA) est l&#39;une des plus mortelles conditions neuropathologiques, conduisant souvent à des dommages ou la mort 1 neurologiques permanentes. Des recherches antérieures ont mis l&#39;accent sur ​​le vasospasme cérébral retardé (DCVS) que l&#39;étiologie primaire des déficits neurologiques associés à la SEP 2. Cependant, les pauvres en général les résultats cliniques des patients souffrant de SEP après le traitement du vasospasme a conduit à une expansion de l&#39;axe de recherche pour inclure les effets de la petite lésion cérébrale (EBI) après SAH 3. Une meilleure compréhension de l&#39;importance à la fois de EBI et DCVS en contribuant à de mauvais résultats cliniques après SAH est essentiel pour le développement de stratégies thérapeutiques plus efficaces. Jusqu&#39;à maintenant, l&#39;injection de sang autologue simple et double dans la grande citerne a été la méthode standard pour SAH induction pour l&#39;étude de DCVS 2-6. Bien que couramment utilisé dans les études antérieures,ce modèle très probablement ne pas reproduire les principaux changements neuropathologiques de la SEP induit EBI 7. En revanche, la perforation endovasculaire est connu pour produire des modifications physiopathologiques aiguës sévères qui reproduisent partiellement les symptômes de l&#39;EBI 7. Ce rapport décrit un nouveau modèle de lapin de SAH conçu pour permettre une enquête de deux EBI et DCVS, ce qui permet une caractérisation plus précise de la pathologie SEP-induite 8-10. Avec la technique décrite, modèle standard grande citerne est adapté en connectant le système artériel de l&#39;artère sous-clavière et la grande citerne via un shunt extracorporelle. La circulation du sang est ainsi liée à la physiologie du lapin et entraîné par la différence de pression entre le sang artériel et de la pression intracrânienne. Le saignement s&#39;arrête lorsque la pression intracrânienne (PIC) est égale à la pression artérielle diastolique et le sang dans le système de shunt coagule. Utilisant l&#39;hôte &amp; #8217; s physiologie réduit examinateur dépendant SAH induction, ce qui conduit à un modèle plus cohérent de SEP qui produit de façon fiable à la fois EBI et phénotypes DCVS 3,8-10.

<table border="0" cellpadding="0" cellspacing="0" width="905">
	<tbody>
		<tr height="34">
			<td height="34" style="height:35px;width:279px;">Name</td>
			<td style="width:236px;">Company</td>
			<td style="width:223px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:279px;">Equipment</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">operation microscope</td>
			<td style="width:236px;">Zeiss, Jena, Germany</td>
			<td colspan="2">Zeiss, OPMI-MD surgical microscope</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:279px;">surgical equipment</td>
			<td style="width:236px;">B. Braun, Germany</td>
			<td style="width:223px;">forceps medical n&#176;5, vessel sciccors 8cm, microclip 4mm</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">respirator</td>
			<td style="width:236px;">Hugo Sachs</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">hair clipper</td>
			<td style="width:236px;">3M Surgical Clipper&#160;</td>
			<td style="width:223px;">&#160;Starter Kit 9667A</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">body warm plate</td>
			<td style="width:236px;">FHC</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:279px;">blood gas analyzer</td>
			<td style="width:236px;">Radiometer, Copenhagen, Denmark</td>
			<td style="width:223px;">ABL 725</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:279px;">cardiac monitoring</td>
			<td style="width:236px;">Camino Multi-Parameter Monitor, Integra, Plainsboro, NJ, US</td>
			<td style="width:223px;">AP-05</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;">software analysis</td>
			<td style="width:236px;">BIOPAC Systems, Inc., Goleta, CA, USA</td>
			<td style="width:223px;">Biopac MP100 and acqKnowledge software,version 3.8.1</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;">software analysis</td>
			<td style="width:236px;">ImagePro Discovery, MediaCybernetics, Silver Spring, MD, USA</td>
			<td style="width:223px;">image-Pro Plus version&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">angiography apparatus</td>
			<td style="width:236px;">DFP 2000 A-Toshiba</td>
			<td style="width:223px;">MIIXR0001EAA</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">ICP monitor</td>
			<td style="width:236px;">Camino Laboratories, San Diego, CA, USA</td>
			<td style="width:223px;">ICP monitor, Model 110-4B</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">blood flow monitor</td>
			<td style="width:236px;">Oxford Optronix Ltd., Oxford, UK</td>
			<td style="width:223px;">CAL KIT microsphere solution</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">laser-Doppler flowmetry fine needle probes&#160;</td>
			<td style="width:236px;">Oxford Optronix Ltd., Oxford, UK</td>
			<td style="width:223px;">MNP110XP, 0.48 mm diameter</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">pressure tube</td>
			<td>B. Braun, Germay</td>
			<td style="width:223px;">PE 1.0 mm &#215; 2.0 mm</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">anesthesia monitor</td>
			<td>GE Medical Systems, Switzerland&#160;</td>
			<td style="width:223px;">Datex S5 Monitor</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:279px;">Material</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">20 G vascular catheter</td>
			<td style="width:236px;">Smiths Medical</td>
			<td style="width:223px;">Jelco i.v. catheter, REF 4057</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">5.5F three-lumen central venous catheter&#160;</td>
			<td style="width:236px;">Connectors, Tagelswangen, Switzerland</td>
			<td>silicone catheter STH-C040</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">22Gx40mm needle&#160;</td>
			<td style="width:236px;">Emergo Group Inc., Netherlands</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">high-speed microdrill</td>
			<td style="width:236px;">Stryker, Solothurn, Switzerland</td>
			<td>5400-15&#160;</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">bone wax</td>
			<td style="width:236px;">Ethicon, Johnson &#38; Johnson,NJ, USA</td>
			<td style="width:223px;">ETHW31G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">bipolar forceps</td>
			<td style="width:236px;">Aesculap, Inc., PA, US</td>
			<td>US349SP&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Ketamin</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Xylazine</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Buprenorphine</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fentanyl</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">transdermal fentanyl matrix patches&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Lidocaine 1%&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">4% papaverin HCl&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Neomycin sulfate&#160;</td>
			<td style="width:236px;">Research Organics Inc., OH, USA</td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Povidone-iodine&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">0.9% sodium chloride</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Iopamidol&#160;</td>
			<td style="width:236px;">Abott Laboratories, IL, USA</td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">3-0 resorbable suture</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td>VCP824G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">5-0 non absorbable suture</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td>8618G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">4-0 polyfilament sutures</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td style="width:223px;">VCP284G</td>
			<td></td>
		</tr>
	</tbody>
</table>

the rabbit blood-shunt model for the study of acute and late sequelae of subarachnoid hemorrhage: technical aspects

Lésion cérébrale précoce et le vasospasme cérébral retardé à la fois contribuer à des résultats défavorables après une hémorragie méningée (SAH). Modèles animaux reproductibles et contrôlables qui simulent les conditions sont actuellement rares. Par conséquent, de nouveaux modèles sont nécessaires afin de reproduire les conditions physiopathologiques humaines résultant de SEP. Ce rapport décrit les nuances techniques d&#39;un modèle de SEP sang-shunt lapin qui permet un contrôle de la pression intracrânienne (PIC). Une dérivation extracorporelle est placée entre le système artériel et l&#39;espace sous-arachnoïdien, ce qui permet examinateur indépendant SAH dans un crâne de fermeture. Étape par étape les instructions de procédure et de l&#39;équipement nécessaire sont décrits, ainsi que les considérations techniques pour produire le modèle de la mortalité et une morbidité minimale. Des détails importants nécessaires pour la création chirurgicale réussie de ce modèle robuste, simple et cohérente SAH lapin ICP-contrôlée sont décrits.

<html> Le modèle de shunt de sang de lapin de SAH décrit dans ce rapport produit EBI dans l&#39;hippocampe (figure 2A, B), le cortex basale (Figure 2A, B), et le système vasculaire cérébral (figure 2C) dès 24 h après la blessure et représente une caractéristique distribution du sang (figure 2D) 8. En outre, le modèle déclenche modérée à des degrés sévères de DCVS sur trois jours après SAH induction (figure 3)...

Watch this Scientific Journal Video about The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects at JoVE.com

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

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.

Le lapin de sang-shunt modèle pour l&#39;étude de aiguë et les séquelles tardives de hémorragie méningée: Aspects techniques

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

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13.5K vues.  University and Bern University Hospital (Inselspital). The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures — 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.

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

Microsurgical Venous Pouch Arterial-Bifurcation Aneurysms in the Rabbit Model: Technical Aspects

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials have shown disappointing long-term results with unacceptable high rates of aneurysm recanalization and delayed aneurysm rupture.2 To overcome these problems, animal experimental studies are crucial for the development of better endovascular devices.3-5
Several animal models in rats, rabbits, canines and swine are available.6-8 Comparisons of the different animal models showed the superiority of the rabbit model with regard to hemodynamics and comparability of the coagulation system and cost-effectiveness.9-11 
The venous pouch arterial bifurcation model in rabbits is formed by a venous pouch sutured into an artificially created true bifurcation of both common carotid arteries (CCA). The main advantage of this model are true bifurcational hemodynamics.12 The major drawbacks are the sofar high microsurgical technical demands and high morbidity and mortality rates of up to 50%.13 These limitations have resulted in less frequent use of this aneurysm model in the recent years. These shortcomings could be overcome with improved surgical procedures and modified peri- and postoperative analgetic management and anticoagulation.14-16 Our techniques reported in this paper demonstrate this optimized technique for microsurgical creation of arterial bifurcation aneurysms.

An optimized technique for the microsurgical creation of arterial bifurcation aneurysms mimicking bifurcation human cerebral aneurysms is described. A venous pouch is sutured into an artificially created true bifurcation of both common carotid arteries. Facilitated microsurgical techniques and aggressive postoperative anticoagulation and analgesia lead to minimized morbidity rates and high aneurysm patency rates.

Microchirurgie veineuse Pouch artériel Bifurcation anévrismes dans le modèle du lapin: Aspects techniques

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials ...

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Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

Biomarqueurs dans un modèle animal pour Révéler Corrélats neuronaux, hématologiques et du comportement de stress post-traumatique

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

NADH Fluorescence Imaging of Isolated Biventricular Working Rabbit Hearts

Since its inception by Langendorff1, the isolated perfused heart remains a prominent tool for studying cardiac physiology2. However, it is not well-suited for studies of cardiac metabolism, which require the heart to perform work within the context of physiologic preload and afterload pressures. Neely introduced modifications to the Langendorff technique to establish appropriate left ventricular (LV) preload and afterload pressures3. The model is known as the isolated LV working heart model and has been used extensively to study LV performance and metabolism4-6. This model, however, does not provide a properly loaded right ventricle (RV). Demmy et al. first reported a biventricular model as a modification of the LV working heart model7, 8. They found that stroke volume, cardiac output, and pressure development improved in hearts converted from working LV mode to biventricular working mode8. A properly loaded RV also diminishes abnormal pressure gradients across the septum to improve septal function. Biventricular working hearts have been shown to maintain aortic output, pulmonary flow, mean aortic pressure, heart rate, and myocardial ATP levels for up to 3 hours8.
When studying the metabolic effects of myocardial injury, such as ischemia, it is often necessary to identify the location of the affected tissue. This can be done by imaging the fluorescence of NADH (the reduced form of nicotinamide adenine dinucleotide)9-11, a coenzyme found in large quantities in the mitochondria. NADH fluorescence (fNADH) displays a near linearly inverse relationship with local oxygen concentration12 and provides a measure of mitochondrial redox state13. fNADH imaging during hypoxic and ischemic conditions has been used as a dye-free method to identify hypoxic regions14, 15 and to monitor the progression of hypoxic conditions over time10.
The objective of the method is to monitor the mitochondrial redox state of biventricular working hearts during protocols that alter the rate of myocyte metabolism or induce hypoxia or create a combination of the two. Hearts from New Zealand white rabbits were connected to a biventricular working heart system (Hugo Sachs Elektronik) and perfused with modified Krebs-Henseleit solution16 at 37 &deg;C. Aortic, LV, pulmonary artery, and left &amp; right atrial pressures were recorded. Electrical activity was measured using a monophasic action potential electrode. To image fNADH, light from a mercury lamp was filtered (350&plusmn;25 nm) and used to illuminate the epicardium. Emitted light was filtered (460&plusmn;20 nm) and imaged using a CCD camera. Changes in the epicardial fNADH of biventricular working hearts during different pacing rates are presented. The combination of the heart model and fNADH imaging provides a new and valuable experimental tool for studying acute cardiac pathologies within the context of realistic physiological conditions.

The objective is to monitor the mitochondrial redox state of isolated hearts within the context of physiologic preload and afterload pressures. A biventricular working rabbit heart model is presented. High spatiotemporal resolution fluorescence imaging of NADH is used to monitor the mitochondrial redox state of epicardial tissue.

NADH imagerie par fluorescence de biventriculaire isolé travail coeurs de lapin

Since its inception by Langendorff1, the isolated perfused heart remains a prominent tool for studying cardiac physiology2. However, it is not well-suited ...

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

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) in rats, in order to identify the initiating events, pathophysiological changes and potential targets for treatment.
Methods: Twenty-eight male Sprague-Dawley rats (250 - 300 g) were arbitrarily assigned to one of two groups - SAH or saline control. Rat subarachnoid hemorrhage in the SAH group (n=15) was induced by double injection of autologous blood, 48 hr apart, into the cisterna magna. Similarly, normal saline (n=13) was injected into the cisterna magna of the saline control group. Rats were sacrificed on day five after the second blood injection and the brains were preserved for histological analysis. The degree of vasospasm was measured using sections of the basilar artery, by measuring the internal luminal cross sectional area using NIH Image-J software. The significance was tested using Tukey/Kramer's statistical analysis.
Results: After analysis of histological sections, basilar artery luminal cross sectional area were smaller in the SAH than in the saline group, consistent with cerebral vasospasm in the former group. In the SAH group, basilar artery internal area (.056 &mu;m &plusmn; 3) were significantly smaller from vasospasm five days after the second blood injection (seven days after the initial blood injection), compared to the saline control group with internal area (.069 &plusmn; 3; p=0.004). There were no mortalities from cerebral vasospasm.
Conclusion: The rat double SAH model induces a mild, survivable, basilar artery vasospasm that can be used to study the pathophysiological mechanisms of cerebral vasospasm in a small animal model. A low and acceptable mortality rate is a significant criterion to be satisfied for an ideal SAH animal model so that the mechanisms of vasospasm can be elucidated 7, 8. Further modifications of the model can be made to adjust for increased severity of vasospasm and neurological exams.

Aneurysmal subarachnoid hemorrhage (SAH) is bleeding that occurs into the subarachnoid space when an aneurysm ruptures. While the morbidity and mortality from this event has been on a decline due to improved treatment approaches, the risk of vasospasm after subarachnoid hemorrhage continues to be the same as it was several years ago. The importance of establishing a comprehensive and reproducible animal model to identify initiating events of cerebral vasospasm has been the focus of research since the first use of rats in an experimental vasospasm model in 1979 by Barry et al. Early work in rats demonstrated that a single injection of autologous blood into the cisterna magna led to acute (within minutes) but not delayed cerebral vasospasm 3, 6, 14. Here we characterize a low mortality SAH rat model that results in reproducible delayed vasospasm.

Un modèle de mortalité faible Rat pour évaluer différée vasospasme cérébral après une hémorragie sous-arachnoïdienne expérimentale

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

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human malignancies.1 Other than surgery for the minority of patients who present with localized disease, there is little or no survival benefit of systemic therapy. Therefore, there is a great need to better understand the biology of NETs, and in particular define new therapeutic targets for patients with nonresectable or metastatic neuroendocrine tumors. 3D cell culture is becoming a popular method for drug screening due to its relevance in modeling the in vivo tumor tissue organization and microenvironment.2,3 The 3D multicellular spheroids could provide valuable information in a more timely and less expensive manner than directly proceeding from 2D cell culture experiments to animal (murine) models.
To facilitate the discovery of new therapeutics for NET patients, we have developed an in vitro 3D multicellular spheroids model using the human NET cell lines. The NET cells are plated in a non-adhesive agarose-coated 24-well plate and incubated under physiological conditions (5% CO2, 37 &deg;C) with a very slow agitation for 16-24 hr after plating. The cells form multicellular spheroids starting on the 3rd or 4th day. The spheroids become more spherical by the 6th day, at which point the drug treatments are initiated. The efficacy of the drug treatments on the NET spheroids is monitored based on the morphology, shape and size of the spheroids with a phase-contrast light microscope. The size of the spheroids is estimated automatically using a custom-developed MATLAB program based on an active contour algorithm. Further, we demonstrate a simple method to process the HistoGel embedding on these 3D spheroids, allowing the use of standard histological and immunohistochemical techniques. 
This is the first report on generating 3D spheroids using NET cell lines to examine the effect of therapeutic drugs. We have also performed histology on these 3D spheroids, and displayed an example of a single drug's effect on growth and proliferation of the NET spheroids. Our results support that the NET spheroids are valuable for further studies of NET biology and drug development.

We present a simple agarose overlay platform to grow 3D multicellular spheroids using neuroendocrine cancer cell lines. This method provides a very convenient way to examine the effect of therapeutic drugs on the neuroendocrine tumor cells. It could also help us establish human neuroendocrine tumor spheroids for cancer therapy.

L&#39;homme neuroendocriniennes lignées cellulaires tumorales comme un modèle tridimensionnel pour l&#39;étude de la thérapie des tumeurs neuro-endocrines humaines

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

Un modèle murin d&#39;implantation d&#39;une endoprothèse dans l&#39;artère carotide pour l&#39;étude de la resténose

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Aspects techniques de la fistule aorto-cave de la souris

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

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.

Un modèle murin d&#39;hémorragie sous-arachnoïdienne

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

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to atherosclerotic plaque development or the rupturing of established plaques, interrupts normal blood flow leading to various morbidities such as myocardial infarction and stroke. In the clinic endovascular procedures such as angioplasty are commonly performed to reopen the lumen. However, these treatments inevitably damage the vessel wall as well as the vascular endothelium, triggering an excessive healing response and the development of a neointimal plaque that extends into the lumen causing vessel restenosis (re-narrowing). Restenosis remains a major cause of failure of endovascular treatments for atherosclerosis. Thus, preclinical animal models of restenosis are vitally important for investigating the pathophysiological mechanisms as well as translational approaches to vascular interventions. Among several murine experimental models, femoral artery wire injury is widely accepted as the most suitable for studies of post-angioplasty restenosis because it closely resembles the angioplasty procedure that injures both endothelium and vessel wall. However, many researchers have difficulty utilizing this model due to its high degree of technical difficulty. This is primarily because a metal wire needs to be inserted into the femoral artery, which is approximately three times thinner than the wire, to generate sufficient injury to induce prominent neointima. Here, we describe the essential surgical details to effectively overcome the major technical difficulties of this model. By following the presented procedures, performing the mouse femoral artery wire injury becomes easier. Once familiarized, the whole procedure can be completed within 20 min.

The mouse femoral artery wire injury model of restenosis is technically challenging. In this protocol we show the key technical details essential for successfully performing wire injury to induce consistent&#160;neointima for studies of restenosis.

Un modèle murin de la resténose artérielle: les aspects techniques de fémur fil blessures

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

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 ...