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

倫理に関する声明：オスの成体Sprague-Dawleyラット（330〜380グラムの重さ）が、このプロトコルで使用されていた。このプロトコルは、LSUの健康科学センター、シュリーブポートで施設内動物管理使用委員会（IACUC）によって承認され、（第8版、全米科学アカデミー「実験動物の管理と使用に関する指針」に準拠している、2011た）。 変性PE-50チューブの調製<ol><li>徐々チューブを柔らかく、手でガス火の上に長さ30cm、PE-50チューブを握り。静かにチューブを伸ばす。 </li><li>デジタルノギス（ 図1）を使用して、延伸管上のポイントを選択し、それをマークし、長さ1cmの先端部（外径：0.30〜0.34ミリメートル）で長さ25cmのセグメントに修正されたPE-50チューブをカット。 </li></ol>同種血の塊の調製<ol><li> 70％N 2 Oおよび30％O中のイソフルランでラット（誘導のための5％、維持2-3％）で麻酔&lt;採血前フェイスマスクによるサブ&gt; 2。つま先のピンチによって麻酔を確認してください。 </li><li>発表された方法4でドナーラットの大腿動脈カニュレーションを行い、20cm長のPE-50チューブに直接大腿動脈の血液を移す。血液を凝固さ、室温で2時間チューブを配置し、その後、4℃で22時間、チューブを保持している。注：ステップ3.1で説明したようにカニューレ挿入は無菌技術を用いて行われる。ドナーラットをさらにサンプリングのために回収する。 </li><li> 50ミリメートルのセグメントに、PE-50チューブをカットし、生理食塩水を含む滅菌ペトリ皿にチューブから血餅を洗い流す。 </li><li>長さ60mmのPE-10チューブに50ミリメートルに長い凝血塊を移し、23Gの針を用いて、通常の生理食塩水を含有する1ミリリットルの注射器に接続された20cm長のPE-50管に、PE-10チューブの両端を接続します（ 図2） 。 </li><li>から血液細胞を除去することができる5分間の他の1つのシリンジからの連続交互運動によって血餅をシフト血餅とフィブリンコアを破壊することなく、血管外凝固血餅の壊れやすいを最小限に抑えます。 </li><li> 40ミリメートル長いセグメントに血餅をカットし、無菌条件下でステップ1.2で説明したように修正されたPE-50カテーテル内に血塊セグメントを転送します。注：すべてのPEチューブはエチレンオキシドで滅菌する。 </li></ol> 3塞栓中大脳動脈閉塞（図3A、B） <ol><li> （15分の最低121℃、15 PSI）オートクレーブにより、すべての手術器具を滅菌する。 70％エタノールを用いて、手術テーブルと関連した手術用機器を消毒。 </li><li>ステップ2.1で説明したようにイソフルランでラットを麻酔。両後脚につま先のピンチを実行することによって、麻酔の深さをテストし、（足を引き抜く）のいずれか観察した運動は、動物が手術を行うのに十分に麻酔をかけないことを示します。麻酔下ながら乾燥を防ぐために、両眼に獣医軟膏の少量を適用します。腹側首と頭の再上の毛皮を剃る皮膚領域を露出させ、電気バリカンでgions。 </li><li>加熱パッド上に仰臥位でラットを置きます。直腸プローブ、コントロールを挿入し、恒温ブランケット制御ユニットを使用して36.5から37.5℃の間の体温を維持する。 </li><li> 70％エタノールに続いて、10％ポビドンヨードで剃毛した皮膚や周囲の毛を消毒。 </li><li>解剖顕微鏡下で首に2センチ長い正中切開を行います。手術野を露出し、右総頸動脈（CCA）、外頚動脈（ECA）、内頸動脈（ICA）、および神経および筋膜（ 図3A）の周囲から遊離翼口蓋動脈（PPA）を解剖するために使用するリトラクター。注：アシスタントが動物を準備するのに役立ちますしない場合は、手術の前に、滅菌手袋を変更してください。 </li><li> （迷走神経を傷つけることなく）、周囲の神経から無料でCCAを解剖し、動脈の下で無菌の5-0絹縫合糸を配置します。スリップノット（＃1）を接続し、小さなHEMOSを使用して縫合糸を把握tatおよび身体に向かって教えたプル。 </li><li> ECAとその2つの枝、後頭動脈（OA）と上甲状腺動脈（STA）を解剖。獣医電気外科ユニットを用いて二つの枝を凝固させる。 ECAの下で無菌の6-0絹縫合糸を2枚配置します。 </li><li>動脈の下に配置された二つのシルク、本体に向かってヘッド（先端）と他に向けて一枚を分離します。 （2＃）の頭に最も近い側に、小さな止血とシルクをつかみ、頭に向かって教えたプルタイトな合字を接続します。後で使用するために他の絹糸で緩い結び目（＃3）を準備します。 </li><li> （迷走神経を傷つけることなく）、周囲の神経からのICAとPPAを解剖。 6-0縫合糸（＃4）PPAを接続します。 ICA（＃5）の周りに6-0縫合糸で引き結びを作る。あるいはまた、微小血管クリップを使用してICAをクリップ。 </li><li> Vannasスタイルの春のはさみとの緊密な（＃2）ルーズ（＃3）合字間のECAに小さな穴（スルー1/2）カット。変性PE-50用浴槽を挿入CCAの分岐に切開して、事前に血液凝固を含む電子。 （＃3）だけで十分な内腔の周りに留置チューブの移動性を維持確保するために緩い結紮糸を締めます。 </li><li>切り株を解放し、ECAとICAの分岐下の切り株を配置するために小さな穴の部位にECAをカット。これは修正されたチューブが簡単にICAにスライドできるようになります。 ICAを開いて、静かに、カテーテルの先端がMCA（分岐から〜17ミリメートル）の原点に到達するまで、ICAにECAの内腔からチューブを進める。注：前の70％エタノールで滅菌した、ECA内管の外面をチューブの先端を挿入し、滅菌生理食塩水で洗浄した。 </li><li> 100μlのハミルトンシリンジ（ 図3B）を使用して、10秒以上の生理食塩水を10μlと一緒に修正されたPE-50カテーテルを通して血塊を注入。 </li><li> 5分後、ECAからカテーテルを撤回。 ECAを結ぶと、CCAを再度開く。首に切開を縫合。 </li></ol> 4。監視局所脳血流量（rCBFの） <ol><li> MCA閉塞の前に、頭蓋骨を露出させ、頭皮が1.2センチ正中切開を行います。歯科スクレーパーおよび滅菌綿スワップで頭蓋骨上の組織を削除します。注：手術の前に、露出した頭皮エリアと周囲の毛を70％エタノールに続いて、10％ポビドンヨードで消毒されています。 </li><li> 2mmの後部と0.7ミリメートルの球状のステンレス製バリ（ 図4）を使用して、ブレグマから5mm横に位置して1.5ミリメートルの直径のバーホールを開けます。 注：無傷の硬膜にしてください。 </li><li> 0.5ミリメートル硬膜表面の上にプローブを配置します。塞栓後の0（ベースライン）、5、15、30、60、90、および120分で脳血流量を監視し、5、15、30、45、およびtPAの静脈内投与後60分で続ける。脳血流の最後の測定後、頭皮切開を縫合によって閉じられている。注：首の切開の前に私たちは、MCAのocclus前ベースラインとして脳血流を測定イオン。ポストMCA閉塞私たちは首の切開の縫合閉鎖後のrCBFを測定します。したがって、頸部切開を無菌に保たれる。 </li></ol> 5。術後ケア<ol><li>脱水を防ぐ、すぐに手術や痛みの軽減のために必要に応じて、すべての6〜12時間後にBuprenex（0.05ミリグラム/ kgで、SC）を注入するために生理食塩水を皮下2.5mlの注入する。 </li><li>イソフルラン麻酔を停止します。 37℃獣医回収室（動物を暖かく保つ）にラットを置き、観察を続ける。ラットが麻酔から回復することは一般的に10分かかります。次に、滅菌したケージに動物を入れるケージにペトリ皿にいくつ濡れる食べ物を置き、動物滅菌室にケージを返す。 </li><li>脳卒中後24時間は、ペントバルビタールナトリウム（100 mg / kg体重、IP）の過剰投与によりラットを安楽死させる。 </li></ol> 6。神経学的欠損スコア<ol><li>塞栓後の2時間前とでBedersonスコアを実行します。グレーディングsを使用0-3のCALEは、先に説明した5：尾によってラットを上げ、0を、床に両方の前肢を拡張し、他の障害を示さない動物（通常の移動）; 1、尾によってラットを高め、対側前肢を曲げ; 2、旋回することなく、横方向のプッシュ（および前肢の屈曲）に対する耐性を減少した。 3、旋回とグレード2と同じ動作。 注：塞栓後の2時間後にBedersonスコア= 0（なし赤字）示さないラットは、さらなる研究から除外されます。 </li></ol>

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We do not have any potential conflicting interests to disclose.

本研究では、MCAの起源は、フィブリンに富む血餅により閉塞されたラットの塞栓MCAO脳卒中モデルを実行するための標準的な方法を、実証した。このモデルの主な利点は：フィブリンに富む血餅によるMCAのステムの閉塞は、ヒトにおける血栓塞栓性脳卒中に似て、塞栓性脳卒中モデルは、線溶療法の前臨床研究を行うのに適しており、このモデルが開発できることMCAによって供給された領域内...

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

脳卒中は米国における死因の第3位であるが、急性脳卒中治療の選択肢は限られて残っている。現在では、血餅を溶解する組み換え組織プラスミノーゲンアクチベーター（tPA）の静脈内注入は、急性虚血性脳卒中のための最も効率的な療法である。しかし、tPAの使用は、その狭い治療窓によって及び脳内出血のリスクの増大によって制限されます。このため、血栓溶解研究するのに適したストロークのモデルが緊急に必要とされている。 中大脳動脈（MCA）は、ほとんどの場合、ヒトでの行程に閉塞した動脈である。この動脈を中心に、虚血性脳卒中の多くの動物モデルが確立されている。現在では、MCAを閉塞することにより齧歯類局所虚血モデルの二つの主要な種類が開発されている：縫合MCAOモデルおよび塞栓MCAOモデル。管腔内縫合技術によってMCAOモデルが広く機構駆動のストロークの研究で使用されてきたが、これらの縫合糸のモデルはない人間のストロークの最大80％が血栓症または塞栓症によって引き起こされるように、ヒトの脳卒中を模倣する。しかし、血の塊を使用して、塞栓性脳卒中モデルは、密接に人間のストロークを模倣し、血栓溶解の研究に適していると考えられる。この塞栓モデルは、最初1992年にオーバーガードら。1によってラットにおいて開発され、さらにZhang らにより特徴付けられた1997年2およびディナポリらによって2006年3。塞栓MCAOは、ますます注目を集めているが、直面する技術的問題がある多くの研究室による。 本稿では、MCA領域内の予測可能な梗塞を開発することができ、成体ラットにおける同種血の塊と塞栓MCAOを製造するための標準的な方法を示しています。この記事で紹介した技術は研究者が脳卒中研究のためのこのモデルを確立するための技術的な問題を克服するのに役立つはずです。

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			<td style="width:135px;">RBMA-310C</td>
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			<td height="21" style="height:21px;width:232px;">Dissecting microscope</td>
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			<td height="23" style="height:23px;width:232px;">Veterinary electrosurgical unit</td>
			<td style="width:135px;">Equipment</td>
			<td style="width:237px;">MACAN Manufacturing Company</td>
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			<td height="20" style="height:20px;width:232px;">23G needle</td>
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			<td height="25" style="height:25px;width:232px;">PE-50 tubing</td>
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			<td style="width:237px;">Becton, Dickinson and Company</td>
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			<td height="20" style="height:20px;width:232px;">5-0 Silk suture</td>
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			<td>517607</td>
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embolic middle cerebral artery occlusion (mcao) for ischemic stroke with homologous blood clots in rats

組み換え組織プラスミノーゲンアクチベーター（tPA）を用いた臨床的には、血栓溶解療法は、急性虚血性脳卒中のための最も効果的な治療のままである。しかし、tPAの使用は、その狭い治療窓によって及び出血性変化のリスクの増大によって制限されます。虚血性脳卒中の治療のための新たな血栓溶解剤や戦略を研究するための適切な脳卒中モデルを開発する緊急の必要性がある。腔内縫合MCAO塞栓MCAO：現在、虚血性脳卒中モデルの2つの主要なタイプはラットおよびマウスにおいて開発されてきた。管腔内縫合技術によってMCAOモデルが広く機構駆動のストロークの研究で使用されてきたが、これらの縫合糸モデルは、臨床状況を模倣せず、血栓溶解の研究には適していないでください。これらのモデルの中では、塞栓MCAOモデルは、密接に人間の虚血性脳卒中を模倣し、血栓溶解療法の前臨床研究に適しています。この塞栓モデルは、最初にOVによるラットで開発されましたergaard らは 1992年の1、さらにZhang らによって特徴付け1997 2。塞栓MCAOがますます注目を集めているが、多くの研究室が直面する技術的な問題がある。血栓溶解研究のための増加のニーズを満たすために、MCA領域内の予測可能な梗塞体積を開発することができ、ラットにおける塞栓MCAOの再現性の高いモデルを提示する。簡単に述べると、修飾されたPE-50チューブを穏やかにMCAの起点に到達​​するカテーテルの先端まで内頸動脈（ICA）の内腔に外頸動脈（ECA）から前進する。カテーテルを介して、単一の相同血餅はMCAの起点に配置される。 MCA閉塞の成功を識別するために、局所脳血流量をモニターし、神経学的欠損および梗塞体積を測定した。本論文で示された技術は、研究者は、脳卒中の研究のためにこのモデルを確立するための技術的な問題を克服するのに役立つはずです。&lt;/ pの&gt;

レーザードップラーフローメトリー（LDF）は脳虚血6,7の誘導の間脳血流量をモニターした。私たちの研究室を含む多くの研究室が成功したMCA閉塞を有する動物を識別するために、局所脳血流を使用しているが、ベースラインのしきい値は、測定部位に関連している研究室間で変化した。以前に説明したように6 LDFのプローブは、2mmの後部とブレグマに対して横5mmと配置されて?...

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.

ラットにおける同種血血栓による虚血性脳卒中のための塞栓中大脳動脈閉塞（MCAO）

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 ビュー.  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. 

ビデオ: 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.

マウスにおけるモデリングのストローク - フィラメントモデルと中大脳動脈閉塞

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

医学

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.

中大脳動脈閉塞のマウスモデル

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.

マウスにおけるレーザードップラー流量計の指針と虚血性脳卒中に対する腔内中大脳動脈閉塞（MCAO）モデル

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.

局所脳虚血への早期虚血耐性を誘導するために適切な前処理刺激として二国間の総頚動脈閉塞

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.

ラットにおけるレーザードップラー血流計の指針と虚血性脳卒中のエンドセリン-1誘発性中大脳動脈閉塞モデル

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.

エンドセリン-1誘発性中大脳動脈閉塞中大脳動脈を可視化するために頭蓋窓を利用

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.

腔内中大脳動脈閉塞のラット脳卒中モデルにおける脳血流モニタリングのための最適化されたシステム

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.

マウスにおけるモデリングのストローク：遠位中大脳動脈の永久的な凝固

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.

持続的な障害と皮質虚血を研究するために高齢ラットにおける一般的な頸動脈閉塞と永久遠位中大脳の実行

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.