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.

ETIK ANALYS: Experiment som involverar djurförsök har utförts i enlighet med de riktlinjer och regler som anges av Landesamt für Gesundheit und Soziales, Berlin, Tyskland (G0298 / 13) och anländer kriterier, som är tillämpligt. För denna studie 10- till 12-veckor gamla C57BL / 6J-möss användes. 1. Lateral Kronisk Kraniell fönster Framställning <ol><li> Utföra anestesi med en subkutan injektion av ketamin (90 mg / kg) och xylazin (10 mg / kg). Test för adekvat sedering med en ...

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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intravital+microscopy+of+the+mouse+brain+microcirculation+using+a+closed+cranial+window.">Intravital microscopy of the mouse brain microcirculation using a closed cranial window.</a> J Vis Exp. (45), (2010).</li><li>Rosell, A., Agin, 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=Distal+occlusion+of+the+middle+cerebral+artery+in+mice:+are+we+ready+to+assess+long-term+functional+outcome.">Distal occlusion of the middle cerebral artery in mice: are we ready to assess long-term functional outcome.</a> Transl Stroke Res. 4 (3), 297-307 (2013).</li><li>Dorand, R. D., Barkauskas, D. S., Evans, T. A., Petrosiute, A., Huang, A. 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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J., Prass, K., Dirnagl, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isoflurane+induced+prolonged+protection+against+cerebral+ischemia+in+mice:+a+redox+sensitive+mechanism.">Isoflurane induced prolonged protection against cerebral ischemia in mice: a redox sensitive mechanism.</a> Neuroreport. 13 (11), 1431-1435 (2002).</li><li>Gertz, K., Priller, 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=Physical+activity+improves+long-term+stroke+outcome+via+endothelial+nitric+oxide+synthase-dependent+augmentation+of+neovascularization+and+cerebral+blood+flow.">Physical activity improves long-term stroke outcome via endothelial nitric oxide synthase-dependent augmentation of neovascularization and cerebral blood flow.</a> Circ Res. 99 (10), 1132-1140 (2006).</li><li>Dirnagl, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bench+to+bedside:+the+quest+for+quality+in+experimental+stroke+research.">Bench to bedside: the quest for quality in experimental stroke research.</a> J Cereb Blood Flow Metab. 26 (12), 1465-1478 (2006).</li></ol>

Stroke är en av de främsta orsakerna till långvarig funktionsnedsättning och död över hela världen en. Utöver akut behandling, undersöka nya metoder och mekanismer för att påskynda och förbättra återhämtningen efter stroke är fortfarande ett utmärkt medicinsk mål 7. Experimentella strokeforskare använder ofta gnagarmodeller av fokal cerebral ischemi. I själva verket, modeller inducerar övergående eller permanent MCAo efterlikna en av de vanligaste typerna av fokal cerebral isc...

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

Fokal cerebral ischemi (dvs ischemisk stroke) kan orsaka allvarliga hjärnskador, vilket leder till en allvarlig förlust av neuronal funktion och följaktligen till en värd av motoriska och kognitiva funktionsnedsättningar. Den höga förekomsten utgör en allvarlig hälsobörda, som stroke är bland de främsta orsakerna till långvarig funktionsnedsättning och död över hela världen en. Återvinning av neuronal funktion är, i de flesta fall endast partiell. Hittills är behandlingsalternativ mycket begränsade, särskilt på grund av den snäva tidsfönster för trombolys 2,3. Fastställande metoder för att påskynda återhämtningen från stroke fortfarande ett utmärkt medicinsk mål; men detta har hindrats av otillräckliga mekaniska insikter i återhämtningsprocessen. Experimentella strokeforskare använder ofta gnagarmodeller av fokal cerebral ischemi. Bortom den akuta fasen, är stroke allt mer fokuserat på subakut och kronisk fas efter cerebral ischemi. De flesta strokeforskare gäller permanent eller transient ocklusion av MCA hos möss eller råttor. Hos patienter, inneslutningar av MCA är bland de vanligaste orsakerna till ischemisk stroke 4. Förutom proximal ocklusion av MCA använder trådmodellen, är kirurgisk ocklusion av distala MCA förmodligen den mest använda modellen i experimentell stroke forskning 5. Ocklusion av en distal (till förgrening av lenticulo-striate artärer) MCA gren reservdelar typiskt striatum och främst drabbar neocortex. Kärlocklusion kan vara permanent eller övergående. Hög reproducerbarhet av skada volym och mycket låga dödstal i förhållande till den långsiktiga resultat är de viktigaste fördelarna med denna modell. Här visar vi hur du utför en kronisk kraniell fönster (CW) förberedelse sidled till sinus sagittalis, och därefter hur man kirurgiskt framkalla en distal slag under fönstret med hjälp av en kraniotomi tillvägagångssätt. Denna metod kan användas för sekventiell avbildning av akuta och kroniska förändringar efter ischemi viaepi-belysning, konfokala laserskanning, och två-foton intravital mikroskopi.

Tidslinjen och representativa resultat visas i figurerna 2 och 3. Hjärn fönster förberedelse, med en liten hjärn fönster i sidled till sinus sagittalis superior (Figur 2 B, C, D) resulterar i en mycket låg dödlighet och sjuklighet när de utförs av en erfaren kirurg. Alla de 10 djur överlevde, och alla kronisk CW kan användas för hög bildkvalitet, även 28 dagar efter operationen. Det var inga problem med sårinfektioner elle...

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

Kirurgisk ocklusion av en distal mellersta cerebral artär gren (MCAo) är en ofta använd modell i experimentell stroke forskning. Detta manuskript beskriver den grundläggande tekniken för permanent MCAo, kombinerat med insättning av en lateral kraniell fönster, vilket ger möjlighet för längs intravital mikroskopi i möss.

Lateral Kronisk Kraniell fönster Framställning Aktiverar<em&gt; In Vivo</em&gt; Observation Efter Distal Middle Cerebral artärocklusion hos möss

Focal cerebral ischemia (i.e., ischemic stroke) may cause major brain injury, leading to a severe loss of neuronal function and consequently to a host of ...

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

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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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Piezo High Accuracy Surgical Osteal Removal PHASOR: A Technique for Improved Cranial Window Surgery in Mice

Piezo hög noggrannhet kirurgiska Osteal borttagning (PHASOR): En teknik för förbättrat kraniala fönster kirurgi hos möss

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

Modellering stroke hos möss: Transient Middle Cerebral Artery Ocklusion via den yttre halsartären

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

Implantation av ett kranialfönster för upprepad in vivo-avbildning hos vakna möss

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

Konstruktion och implementering av kolfibermikroelektrodmatriser för kroniska och akuta in vivo-inspelningar

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

En modell för encefalomyosynangios behandling efter mellersta cerebral artär ocklusion-inducerad stroke hos möss

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 Bredfälts- och tvåfotonkalciumavbildning från en mus med ett stort kranialfönster

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

Bedömning av termiska skador från robotborrad kraniotomi för kranial fönsterkirurgi hos möss

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

En modifierad transkraniell ocklusionsmodell för mellersta hjärnartären för att studera strokeresultat hos åldrade möss

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