La recherche actuelle est financé par la Fondation de recherche Solorz-Zak.

Toutes les interventions chirurgicales ont été soumis à un examen éthique et approuvés par le gouvernement de Haute-Bavière (numéro de référence: 55.2-1-54-2532.3-13-13 et -2532-136-11). Les animaux sont mâles C57BL / 6 avec un poids corporel d&#39;environ 25 g. Une. Préparation des animaux <ol><li> Induire une anesthésie en plaçant la souris dans une chambre. Rincer la chambre avec 5% d&#39;isoflurane que l&#39;animal perd conscience. </li><li> Injecter des anesthésiques par voie intrapéritonéale prémélangés: fentanyl (0,05 mg / kg), le midazolam (5 mg / kg) et la médétomidine (0,5 mg / kg). Vérifiez réflexes avant et régulièrement pendant la procédure. Réinjecter un tiers du montant initial horaire pour maintenir l&#39;anesthésie. </li><li> Intuber orotracheally avec un tube d&#39;un cathéter veineux G 11 20. Pour intubation fixer l&#39;animal sur une plate-forme inclinée (30 °), rétracter la languette avec une pince pliées, de visualiser les cordes vocales sous un microscope d&#39;opération et insérer le tube dans la trachéependant l&#39;inspiration. </li><li> Placez la souris dans une position couchée et vérifier le positionnement correct du tube avec un morceau de coton ou un microcapnograph. </li><li> Connecter le tube d&#39;intubation au respirateur. Ventiler la souris avec l&#39;air ambiant supplémenté avec 25% d&#39;oxygène avec une fréquence de 180 à 220 respirations / min et un volume d&#39;éjection de 200 à 250 pi. </li><li> Connecter le tube d&#39;intubation à la microcapnograph. Maintenir la fin d&#39;expiration pCO 2 à 30 mm de Hg en ajustant la fréquence de ventilation. </li><li> Insérer une sonde de température rectale et à placer l&#39;animal sur un coussin chauffant pour maintenir 37 ° C de la température corporelle. </li><li> Appliquer le capteur de pouls annulaire sur la patte arrière droite. </li><li> Ouvrez la peau au-dessus du crâne avec une paire de ciseaux. L&#39;incision doit être d&#39;environ 0,5 cm de long et entre l&#39;oreille et l&#39;œil. </li><li> Disséquer le muscle temporal gauche avec un scalpel à partir de l&#39;os temporal. </li><li> Collez le débitmètre laser Doppler (LDF) de la sonde surl&#39;os temporal gauche. Tenir la sonde dans une position fixe jusqu&#39;à ce que la colle durcisse. </li><li> Percez un trou d&#39;environ 1,5 mm de diamètre avec une fraise dentaire dans l&#39;os temporal gauche. Refroidir l&#39;os avec une solution saline pour prévenir les dommages de la chaleur. </li><li> Insérez la sonde ICP dans la cavité crânienne. Pousser en avant comme le dos que possible pour éviter des dommages aux tissus du cerveau et une hémorragie. </li><li> Si la sonde est à la bonne position, fixer et sceller avec du ciment. Laissez le ciment sécher pendant 5 min. </li><li> Retournez la souris avec soin pour une position couchée. </li><li> Pour la surveillance continue de la pression artérielle, cathétériser l&#39;artère fémorale gauche. </li><li> Connecter le cathéter fémoral au dispositif de surveillance de la pression sanguine. </li></ol> 2. SAH induction <ol><li> Ouvrez la peau avec une paire de ciseaux de sternum au menton (2 cm). Disséquer le tissu conjonctif et carrément pousser les glandes salivaires de côté. </li><li> Exposer l&#39;artère carotide commune gauche (CCA) et le mobiliser. Préserver l&#39;nerf vague, qui se déroule dans la même gaine de tissu conjonctif comme le CCA. Déplacez crânienne et exposer et mobiliser l&#39;ICA et la Cour des comptes en utilisant la même technique. </li><li> Ligaturer le CEA autant crânienne que possible. </li><li> Convenir deux autres ligatures pour filament autour de la CEA. </li><li> Obstruer la CCA et la CIA temporairement microclips. Placez les microclips avec un applicateur de microclip. Assurez-vous que les clips sont correctement appliquées en les tirant doucement le dos. </li><li> Coupez un trou pour l&#39;insertion du filament dans le CEA avec un navire ciseaux. </li><li> Insérez un filament Prolene 5-0 avec 12 mm de longueur dans le CEA. </li><li> Fermer le site d&#39;insertion avec une ligature préétabli. </li><li> Retirez les microclips avec un applicateur de microclip de la CCA et la CIA. </li><li> Avancer le filament avec une pince dans l&#39;ICA jusqu&#39;à la PIC augmente. Une hausse soudaine de l&#39;ICP indique saignement induction. </li><li> Retirer le filament immédiatement et ligaturer le CEA en fermant les deux prligatures earranged consécutivement. Cela empêche le saignement sur le site d&#39;insertion. </li><li> Suturer la plaie de la peau. </li><li> Surveiller les paramètres physiologiques de l&#39;animal pour un autre 20 min. </li><li> Retirer ICP et les sondes de FDL et suturer la plaie de la peau. </li></ol> 3. Fin de l&#39;expérience <ol><li> Perfuser l&#39;animal transcardiaque avec 20 ml d&#39;une solution saline (à température ambiante), suivi par 20 ml de PFA à 4% dans du PBS (4 ° C). </li><li> Disséquer le cerveau sur le crâne. Couper le crâne sur la ligne médiane et entre les cavités orbitales. Puis peler l&#39;os du cerveau. </li><li> Évaluer la distribution du sang dans l&#39;espace sous-arachnoïdien. </li></ol> 4. Considérations en cas de chirurgie de survie (non représenté dans la vidéo) <ol><li> Injecter Carprofen (4 mg / kg par voie sous cutanée) pour l&#39;analgésie postopératoire directement après induction de l&#39;anesthésie. Au cours de la période d&#39;observation postopératoire carprofène (4 mg / kg sous-cutanée) est injecté à tous les 24 h. </li&gt; <li> Au lieu de la mesure de la pression artérielle invasive font usage d&#39;un système de surveillance de pression sanguine non invasive. </li><li> Pour la terminaison de l&#39;anesthésie injection sous-cutanée, les agents antagonistes: la naloxone (1,2 mg / kg), le flumazénil (0,5 mg / kg) et de l&#39;atipamézole (2,5 mg / kg). </li><li> Extubation l&#39;animal. </li><li> Après avoir repris des réflexes mettre l&#39;animal dans une chambre préchauffé. Garder l&#39;animal à environ 32 ° C pendant 24 heures. </li><li> Les animaux sont contrôlés régulièrement pour la respiration spontanée et leur état général durant les premières heures après la chirurgie. Si la tige de cerveau est affecté animaux ont des problèmes respiratoires et doivent être euthanasiés. </li><li> Les animaux sont contrôlés quotidiennement pour leur état ​​général et le poids du corps ainsi que des déficits neurologiques et sensoriels 15. </li></ol>

<ol>
	<li>Cahill, J., Zhang, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subarachnoid+hemorrhage:+is+it+time+for+a+new+direction.">Subarachnoid hemorrhage: is it time for a new direction.</a> Stroke. 40, 86-87 (2009).</li><li>van Gijn, J., Kerr, R. S., Rinkel, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subarachnoid+haemorrhage.">Subarachnoid haemorrhage.</a> Lancet. 369, 306-318 (2007).</li><li>Lin, C. L., 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=A+murine+model+of+subarachnoid+hemorrhage-induced+cerebral+vasospasm.">A murine model of subarachnoid hemorrhage-induced cerebral vasospasm.</a> J. Neurosci. Methods. 123, 89-97 (2003).</li><li>Altay, 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=A+novel+method+for+subarachnoid+hemorrhage+to+induce+vasospasm+in+mice.">A novel method for subarachnoid hemorrhage to induce vasospasm in mice.</a> J. Neurosci. Methods. 183, 136-140 (2009).</li><li>Feiler, S., Friedrich, B., Scholler, K., Thal, S. C., Plesnila, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Standardized+induction+of+subarachnoid+hemorrhage+in+mice+by+intracranial+pressure+monitoring.">Standardized induction of subarachnoid hemorrhage in mice by intracranial pressure monitoring.</a> J. Neurosci. Methods. 190, 164-170 (2010).</li><li>Bederson, J. B., Germano, I. M., Guarino, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+blood+flow+and+cerebral+perfusion+pressure+in+a+new+noncraniotomy+model+of+subarachnoid+hemorrhage+in+the+rat.">Cortical blood flow and cerebral perfusion pressure in a new noncraniotomy model of subarachnoid hemorrhage in the rat.</a> Stroke. 26, 1086-1091 (1995).</li><li>Veelken, J. A., Laing, R. J., Jakubowski, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Sheffield+model+of+subarachnoid+hemorrhage+in+rats.">The Sheffield model of subarachnoid hemorrhage in rats.</a> Stroke. 26, 1279-1283 (1995).</li><li>Kamii, H., 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=Amelioration+of+vasospasm+after+subarachnoid+hemorrhage+in+transgenic+mice+overexpressing+CuZn-superoxide+dismutase.">Amelioration of vasospasm after subarachnoid hemorrhage in transgenic mice overexpressing CuZn-superoxide dismutase.</a> Stroke. 30, 867-871 (1999).</li><li>Parra, A., 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=Mouse+model+of+subarachnoid+hemorrhage+associated+cerebral+vasospasm:+methodological+analysis.">Mouse model of subarachnoid hemorrhage associated cerebral vasospasm: methodological analysis.</a> Neurol. Res. 24, 510-516 (2002).</li><li>Broderick, J. P., Brott, T. G., Duldner, J. E., Tomsick, T., Leach, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Initial+and+recurrent+bleeding+are+the+major+causes+of+death+following+subarachnoid+hemorrhage.">Initial and recurrent bleeding are the major causes of death following subarachnoid hemorrhage.</a> Stroke. 25, 1342-1347 (1994).</li><li>Thal, S. C., Plesnila, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+intraoperative+monitoring+of+blood+pressure+and+arterial+pCO2+during+surgical+anesthesia+in+mice.">Non-invasive intraoperative monitoring of blood pressure and arterial pCO2 during surgical anesthesia in mice.</a> J. Neurosci. Methods. 159, 261-267 (2007).</li><li>Hockel, K., Trabold, R., Scholler, K., Torok, E., Plesnila, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impact+of+anesthesia+on+pathophysiology+and+mortality+following+subarachnoid+hemorrhage+in+rats.">Impact of anesthesia on pathophysiology and mortality following subarachnoid hemorrhage in rats.</a> Exp. Transl. Stroke Med. 4, 5 (2012).</li><li>Wang, Z., Schuler, B., Vogel, O., Arras, M., Vogel, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+is+the+optimal+anesthetic+protocol+for+measurements+of+cerebral+autoregulation+in+spontaneously+breathing+mice?.">What is the optimal anesthetic protocol for measurements of cerebral autoregulation in spontaneously breathing mice?.</a> Exp. Brain Res. 207, 249-258 (2010).</li><li>Schwartz, A. Y., Masago, A., Sehba, F. A., Bederson, J. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+models+of+subarachnoid+hemorrhage+in+the+rat:+a+refinement+of+the+endovascular+filament+model.">Experimental models of subarachnoid hemorrhage in the rat: a refinement of the endovascular filament model.</a> J. Neurosci. Methods. 96, 161-167 (2000).</li><li>Feiler, S., Plesnila, N., Thal, S. C., Zausinger, S., Scholler, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+matrix+metalloproteinase-9+to+cerebral+edema+and+functional+outcome+following+experimental+subarachnoid+hemorrhage.">Contribution of matrix metalloproteinase-9 to cerebral edema and functional outcome following experimental subarachnoid hemorrhage.</a> Cerebrovasc. Dis. 32, 289-295 (2011).</li></ol>

Les auteurs ont rien à révéler.

Les options de traitement après HSA sont rares et la plupart du temps inefficaces. Par conséquent, la physiopathologie de lésions cérébrales post-hémorragique doit encore être compris afin d&#39;identifier de nouvelles cibles thérapeutiques et de développer de nouvelles approches thérapeutiques. Standardisés et des modèles animaux et reproductibles chez les animaux génétiquement modifiés, à savoir les souris, sont cruciales pour ces enquêtes. Le modèle CWP est devenu un modèle largement utili...

SAH est le sous-type de course avec le résultat le moins bénéfique pour les patients: 40% des patients meurent dans le mois suivant le saignement 1 et survivants ont rarement un résultat clinique favorable. La grande majorité des SEP spontanées (80%) sont causés par la rupture des anévrismes intracrâniens qui sont principalement situés le long de la partie antérieure et artère communicante postérieure, l&#39;artère basilaire, et de l&#39;artère cérébrale moyenne (MCA) 2. Ces anévrismes sont difficiles à modéliser chez l&#39;animal et par conséquent des modèles animaux de SEP sont effectuées par injection de sang dans l&#39;espace sous-arachnoïdien ventricules cérébraux / ou par perforation endovasculaire d&#39;un vaisseau sous-arachnoïdienne. Injection de sang autologue dans la grande citerne est facile à réaliser et reproductible que le volume de sang peut être commandé directement 3. Malheureusement, certains aspects de la physiopathologie SEP, par exemple lela lésion du vaisseau, ne peut être modélisée par cette procédure. Une autre approche technique pour l&#39;induction de SAH est l&#39;ouverture d&#39;une veine intracisternale 4. Toutefois, le CWP intraluminal à la succursale MCA semble être la procédure que les modèles de la physiopathologie chez l&#39;homme le plus étroitement 5. La méthode a été développée et décrite pour la première chez le rat par Bederson et collègues, et en même temps par Veelken et collègues 6,7. Plus tard, le modèle de perforation intraluminal a été adapté aux souris 8,9. Un filament est inséré dans l&#39;artère carotide externe (CEA) et avancé à la base du crâne par l&#39;artère carotide interne (ACI). A l&#39;endroit de la MCA de branchement du filament perfore la cuve et provoque un saignement dans l&#39;espace sous-arachnoïdien de la base du crâne. Le sang distribue ensuite dans l&#39;espace sous-arachnoïdien restant le long des fissures et des vaisseaux sanguins. Saignement est arrêté par la formation de caillots sur le site de perforation, mais rebleedings, which sont souvent préjudiciable chez les patients 10, peuvent se produire. Par conséquent, le modèle de filament endovasculaire est devenu un modèle SAH largement utilisé au cours des dernières années. L&#39;inconvénient le plus souvent mentionné le modèle filament de perforation est que le volume de saignement peut pas être contrôlée directement et peut donc être variable. Cette variabilité peut être significativement réduite par un contrôle strict de la physiologie animale et post-hémorragique ICP. Les souris ont le grand avantage d&#39;un grand nombre de souches génétiquement modifiées sont disponibles. Toutefois, en raison de leur petite taille les procédures chirurgicales ont tendance à être plus complexe que dans les plus grandes espèces, par exemple les rats ou les lapins. Par conséquent, la mise à l&#39;échelle des techniques développées pour les rats à des souris souvent n&#39;entraîne pas les résultats escomptés, par exemple que les souris ont une des techniques non invasives très limitées de poids de corps et de volume de sang pour la pression sanguine et l&#39;analyse des gaz du sang ainsi que pour la saturation de l&#39;hémoglobine et de la surveillance de la fréquence cardiaquedoivent être appliquées dès que possible. Par conséquent, le but de la présente publication est de décrire le modèle de perforation de filament pour SAH chez les souris et pour montrer comment ce modèle peut être effectuée d&#39;une manière normalisée et hautement reproductible.

<table border="1">
	<tbody>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Equipment</td>
		</tr>
		<tr>
			<td>operation microscope</td>
			<td>Leica</td>
			<td>KL2500</td>
			<td></td>
		</tr>
		<tr>
			<td>isoflurane vaporizer</td>
			<td>Harvard Instruments</td>
			<td>Continuous Flow Vaporizer</td>
			<td></td>
		</tr>
		<tr>
			<td>respirator</td>
			<td>Hugo Sachs</td>
			<td>Minivent 845</td>
			<td></td>
		</tr>
		<tr>
			<td>microcapnograph</td>
			<td>Hugo Sachs</td>
			<td>Type 340</td>
			<td></td>
		</tr>
		<tr>
			<td>temperature controller</td>
			<td>FHC</td>
			<td>DC Temperature Controller</td>
			<td></td>
		</tr>
		<tr>
			<td>dental drill</td>
			<td>Paggen</td>
			<td>Labset- N</td>
			<td></td>
		</tr>
		<tr>
			<td>ICP monitor</td>
			<td>Codman</td>
			<td>ICP monitor</td>
			<td></td>
		</tr>
		<tr>
			<td>blood pressure monitor</td>
			<td>AD Instruments</td>
			<td>Bridge Amp FE221</td>
			<td></td>
		</tr>
		<tr>
			<td>syringe pump</td>
			<td>World Precision Instruments</td>
			<td>SP101IZ</td>
			<td></td>
		</tr>
		<tr>
			<td>pulsoximeter</td>
			<td>Kent Scientific</td>
			<td>MouseSTAT</td>
			<td></td>
		</tr>
		<tr>
			<td>LDF</td>
			<td>Perimed</td>
			<td>Periflux 5000</td>
			<td></td>
		</tr>
		<tr>
			<td>analog data monitor</td>
			<td>AD Instruments</td>
			<td>Power Lab 16/35</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Material</td>
		</tr>
		<tr>
			<td>cement for ICP probe fixation</td>
			<td>Speiko</td>
			<td>Carboxylate cement</td>
			<td></td>
		</tr>
		<tr>
			<td>glue for LDF probe fixation</td>
			<td>Bob Smith Industries</td>
			<td>Cyanoacrylate glue (Maxi Cure and Insta Set)</td>
			<td></td>
		</tr>
		<tr>
			<td>venous catheter</td>
			<td>Johnson &#38; Johnson</td>
			<td>Jelco winged i.v. catheter; REF 4076</td>
			<td>modified intubation tube</td>
		</tr>
		<tr>
			<td>tubing for femoral catheter</td>
			<td>Smiths Medical</td>
			<td>Fine Bore Polythene Tubing; ID 0.28 mm OD 0.61 mm; REF 800/100/100</td>
			<td>cut to 30 cm length</td>
		</tr>
		<tr>
			<td>filament for vessel perforation</td>
			<td>Ethicon</td>
			<td>Prolene 5-0</td>
			<td>cut to 12 mm length</td>
		</tr>
		<tr>
			<td>surgical equipment</td>
			<td>Fine Scientific Instruments</td>
			<td>forceps medical #5, vessel scissors 8 cm, microclip 4 mm jaw</td>
			<td></td>
		</tr>
	</tbody>
</table>

a murine model of subarachnoid hemorrhage

Dans cette publication vidéo un modèle de souris standardisée de l&#39;hémorragie sous-arachnoïdienne (HSA) est présenté. Saignement est induite par Cercle endovasculaire de Willis perforation (CWP) et prouvé par la pression intracrânienne (PIC) de surveillance. De ce fait une répartition homogène de sang dans les espaces sous-arachnoïdiens entourant la circulation artérielle et des fissures du cervelet est atteint. Physiologie animale est maintenu par intubation, ventilation mécanique, et la surveillance de divers paramètres physiologiques et cardiovasculaires continue en ligne: la température du corps, la pression artérielle systémique, la fréquence cardiaque et la saturation de l&#39;hémoglobine. De ce fait la pression de perfusion cérébrale peut être surveillé étroitement résultant en un volume variable de moins de sang extravasé. Cela permet une meilleure standardisation des endovasculaire perforation de filament dans les souris et rend l&#39;ensemble du modèle hautement reproductible. Ainsi, il est facilement disponible pour les études pharmacologiques et physiopathologiques de type sauvage et génétiquely modifié souris.

Mortalité Une fois la technique de la chirurgie est maîtrisé la procédure ne provoque pas de mortalité peropératoire. Saignement peut également être réalisé dans pratiquement tous les animaux. La mortalité postopératoire est de 30-40%, la plupart des animaux meurent au jour 1 après la chirurgie (Figure 5). ICP valeurs après SAH Le PIC avant la saignée est d&#39;environ 4 mmHg. Saignement des résultats dans u...

Watch this Scientific Journal Video about A Murine Model of Subarachnoid Hemorrhage at JoVE.com

A Murine Model of Subarachnoid Hemorrhage

Un modèle de souris standardisée de l&#39;hémorragie sous-arachnoïdienne par intraluminal cercle de Willis perforation est décrit. perforation du navire et méningée saignements sont contrôlés par la surveillance de la pression intracrânienne. En outre différents paramètres vitaux sont enregistrées et contrôlées pour maintenir des conditions physiologiques.

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-subarachnoid-hemorrhage

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19.5K vues.  University of Munich Medical Center.  Un modèle de souris standardisée de l'hémorragie sous-arachnoïdienne par intraluminal cercle de Willis perforation est décrit. perforation du navire et méningée saignements sont contrôlés par la surveillance de la pression intracrânienne. En outre différents paramètres vitaux sont enregistrées et contrôlées pour maintenir des conditions physiologiques. 

Vidéo: A Murine Model of Subarachnoid Hemorrhage

A Murine Model of Muscle Training by Neuromuscular Electrical Stimulation

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional capacity. Transcutaneous surface stimulation of skeletal muscle involves a current flow between a cathode and an anode, thereby inducing excitement of the motor unit and the surrounding muscle fibers. 
NMES is an attractive modality to evaluate skeletal muscle adaptive responses for several reasons. First, it provides a reproducible experimental model in which physiological adaptations, such as myofiber hypertophy and muscle strengthening6, angiogenesis7-9, growth factor secretion9-11, and muscle precursor cell activation12 are well documented. Such physiological responses may be carefully titrated using different parameters of stimulation (for Cochrane review, see 13). In addition, NMES recruits motor units non-selectively, and in a spatially fixed and temporally synchronous manner14, offering the advantage of exerting a treatment effect on all fibers, regardless of fiber type. Although there are specified contraindications to NMES in clinical populations, including peripheral venous disorders or malignancy, for example, NMES is safe and feasible, even for those who are ill and/or bedridden and for populations in which rigorous exercise may be challenging. 
Here, we demonstrate the protocol for adapting commercially available electrodes and performing a NMES protocol using a murine model. This animal model has the advantage of utilizing a clinically available device and providing instant feedback regarding positioning of the electrode to elicit the desired muscle contractile effect. For the purpose of this manuscript, we will describe the protocol for muscle stimulation of the anterior compartment muscles of a mouse hindlimb.

A murine model of neuromuscular electrical stimulation (NMES), a safe and inexpensive clinical modality, to the anterior compartment muscles is described. This model has the advantage of modifying a readily available clinical device for the purpose of eliciting targeted and specific muscle contractions in mice.

Un modèle murin de la formation du muscle par stimulation neuromusculaire électrique

Neuromuscular electrical stimulation (NMES) is a common clinical modality that is widely used to restore1, maintain2 or enhance3-5 muscle functional ...

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Médecine

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) ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

Modèle murin de la cicatrisation

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

A New Murine Model of Endovascular Aortic Aneurysm Repair

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of Abdominal aortic aneurysm (AAA) pathophysiology. There is no abdominal aortic aneurysm endograft exclusion model cheap and reproducible, which would allow deep investigations of AAA before and after treatment. We hereby describe how to induce, and then to exclude with a covered coronary stentgraft an abdominal aortic aneurysm in a rat. The well known elastase induced AAA model was first reported in 19901 in a rat, then described in mice2. Elastin degradation leads to dilation of the aorta with inflammatory infiltration of the abdominal wall and intra luminal thrombus, matching with human AAA. Endovascular exclusion with small covered stentgraft is then performed, excluding any interactions between circulating blood and the aneurysm thrombus. Appropriate exclusion and stentgraft patency is confirmed before euthanasia by an angiography thought the left carotid artery. Partial control of elastase diffusion makes aneurysm shape different for each animal. It is difficult to create an aneurysm, which will allow an appropriate length of aorta below the aneurysm for an easy stentgraft introduction, and with adequate proximal and distal neck to prevent endoleaks. Lots of failure can result to stentgraft introduction which sometimes lead to aorta tear with pain and troubles to stitch it, and endothelial damage with post op aorta thrombosis. Giving aspirin to rats before stentgraft implantation decreases failure rate without major hemorrhage. Clamping time activates neutrophils, endothelium and platelets, and may interfere with biological analysis.

Histological and biochemical modifications after aneurysm endograft exclusion are still unclear. We describe a new model of endograft implantation on a murine aneurysm. Through thrombus analysis with and without persistant circulating blood stream interface, and new endovascular biomaterials evaluation, this model will help to better understand endovascular aneurysm exclusion pathobiology.

Un nouveau modèle murin de endovasculaire anévrisme de l&#39;aorte réparation

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of ...

Intrastriatal Injection of Autologous Blood or Clostridial Collagenase as Murine Models of Intracerebral Hemorrhage

Intracerebral hemorrhage (ICH) is a common form of cerebrovascular disease and is associated with significant morbidity and mortality. Lack of effective treatment and failure of large clinical trials aimed at hemostasis and clot removal demonstrate the need for further mechanism-driven investigation of ICH. This research may be performed through the framework provided by preclinical models. Two murine models in popular use include intrastriatal (basal ganglia) injection of either autologous whole blood or clostridial collagenase. Since, each model represents distinctly different pathophysiological features related to ICH, use of a particular model may be selected based on what aspect of the disease is to be studied. For example, autologous blood injection most accurately represents the brain&#39;s response to the presence of intraparenchymal blood, and may most closely replicate lobar hemorrhage. Clostridial collagenase injection most accurately represents the small vessel rupture and hematoma evolution characteristic of deep hemorrhages. Thus, each model results in different hematoma formation, neuroinflammatory response, cerebral edema development, and neurobehavioral outcomes. Robustness of a purported therapeutic intervention can be best assessed using both models. In this protocol, induction of ICH using both models, immediate post-operative demonstration of injury, and early post-operative care techniques are demonstrated. Both models result in reproducible injuries, hematoma volumes, and neurobehavioral deficits. Because of the heterogeneity of human ICH, multiple preclinical models are needed to thoroughly explore pathophysiologic mechanisms and test potential therapeutic strategies.

Preclinical models of intracerebral hemorrhage are utilized to mimic certain aspects of clinical disease. Thus, mechanisms of injury and potential therapeutic strategies may be explored. In this protocol, two models of intracerebral hemorrhage are described, intrastriatal (basal ganglia) injections of autologous blood or collagenase.

Injection intrastriatale de sang autologue ou Clostridium collagénase comme modèles murins de Hémorragie intracérébrale

Intracerebral hemorrhage (ICH) is a common form of cerebrovascular disease and is associated with significant morbidity and mortality. Lack of effective ...

A Preclinical Murine Model of Hepatic Metastases

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, murine pancreatic tumor model of hepatic metastases via a hemispleen injection of syngeneic murine pancreatic tumor cells is described. This model mimics many of the clinical conditions in patients with metastatic disease to the liver. Mice consistently develop metastases in the liver allowing for investigation of the metastatic process, experimental therapy testing, and tumor immunology research.

A preclinical, murine model of hepatic metastases performed via a hemispleen injection technique.

Un préclinique modèle murin de métastases hépatiques

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

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

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

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

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 ...

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive nephropathy induced by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). In the irreversible UUO model, the ureter may be obstructed for variable periods of time in order to induce increasingly severe renal inflammation and interstitial fibrotic scarring. In the reversible R-UUO model the ureter is obstructed to induce hydronephrosis, tubular dilation and inflammation. After a suitable period of time the ureteric obstruction is then surgically reversed by anastomosis of the severed previously obstructed ureter to the bladder in order to allow complete decompression of the kidney and restoration of urinary flow to the bladder. The irreversible UUO model has been used to investigate various aspects of renal inflammation and scarring including the pathogenesis of disease and the testing of potential anti-inflammatory or anti-fibrotic therapies. The more challenging model of R-UUO has been used by some investigators and does offer significant research potential as it allows the study of inflammatory and immune processes and tissue remodeling in an injured and scarred kidney following the removal of the injurious stimulus. As a result, the R-UUO model offers investigators the opportunity to explore the resolution of kidney inflammation together with key aspects of tissue repair. These experimental models are of relevance to human disease as patients often present with obstruction of the renal tract that requires decompression and are commonly left with significant residual kidney impairment that has no current treatment options and may lead to eventual end stage kidney failure.

The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

Un modèle murin d&#39;irréversible et réversible unilatérale obstruction urétérale

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information about this disease. The human orthotopic prostate cancer xenograft mouse model provides a useful alternative approach for understanding the specific interactions between genetically and molecularly altered tumor cells, their organ microenvironment, and for evaluation of efficacy of therapeutic regimens. This is a well characterized model designed to study the molecular events of primary tumor development and it recapitulates the early events in the metastatic cascade prior to embolism and entry of tumor cells into the circulation. Thus it allows elucidation of molecular mechanisms underlying the initial phase of metastatic disease. In addition, this model can annotate drug targets of clinical relevance and is a valuable tool to study prostate cancer progression. In this manuscript we describe a detailed procedure to establish a human orthotopic prostate cancer xenograft mouse model.

Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

Pré-clinique Orthotopique modèle murin de cancer de la prostate humaine

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Integrated Compensatory Responses in a Human Model of Hemorrhage

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

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