Name: lateral chronic cranial window preparation enables in vivo observation following distal middle cerebral artery occlusion in mice
Uploaded: 2016-12-29T13:00:00
Description: Watch this Scientific Journal Video about Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice at JoVE.com

VP is a participant in the Charit&#233; Clinical Scientist Program, funded by the Charit&#233; - Universit&#228;tsmedizin Berlin and the Berlin Institute of Health. TB is an SNSF PostDoc Mobility fellow. The authors receive grant support from EinsteinStiftung/A-2012-153 to PV.

DECLARACIÓN DE ÉTICA: Los experimentos con seres animales se realizaron de acuerdo con las directrices y normas establecidas por Landesamt für Gesundheit und Soziales, Berlín, Alemania (G0298 / 13) y los criterios llegar, según sea el caso. Para este estudio, de 10 a 12 semanas de edad, machos C57BL / 6J fueron utilizados. 1. lateral crónica craneal Ventana Preparación <ol><li> Realizar la anestesia con una inyección subcutánea de ketamina (90 mg / kg) y xilazina (10 mg / kg). Pru...

<ol>
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Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+intravital+thinned+skull+and+cranial+window+approaches+to+study+CNS+immunobiology+in+the+mouse+cortex.">Comparison of intravital thinned skull and cranial window approaches to study CNS immunobiology in the mouse cortex.</a> Intravital. 3 (2), (2014).</li><li>Balkaya, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+post-stroke+behavior+in+mouse+models+of+focal+ischemia.">Assessing post-stroke behavior in mouse models of focal ischemia.</a> J Cereb Blood Flow Metab. 33 (3), 330-338 (2013).</li><li>Balkaya, M., Kr&#246;ber, J., Gertz, K., Peruzzaro, S., Endres, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+long-term+functional+outcome+in+a+murine+model+of+mild+brain+ischemia.">Characterization of long-term functional outcome in a murine model of mild brain ischemia.</a> J Neurosci Methods. 213 (2), 179-187 (2013).</li><li>Freret, T., Bouet, V., 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=Behavioral+deficits+after+distal+focal+cerebral+ischemia+in+mice:+Usefulness+of+adhesive+removal+test.">Behavioral deficits after distal focal cerebral ischemia in mice: Usefulness of adhesive removal test.</a> Beh Neurosci. 123 (1), 224-230 (2009).</li><li>Liu, S., Zhen, G., Meloni, B. P., Campbell, K., Winn, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RODENT+STROKE+MODEL+GUIDELINES+FOR+PRECLINICAL+STROKE+TRIALS+(1ST+EDITION).">RODENT STROKE MODEL GUIDELINES FOR PRECLINICAL STROKE TRIALS (1ST EDITION).</a> J Exp Stroke Trans Med. 2 (2), 2-27 (2009).</li><li>Florian, B., Vintilescu, 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=Long-term+hypothermia+reduces+infarct+volume+in+aged+rats+after+focal+ischemia.">Long-term hypothermia reduces infarct volume in aged rats after focal ischemia.</a> Neurosci Lett. 438 (2), 180-185 (2008).</li><li>Noor, R., Wang, C. X., Shuaib, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+hyperthermia+on+infarct+volume+in+focal+embolic+model+of+cerebral+ischemia+in+rats.">Effects of hyperthermia on infarct volume in focal embolic model of cerebral ischemia in rats.</a> Neurosci Lett. 349 (2), 130-132 (2003).</li><li>Barber, P. A., Hoyte, L., Colbourne, F., Buchan, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Temperature-regulated+model+of+focal+ischemia+in+the+mouse:+a+study+with+histopathological+and+behavioral+outcomes.">Temperature-regulated model of focal ischemia in the mouse: a study with histopathological and behavioral outcomes.</a> Stroke. 35 (7), 1720-1725 (2004).</li><li>Shin, H. 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Accidente cerebrovascular es una de las principales causas de discapacidad a largo plazo y la muerte en todo el mundo 1. Más allá de tratamiento agudo, la investigación de nuevos enfoques y mecanismos para acelerar y mejorar la recuperación después del accidente cerebrovascular sigue siendo un objetivo primordial médica 7. investigadores ictus experimentales emplean con frecuencia modelos de roedores de isquemia cerebral focal. De hecho, los modelos que inducen transitoria o permanente MCAO i...

Stroke is among the principal causes of long-term disability and death worldwide1, coming second after coronary heart disease. In addition, stroke is the primary cause of long-term disability, underscoring its tremendous socioeconomic impact6-8. Beyond acute treatment, investigating new approaches and mechanisms to accelerate and enhance recovery after stroke remains a prime medical goal7. 
In the last few decades, data from experimental stroke research has contributed substantially to understanding the complex pathophysiological cascades triggered by ischemia9,10. Excitotoxicity, apoptosis, peri-infarct depolarization, and inflammation have been identified as the most relevant mediators of cell death following focal cerebral ischemia. Moreover, using animal models of cerebral ischemia, important concepts, diagnostic modalities, and therapeutic approaches have been developed and validated (e.g., "penumbra" and thrombolysis)11.
The availability of experimental stroke models, combined with non-invasive imaging modalities (e.g., magnetic resonance imaging (MRI), computed tomography, or laser speckle contrast analysis), enables the researcher to investigate hyperacute and chronic pathophysiological changes induced by the ischemic insult in a longitudinal manner12. Along with studying the spatiotemporal profile of the evolving lesion, changes resembling neuronal plasticity can be investigated and correlated to functional outcomes and histological findings. Within the last few years, further methodological advances have been made using the combination of cerebral ischemia models and in vivo microscopy via cranial windows13. These new techniques allow investigators to analyze the neurovascular unit at the cellular and molecular level, with great analytic power in the acute, subacute, and chronic phases following focal cerebral ischemia14. Moreover, in vivo microscopy imaging of microcirculatory dynamics has revealed novel aspects of cerebral microvasculature function and angioarchitecture, with significant pathophysiological relevance15-17. 
In this protocol, we present how to perform a chronic CW preparation lateral to the sagittal sinus and how to surgically induce a distal stroke underneath the window. This mouse model can be applied to sequential imaging of acute, subacute, and chronic changes following focal cerebral ischemia via epi-illuminating, confocal laser scanning, and two-photon intravital microscopy.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Binocular surgical microscope</td>
			<td>Zeiss</td>
			<td>Stemi 2000 C</td>
			<td></td>
		</tr>
		<tr>
			<td>Light source for microscope</td>
			<td>Zeiss</td>
			<td>CL 6000 LED</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad with rectal probe</td>
			<td>FST</td>
			<td>21061-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotactic frame</td>
			<td>Kopf</td>
			<td>Model 930</td>
			<td></td>
		</tr>
		<tr>
			<td>Anaethesia system for isoflurane</td>
			<td>Draeger</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Abott</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont forceps #5</td>
			<td>FST</td>
			<td>11251-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont forceps #7</td>
			<td>FST</td>
			<td>11271-30</td>
			<td></td>
		</tr>
		<tr>
			<td>Bipolar Forceps</td>
			<td>Erbe</td>
			<td>20195-501</td>
			<td></td>
		</tr>
		<tr>
			<td>Bipolar Forceps&#160;</td>
			<td colspan="2">Erbe&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 20195-022</td>
			<td></td>
		</tr>
		<tr>
			<td>Microdrill</td>
			<td colspan="2">FST&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; 18000-17&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>FST</td>
			<td>12010-14</td>
			<td></td>
		</tr>
		<tr>
			<td>5-0 silk suture</td>
			<td>Feuerstein, Suprama</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>7-0 silk suture</td>
			<td>Feuerstein,Suprama</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>8-0 silk suture</td>
			<td>Feuerstein, Suprama</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Veterinary Recovery Chamber</td>
			<td>Peco Services</td>
			<td>V1200</td>
			<td></td>
		</tr>
	</tbody>
</table>

lateral chronic cranial window preparation enables in vivo observation following distal middle cerebral artery occlusion in mice

Isquemia cerebral focal (es decir, accidente cerebrovascular isquémico) puede causar lesión cerebral importante, lo que lleva a una pérdida grave de la función neuronal y por lo tanto a un anfitrión de motor y discapacidades cognitivas. Su alta prevalencia supone una grave carga para la salud, como accidente cerebrovascular es una de las principales causas de discapacidad a largo plazo y la muerte en todo el mundo 1. Recuperación de la función neuronal es, en la mayoría de los casos, sólo parcial. Hasta ahora, las opciones de tratamiento son muy limitadas, en particular, debido a la estrecha ventana de tiempo para la trombólisis 2,3. La determinación de métodos para acelerar la recuperación del accidente cerebrovascular sigue siendo un objetivo primordial médica; sin embargo, esto se ha visto obstaculizada por la insuficiencia de conocimientos mecánicos en el proceso de recuperación. investigadores ictus experimentales emplean con frecuencia modelos de roedores de isquemia cerebral focal. Más allá de la fase aguda, investigación del accidente cerebrovascular se centra cada vez más en la fase subaguda y crónica después de una isquemia cerebral. La mayoría de investigadores aplican ictus permanente o tranoclusión de la MCA sient en ratones o ratas. En los pacientes, las oclusiones de la MCA se encuentran entre las causas más frecuentes de accidente cerebrovascular isquémico 4. Además de la oclusión proximal de la MCA mediante el modelo de filamento, oclusión quirúrgica de la MCA distal es probablemente el modelo más utilizado en la investigación del accidente cerebrovascular experimental 5. La oclusión de una distal (a la ramificación de las arterias lenticulo-estriados) rama MCA normalmente detiene el cuerpo estriado y afecta principalmente a la neocorteza. oclusión de los vasos puede ser permanente o transitoria. Alta reproducibilidad del volumen de la lesión y muy bajas tasas de mortalidad con respecto a los resultados a largo plazo son las principales ventajas de este modelo. Aquí, demostramos cómo llevar a cabo una preparación lateral crónica ventana del cráneo (CW) al seno sagital, y después cómo quirúrgicamente inducir un accidente cerebrovascular distal debajo de la ventana usando un enfoque craneotomía. Este enfoque se puede aplicar para la imagen secuencial de los cambios agudos y crónicos después de la isquemia a través deepi-iluminación, láser confocal de barrido, y de dos fotones microscopía intravital.

La línea de tiempo y los resultados representativos se muestran en las figuras 2 y 3. La preparación de ventana del cráneo, con una pequeña ventana lateral craneal al seno sagital superior (Figura 2 B, C, D) se traduce en una tasa de mortalidad y morbilidad muy baja cuando se lleva a cabo por un cirujano experimentado. Todos los 10 animales sobrevivieron, y todos CW crónica se podría utilizar para obtener imágenes de alta calidad,...

Watch this Scientific Journal Video about Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice at JoVE.com

Lateral Chronic Cranial Window Preparation Enables In Vivo Observation Following Distal Middle Cerebral Artery Occlusion in Mice

oclusión quirúrgica de una rama distal de la arteria cerebral media (MCAO) es un modelo frecuentemente usado en investigación del accidente cerebrovascular experimental. Este manuscrito describe la técnica básica de la MCAO permanente, combinado con la inserción de una ventana craneal lateral, que ofrece la oportunidad para microscopía intravital longitudinal en ratones.

Ventana lateral crónica craneal Preparación Activa<em&gt; En Vivo</em&gt; Observación Siguiendo distal de la arteria cerebral media oclusión en ratones

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Modelo de isquemia cerebral focal por oclusión endovascular de sutura de la arteria cerebral media en la rata

Stroke is the leading cause of disability and the third leading cause of death in adults worldwide1. In human stroke, there exists a highly variable ...

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Multiphoton microscopy has been widely adapted for imaging neurons in vivo. Repeated imaging requires implantation of a cranial window or repeated ...

Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery

Modelado del accidente cerebrovascular en ratones: oclusión transitoria de la arteria cerebral media a través de la arteria carótida externa

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options ...

Implantation of a Cranial Window for Repeated In Vivo Imaging in Awake Mice

Implantation of a Cranial Window for Repeated In Vivo Imaging in Awake Mice

Implantación de una ventana craneal para imágenes repetidas in vivo en ratones despiertos

To fully understand the cellular physiology of neurons and glia in behaving animals, it is necessary to visualize their morphology and record their ...

Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings

Construction and Implementation of Carbon Fiber Microelectrode Arrays for Chronic and Acute In Vivo Recordings

Construcción e implementación de matrices de microelectrodos de fibra de carbono para grabaciones in vivo crónicas y agudas

Multichannel electrode arrays offer insight into the working brain and serve to elucidate neural processes at the single-cell and circuit levels. ...

A Model for Encephalomyosynangiosis Treatment after Middle Cerebral Artery Occlusion-Induced Stroke in Mice

Un modelo para el tratamiento de la encefalomiosinangiosis después del accidente cerebrovascular inducido por la oclusión de la arteria cerebral media en ratones

There is no effective treatment available for most patients suffering with ischemic stroke, making development of novel therapeutics imperative. The ...

In Vivo Wide-Field and Two-Photon Calcium Imaging from a Mouse Using a Large Cranial Window

In Vivo Wide-Field and Two-Photon Calcium Imaging from a Mouse Using a Large Cranial Window

In vivo Imágenes de calcio de campo amplio y dos fotones de un ratón usando una ventana craneal grande

Wide-field calcium imaging from the mouse&#39;s neocortex allows one to observe cortex-wide neural activity related to various brain functions. On the ...

Assessment of Thermal Damage from Robot-Drilled Craniotomy for Cranial Window Surgery in Mice

Evaluación del daño térmico de la craneotomía perforada por robot para la cirugía de ventana craneal en ratones

Cranial window surgery allows for the imaging of brain tissue in live mice with the use of multiphoton or other intravital imaging techniques. However, ...

Establishing a Transcranial Middle Cerebral Artery Occlusion Mice Model

A Modified Transcranial Middle Cerebral Artery Occlusion Model to Study Stroke Outcomes in Aged Mice

Innovative Techniques and Future Directions in Stroke Research (Video) | JoVE

Un modelo de oclusión transcraneal modificado de la arteria cerebral media para estudiar los resultados del accidente cerebrovascular en ratones ancianos

In experimental stroke research, middle cerebral artery occlusion (MCAO) with an intraluminal filament is widely used to model ischemic stroke in mice. ...