この作品は、NIHの助成金NS057195、UCSF REAC賞、そしてケント州立大学のスタートアップ資金WHチョウにによってサポートされていました。

<html> 1。 MCAO方法このプロトコルは、実験動物の使用のための健康ガイドラインの国立研究所で動物実験によって承認され、UCSFとケント州立大学で委員会を使用して、遵守された。 <ol><li> 20mmのセグメントに5から0モノフィラメント縫合糸（ハーバード装置、ホリストン、MA）をカット。 cauterizer（ブレーントリーサイエンティフィック社、ブレーンツリー、MA）の近くに加熱することにより、各セグメントの先端を丸める。マイクロメートル（応用イメージ社、ロチェスター、NY）を用いて先端の直径を測定する。我々は25-30 gの体重を持つマウスのために0.21〜0.22ミリメートルの最後の先端径との縫合糸を使用してください</li><li>オートクレーブ（最小121 ° C、15 PSI、15分間）により、すべての手術器具を滅菌する。 70％エタノールを使用して手術のテーブルおよび関連機器をサニタイズ。 </li><li> V - 10麻酔システムを使用して、OはO 2 / 70％N 2 30％のイソフルラン5％8月12日週齢のマウス（25〜30グラム）（Aerrane、バクスター、ディアフィールド、イリノイ州）の麻酔（VetEquip株式会社。、プレザントン、カリフォルニア州）。麻酔の誘導後、イソフルランのレベルを低下させ、1.5％で、それを維持する。 </li><li>加熱パッド上に仰臥位にマウスを置きます。直腸プローブを挿入し、℃、TR - 200恒温温度システム（ファイン科学ツール（株）、フォスターシティー、CA）を使用して36.5から37.5の間に体温を監視し、維持する。 </li><li>肌を露出するために電気バリカン（ブレーントリーの科学）と頸部腹側の領域の上に毛皮を剃る。 70％エタノールつのアプリケーションを使用して手術部位を消毒する。 </li><li>ステレオ解剖顕微鏡下で（ニコン、日本）、首に1センチの正中切開を行います。手術野を公開し、右総頸動脈（CCA）、外頸動脈（ECA）、および内頸動脈（ICA）を識別するために、リトラクター（ブレーントリーサイエンティフィック）を使用してください。慎重に神経や筋膜の周囲から無料の動脈を詳細に分析します。 </li><li>さらに遠位にECAを摘出し、バイポーラ凝固剤（ハワードインストゥルメント社、アラバマ州タスカルーサ）を使用してECAとその上甲状腺動脈（STA）分岐を凝固。凝固したセグメントでECAとSTAをカット。 </li><li>大まかにECAの切り株2箇所を8から0絹縫合糸を結ぶ。 ECAとICAにCCAの分岐部に血管クランプ（ファイン科学のツール）を適用します。 </li><li> Vannas -スタイル春のはさみ（ファイン科学のツール）とECA切り株の端の小さな切開を加えます。先端に丸みを帯びた5から0モノフィラメント縫合糸の長さを測定し記録する。切開に縫合を挿入し、クランプに進みます。だけで十分な確保はまだで、住居のモノフィラメント縫合糸の機動性を維持するために内腔2箇所を絹の縫合糸を締めます。 </li><li>分岐からクランプを外します。ゆっくりとMCAの起源を閉塞するためにCCAの分岐を超えて9〜10ミリメートルの距離にICAにECAの内腔からモノフィラメント縫合糸を進める。手術時間は約30-45分です。 </li><li>首に切開を縫合し、麻酔からの回復、そしてケージに戻すために35 ° C看護のボックスにマウスを置きます。それは一般的に麻酔から回復するためにマウスの5〜10分かかります。過渡MCAOを実行するには、研究者は、通常の間に0.5〜2時間をマウスを再度麻酔し、一定期間後にECAの切り株に戻って縫合を撤回することができます</li><li> MCAOの誘導24時間後には、イソフルラン5％、マウスを麻酔し、頚椎脱臼によってそれを安楽死させる。マウスの首を切ると脳を収集する。氷の上で脳の行列（ブレーントリーサイエンティフィック）を持つ4 2mmのスライスにcoronally脳をスライスして。梗塞の大きさと範囲を決定するために室温で20分間、1 × PBS中の2％2,3,5 - トリフェニル塩化（TTC）（Sigma - Aldrich社）での脳切片をインキュベートします。イメージングまで中立を4でホルマリン溶液を（Sigma - Aldrich）をバッファリング10％の脳のスライス° Cを修正。 Joveのプロトコル955（で説明されているように心筋梗塞の程度を定量化することができる<a href="http://www.jove.com/index/Details.stp?ID=955">http://www.jove.com/index/Details.stp?ID=955</a> ）9。 </li></ol> 2。代表的な結果 MCAOによって生成された梗塞は、線条体と背外側皮質に見られる。線条体は大脳皮質よりも虚血の影響を受けやすくなります。 MCAOの一時間以上は、線条体と皮質の両方を損傷する恐れながら、MCAOの30分は、唯一の線条体の梗塞が生成されます。 24時間永久MCAO後、合計梗塞の割合は、±半球の5％〜40であり、手術後の死亡率は約10％です。過度の出血が手術中に発生した場合、我々はさらに研究からマウスを除外する、操作時間が90分を超えると、マウスは15分以内に麻酔からの回復に失敗する、または出血は、脳のスライスにまたは剖検時にウィリス動脈輪の根元に発見され。 61/2761fig1.jpg&quot;altが=&quot;図1&quot;/&gt; 永久MCAOの24時間後のTTC染色した脳切片の図1。代表画像（コロナルレベル1-4）。壊死領域でそのような酵素活性の欠如に起因する白のまま組織TTCを生体内で酵素的に色が赤になっている1,3,5 - triphenylformazan（TPF）、、にデヒドロゲナーゼによって低減されます。そのため、梗塞の領域はTPFへのTTCの変換が不足しているため、その白い色で識別することができます。注：これは可能な限り熱と光から染色した切片を保護TTCが不安定にやや熱と光です。

<ol>
	<li>Howard, G., Howard, V. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Distribution of stroke: hetrogeneity of stroke by age, and sex. , 3-12 (2004).</li><li>Thom, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Heart+disease+and+stroke+statistics--2006+update:+a+report+from+the+American+Heart+Association+Statistics+Committee+and+Stroke+Statistics+Subcommittee.">Heart disease and stroke statistics--2006 update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee.</a> Circulation. 113, e85-e151 (2006).</li><li>Mohr, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Middle cerebral artery disease. , 123-166 (2004).</li><li>Longa, E. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reversible+middle+cerebral+artery+occlusion+without+craniectomy+in+rats.">Reversible middle cerebral artery occlusion without craniectomy in rats.</a> Stroke. 20, 84-91 (1989).</li><li>Chou, W. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neutrophil+protein+kinase+Cdelta+as+a+mediator+of+stroke-reperfusion+injury.">Neutrophil protein kinase Cdelta as a mediator of stroke-reperfusion injury.</a> J. Clin. Invest. 114, 49-56 (2004).</li><li>Hudgins, W. R., Garcia, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+electrocautery,+atmospheric+exposure,+and+surgical+retraction+on+the+permeability+of+the+blood-brain-barrier.">The effect of electrocautery, atmospheric exposure, and surgical retraction on the permeability of the blood-brain-barrier.</a> Stroke. 1, 375-380 (1970).</li><li>Carmichael, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+of+focal+stroke:+size,+mechanism,+and+purpose.">Rodent models of focal stroke: size, mechanism, and purpose.</a> NeuroRx. 2, 396-409 (2005).</li><li>Durukan, A., Tatlisumak, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+ischemic+stroke:+overview+of+major+experimental+rodent+models,+pathophysiology,+and+therapy+of+focal+cerebral+ischemia.">Acute ischemic stroke: overview of major experimental rodent models, pathophysiology, and therapy of focal cerebral ischemia.</a> Pharmacol. Biochem. Behav. 87, 179-197 (2007).</li><li>Taniguchi, H., Andreasson, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hypoxic-ischemic+encephalopathy+model+of+perinatal+ischemia.">The hypoxic-ischemic encephalopathy model of perinatal ischemia.</a> J Vis Exp. , (2008).</li></ol>

我々は、開示する潜在的な利害対立を持っていない。

マウスのMCAOは一般にヒトで局所性脳虚血をモデル化するために使用されます。脳卒中の研究のためのマウスの使用は、トランスジェニックやノックアウト株の可用性をより頻繁になっています。プロトコルで注意するいくつかの重要な詳細があります。 <ol><li>それは手術中にマウスの体温を維持するために不可欠であり、それは完全に麻酔から回復する前に。体温は、梗塞の程度に影響があり、低体温が低下し、温熱療法は、梗塞のサイズが大きくなります。 </li><li>公開し、CCAとECAを隔離し、梗塞サイズと減少生存率を増加させることができる近くの迷走神経や気管を、損傷を避ける。 </li><li> 10mm以上の分岐を通過モノフィラメント縫合糸を挿入しないでください。遠く挿入縫合糸は、脳内出血で前大脳動脈とその結果を穿孔することができます。縫合糸は、分岐点を超えて9〜10ミリメートル程度進んでいるときに抵抗が感じられる。この問題が発生した場合、研究者は、縫合糸を進める停止し、距離を確認する必要があります。 </li></ol>

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<td>Isoflurane</td>
<td>Chemical	</td>
<td>Aerrane, Baxter</td>
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mouse model of middle cerebral artery occlusion

脳卒中は米国1の最も一般的な致命的な神経疾患です。脳の血管の閉塞（虚血性脳卒中）2からのストロークの大多数（88％）の結果。ほとんどの虚血性脳卒中（〜80％）中大脳動脈（MCA）の3地域で行われるため、開発されている多くの動物ストロークモデルは、この動脈に焦点を当てている。中大脳動脈閉塞（MCAO）の管腔内モノフィラメントモデルは、外頸動脈に外科的にフィラメントの挿入と先端がMCAの起源を閉塞するまで停止の結果、内頸動脈（ICA）に前方にスレッドを含む血流とMCA領域4のその後の脳梗塞の。手法は永続的または一時的な閉塞5をモデル化するために使用することができます。縫合糸は、一定のインターバル後に削除されている場合（30分、1時間、または2時間）、再灌流が（一時的なMCAO）が達成され、フィラメントはその場に放置されている場合（24時間）の手順では、恒久的なMCAOのモデルとして適しています。 。このテクニックはクレイニエクトミ、頭蓋内圧と温度6に影響を与える可能性のある頭蓋骨の一部を、削除するには、脳外科的処置を必要としません。それは、ラットやマウス7,8の永久および一時的な局所脳虚血を模倣するために最も頻繁に使用される方法となっている。脳梗塞の程度を評価するために、我々は、虚血脳組織9を識別するために、2,3,5 -塩化トリフェニルテトラゾリウム（TTC）での脳切片を染色。このビデオでは、我々はMCAO法およびTTC染色により梗塞サイズの測定を示しています。

Watch this Scientific Journal Video about Mouse Model of Middle Cerebral Artery Occlusion at JoVE.com

Mouse Model of Middle Cerebral Artery Occlusion

我々はビデオで腔内モノフィラメントを使用して成体マウスの中大脳動脈閉塞を製造する方法を示しています。我々はまた、2,3,5 - 塩化トリフェニルテトラゾリウム（TTC）染色による脳梗塞の程度を評価する方法を示します。

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 ...

mouse-model-of-middle-cerebral-artery-occlusion

Research

JoVE Journal

Medicine

77.3K ビュー.  Ernest Gallo Clinic and Research Center, University of California, San Francisco. 我々はビデオで腔内モノフィラメントを使用して成体マウスの中大脳動脈閉塞を製造する方法を示しています。我々はまた、2,3,5 - 塩化トリフェニルテトラゾリウム（TTC）染色による脳梗塞の程度を評価する方法を示します。

ビデオ: Mouse Model of Middle Cerebral Artery Occlusion

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

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 ...

The Application Of Permanent Middle Cerebral Artery Ligation in the Mouse

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 develop suitable animal models to study the events that unfold during ischemic insult. These techniques include transient or permanent, focal or global ischemia models using many different animal models, with the most common being rodents.
The permanent MCA ligation method which is also referred as pMCAo in the literature is used extensively as a focal ischemia model in rodents 1-6. This method was originally described for rats by Tamura et al. in 1981 7. In this protocol a craniotomy was used to access the MCA and the proximal regions were occluded by electrocoagulation. The infarcts involve mostly cortical and sometimes striatal regions depending on the location of the occlusion. This technique is now well established and used in many laboratories 8-13. Early use of this technique led to the definition and description of &#x201C;infarct core&#x201D; and &#x201C;penumbra&#x201D; 14-16, and it is often used to evaluate potential neuroprotective compounds 10, 12, 13, 17. Although the initial studies were performed in rats, permanent MCA ligation has been used successfully in mice with slight modifications 18-20 .
This model yields reproducible infarcts and increased post-survival rates. Approximately 80% of the ischemic strokes in humans happen in the MCA area 21 and thus this is a very relevant model for stroke studies. Currently, there is a paucity of effective treatments available to stroke patients, and thus there is a need for good models to test potential pharmacological compounds and evaluate physiological outcomes. This method can also be used for studying intracellular hypoxia response mechanisms in vivo. 
Here, we present the MCA ligation surgery in a C57/BL6 mouse. We describe the pre-surgical preparation, MCA ligation surgery and 2,3,5 Triphenyltetrazolium chloride (TTC) staining for quantification of infarct volumes.

Middle cerebral artery (MCA) ligation is a technique to study focal cerebral ischemia in animal models. In this method, the middle cerebral artery is exposed by craniotomy and ligated by cauterization. This method gives highly reproducible infarct volumes and increased post-operative survival rates compared to other methods available.

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 ...

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 ...

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

Clinically, thrombolytic therapy with use of recombinant tissue plasminogen activator (tPA) remains the most effective treatment for acute ischemic stroke. However, the use of tPA is limited by its narrow therapeutic window and by increased risk of hemorrhagic transformation. There is an urgent need to develop suitable stroke models to study new thrombolytic agents and strategies for treatment of ischemic stroke. At present, two major types of ischemic stroke models have been developed in rats and mice: intraluminal suture MCAO and embolic MCAO. Although MCAO models via the intraluminal suture technique have been widely used in mechanism-driven stroke research, these suture models do not mimic the clinical situation and are not suitable for thrombolytic studies. Among these models, the embolic MCAO model closely mimics human ischemic stroke and is suitable for preclinical investigation of thrombolytic therapy. This embolic model was first developed in rats by Overgaard et al.1 in 1992 and further characterized by Zhang et al. in 19972. Although embolic MCAO has gained increasing attention, there are technical problems faced by many laboratories. To meet increasing needs for thrombolytic research, we present a highly reproducible model of embolic MCAO in the rat, which can develop a predictable infarct volume within the MCA territory. In brief, a modified PE-50 tube is gently advanced from the external carotid artery (ECA) into the lumen of the internal carotid artery (ICA) until the tip of the catheter reaches the origin of the MCA. Through the catheter, a single homologous blood clot is placed at the origin of the MCA. To identify the success of MCA occlusion, regional cerebral blood flow was monitored, neurological deficits and infarct volumes were measured. The techniques presented in this paper should help investigators to overcome technical problems for establishing this model for stroke research.

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 ...

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

Experimental animal models of stroke are invaluable tools for understanding stroke pathology and developing more effective treatment strategies. A 2 week protocol for repetitive hypoxic preconditioning (RHP) induces long-term protection against central nervous system (CNS) injury in a mouse model of focal ischemic stroke. RHP consists of 9 stochastic exposures to hypoxia that vary in both duration (2 or 4 hr) and intensity (8% and 11% O2). RHP reduces infarct volumes, blood-brain barrier (BBB) disruption, and the post-stroke inflammatory response for weeks following the last exposure to hypoxia, suggesting a long-term induction of an endogenous CNS-protective phenotype. The methodology for the dual quantification of infarct volume and BBB disruption is effective in assessing neurovascular protection in mice with RHP or other putative neuroprotectants. Adult male Swiss Webster mice were preconditioned by RHP or duration-equivalent exposures to 21% O2 (i.e. room air). A 60 min transient middle cerebral artery occlusion (tMCAo) was induced 2 weeks following the last hypoxic exposure. Both the occlusion and reperfusion were confirmed by transcranial laser Doppler flowmetry. Twenty-two hr after reperfusion, Evans Blue (EB) was intravenously administered through a tail vein injection. 2 hr later, animals were sacrificed by isoflurane overdose and brain sections were stained with 2,3,5- triphenyltetrazolium chloride (TTC). Infarcts volumes were then quantified. Next, EB was extracted from the tissue over 48 hr to determine BBB disruption after tMCAo. In summary, RHP is a simple protocol that can be replicated, with minimal cost, to induce long-term endogenous neurovascular protection from stroke injury in mice, with the translational potential for other CNS-based and systemic pro-inflammatory disease states.

This protocol describes repetitive hypoxic preconditioning, or brief exposures to systemic hypoxia that reduce infarct volumes and blood-brain barrier disruption following transient middle cerebral artery occlusion in mice. It also details dual quantification of infarct volume and blood-brain barrier disruption after stroke to assess the efficacy of neurovascular protection.

マウスにおける反復低酸素プレコンディショニングと過渡中大脳動脈閉塞後に神経血管保護の定量

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

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.

マウスの中大脳動脈閉塞再灌流誘発性の脳卒中のための外科的アプローチ

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 ...

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 ...