手術手技は、もともとはジョージア医科大学の博士ウィリアムD.ヒルの研究室に買収された。また、作者は解剖のカメラの使用のためにデビッドA. RempeとランダPriftiを感謝したいと思います。この研究はNIH NS041744、NS051279、F31 NS064700とAHA 30815697Dによってサポートされていました。

このプロトコルは、研究における動物の倫理的な使用（UCAR）に捧げられたロチェスター委員会の大学により承認された。無菌技術は、プロトコル中に従うべきである。滅菌手袋とマスクを使用する必要があります。 すべての機器、材料、化学物質、およびプロトコル中に使用されるツールは、表1で説明されています。 1。術前準備<ol><li>最初の24時間、術後の期間中のすべての3から5時間後、手術直後、手術前にブプレノルフィン（0.05mg/kg）2時間で皮下マウスを注入して。 </li><li>オートクレーブですべての手術器具、ガーゼスポンジ、そして綿の先端のアプリケーターを滅菌する。使用前に滅菌ガーゼの右上に手術して空気乾燥時に70％エタノールで手術器具を保管してください。 70％エタノールで作業領域を吹き付けます。 </li><li>麻酔気化器を使用して3％イソフルラン- 20％の酸素ガスの混合物で、マウスを麻酔。流量計で酸素の量を調整します。つま先や尾のピンチ（彼らが応答しなくなるはず）で麻酔のレベルをテストします。 2％（v / v）をイソフルランで麻酔のレベルを維持する。 </li><li>マウスの目に人工涙液を適用し、手術中に目を損傷しないように注意してください。側臥位で、その右側にマウスを置きます。 </li><li>左眼と動物のバリカンを使用して、左耳の付け根の間の左側の領域を剃る。 betadine液と70％エタノール綿の先端のアプリケーターを使用し、軽く面積をこすりで交互にエリアを清め。必要に応じて繰り返します。 </li><li> 37℃で体温を維持するために直腸プローブに接続された加熱パッド℃の上にマウスを置きますミネラルオイルを使用して直腸プローブを挿入します。 </li><li>顕微鏡下で、その右側にマウスを置き、テープで固定します。ガーゼのスポンジでウィンドウを切り取り、外科領域をカバー。 </li></ol> 2。外科手術とMCAライゲーション<ol><li>左眼と素晴らしいストレートハサミを使用して、左耳の付け根の間に縦切開を加えます。外科領域を開いておくための湾曲した止血を使用してください。 </li><li>春のはさみを使って、側頭筋上に水平方向の切開を行い、少しゆっくりと鉗子で引っ張って頭蓋骨から側頭筋を分離。 </li><li>湾曲鉗子で顎骨を保持しながら18G針を用いて頬骨弓と側頭鱗骨の接合部に小さな側頭下開頭術を行います。 </li><li>骨Rongeursと頭蓋骨の小さ​​な断片を除去することによりMCAを公開。頬骨弓と眼窩内容は、このプロセス中に破損してはならない。組織の過度の乾燥は、このプロセス中に発生した場合は、綿の先端のアプリケーターで滅菌PBSを適用します。 </li><li>小血管cauterizerを使用してMCAの遠位部分をライゲーションする。 </li><li>元の位置に戻して側頭筋を置き、外科5から0ナイロン縫合糸で切開部位を閉じる。 </li><li>麻酔を中止し、直腸プローブを削除します。皮下ブプレノルフィン（0.05mg/kg）の2番目の用量でマウスに注入する。加熱パネルで37℃に保温したケージにマウスを返します。 </li><li>密接に不快感（食欲減退/水消費量、猫背の姿勢、増加した呼吸、髪をピーロは、建てられた）のための次の24時間マウスを監視する。最高24時間までブプレノルフィン皮下すべての3から5時間後に手術とマウスを注入する。この期間中に回復ゲルでマウスを提供しています。 </li></ol> 3。 TTC染色と一回拍出量の測定<ol><li>手術24時間後には、深く、マウスを麻酔transcardially 15分間、その後10分間、4％パラホルムアルデヒド溶液で使用することによりリン酸塩のTTC（w / v）の緩衝生理食塩水4％（PBS）でマウスを灌流培地流量でミニ蠕動ポンプ。脳を取り出して、一晩4％パラホルムアルデヒド溶液に入れてください。 </li><li>セクショニングのブロックに脳を置きます。かみそりの刃を使用して1mm厚のスライスに脳をスライスして。 </li><li>ミリメートル規模の定規の横のスライスを置きます。解剖顕微鏡に接続したデジタルカメラを使用してスライスを撮影する。 </li><li>画像Jソフトウェア（http://rsbweb.nih.gov/ij/）を使用して対側のサイトの総面積から同側のサイトの非梗塞領域を差し引くことにより、ストロークの領域を計算します。スライス22のストロークの領域を積み重ねることによって一回拍出量を計算します。 </li></ol> 4。画像Jソフトウェアを使用する方向<ol><li>ファイルメニューをクリックして画像Jソフトウェアで分析する画像ファイルを開きます。 </li><li>プログラムで&quot;直線&quot;タブをクリックします。ルーラー上の2つのマージンの間に直線を描画します。 &quot;分析&quot;メニューをクリックし、&quot;セットの規模&quot;を選択します。 1と既知の距離、mmの長さの単位を設定します。 </li><li> &quot;フリーハンドサークル&quot;タブをクリックします。対側半球の輪郭を円を描く。 &quot;分析&quot;メニューをクリックし、&quot;尺度&quot;を選択します。計算画面では、描画した円の面積を示すポップアップ表示されます。 </li><li>同側半球を除くストローク（白）周辺に別の円を描きます。として、ステップ4.3で示された領域を測定します。新しい値が計算ウィンドウに追加されます。 1番目と2番目の値の差は、以下の数式と同様に示される梗塞の領域を表します。 </li><li>ステップ4.2から4.4までを繰り返すことによって、各スライスのストロークの領域を計算します。各スライス（ΣAn）で計算されたストローク領域の総和を取る。これは、各スライスの厚さは1mmであるという事実を知って一回拍出量を表しています。 ΣAn個の x 1mmの（各スライスの厚さ）=ストローク量</li></ol> 5。代表的な結果： マウスで恒久的なMCAの結紮によって得られた梗塞は主に皮質です。しかし、MCAは、レンズ核線条体枝の近位に連結している場合皮質下病変を得ることができる。 MCA結紮後のストローク量が10mm ³から35mm ³ 19、23に変わる可能性があります。 MoyanovaらのTTC染色で計算されたストロークボリューム。Filianoら23のMRI画像から決定ストローク量が35mm ³に20ミリメートル³の間にある間、19は 22ミリメートル³に10ミリメートル³の間です。多分ライゲーションの正確な位置やストローク量を測定するために使用される様々な方法でこれらの違いの可能な理由。 図1では、野生型C57/BL6マウスにおけるTTC染色した脳の24時間後に永久的なMCAの結紮が表示されます。脳卒中サイトは外観（図1A、B）に白です。図1AおよびBには、さまざまな角度から梗塞部位を示しています。図2は、後に前方からTTC染色した脳卒中脳の1mm厚のスライス（図2A - G）を示しています。梗塞体積は、24時間後に手術を算出した。梗塞の体積は、〜23ミリメートル³です。 <img src="/files/ftp_upload/3039/3039fig1.jpg" alt="図1"/> 図1。脳卒中サイトの表現。AB、TTC ​​は、MCAの結紮後の全脳の画像、24時間を染色した。白い部分は梗塞を表します。 <img src="/files/ftp_upload/3039/3039fig2.jpg" alt="図2"/> 図2。代表的な結果。（&gt; G）、TTC染色した1mmの脳切片、MCAL 24時間後。スライスは、後に前から整列されます。白い部分は梗塞を表します。

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利害の衝突は宣言されません。

永久的なMCAのライゲーション法は再現性の高い梗塞量と利用可能な他の方法に比べて増加した術後の生存率を与える。手続きの容易さと、短い期間（〜30分）は、さらに実践的なもの。法が広く、マウスとラットの両方で使用されています。 この手法は、実体顕微鏡下低侵襲手術が必要になります。そのため、顕微鏡下で動作し、成功した開頭術を完璧に経験が不可欠で...

<table border="1"><tbody><tr><th> NAME </th><th>会社とカタログ番号</th></tr><tr><td> A.ソリューションと化学物質 </td><td></td></tr><tr><td>ポビドンヨード溶液、手術スクラブ</td><td> Aplicare、82から209 </td></tr><tr><td> 70％エタノール</td><td> Koptec、V1001 </td></tr><tr><td> 2,3,5塩化トリフェニルテトラゾリウム（TTC） </td><td>シグマ、T8877 </td></tr><tr><td> 4％パラホルムアルデヒド</td><td>電子顕微鏡の科学、159SP </td></tr><tr><td> 7.4：リン酸緩衝生理食塩水、pHをバッファ</td><td>行い、研究室で滅菌。 </td></tr><tr><td>鉱油</td><td> PML Microbiologicals、R6570 </td></tr><tr><td>イソフルラン</td><td>バトラーカンパニー、029405 </td></tr><tr><td>ブプレノルフィン（0.3mg/ml） </td><td>ホスピーラ、NDC 0409-2012-32 </td></tr><tr><td>人工涙液</td><td>バトラーカンパニー、007312 </td></tr><tr><td>リカバリージェル</td><td>明確なH 2 O、72-01-5022 </td></tr><tr><td> B.手術材料と機器 </td><td></td></tr><tr><td> 10〜12週古いC57/BL6マウス、 </td><td>コロニーは、大学のビバリウムで飼育された</td></tr><tr><td> 100％の酸素タンク</td><td> Airgasの</td></tr><tr><td>麻酔気化器と流量計</td><td> Surgivet、モデル100 </td></tr><tr><td>直腸プローブによる加熱パッド</td><td>ファイン科学ツール、TR200 </td></tr><tr><td>解剖顕微鏡</td><td>ツァイス、STEMI 2000 </td></tr><tr><td>暖房パネル</td><td>ペトコサービス株式会社0307-013 </td></tr><tr><td>小血管Cauterizerセット</td><td>ファイン科学ツール、18000〜00 </td></tr><tr><td>ミニ蠕動ポンプ</td><td>ハーバード装置、MPII </td></tr><tr><td> 1ミリリットルの注射器</td><td> BD、329650 </td></tr><tr><td> 18Gの針</td><td>ベクトンディッキンソン、305196 </td></tr><tr><td>外科5.0ナイロン縫合糸</td><td> Ethilon、698H </td></tr><tr><td>ブロックをセクショニング</td><td>ケントサイエンティフィック、1ミリメートル</td></tr><tr><td>単一のエッジかみそりの刃</td><td>電子顕微鏡の科学、71962 </td></tr><tr><td>ラベルテープ</td><td> Scienceware、ベルアート製品F13463 - 2075 </td></tr><tr><td>ガーゼスポンジ</td><td>ケンドールキュリティ2252、ジョンソンアンドジョンソン6415 </td></tr><tr><td>綿のヒントアプリケーター</td><td>ピューリタン806 - WCL </td></tr><tr><td> C.手術ツール </td><td></td></tr><tr><td>外科春のはさみ</td><td>ファイン科学ツール15003〜08 </td></tr><tr><td> Microdissecting鉗子は、角度</td><td>ファイン科学のツール45分の5 </td></tr><tr><td> Microdissecting鉗子の湾曲</td><td>ファイン科学ツール5のTi </td></tr><tr><td>ファインストレートはさみ</td><td>ファイン科学ツール14568〜12 </td></tr><tr><td>湾曲した骨の骨鉗子</td><td>ファイン科学ツール16021〜14 </td></tr><tr><td>ハートマンの止血剤は、湾曲した</td><td>ファイン科学ツール13003〜10 </td></tr></tbody></table> 表1。MCAのライゲーションの手続きのために使用される材料、ツール、および化学物質のリスト。 A.ソリューションと化学、B.手術材料と機器、C.外科ツール。

the application of permanent middle cerebral artery ligation in the mouse

局所脳虚血は、患者に見られる脳卒中の最も一般的なタイプのひとつです。臨床的意義による虚血性傷害の間に展開するイベントを研究するために適切な動物モデルを開発するための長期の努力がなされている。これらの技術は、最も一般的なげっ歯類で、多くの異なる動物モデルを用いて一時的または永続的な、局所またはグローバル虚血モデルが含まれています。 また、文献で​​pMCAoと呼ばれる永久的なMCAのライゲーション法は、げっ歯類1-6の局所的虚血モデルとして広く使用されています。このメソッドは当​​初、田村らによりラットに記述されていた。 1981年7インチこのプロトコルでは開頭はMCAにアクセスするために使用され、近位領域は、電気凝固法によって閉塞させた。梗塞は閉塞の位置に応じて、主に皮質と時々線条体領域を含む。この手法は、今や十分に確立し、多くの研究室8-13に使用されています。この技術の早期使用は、&quot;梗塞コア&quot;と&quot;半影&quot;14から16の定義と記述につながった、とそれはしばしば潜在的な神経保護化合物10、12、13、17を評価するために使用されます。初期の研究はラットで行われたが、恒久的なMCAの結紮は、若干の修正18から20を有するマウスで正常に使用されています。 このモデルは、再現性梗塞と増加後の生存率が得られます。ヒトの虚血性脳卒中の約80％は、MCA領域21で発生するので、これは、脳卒中の研究のために非常に関連したモデルです。現在、脳卒中患者が利用できる効果的な治療の不足があるため、潜在的な薬理学的化合物をテストし、生理学的アウトカムを評価する良いモデルが必要である。このメソッドは、 生体内でも細胞内の低酸素応答のメカニズムを研究するために使用することができます。 ここで、我々は、C57/BL6マウスにおけるMCAの結紮術を紹介。我々は術前準備、MCAの結紮術および梗塞体積の定量化のための2,3,5塩化トリフェニルテトラゾリウム（TTC）染色を説明します。

Watch this Scientific Journal Video about The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse at JoVE.com

The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

中大脳動脈（MCA）ライゲーションは、動物モデルでの局所脳虚血を研究する技術です。この方法では、中大脳動脈は開頭術によって公開されており、焼灼によって連結した。このメソッドは、非常に再現性の梗塞量と利用可能な他の方法に比べて増加した術後の生存率を与える。

マウスの永久中大脳動脈結紮の応用

Focal cerebral ischemia is among the most common type of stroke seen in patients. Due to the clinical significance there has been a prolonged effort to ...

the-application-permanent-middle-cerebral-artery-ligation

Research

JoVE Journal

Medicine

19.5K ビュー.  University of Rochester. 中大脳動脈（MCA）ライゲーションは、動物モデルでの局所脳虚血を研究する技術です。この方法では、中大脳動脈は開頭術によって公開されており、焼灼によって連結した。このメソッドは、非常に再現性の梗塞量と利用可能な他の方法に比べて増加した術後の生存率を与える。

ビデオ: The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

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 ...

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 ...

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow (ischemia) 1. Middle cerebral artery (MCA) delivers a great majority of blood to the lateral surface of the cortex 2, is the most common site of human stroke 3, and ischemia within its territory can result in extensive dysfunction or death 1,4,5. Survivors of ischemic stroke often suffer loss or disruption of motor capabilities, sensory deficits, and infarct. In an effort to capture these key characteristics of stroke, and thereby develop effective treatment, a great deal of emphasis is placed upon animal models of ischemia in MCA.
Here we present a method of permanently occluding a cortical surface blood vessel. We will present this method using an example of a relevant vessel occlusion that models the most common type, location, and outcome of human stroke, permanent middle cerebral artery occlusion (pMCAO). In this model, we surgically expose MCA in the adult rat and subsequently occlude via double ligature and transection of the vessel. This pMCAO blocks the proximal cortical branch of MCA, causing ischemia in all of MCA cortical territory, a large portion of the cortex. This method of occlusion can also be used to occlude more distal portions of cortical vessels in order to achieve more focal ischemia targeting a smaller region of cortex. The primary disadvantages of pMCAO are that the surgical procedure is somewhat invasive as a small craniotomy is required to access MCA, though this results in minimal tissue damage. The primary advantages of this model, however, are: the site of occlusion is well defined, the degree of blood flow reduction is consistent, functional and neurological impairment occurs rapidly, infarct size is consistent, and the high rate of survival allows for long-term chronic assessment.

Permanent Cerebral Vessel Occlusion via Double Ligature and Transection

We describe a highly reproducible method for the permanent occlusion of a rodent major cerebral blood vessel. This technique can be accomplished with very little peripheral damage, minimal blood loss, a high rate of long-term survival, and consistent infarct volume commensurate with the human clinical population.

永久的な脳血管閉塞<em&gt;経由</em&gt;ダブルリガチャーと離断

Stroke is a leading cause of death, disability, and socioeconomic loss worldwide. The majority of all strokes result from an interruption in blood flow ...

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 ...

Permanent Ligation of the Left Anterior Descending Coronary Artery in Mice: A Model of Post-myocardial Infarction Remodelling and Heart Failure

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It is characterized by fluid retention, shortness of breath, and fatigue, in particular on exertion. Heart failure is a growing public health problem, the leading cause of hospitalization, and a major cause of mortality. Ischemic heart disease is the main cause of heart failure.
Ventricular remodelling refers to changes in structure, size, and shape of the left ventricle. This architectural remodelling of the left ventricle is induced by injury (e.g., myocardial infarction), by pressure overload (e.g., systemic arterial hypertension or aortic stenosis), or by volume overload. Since ventricular remodelling affects wall stress, it has a profound impact on cardiac function and on the development of heart failure. A model of permanent ligation of the left anterior descending coronary artery in mice is used to investigate ventricular remodelling and cardiac function post-myocardial infarction. This model is fundamentally different in terms of objectives and pathophysiological relevance compared to the model of transient ligation of the left anterior descending coronary artery. In this latter model of ischemia/reperfusion injury, the initial extent of the infarct may be modulated by factors that affect myocardial salvage following reperfusion. In contrast, the infarct area at 24 hr after permanent ligation of the left anterior descending coronary artery is fixed. Cardiac function in this model will be affected by 1) the process of infarct expansion, infarct healing, and scar formation; and 2) the concomitant development of left ventricular dilatation, cardiac hypertrophy, and ventricular remodelling.
Besides the model of permanent ligation of the left anterior descending coronary artery, the technique of invasive hemodynamic measurements in mice is presented in detail.

Heart failure is the leading cause of hospitalization and a major cause of mortality. A model of permanent ligation of the left anterior descending coronary artery in mice is applied to investigate ventricular remodelling and cardiac dysfunction post-myocardial infarction. The technique of invasive hemodynamic measurements in mice is presented.

マウスの左前下冠動脈の動脈の常設ライゲーション：心筋梗塞後のリモデリングおよび心不全のモデル

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It ...

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, ...

Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature and mature brain. Stroke occurs commonly in the perinatal period in humans, and transient ischemia-reperfusion is the most common form of stroke in neonates. The reperfusion phase is a critical component of injury progression, which occurs over a period of days to weeks, and of the endogenous response to injury. This postnatal day 10 (p10) rat model of transient middle cerebral artery occlusion (tMCAO) creates a unilateral, non-hemorrhagic focal ischemia-reperfusion injury that can be utilized to study the mechanisms of focal injury and repair in the full-term-equivalent brain. The injury pattern that is produced by tMCAO is consistent and highly reproducible and can be confirmed with MRI or histological analyses. The severity of injury can be manipulated through changes in occlusion time and other methods that will be discussed.

Neonatal stroke is a significant cause of early brain injury requiring a translational model with consistent focal injury patterns and high reproducibility in order to enable study. This study describes the detailed surgical procedure for creating a non-hemorrhagic, unilateral focal ischemia-reperfusion injury in full-term-equivalent rodents.

P10ラットにおける新生児脳卒中の一過性中大脳動脈閉塞モデル

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature ...

Middle Cerebral Artery Occlusion Allowing Reperfusion via Common Carotid Artery Repair in Mice

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue plasminogen activator (tPA) as an approved drug treatment to target ischemic strokes. Current research in the field of ischemic stroke focuses on better understanding the pathophysiology of stroke, to develop and investigate novel pharmaceutical targets. Reliable experimental stroke models are crucial for the progression of potential treatments. The middle cerebral artery occlusion (MCAO) model is clinically relevant and the most frequently used surgical model of ischemic stroke in rodents. However, the outcomes of this model, such as lesion volume, are associated with high levels of variability, particularly in mice. The alternative MCAO model described here allows the reperfusion of the common carotid artery (CCA) and the increased perfusion of the middle cerebral artery (MCA) territory, using a tissue pad with fibrinogen-based sealant to repair the vessel, and the improved welfare of the mice by avoiding external carotid artery (ECA) ligation. This reduces the reliance on the Circle of Willis, which is known to be highly anatomically variable in mice. Representative data show that using this alternative surgical approach decreases the variability in lesion volumes between the traditional MCAO approach and the alternative approach described here.

Intraluminal filament occlusion of the middle cerebral artery is the most frequently used in vivo model of experimental stroke in rodents. An alternative surgical approach to allow common carotid artery repair is performed here, which allows the reperfusion of the common carotid artery and a full reperfusion to the middle cerebral artery territory.

マウスにおける一般的な頸動脈の修理で再灌流を許可する中大脳動脈閉塞

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue ...