This work was supported by National Institutes of Health grants HL087990 (G.L.).

Declaración de Ética: ratas macho adultas Sprague-Dawley (330-380 g de pesaje) se utilizaron en este protocolo. Este protocolo fue aprobado por el Cuidado de Animales y el empleo Comisión Institucional (IACUC) en la LSU Health Science Center-Shreveport y está en conformidad con la Guía para el Cuidado y Uso de Animales de Laboratorio &quot;(octava edición, la Academia Nacional de Ciencias, 2011 ). 1 Preparación de modificación Tubo PE-50 <ol><li> Mantenga un largo tubo PE-50 de 30 cm por encima de una estufa de gas con las manos, suavizar gradualmente el tubo. Estire suavemente el tubo. </li><li> Seleccionar un punto en el tubo estirado usando un calibre digital (Figura 1), marcarlo, y cortar el tubo de PE-50 modificado en un segmento de 25 cm de largo con una punta de 1 cm de largo (diámetro exterior: ,30-,34 mm). </li></ol> 2 Preparación de homólogos coágulos sanguíneos <ol><li> Anestesiar la rata con isoflurano (5% para la inducción, 2-3% para mantenimiento) en el 70% de N 2 O y 30% O &lt;sub&gt; 2 por una mascarilla antes de la recogida de sangre. Confirmar la anestesia por una pizca dedo del pie. </li><li> Realizar canulación arterial femoral de una rata donante con método publicado 4 y transferir sangre de la arteria femoral directamente en un 20 cm de largo PE-50 tubos. Colocar el tubo durante 2 horas a temperatura ambiente para coagular la sangre, y después retener el tubo durante 22 horas a 4 ° C. Nota: La canulación se realiza utilizando una técnica aséptica como se describe en el paso 3.1. La rata donante se recupera para su posterior muestreo. </li><li> Cortar el tubo de PE-50 en segmentos de 50 mm y enjuagar el coágulo desde el tubo en una placa de Petri estéril que contiene solución salina normal. </li><li> La transferencia de los 50 mm de largo en un coágulo de 60 mm de largo tubo de PE-10 y conectar cada extremo del tubo de PE-10 a un 20 cm de largo PE-50 tubo conectado a la jeringa de 1 ml que contiene solución salina normal con 23 T de la aguja (Figura 2) . </li><li> Desplazar el coágulo mediante movimiento alternativo continuo de una jeringa a la otra durante 5 min, lo que puede eliminar las células de sangre deel coágulo y minimizar el frágil del coágulo coagulada extravascular, sin interrumpir el núcleo de fibrina. </li><li> Cortar el coágulo en un segmento de 40 mm de largo y transferir el segmento de coágulo en un catéter PE-50 modificado como se describe en el paso 1.2 en condiciones asépticas. Nota: Todos los tubos PE se esteriliza con óxido de etileno. </li></ol> 3. embólico arteria cerebral media oclusión (Figura 3A, B) <ol><li> Esterilizar todas las herramientas quirúrgicas en autoclave (mínimo 121 ° C, 15 PSI, durante 15 min). Desinfectar la mesa de cirugía y equipo quirúrgico asociado utilizando etanol al 70%. </li><li> Anestesiar la rata con isoflurano como se describe en el paso 2.1. Pon a prueba la profundidad de la anestesia mediante la realización de una pizca dedo del pie en ambos pies traseros, y cualquier movimiento observado (la retirada de la pata) indica que el animal no es suficientemente anestesiado para hacer la cirugía. Aplique una pequeña cantidad de ungüento veterinario en ambos ojos para evitar la sequedad, mientras que bajo anestesia. Afeitarse la piel en el cuello y la cabeza ventral reregiones con maquinilla eléctrica para exponer las áreas de la piel. </li><li> Coloque la rata en la posición supina sobre una almohadilla térmica. Inserte una sonda rectal, controlar y mantener la temperatura corporal entre 36,5 a 37,5 ° C utilizando una unidad de control manta homeotérmica. </li><li> Desinfectar la piel afeitada y piel circundante con un 10% de povidona-yodo seguido de etanol al 70%. </li><li> Hacer una incisión de 2 cm de largo en el cuello con un microscopio de disección. Utilice separadores para exponer el campo quirúrgico y disecar la arteria carótida común derecha (CCA), la arteria carótida externa (ACE), la arteria carótida interna (ACI), y la arteria pterigopalatino (PPA) libres de los nervios circundantes y fascia (Figura 3A). Nota: Antes de la cirugía, cambiar los guantes estériles si no hay asistente ayuda a preparar el animal. </li><li> Diseccionar la CCA libres de los nervios que lo rodean (sin dañar el nervio vago) y colocar una sutura de seda 5-0 estéril debajo de la arteria. Ate un nudo corredizo (# 1) y captar la sutura utilizando unos pequeños: hemostat y tire enseñó hacia el cuerpo. </li><li> Diseccionar la ECA y sus dos ramas, la arteria occipital (OA) y la arteria tiroidea superior (STA). Coagular dos ramas utilizando una unidad de electrocirugía veterinaria. Coloque dos trozos de 6-0 suturas de seda estériles en el ACA. </li><li> Separar las dos sedas colocados debajo de la arteria, de una sola pieza hacia la cabeza (extremo distal) y el otro hacia el cuerpo. Ate una ligadura apretada (# 2) en el lado más cercano a la cabeza, agarre la seda con pequeñas pinzas y tire enseñado hacia la cabeza. Preparar un nudo flojo (# 3) con la otra seda para su uso posterior. </li><li> Diseccionar el ICA y PPA de los nervios que lo rodean (sin dañar el nervio vago). Ate la PPA con una sutura 6-0 (# 4). Haga un nudo corredizo con 6-0 sutura alrededor del ICA (# 5). Alternativamente, una pinza de la ICA usando un clip microvascular. </li><li> Corte un agujero pequeño (1/2 a través) en el ACE entre la apretada (# 2) y (# 3) ligaduras sueltas con unas tijeras de primavera al estilo Vannas. Inserte la PE-50 modificado bañerae que contiene coágulo de sangre dentro de la incisión y avanzar a la bifurcación de la CCA. Apriete la ligadura floja (# 3) alrededor de la luz lo suficiente como para asegurar preservar la movilidad del tubo-en la vivienda. </li><li> Cortar la ECA en el lugar del pequeño agujero para liberar el muñón y colocar el muñón por debajo de la bifurcación de la ECA y el ICA; esto permitirá que el tubo modificado deslice fácilmente en el ICA. Abra la ACI y avanzar suavemente el tubo del lumen de la ECA en el ICA hasta que la punta del catéter alcanza el origen de la MCA (~ 17 mm de la bifurcación). Nota: Antes de insertar la punta del tubo en el ECA, estéril la superficie exterior del tubo con etanol al 70%, y luego se lava con solución salina normal estéril. </li><li> Inyectar el coágulo a través de la PE-50 modificado catéter junto con 10 l de solución salina durante 10 s usando una jeringa Hamilton de 100 l (Figura 3B). </li><li> Retire la sonda de la ECA 5 minutos más tarde. Ate la ECA y volver a abrir CCA. Suturar la incisión en el cuello. </li></ol> 4. Supervisión del flujo sanguíneo cerebral regional (rCBF) <ol><li> Antes de la oclusión de la MCA, hacer una incisión de línea media 1.2 cm de longitud en el cuero cabelludo para exponer el hueso del cráneo. Quite los tejidos del hueso del cráneo con un raspador dental y swaps de algodón estériles. Nota: Antes de la cirugía, la zona del cuero cabelludo y la piel circundante expuesta se desinfectan con un 10% de povidona-yodo seguido de etanol al 70%. </li><li> Perforar un agujero de trépano de diámetro 1,5 mm situado en 2 mm posterior y 5 mm lateral al bregma usando una fresa de 0,7 mm esférica de acero inoxidable (Figura 4). NOTA: Mantenga la duramadre intacta. </li><li> Coloque la sonda de 0,5 mm por encima de la superficie dura. Monitorear la rCBF a 0 (línea de base), 5, 15, 30, 60, 90, y 120 min después de la embolización y continuar en 5, 15, 30, 45, y 60 min después de la administración intravenosa de tPA. Después de la última medición de rCBF, la incisión del cuero cabelludo se cierra mediante sutura. Nota: Antes de la incisión en el cuello medimos rCBF como línea de base antes de MCA Occlusde iones. Mensaje MCA oclusión medimos rCBF después del cierre de sutura de incisión del cuello. Por lo tanto, la incisión en el cuello se mantiene estéril. </li></ol> Cuidado 5. postoperatoria <ol><li> Inyectar 2,5 ml de solución salina por vía subcutánea para evitar la deshidratación e inyectar Buprenex (0,05 mg / kg, SC) inmediatamente después de la cirugía y cada 6-12 horas según sea necesario para aliviar el dolor. </li><li> Deje de anestesia con isoflurano. Coloque la rata en una cámara de recuperación veterinaria 37 ° C (mantener animales caliente) y mantener la observación. Generalmente se tarda 10 min para la rata para recuperarse de la anestesia. Luego, se coloca el animal en una jaula esterilizada, coloque un poco de comida humedecida en una placa de Petri en la jaula, y volver a la jaula para sala de esterilización de animales. </li><li> 24 hr después del accidente cerebrovascular, la rata eutanasia con una sobredosis de pentobarbital sódico (100 mg / kg de peso corporal, ip). </li></ol> 6. neurológico de Déficit de Score <ol><li> Realizar la puntuación Bederson antes ya las 2 horas después de la embolización. Use un s de calificacióncale de 0-3 como se ha descrito previamente 5: 0, elevando la rata por la cola, el animal se extiende ambas patas delanteras en el suelo y no exhiben otros déficits (movimiento normal); 1, el aumento de la rata por la cola, flexionar la extremidad anterior contralateral; 2, disminución de la resistencia al empuje lateral (y la flexión de la extremidad anterior) y sin vueltas; 3, el mismo comportamiento que el grado 2 con círculos. NOTA: Las ratas que muestran Bederson puntuación = 0 (sin déficit) a las 2 horas después de la embolización se excluyen de mayor estudio. </li></ol>

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We do not have any potential conflicting interests to disclose.

En este estudio, hemos demostrado un método estándar para la realización de un modelo de accidente cerebrovascular embólico MCAO en la rata, en el que el origen de la MCA se ocluye por un coágulo rico en fibrina. La principal ventaja de este modelo es: la oclusión del tronco de MCA con un rico en fibrina del coágulo de sangre es similar al ictus tromboembólico en los seres humanos, el modelo de accidente cerebrovascular embólico es adecuada para realizar investigación preclínica de la terapia fibrinolítica, ...

A correction was made to <a href="/video/51956">Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats</a>. The institution information was updated.
The institution "Louisiana State University Health Science Center" was changed to "Louisiana State University Health Science Center, Shreveport".

A correction was made to <a href="/video/51956">Embolic Middle Cerebral Artery Occlusion (MCAO) for Ischemic Stroke with Homologous Blood Clots in Rats</a>. The institution information was updated.

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

El accidente cerebrovascular es la tercera causa principal de muerte en los Estados Unidos, pero las opciones de tratamiento para el accidente cerebrovascular agudo permanece limitada. En la actualidad, la infusión intravenosa de activador de plasminógeno tisular recombinante (tPA) para disolver los coágulos de sangre es la terapia más eficaz para el accidente cerebrovascular isquémico agudo. Sin embargo, el uso de tPA está limitada por su estrecha ventana terapéutica y por el aumento de riesgo de hemorragia intracerebral. Por lo tanto, se necesita urgentemente un modelo de accidente cerebrovascular adecuado para la investigación trombolítico. La arteria cerebral media (MCA) es la arteria ocluida más a menudo en el accidente cerebrovascular en los seres humanos. Centrado en esta arteria, se han establecido muchos modelos animales de accidente cerebrovascular isquémico. En la actualidad, se han desarrollado dos tipos principales de modelos de isquemia focal de roedores por la oclusión de la MCA: sutura modelo MCAO y embólico MCAO modelo. Aunque los modelos MCAO a través de la técnica de sutura intraluminal han sido ampliamente utilizados en mecanismo impulsado investigación del accidente cerebrovascular, estos modelos de sutura no lo hacenderrame cerebral humano imitador, como hasta el 80% de los ictus humanos son causados ​​por trombosis o embolia. Sin embargo, embólico modelo de accidente cerebrovascular usando coágulos de sangre imita estrechamente derrame cerebral humano y se considera adecuada para el estudio trombolítico. Este modelo embólico fue desarrollado por primera vez en ratas por Overgaard et al. 1 en 1992 y se caracteriza por Zhang et al. En 1997 2 y por Dinapoli et al. En 2006 3. Aunque embólico MCAO ha ganado cada vez más atención, hay problemas técnicos que enfrentan por muchos laboratorios. En este artículo, se demuestra un método estándar para la producción de embólico MCAO con coágulos de sangre homóloga en la rata adulta, lo que puede desarrollar un infarto predecible dentro del territorio de la ACM. Las técnicas presentadas en este artículo deben ayudar a los investigadores a superar los problemas técnicos que permitan determinar este modelo para la investigación de un accidente cerebrovascular.

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		<tr height="24">
			<td height="24" style="height:24px;width:232px;">1ml syringe</td>
			<td style="width:135px;">Consumable</td>
			<td style="width:237px;">Becton, Dickinson and Company</td>
			<td style="width:135px;">309659</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">23G needle</td>
			<td style="width:135px;">Consumable</td>
			<td style="width:237px;">Becton, Dickinson and Company</td>
			<td style="width:135px;">305143</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:232px;">30G needle</td>
			<td style="width:135px;">Consumable</td>
			<td style="width:237px;">Becton, Dickinson and Company</td>
			<td style="width:135px;">305106</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:232px;">PE-50 tubing</td>
			<td style="width:135px;">Consumable</td>
			<td style="width:237px;">Becton, Dickinson and Company</td>
			<td style="width:135px;">427517</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;width:232px;">PE-10 tubing</td>
			<td style="width:135px;">Consumable</td>
			<td style="width:237px;">Becton, Dickinson and Company</td>
			<td style="width:135px;">427400</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:232px;">6-0 Silk suture</td>
			<td style="width:135px;">Consumable</td>
			<td>Harvard apparatus</td>
			<td style="width:135px;">723287</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:232px;">5-0 Silk suture</td>
			<td style="width:135px;">Consumable</td>
			<td>Harvard Apparatus</td>
			<td>517607</td>
		</tr>
	</tbody>
</table>

embolic middle cerebral artery occlusion (mcao) for ischemic stroke with homologous blood clots in rats

Terapia Clínicamente, trombolítico con el uso del activador del plasminógeno tisular recombinante (tPA) sigue siendo el tratamiento más eficaz para el accidente cerebrovascular isquémico agudo. Sin embargo, el uso de tPA está limitada por su estrecha ventana terapéutica y por el aumento de riesgo de transformación hemorrágica. Hay una necesidad urgente de desarrollar modelos de carrera adecuados para estudiar nuevos agentes trombolíticos y estrategias para el tratamiento del accidente cerebrovascular isquémico. En la actualidad, hay dos tipos principales de modelos de accidente cerebrovascular isquémico se han desarrollado en ratas y ratones: MCAO sutura intraluminal y embólico MCAO. Aunque los modelos MCAO a través de la técnica de sutura intraluminal han sido ampliamente utilizados en mecanismo impulsado investigación del accidente cerebrovascular, estos modelos de sutura no imitan la situación clínica y no son adecuados para estudios trombolíticos. Entre estos modelos, el modelo de MCAO embólico imita estrechamente accidente cerebrovascular isquémico humano y es adecuado para la investigación preclínica de la terapia trombolítica. Este modelo embólico fue desarrollado por primera vez en ratas por Overgaard et al. 1 en 1992 y caracterizado además por Zhang et al. en 1997 2. Aunque embólico MCAO ha ganado cada vez más atención, hay problemas técnicos que enfrentan muchos laboratorios. Para satisfacer las crecientes necesidades de investigación trombolítico, se presenta un modelo altamente reproducible de embólico MCAO en ratas, lo que puede desarrollar un volumen de infarto predecible dentro del territorio de la ACM. En resumen, un PE-50 tubo modificado se avanza suavemente desde la arteria carótida externa (ACE) en el lumen de la arteria carótida interna (ACI) hasta que la punta del catéter alcanza el origen de la MCA. A través del catéter, un solo coágulo de sangre homóloga se coloca en el origen de la MCA. Para identificar el éxito de oclusión de la ACM, el flujo sanguíneo cerebral regional se controló, se midieron los déficits neurológicos y los volúmenes de infarto. Las técnicas que se presentan en este documento deben ayudar a los investigadores a superar los problemas técnicos para el establecimiento de este modelo para la investigación de un accidente cerebrovascular. &lt;/ P&gt;

Flujometría láser Doppler (LDF) se utilizó para controlar rCBF durante la inducción de la isquemia cerebral 6,7. Muchos laboratorios incluyendo nuestro laboratorio han estado utilizando rCBF para identificar animales con éxito la oclusión MCA, pero los umbrales de la línea de base variaron entre los laboratorios que están relacionados con el sitio de medición. La sonda de la LDF se coloca en 2 mm posterior y 5 mm lateral a la bregma como se ha descrito previamente 6. rCBF se monitorizó a ...

Watch this Scientific Journal Video about Embolic Middle Cerebral Artery Occlusion MCAO for Ischemic Stroke with Homologous Blood Clots in Rats at JoVE.com

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 ...

embolic-middle-cerebral-artery-occlusion-mcao-for-ischemic-stroke

Research

JoVE Journal

Medicine

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

26.8K vistas.  Louisiana State University Health Science Center, Shreveport. 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. 

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

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

Mouse Model of Middle Cerebral Artery Occlusion

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

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

Modelo de ratón de la oclusión arteria cerebral media

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

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 ...

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

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

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

Utilizando una ventana craneal para visualizar la arteria cerebral media Durante la endotelina-1 inducida por oclusión de la arteria cerebral media

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

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

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

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

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 ...

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 ...

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

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

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

Realización Permanente distal cerebral media carótida común con oclusión de la arteria en ratas de edad avanzada para el Estudio de la isquemia cortical con sostenida de la discapacidad

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

Surgical Approach for Middle Cerebral Artery Occlusion and Reperfusion Induced Stroke in Mice

Stroke is a leading cause of death worldwide and continues to be one of the major causes of long-term adult disabilities. About 87% of strokes are ischemic in origin and occur in the territory of the middle cerebral artery (MCA). Currently the only Food and Drug Administration (FDA) approved drug for the treatment of this devastating disease is tissue plasminogen activator (tPA). However, tPA has a small therapeutic window for administration (3 - 6 hr), and is only effective in 4% of the patients who actually receive it. Current research focuses on understanding the pathophysiology of stroke in order to find potential therapeutic targets. Thus, reliable models are crucial, and the MCA occlusion (MCAo) model (also termed the intraluminal filament or suture model) is deemed to be the most clinically relevant surgical model of ischemic stroke, and is fairly non-invasive and easily reproducible. Typically the MCAo model is used with rodents, especially with mice due to all the genetic variations available for this species. Here we describe (and present in the video) how to successfully perform the MCAo model (with reperfusion) in mice to generate reliable and reproducible data.

In order to understand the pathophysiology of stroke, it is important to use reliable models. This paper will describe one of the most frequently used stroke models in mice, termed the middle cerebral artery occlusion (MCAo) model (also termed the intraluminal filament or suture model) with reperfusion.