This study was supported by the NIH grant NS074559 (to C. K.). We thank all collaborators who contributed to our research articles that the present methodology report is based upon.

Este protocolo ha sido aprobado por el Cuidado y Uso de Animales Comité Institucional (IACUC) de la Universidad de Emory y sigue los Institutos Nacionales de la Salud Guía para el Cuidado y Uso de Animales de Laboratorio. 1. Configuración <ol><li> Preparar el lecho quirúrgico en el cojín de calentamiento conectada con bomba de calor a 37 ° C durante al menos 15 minutos antes de la cirugía. Coloque un rollo de cuello con el cañón de 3 ml jeringa en el lecho quirúrgico. Preparar el gas de la anestesia con isoflurano 2% en el aire médica. </li><li> Preparar fórceps tratado al autoclave, tijeras, tenedores micro agujas, pinzas hemostáticas, hisopos de algodón y suturas. Preparar adhesivo tisular y ungüento para los ojos. </li><li> Configurar el sistema de hipoxia y controladores de temperatura con lámpara de calefacción y la sonda rectal. Preparar gas hipoxia con 2% de isoflurano en 7,5% de O 2 equilibrada por el 92,5% de N 2. </li><li> Una hora antes de la cirugía, los ratones se analgesized por inyección subcutánea de una liberación lenta Meloxicam (4,0 mg / kg). </li></ol> &lt;p class = &quot;jove_title&quot;&gt; 2. Transient cerebral hipoxia-isquemia (Figura 1B) <ol><li> Anestesiar C57BL macho 10-13 semanas de edad / 6 ratones con un peso de 22 a 30 g en la cámara de inducción de la anestesia con isoflurano 3% hasta que el animal no responde a la compresión de los pies, y luego quitar el pelo en el cuello derecha usando una máquina de afeitar electrónica. </li><li> Coloca los ratones en el lecho quirúrgico conectado con 2% de isoflurano en el aire médica a un caudal de 2 l / min. Extremidades anteriores Secure estiradas a lo largo rollo de cuello a los lados con cinta médica. </li><li> Limpie el sitio de la cirugía para la incisión con betadine seguido por el alcohol y luego hisopos de algodón. </li><li> Bajo el microscopio de disección, hacer una incisión de 0,5 cm derecha del cuello uterino usando un fórceps rectos y un micro tijeras aproximadamente 0,2 cm lateral de la piel en la línea media. </li><li> Use un par de pinzas dentadas finas de separar la fascia y el tejido para exponer la arteria carótida común derecha (RCCA). Separar cuidadosamente el RCCA desde el nervio vagal usando un par de pinzas lisas finas. </li><li> Vivo nudo dos precortadas 5-0 sutura de seda (desmontable) del RCCA, y luego coser la piel utilizando 4-0 Nylon sutura monofilamento (Figura 1C). </li><li> Aplicar pomada ocular en ambos ojos para evitar la sequedad. </li><li> Transferir rápidamente los ratones al sistema de hipoxia y poner la nariz y la boca en la cara-máscara con 2% de isoflurano en 7,5% O 2 a un caudal de 0,5-1 L / min durante 30 min. <ol><li> Durante la hipoxia, utilizar reguladores de temperatura con lámparas de calor para controlar la temperatura rectal a 37,5 ± 0,5 ° C. Monitorear la frecuencia respiratoria en 80 a 120 respiraciones / min. El mantenimiento de la temperatura corporal por encima de 37 ° C durante la hipoxia es importante crear infarto cerebral coherente. Frecuencia respiratoria baja por lo general ocurre después de 20 min de hipoxia. Retire la mascarilla y permitir el suministro de aire normal si la frecuencia respiratoria cae por debajo de 40. Esto se lleva a 1-2 minutos y no cuenta en la duración hipoxia 30 min. </li></ol></li><li> Después de la hipoxia, transferirlos ratones a una cama quirúrgica y suelte las dos suturas de RCCA. Cierre la herida con pegamento tisular y, a continuación, volver a la jaula de los ratones. Excluir a los animales si tanto de dos nudos vivos se liberan de forma inesperada después de la hipoxia. </li><li> Monitorear los ratones para 5-10 minutos para recuperarse de la hipoxia y la anestesia. Coloque la comida humedecida en la jaula y devolverlo al centro de cuidado de los animales. Nota: Los animales que presenten leves a comportamientos en circuito severa a las 24 horas después de THI se correlacionan con infracción cerebro. La mayoría de los animales con síntomas convulsivos mueren antes de que el punto de tiempo de 24 horas después de THI. </li></ol> 3. láser Speckle de contraste de imagen Nota: Aunque este no es un procedimiento esencial del modelo THI, un láser speckle sistema de imágenes de contraste de dos dimensiones 16 puede ser utilizado para caracterizar los cambios de flujo sanguíneo cerebral (CBF) durante o después de la hipoxia transitoria-isquemia. Para documentar las alteraciones de la CBF bajo THI, ficha inmediatamente después del step 2.6. Alternativamente, para comparar la recuperación CBF después THI insulto, estos procedimientos se pueden realizar después de la etapa 2.10. <ol><li> Publicar un ratón anestesiado en decúbito prono y realizar una incisión de 1 cm de largo en el cuero cabelludo con el cráneo impresionados pero sin abrir. </li><li> Monitorear CBF en ambos hemisferios cerebrales bajo un generador de imágenes del flujo sanguíneo de acuerdo con el protocolo del fabricante y empezar a grabar el flujo sanguíneo cerebral inmediatamente después de la cirugía CCAO (paso 2.6). Continuar durante 50 minutos. </li><li> Mostrar imagen CBF con unidades arbitrarias en una paleta de 16 colores y analizar en tiempo real las regiones seleccionadas utilizando el software MoorFLPI siguiendo las instrucciones del fabricante (Figura 2). </li><li> Después de grabar la imagen CBF, cierre el cuero cabelludo con adhesivo tisular y devolver el animal a la jaula. </li></ol> 4. Administración de tPA <ol><li> Inyectar animales en la vena de la cola con el disolvente o 10 mg / kg de tPA recombinante (220-300 y# 956; l de 1 mg / m tPA) a las 0,5, 1, o 4 horas después de tCCAo más hipoxia (Figura 4). </li></ol> Detección de Daño 5. cerebro con varios diferentes opciones Nota: Para recoger muestras de cerebro, la eutanasia a los ratones a los 1, 4 ó 24 horas después de THI. <ol><li> Realizar cuantificación del volumen de infarto por in vivo cloruro de 2,3,5-trifeniltetrazolio método (TTC) a las 24 horas después de THI insulto como se describe anterior. 17 <ol><li> Intraperitoneal inyectar animales con solución 1,4 M manitol (~ 0,1 ml / g de peso corporal) 30 minutos antes de la perfusión transcardial. Ratones perfundir transcardial con PBS seguido por 10 ml de 2% TTC. </li><li> Retire el cerebro de los animales con los instrumentos quirúrgicos después de 10 minutos y colocar en paraformaldehído al 4% para la fijación durante la noche y en la sección 1 mm de espesor con un vibratome. </li><li> Toma una serie de cuatro diapositivas cerebro seccionado por microscopio digital y quantify el volumen del infarto como la relación de la zona infartada (área blanca en el lado derecho) a la zona de la intacto, hemisferio contralateral usando el software ImageJ. </li></ol></li><li> Alternativamente, lleve a cabo la formación de trombosis por inmunofluorescencia en 1 hora después de THI insulto. <ol><li> Congelar el cerebro fija en compuesto OCT y la sección de los cerebros de 12 micras de espesor utilizando un criostato. </li><li> Incubar el portaobjetos cerebro con anticuerpo de conejo anti-fibrinógeno (1: 100) siguiendo por cabra anti-conejo Alexa 488 Fluro colorante (1: 200) para observar la fluorescencia en un microscopio fluorescente. </li></ol></li><li> Alternativamente, realice la obstrucción buque por inyección en la vena de la cola de 100 l 2% colorante azul de Evans en 4 horas después de THI insulto. <ol><li> La eutanasia a los ratones y rápidamente cortar la cabeza para eliminar cerebros en paraformaldehído al 4% después de la inyección de azul de Evans. Nota: Se necesita 5-10 min para Evans circulación azul con el color azul de tanto delanteras y traseras extremidades. </li><li> SECTIen cerebros fijos a 100 micras de espesor usando un micrótomo de deslizamiento y observar la fluorescencia utilizando un filtro de emisión de 680 nm en un microscopio fluorescente. </li></ol></li></ol>

<ol>
	<li>Broderick, J. P., Hacke, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+acute+ischemic+stroke:+Part+I:+recanalization+strategies.">Treatment of acute ischemic stroke: Part I: recanalization strategies.</a> Circulation. 106 (12), 1563-1569 (2002).</li><li>Hossmann, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathophysiological+basis+of+translational+stroke+research.">Pathophysiological basis of translational stroke research.</a> Folia Neuropathol. 47 (3), 213-227 (2009).</li><li>Hossmann, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+two+pathophysiologies+of+focal+brain+ischemia:+implications+for+translational+stroke+research.">The two pathophysiologies of focal brain ischemia: implications for translational stroke research.</a> J. Cereb. Blood Flow Metab. 32 (7), 1310-1316 (2012).</li><li>Macrae, I. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preclinical+stroke+research--advantages+and+disadvantages+of+the+most+common+rodent+models+of+focal+ischaemia.">Preclinical stroke research--advantages and disadvantages of the most common rodent models of focal ischaemia.</a> Br. J. Pharmacol. 164 (4), 1062-1078 (2011).</li><li>Niessen, F., Hilger, T., Hoehn, M., Hossmann, K. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differences+in+clot+preparation+determine+outcome+of+recombinant+tissue+plasminogen+activator+treatment+in+experimental+thromboembolic+stroke.">Differences in clot preparation determine outcome of recombinant tissue plasminogen activator treatment in experimental thromboembolic stroke.</a> Stroke. 34 (8), 2019-2024 (2003).</li><li>Orset, C., 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+in+situ+thromboembolic+stroke+and+reperfusion.">Mouse model of in situ thromboembolic stroke and reperfusion.</a> Stroke. 38 (10), 2771-2778 (2007).</li><li>Watson, B. D., Dietrich, W. D., Prado, R., Ginsberg, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Argon+laser-induced+arterial+photothrombosis.+Characterization+and+possible+application+to+therapy+of+arteriovenous+malformations.">Argon laser-induced arterial photothrombosis. Characterization and possible application to therapy of arteriovenous malformations.</a> J. Neurosurgery. 66 (5), 748-754 (1987).</li><li>Sun, Y. Y., 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=Synergy+of+combined+tPA-edaravone+therapy+in+experimental+thrombotic+stroke.">Synergy of combined tPA-edaravone therapy in experimental thrombotic stroke.</a> PLoS One. 9, e98807 (2014).</li><li>Levine, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anoxic-ischemic+encephalopathy+in+rats.">Anoxic-ischemic encephalopathy in rats.</a> Am. J. Pathol. 36, 1-17 (1960).</li><li>Rice, J. E. 3. r. d., Vannucci, R. C., Brierley, 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=The+influence+of+immaturity+on+hypoxic-ischemic+brain+damage+in+the+rat.">The influence of immaturity on hypoxic-ischemic brain damage in the rat.</a> Annals Neurol. 9 (2), 131-141 (1981).</li><li>Vannucci, S. J., 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=Experimental+stroke+in+the+female+diabetic,+db/db,+mouse.">Experimental stroke in the female diabetic, db/db, mouse.</a> J. Cereb. Blood Flow Metab. 21 (2), 52-60 (2001).</li><li>Adhami, F., 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=Cerebral+ischemia-hypoxia+induces+intravascular+coagulation+and+autophagy.">Cerebral ischemia-hypoxia induces intravascular coagulation and autophagy.</a> Am. J. Pathol. 169 (2), 566-583 (2006).</li><li>Shereen, 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=Ex+vivo+diffusion+tensor+imaging+and+neuropathological+correlation+in+a+murine+model+of+hypoxia-ischemia-induced+thrombotic+stroke.">Ex vivo diffusion tensor imaging and neuropathological correlation in a murine model of hypoxia-ischemia-induced thrombotic stroke.</a> J. Cereb. Blood Flow Metab. 31 (4), 1155-1169 (2011).</li><li>Michaud, J. P., Pimentel-Coelho, P. M., Tremblay, Y., Rivest, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+Ly6C+low+monocytes+after+cerebral+hypoxia-ischemia+in+adult+mice.">The impact of Ly6C low monocytes after cerebral hypoxia-ischemia in adult mice.</a> J. Cereb. Blood Flow Metab. 34 (7), e1-e9 (2014).</li><li>Zoppo, 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=Virchow's+triad:+the+vascular+basis+of+cerebral+injury.">Virchow's triad: the vascular basis of cerebral injury.</a> Rev. Neurol. Dis. 5, 12-21 (2008).</li><li>Dunn, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser+speckle+contrast+imaging+of+cerebral+blood+flow.">Laser speckle contrast imaging of cerebral blood flow.</a> Annals Biomed. Eng. 40 (2), 367-377 (2012).</li><li>Sun, Y. Y., Yang, D., Kuan, C. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mannitol-facilitated+perfusion+staining+with+2,3,5-triphenyltetrazolium+chloride+(TTC)+for+detection+of+experimental+cerebral+infarction+and+biochemical+analysis.">Mannitol-facilitated perfusion staining with 2,3,5-triphenyltetrazolium chloride (TTC) for detection of experimental cerebral infarction and biochemical analysis.</a> J. Neurosci. Methods. 203 (1), 122-129 (2012).</li><li>Lozano, R., 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=Global+and+regional+mortality+from+235+causes+of+death+for+20+age+groups+in+1990+and+2010:+a+systematic+analysis+for+the+Global+Burden+of+Disease+Study.">Global and regional mortality from 235 causes of death for 20 age groups in 1990 and 2010: a systematic analysis for the Global Burden of Disease Study.</a> Lancet. 380 (9859), 2095-2128 (2010).</li><li>Murray, C. J., 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=Disability-adjusted+life+years+(DALYs)+for+291+diseases+and+injuries+in+21+regions,+1990-2010:+a+systematic+analysis+for+the+Global+Burden+of+Disease+Study.">Disability-adjusted life years (DALYs) for 291 diseases and injuries in 21 regions, 1990-2010: a systematic analysis for the Global Burden of Disease Study.</a> Lancet. 380 (9859), 2197-2223 (2012).</li><li>Dirnagl, U., Macleod, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stroke+research+at+a+road+block:+the+streets+from+adversity+should+be+paved+with+meta-analysis+and+good+laboratory+practice.">Stroke research at a road block: the streets from adversity should be paved with meta-analysis and good laboratory practice.</a> Br. J. Pharm. 157 (7), 1154-1156 (2009).</li><li>Dirnagl, U., 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+concerted+appeal+for+international+cooperation+in+preclinical+stroke+research.">A concerted appeal for international cooperation in preclinical stroke research.</a> Stroke. 44 (6), 1754-1760 (2013).</li><li>Khatri, P., 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=Revascularization+end+points+in+stroke+interventional+trials:+recanalization+versus+reperfusion+in+IMS-I.">Revascularization end points in stroke interventional trials: recanalization versus reperfusion in IMS-I.</a> Stroke. 36 (11), 2400-2403 (2005).</li><li>Rosenberg, R. D., Aird, W. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular-bed--specific+hemostasis+and+hypercoagulable+states.">Vascular-bed--specific hemostasis and hypercoagulable states.</a> New Eng. J. Med. 340 (20), 1555-1564 (1999).</li><li>Majid, 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=Differences+in+vulnerability+to+permanent+focal+cerebral+ischemia+among+3+common+mouse+strains.">Differences in vulnerability to permanent focal cerebral ischemia among 3 common mouse strains.</a> Stroke. 31 (11), 2707-2714 (2000).</li></ol>

El ictus es un problema de salud importante de creciente importancia para cualquier sociedad con una población que envejece. A nivel mundial, el accidente cerebrovascular es la segunda causa principal de la muerte con un estimado de 5.9 millones de eventos fatales en 2010, lo que equivale al 11,1% de todas las muertes 18. El accidente cerebrovascular es también la tercera causa de discapacidad años de vida ajustados (AVAD) perdidos a nivel mundial en 2010, pasando de la quinta posición en 1990 19.</s...

Trombolisis y recanalización es la terapia más eficaz de accidente cerebrovascular isquémico agudo en la práctica clínica 1. Sin embargo, la mayoría de la investigación preclínica neuroprotección se realizó en un modelo transitorio mecánico obstrucción (oclusión de la arteria cerebral media de sutura intraluminal) que produce la rápida recuperación del flujo sanguíneo cerebral después de la retirada de la oclusión vascular y muestra poco a ningún beneficio por tPA trombólisis. Se ha sugerido que la dudosa elección de los modelos de accidente cerebrovascular contribuyó, al menos en parte, a la dificultad en la traducción de la terapia neuroprotectora a los pacientes 2,3. Por lo tanto, hay una demanda creciente para el empleo de modelos de ictus tromboembólico tPA-respuesta en la investigación preclínica, pero estos modelos también tienen problemas técnicos (ver Discusión) 4-7. Aquí se describe un nuevo modelo basado en accidente cerebrovascular trombótico (THI) unilateral insulto hipóxico-isquémico transitorio y sus respuestas a la terapia intravenosa tPA 8. El modelo de carrera THI fue desarrollado sobre la base del procedimiento de Levine (ligadura permanente de la arteria carótida común unilateral seguido por la exposición a la hipoxia transitoria en una cámara) que se inventó para los experimentos con ratas adultas en 1960 9. El procedimiento original Levine desvaneció en la oscuridad, porque sólo produce daño cerebral variable, pero el mismo insulto causado neuropatología consistente en crías de roedores cuando fue reintroducido por Roberto Vannucci y sus colegas como un modelo de neonatal encefalopatía hipóxico-isquémica (EHI) en 1981 10. En los últimos años, algunos investigadores re-adaptar el modelo Levine-Vannucci a ratones adultos mediante el ajuste de la temperatura en la cámara hipóxica 11. Es plausible que las lesiones cerebrales inconsistentes en el procedimiento original Levine pueden surgir de la fluctuación de las temperaturas corporales de roedores adultos en la cámara hipóxica. Para probar esta hipótesis, hemos modificado el procedimiento Levine mediante la administración de gas hipóxicoa través de una mascarilla, mientras se mantiene la temperatura del núcleo de roedores a 37 ° C sobre la mesa quirúrgica 12. Como era de esperar, el control estricto de la temperatura corporal aumenta considerablemente la reproducibilidad de la patología cerebral inducida por HI. El insulto HI también provoca la coagulación, la autofagia y grises y lesiones 13 la sustancia blanca. Otros investigadores también han utilizado el modelo HI para investigar después del accidente cerebrovascular respuestas inflamatorias 14. Una característica única del modelo de accidente cerebrovascular HI es que sigue de cerca la tríada de la Virchow de la formación de trombos, incluida la estasis del flujo de sangre, lesión endotelial (por ejemplo, debido al estrés oxidativo inducido-HI), y la hipercoagulabilidad (activación de las plaquetas HI inducida) ( Figura 1 A) 15. Como tal, el modelo HI puede capturar algunos mecanismos fisiopatológicos relacionados con el accidente cerebrovascular isquémico en el mundo real. Con esta idea en mente, hemos perfeccionado aún más el modelo HI con ligadura reversible de la ONUarteria carótida común ilateral (por lo tanto, para crear un insulto HI transitorio), y se ensayó sus respuestas a tPA trombolisis con o sin edavarona. Edaravona es un eliminador de radicales libres ya aprobado en Japón para tratar el ictus isquémico dentro de las 24 horas de aparición 9. Nuestros experimentos mostraron que lo más breve 30 min transitoria HI desencadena el infarto trombótico, y que combinado tratamiento con tPA-edaravona confiere beneficios sinérgicos 8. Aquí se describen los procedimientos quirúrgicos detalladas y consideraciones metodológicas del modelo de THI, que pueden ser utilizados para optimizar los tratamientos de reperfusión de accidente cerebrovascular isquémico agudo.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>adult male mice</td>
			<td>Charles River</td>
			<td>C57BL/6&#160;</td>
			<td>10~13 weeks old (22~30 g)</td>
		</tr>
		<tr>
			<td>Mobile Laboratory Animal Anesthesia System</td>
			<td>VetEquip</td>
			<td>901807</td>
			<td>anesthesia</td>
		</tr>
		<tr>
			<td>Medical air (Compressed) air tank</td>
			<td>Airgas</td>
			<td>UN1002</td>
			<td>anesthesia</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Piramal Healthcare</td>
			<td>NDC 66794-013-25</td>
			<td>anesthesia</td>
		</tr>
		<tr>
			<td>Multi-Station Lab Animal AnesthesiaSystem</td>
			<td>Surgivet</td>
			<td>V703501</td>
			<td>hypoxia system</td>
		</tr>
		<tr>
			<td>7.5% O2 balanced by 92.5% N2 tank</td>
			<td>Airgas</td>
			<td>UN1956</td>
			<td>hypoxia system</td>
		</tr>
		<tr>
			<td>Temperature Controller with heating lamp&#160;</td>
			<td>Cole Parmer&#160;</td>
			<td>EW-89000-10</td>
			<td>temperature controllers</td>
		</tr>
		<tr>
			<td>Rectal probe</td>
			<td>Cole Parmer&#160;</td>
			<td>NCI-00141PG</td>
			<td>temperature controllers</td>
		</tr>
		<tr>
			<td>Dissecting microscope&#160;</td>
			<td>Olympus&#160;</td>
			<td>SZ40</td>
			<td>surgical setup</td>
		</tr>
		<tr>
			<td>Heat pump with warming pad</td>
			<td>Gaymar&#160;</td>
			<td>TP700</td>
			<td>surgical setup</td>
		</tr>
		<tr>
			<td>Fine curved forceps (serrated)</td>
			<td>FST</td>
			<td>11370-31</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>Fine curved forceps (smooth)</td>
			<td>FST</td>
			<td>11373-12</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>micro scissors</td>
			<td>FST</td>
			<td>15000-03</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>micro needle holders</td>
			<td>FST</td>
			<td>12060-01</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>Halsted-Mosquito hemostats</td>
			<td>FST</td>
			<td>13008-12</td>
			<td>surgical instrument</td>
		</tr>
		<tr>
			<td>5-0 silk suture&#160;</td>
			<td>Harvard Apparatus</td>
			<td>624143</td>
			<td>surgical supplies</td>
		</tr>
		<tr>
			<td>4-0 Nylon monofilament suture</td>
			<td>LOOK</td>
			<td>766B</td>
			<td>surgical supplies</td>
		</tr>
		<tr>
			<td>Tissue glue</td>
			<td>Abbott Laboratories</td>
			<td>NC9855218</td>
			<td>surgical supplies</td>
		</tr>
		<tr>
			<td>Puralube Vet ointment</td>
			<td>Fisher</td>
			<td>NC0138063&#160;</td>
			<td>eye dryness prevention&#160;</td>
		</tr>
		<tr>
			<td>MoorFLPI-2 blood flow imager</td>
			<td>Moor</td>
			<td>780-nm laser source</td>
			<td>Laser Speckle Contrast Imaging</td>
		</tr>
		<tr>
			<td>Mannitol</td>
			<td>Sigma</td>
			<td>M4125</td>
			<td>in-vivo TTC</td>
		</tr>
		<tr>
			<td>2,3,5-triphenyltetrazolium chloride (TTC)&#160;</td>
			<td>Sigma</td>
			<td>T8877</td>
			<td>in-vivo TTC</td>
		</tr>
		<tr>
			<td>Vibratome</td>
			<td>Stoelting</td>
			<td>51425</td>
			<td>brain section for in-vivo TTC&#160;</td>
		</tr>
		<tr>
			<td>Digital microscope</td>
			<td>Dino-Lite</td>
			<td>AM2111</td>
			<td>whole-braina imaging</td>
		</tr>
		<tr>
			<td>O.C.T compound</td>
			<td>Sakura Finetek</td>
			<td>4583</td>
			<td></td>
		</tr>
		<tr>
			<td>goat anti-rabbit Alexa Fluro 488</td>
			<td>Invitrogen</td>
			<td>A11008</td>
			<td>Immunohistochemistry</td>
		</tr>
		<tr>
			<td>Cryostat</td>
			<td>Vibratome</td>
			<td>ultrapro 5000</td>
			<td>brain section for IHC</td>
		</tr>
		<tr>
			<td>Evans blue</td>
			<td>Sigma</td>
			<td>E2129</td>
			<td>Detecting vascular perfusion</td>
		</tr>
		<tr>
			<td>Microtome</td>
			<td>Electron Microscopy Sciences</td>
			<td>5000</td>
			<td>brain section for histology</td>
		</tr>
		<tr>
			<td>Avertin (2, 2, 2-Tribromoethanol)</td>
			<td>Sigma</td>
			<td>T48402</td>
			<td>euthanasia</td>
		</tr>
		<tr>
			<td>Fluorescent microscope</td>
			<td>Olympus</td>
			<td>DP73</td>
			<td></td>
		</tr>
	</tbody>
</table>

a thrombotic stroke model based on transient cerebral hypoxia-ischemia

Investigación Stroke ha sufrido muchos reveses en la traducción de las terapias neuroprotectoras en la práctica clínica. En contraste, la terapia del mundo real (tPA trombolisis) rara vez produce beneficios en modelos experimentales basados ​​en la oclusión mecánica, que dominan la investigación preclínica derrame cerebral. Esta división entre el banco y la cabecera sugiere la necesidad de emplear modelos tPA-sensibles en la investigación preclínica derrame cerebral. Para este fin, un modelo de accidente cerebrovascular trombótico simple y tPA-reactiva se inventó y se describe aquí. Este modelo consiste en la oclusión transitoria de la arteria carótida común unilateral y la entrega de 7,5% de oxígeno a través de una máscara facial en ratones adultos durante 30 min, manteniendo la temperatura rectal de los animales a 37,5 ± 0,5 ° C. Aunque la ligadura reversible de la arteria carótida unilateral o hipoxia cada suprime el flujo sanguíneo cerebral sólo transitoriamente, la combinación de ambos insultos causada duradera déficit de reperfusión, la fibrina y la deposición de plaquetas, y gran INFARct en el territorio de la arteria cerebral media suministrado. Es importante destacar que la cola de la vena inyección de tPA recombinante en 0,5, 1, o 4 horas después de la THI (10 mg / kg) proporcionó una reducción dependiente del tiempo de la tasa de mortalidad y el tamaño del infarto. Este nuevo modelo de accidente cerebrovascular es simple y puede ser estandarizado a través de laboratorios para comparar los resultados experimentales. Además, se induce la trombosis sin craniectomía o introducir émbolos preformadas. Teniendo en cuenta estas ventajas únicas, el modelo THI es una adición útil al repertorio de la investigación preclínica derrame cerebral.

Bidimensional de imágenes de contraste de moteado de láser (LSCI) 16 se utilizó para comparar las alteraciones del flujo sanguíneo cerebral (CBF) por la oclusión transitoria de la carótida 30 min unilateral (tCCAO), la exposición 30 min a la hipoxia (7,5% de oxígeno), y 30 min unilateral carotídea ligadura en condiciones de hipoxia (THI). Este experimento reveló que tCCAO bajo normoxia suprimió la CBF en el hemisferio carótida se ligó a ~ 50% del valor de línea de base, que se recuperó rápidam...

Watch this Scientific Journal Video about A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia at JoVE.com

A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia

Thromboembolic stroke models are vital tools for optimizing the recanalization therapy. Here we report a murine thrombotic stroke model based on transient cerebral hypoxic-ischemic (tHI) insult, which triggers thrombosis and infarction, and responds favorably to tissue plasminogen activator (tPA)-mediated fibrinolysis in a therapeutic window similar to those in stroke patients.

Un accidente cerebrovascular trombótico Basado En transitoria cerebral por hipoxia-isquemia Modelo

Stroke research has endured many setbacks in translating neuroprotective therapies into clinical practice. In contrast, the real-world therapy (tPA ...

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JoVE Journal

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14.7K vistas.  The Emory University School of Medicine. Thromboembolic stroke models are vital tools for optimizing the recanalization therapy. Here we report a murine thrombotic stroke model based on transient cerebral hypoxic-ischemic (tHI) insult, which triggers thrombosis and infarction, and responds favorably to tissue plasminogen activator (tPA)-mediated fibrinolysis in a therapeutic window similar to those in stroke patients. 

Video: A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia

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

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

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

Modelado de accidente cerebrovascular en ratones - Oclusión arteria cerebral media con el modelo de filamento

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

Investigación

Medicina

Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion 
in the United States in 20091,2. The intraluminal middle cerebral artery occlusion (MCAO) model was developed by Koizumi4 in 1986 to simulate this 
impactful human pathology in the rat. A modification of the MCAO method was later presented by Longa3. Both techniques have been widely used to identify 
molecular mechanisms of brain injury resulting from ischemic stroke and potential therapeutic modalities5. This relatively noninvasive method in rats has been 
extended to use in mice to take advantage of transgenic and knockout strains6,7. To model focal cerebral ischemia, an intraluminal suture is advanced via 
the internal carotid artery to occlude the base of the MCA. Retracting the suture after a specified period of time mimics spontaneous reperfusion, but the 
suture can also be permanently retained. This video will be demonstrating the two major approaches for performing intraluminal MCAO procedure in mice in a 
stepwise fashion, as well as providing insights for potential drawbacks and pitfalls. The ischemic brain tissue will subsequently be stained by 
2,3,5-triphenyltetrazolium chloride (TTC) to evaluate the extent of cerebral infarction8.

Intraluminal Middle Cerebral Artery Occlusion MCAO Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice

The intraluminal middle cerebral artery occlusion (MCAO) model is the most frequent used model among experimental ischemic stroke models. Here we will demonstrate the entire model in detail with the guide of Laser Doppler flowmetry, and its representative results.

Intraluminal la arteria cerebral media de oclusión (MCAO) Modelo para el ictus isquémico con Orientación flujometría láser Doppler en ratones

Stroke is the third leading cause of death and the leading cause of disability in the world, with an estimated cost of near $70 billion 
in the United ...

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

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

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

Bilateral Común oclusión de la arteria carótida como un estímulo de preacondicionamiento adecuado para inducir la tolerancia isquémica precoz de isquemia cerebral focal

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

Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in 20091, 2. Several models of cerebral ischemia have been developed to mimic the human condition of stroke. It has been suggested that up to 80% of all strokes result from ischemic damage in the middle cerebral artery (MCA) area3. In the early 1990s, endothelin-1 (ET-1) 4 was used to induce ischemia by applying it directly adjacent to the surface of the MCA after craniotomy. Later, this model was modified 5 by using a stereotaxic injection of ET-1 adjacent to the MCA to produce focal cerebral ischemia. The main advantages of this model include the ability to perform the procedure quickly, the ability to control artery constriction by altering the dose of ET-1 delivered, no need to manipulate the extracranial vessels supplying blood to the brain as well as gradual reperfusion rates that more closely mimics the reperfusion in humans5-7. On the other hand, the ET-1 model has disadvantages that include the need for a craniotomy, as well as higher variability in stroke volume8. This variability can be reduced with the use of laser Doppler flowmetry (LDF) to verify cerebral ischemia during ET-1 infusion. Factors that affect stroke variability include precision of infusion and the batch of the ET-1 used6. Another important consideration is that although reperfusion is a common occurrence in human stroke, the duration of occlusion for ET-1 induced MCAO may not closely mimic that of human stroke where many patients have partial reperfusion over a period of hours to days following occlusion9, 10. This protocol will describe in detail the ET-1 induced MCAO model for ischemic stroke in rats. It will also draw attention to special considerations and potential drawbacks throughout the procedure.

Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats. In addition, important considerations, advantages, and shortcomings of this model are discussed.

La endotelina-1 inducida por la arteria cerebral media oclusión Modelo para el ictus isquémico con Orientación flujometría láser Doppler en la rata

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in ...

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

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

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

Sistema optimizado para la monitorización de la perfusión cerebral en el modelo de rata de Carrera intraluminal oclusión arteria cerebral media

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

A Murine Model of Subarachnoid Hemorrhage

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

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

Un modelo murino de hemorragia subaracnoidea

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

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As such, primary or &#39;neuronal-like&#39; cell culture systems are commonly utilized. While&#160;these systems are relatively easy to work with, and are useful model systems in which various functional outcomes (such as cell death) can be readily quantified, the examined outcomes and pathways in cultured immature neurons (such as excitotoxicity-mediated cell death pathways) are not necessarily the same as those observed in mature brain, or in intact tissue. Therefore, there is the need to develop models in which cellular mechanisms in mature neural tissue can be examined. We have developed an in vitro technique that can be used to investigate a variety of molecular pathways in intact nervous tissue. The technique described herein utilizes rat cortical tissue, but this technique can be adapted to use tissue from a variety of species (such as mouse, rabbit, guinea pig, and chicken) or brain regions (for example, hippocampus, striatum, etc.). Additionally, a variety of stimulations/treatments can be used (for example, excitotoxic, administration of inhibitors, etc.). In conclusion, the brain slice model described herein can be used to examine a variety of molecular mechanisms involved in excitotoxicity-mediated brain injury.

Excitotoxic Stimulation of Brain Microslices as an In vitro Model of Stroke

We have developed a brain slice model which can be used to examine molecular mechanisms involved in excitotoxicity-mediated brain injury.&#160;This technique generates viable mature brain tissue and reduces animal numbers required for experimentation, whilst keeping the neuronal circuitry, cellular interactions, and postsynaptic compartments partly intact.

Excitotóxica Estimulación del cerebro Microslices como<em&gt; In vitro</em&gt; Modelo de Carrera

Examining molecular mechanisms involved in neuropathological conditions, such as ischemic stroke, can be difficult when using whole animal systems. As ...

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

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

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

Carrera Modelado en ratones: Coagulación permanente de la arteria cerebral media Distal

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

Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic stroke. However, the use of tPA is limited by its narrow therapeutic window and by increased risk of hemorrhagic transformation. There is an urgent need to develop suitable stroke models to study new thrombolytic agents and strategies for treatment of ischemic stroke. At present, two major types of ischemic stroke models have been developed in rats and mice: intraluminal suture MCAO and embolic MCAO. Although MCAO models via the intraluminal suture technique have been widely used in mechanism-driven stroke research, these suture models do not mimic the clinical situation and are not suitable for thrombolytic studies. Among these models, the embolic MCAO model closely mimics human ischemic stroke and is suitable for preclinical investigation of thrombolytic therapy. This embolic model was first developed in rats by Overgaard et al.1 in 1992 and further characterized by Zhang et al. in 19972. Although embolic MCAO has gained increasing attention, there are technical problems faced by many laboratories. To meet increasing needs for thrombolytic research, we present a highly reproducible model of embolic MCAO in the rat, which can develop a predictable infarct volume within the MCA territory. In brief, a modified PE-50 tube is gently advanced from the external carotid artery (ECA) into the lumen of the internal carotid artery (ICA) until the tip of the catheter reaches the origin of the MCA. Through the catheter, a single homologous blood clot is placed at the origin of the MCA. To identify the success of MCA occlusion, regional cerebral blood flow was monitored, neurological deficits and infarct volumes were measured. The techniques presented in this paper should help investigators to overcome technical problems for establishing this model for stroke research.

Embolic Middle Cerebral Artery Occlusion MCAO for Ischemic Stroke with Homologous Blood Clots in Rats

In this paper, we demonstrate a standard method for producing an embolic middle cerebral artery occlusion with homologous blood clots (fibrin-rich) in adult rat. This model closely mimics human ischemic stroke and is suitable for preclinical study of thrombolytic therapy for ischemic stroke.

Arteria embólico cerebral media oclusión (MCAO) para el ictus isquémico con homólogas coágulos de sangre en las ratas

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic ...

Simultaneous PET/MRI Imaging During Mouse Cerebral Hypoxia-ischemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics disturbance in affected cells. Diffusion weighted magnetic resonance imaging (MRI) identifies regions that are damaged, potentially irreversibly, by hypoxia-ischemia. Alterations in glucose utilization in the affected tissue may be detectable by positron emission tomography (PET) imaging of 2-deoxy-2-(18F)fluoro-&#7429;-glucose ([18F]FDG) uptake. Due to the rapid and variable nature of injury in this animal model, acquisition of both modes of data must be performed simultaneously in order to meaningfully correlate PET and MRI data. In addition, inter-animal variability in the hypoxic-ischemic injury due to vascular differences limits the ability to analyze multi-modal data and observe changes to a group-wise approach if data is not acquired simultaneously in individual subjects. The method presented here allows one to acquire both diffusion-weighted MRI and [18F]FDG uptake data in the same animal before, during, and after the hypoxic challenge in order to interrogate immediate physiological changes.

The method presented here uses simultaneous positron emission tomography and magnetic resonance imaging. In the cerebral hypoxia-ischemia model, dynamic changes in diffusion and glucose metabolism occur during and after injury. The evolving and irreproducible damage in this model necessitates simultaneous acquisition if meaningful multi-modal imaging data are to be acquired.

Simultánea de imágenes PET / RM Durante cerebral del ratón hipoxia-isquemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics ...