Name: induction of ischemic stroke and ischemia-reperfusion in mice using the middle artery occlusion technique and visualization of infarct area
Uploaded: 2017-02-02T12:30:00
Description: Watch this Scientific Journal Video about Induction of Ischemic Stroke and Ischemia-reperfusion in Mice Using the Middle Artery Occlusion Technique and Visualization of Infarct Area at JoVE.com

We would like to thank Dr. Lei Chen (Icahn School of Medicine at Mount Sinai, NY) who first established this model in our laboratory. Supported in part by HL126559, DA039576, MH098891, MH63022, MH072567, DA027569, and NSC 2015/17/B/NZ7/02985. Dr. Luc Bertrand is supported in part by a postdoctoral fellowship from the American Heart Association (16POST31170002).

Alla förfaranden godkändes av University of Miami Institutional Animal Care och användning kommittén (IACUC) i enlighet med National Institutes of Health (NIH) riktlinjer. Användningen av steril utrustning och aseptisk teknik krävs. 1. Förbereda Ocklusion Suture <ol><li> Använder en sutur av 0,21 mm diameter för möss mellan 20-25 g och 0,23 mm för möss mellan 25 - 35 g kroppsvikt. Valet av typ av suturer för kortex förfarande baseras på djurens vikt. </li><li> Med hjälp av ...

<ol>
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Den framgångsrika användningen av den beskrivna MCAO-metoden är mycket beroende av en förståelse av det cerebrala blodflödet anatomi. Eftersom korrekt placering av suturen är svårt att urskilja på grund av bristen på direkta visuella ledtrådar, är upprepade praktiken viktigt att behärska förfarandet innan du använder den för undersökande studier. Slagvolym bör analyseras för att säkerställa jämna resultat. Tillsatsen av en laser Doppler-system kan hjälpa till att bestämma den framgångsrika ocklu...

Studiet av hjärt-kärlsjukdom, såsom stroke, förlitar sig på användningen av in vivo-modeller. För att förstå det finns en möjlighet av ischemi, läkemedelstoxicitet, och / eller behandling, finns det ett behov av att använda en lämplig, standardiserad, tillförlitlig och reproducerbar modell av sjukdomen, vilket gör det möjligt för jämförande studier mellan behandlingsgrupperna. I detta manuskript, använder vi möss, med tanke på att det finns ett stort antal transgena möss och standardiserade bedömningsmodeller. Kratta poäng för att bedöma motor- och beteende underskott efter experimentell ischemisk stroke och efter återhämtning har utvecklats ett, två. Flera ischemisk stroke modeller finns tillgängliga, såsom fullständig global cerebral ischemi, ofullständig global ischemi, multifokal cerebral ischemi och fokal cerebral ischemi. Den senare gruppen är också den kategori av stroke mest utbredd hos patienter. Majoriteten av eveNTS initieras genom bildandet av en embolisk eller trombotisk ocklusion vid eller nära mitten cerebral artär (MCA). Med tanke på dessa parametrar, den modell som presenteras nära efterliknar sjukdom etiologi av mänskliga stroke och gör resultat som erhållits i högsta grad relevant 3. Ändå har översättningen av upptäckter från djurmodeller till behandling av sjukdomar hos människor visat sig vara en utmaning. Hittills har endast användningen av trombolytisk vävnadsplasminogenaktivator godkänts för behandling av akut ischemisk stroke 4. Bland modeller av fokal cerebral ischemi i mus, bakre cerebral cirkulation stroke modell och cerebral venös trombos modell är mycket invasiva, vilket minskar deras användbarhet och begränsa utbudet av analyser som kan utföras. Emellertid kan andra tekniker, såsom den embolisk modellen, photothrombosis modell, endotelin-1 inducerad stroke modell och intraluminal sutur arteria cerebri media ocklusion (MCAO) modell, är tillgängliga för användning utan sådana begränsningar. Den kortex modellen är en teknik som beskrivs i detta protokoll. Det erbjuder en tillförlitlig metod för att inducera fokal cerebral ischemi som lätt kan reperfusion och utförs i en hög genomströmning sätt. Det finns två metoder för denna modell, nämligen Zea-Longa och Koizumi metoder. De skiljer sig något i det sätt på vilket ocklusion suturen är insatt i kärlsystemet. I Zea-Longa teknik är suturen införas via den externa karotisartären 5. Tekniken som presenteras här är modifierad från den Koizumi metod i vilken den ockluderande suturen är insatt via den gemensamma halsartären 6. Den kortex modellen har framgångsrikt tillämpats för att utvärdera olika händelser som inträffar under ischemisk stroke. Efter reperfusion, kan observeras hjärnödem tillsammans med nedbrytning av blod-hjärnbarriären. Peak nervcellsdöd brukar observeras vid 24 h; Men, åter detvänder sig till baslinjenivåer efter 7 dagar 7. Hos människor, kön och ålder är viktiga variabler vid fastställandet stroke resultatet är detta också observerats hos möss och råttor 8, 9, 10. Flera publikationer har använt MCAO modell för att visa behandlingseffektiviteten 11, 12, 13, 14.

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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">MCAO suture 0.23mm</td>
			<td style="width:149px;">Doccol</td>
			<td style="width:119px;">702345PK5Re</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">MCAO suture 0.21mm</td>
			<td style="width:149px;">Doccol</td>
			<td style="width:119px;">702145PK5Re</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Silver pen</td>
			<td style="width:149px;">staples</td>
			<td style="width:119px;">503205</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Anesthesia machine</td>
			<td style="width:149px;">Vetequip</td>
			<td style="width:119px;">901806</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Surgical scissors</td>
			<td style="width:149px;">Fine science tool</td>
			<td style="width:119px;">14558-09</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Surgical forceps straight tip</td>
			<td style="width:149px;">Fine science tool</td>
			<td style="width:119px;">00108-11</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Surgical forceps angled tip</td>
			<td style="width:149px;">Fine science tool</td>
			<td style="width:119px;">00109-11</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Spring scissors</td>
			<td style="width:149px;">Fine science tool</td>
			<td style="width:119px;">15000-08</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Nylon suture</td>
			<td style="width:149px;">Braintree scintific</td>
			<td style="width:119px;">SUT-S 104</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Closing suture</td>
			<td style="width:149px;">VWR</td>
			<td style="width:119px;">95057-036</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Isoflurane</td>
			<td style="width:149px;">Piramal</td>
			<td style="width:119px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">2,3,5-Triphenyltetrazolium chloride</td>
			<td style="width:149px;">FisherSci</td>
			<td style="width:119px;">50-121-8005</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Brain block</td>
			<td style="width:149px;">Braintree scintific</td>
			<td style="width:119px;">BS-A 5000C</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Cryostat blade</td>
			<td style="width:149px;">VWR</td>
			<td style="width:119px;">89202-606</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Optional:</td>
			<td style="width:149px;"></td>
			<td style="width:119px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Periflux Laser doppler system</td>
			<td style="width:149px;">Perimed</td>
			<td style="width:119px;">Periflux 5000</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:276px;">Monitoring unit</td>
			<td style="width:149px;">Perimed</td>
			<td style="width:119px;">PF 5010 - LDPM</td>
		</tr>
	</tbody>
</table>

induction of ischemic stroke and ischemia-reperfusion in mice using the middle artery occlusion technique and visualization of infarct area

Cerebrovascular disease is highly prevalent in the global population and encompasses several types of conditions, including stroke. To study the impact of stroke on tissue injury and to evaluate the effectiveness of therapeutic interventions, several experimental models in a variety of species were developed. They include complete global cerebral ischemia, incomplete global ischemia, focal cerebral ischemia, and multifocal cerebral ischemia. The model described in this protocol is based on the middle cerebral artery occlusion (MCAO) and is related to the focal ischemia category. This technique produces consistent focal ischemia in a strictly defined region of the hemisphere and is less invasive than other methods. The procedure described is performed on mice, given the availability of several genetic variants and the high number of tests standardized for mice to aid in the behavioral and neurodeficit evaluation.

Insättningsvägen för ocklusion suturen demonstreras i figur 1. Den kortex sutur ska dirigeras till ocklusion området, bifurcating i ICA. Framgångsrik ocklusion av MCA kommer att leda till vävnadsskada, synliga genom TTC färgning. Figur 2 presenterar bilder av färgning från sham behandlat djur (figur 2A) och från en 60 min MCAO ischemireperfusion djur (färgning vid 90 min eller 24 h efter tilltäppning, figur 2B).</stro...

Watch this Scientific Journal Video about Induction of Ischemic Stroke and Ischemia-reperfusion in Mice Using the Middle Artery Occlusion Technique and Visualization of Infarct Area at JoVE.com

Induction of Ischemic Stroke and Ischemia-reperfusion in Mice Using the Middle Artery Occlusion Technique and Visualization of Infarct Area

We describe a mouse model of stroke induced by the occlusion of the middle cerebral artery using a silicone coated suture. The protocol can be applied to induce permanent occlusion or a temporary ischemia, followed by reperfusion.

Induktion av ischemisk stroke och ischemi-reperfusion i möss med användning av USA artärocklusion Teknik och visualisering av Infarkt Area

Cerebrovascular disease is highly prevalent in the global population and encompasses several types of conditions, including stroke. To study the impact of ...

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

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Musmodell av Middle Cerebral artärocklusion

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

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

Intraluminal Middle Cerebral artärocklusion (kortex) Modell för ischemisk stroke med Laser Doppler Flowmetry Vägledning hos möss

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

Bilateralt gemensamma halspulsådern Ocklusion som en fullgod Konditioneringscykel Stimulus att framkalla tidig ischemisk tolerans till Focal cerebral ischemi

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

Endotelin-1 inducerad Middle cerebral artärocklusion modell för ischemisk stroke med laser Doppler Flowmetry Vägledning i råtta

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

Använda en Kranial fönster för att visualisera den mellersta cerebrala artären under Endotelin-1 inducerad Middle cerebral artärocklusion

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

Optimerat system för cerebral perfusion övervakning i råtta Stroke Modell av Intraluminal Middle cerebral artärocklusion

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

Modellering stroke hos möss: Permanent Koagulation av den distala 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 ...

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

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

Emboli Middle Cerebral artärocklusion (kortex) för ischemisk stroke med Homologa Blodproppar i råttor

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

Quantification of Neurovascular Protection Following Repetitive Hypoxic Preconditioning and Transient Middle Cerebral Artery Occlusion in Mice

Kvantifiering av neurovaskulär skydd Efter Repetitiva Hypoxisk Förkonditionering och Transient Middle Cerebral artärocklusion hos möss

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week ...