Name: a thrombotic stroke model based on transient cerebral hypoxia-ischemia
Uploaded: 2015-08-18T13:00:00.000+00:00
Duration: 6 min 1 s
Description: Watch this Scientific Journal Video about A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia at JoVE.com

This study was supported by the NIH grant NS074559 (to C. K.). We thank all collaborators who contributed to our research articles that the present methodology report is based upon.

このプロトコルは、エモリー大学の施設内動物管理使用委員会（IACUC）によって承認され、ケアと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に追従されます。 1.セットアップ<ol><li>手術の前に少なくとも15分間、37℃で熱ポンプに接続された温暖化のパッドの上に手術台を準備します。手術台の上に3ミリリットルの注射器のバレルを使用して、首のロールを配置します。医療空気中2％イソフルランで麻酔ガスを準備します。 </li><li>オートクレーブした鉗子、はさみ、マイクロニードルホルダー、止血剤、綿棒や縫合糸を準備します。組織接着剤や眼軟膏を準備します。 </li><li>加熱ランプおよび直腸プローブと低酸素システムと温度コントローラを設定します。 92.5％N 2でバランス7.5％のO 2中の2％イソフルランで低酸素ガスを準備します。 </li><li>一時間手術前に、マウスを徐放メロキシカム（4.0ミリグラム/ kg）を皮下注射することによりanalgesizedされています。 </li></ol> &lt;Pクラス= &quot;jove_title&quot;&gt; 2。一過性脳低酸素虚血（図1B） <ol><li>動物が足のスクイズに応答しなくなるまで、3％イソフルランで麻酔導入室で22〜30グラムの重さの10〜13週齢の雄のC57BL / 6マウスを麻酔し、その後電子シェーバーを使用して、右の首の毛を削除します。 </li><li> 2 L / minの流量で医療空気中の2％イソフルランで接続された手術台の上にマウスを置きます。セキュア前肢は、医療テープを使用して、両側にネックロールに沿って伸ばし。 </li><li>アルコール、その後綿棒続いベタジンで切開するための外科手術部位を清掃してください。 </li><li>解剖顕微鏡下で、まっすぐ鉗子と約0.2センチ正中皮膚からの横方向のマイクロはさみを使用して0.5センチメートル右頸部切開を行います。 </li><li>右総頸動脈（RCCA）を露出させるために筋膜と組織を引き離すために微細な鋸歯状の鉗子のペアを使用してください。慎重に細かいなめらかなピンセットを使用して、迷走神経からRCCAを分離します。 </li><li>ライブRCCA上の2つのプレカット5-0絹縫合糸（解除可能）を結び目、その後、4-0ナイロンモノフィラメント縫合糸（ 図1C）を用いて皮膚を縫い合わせます。 </li><li>乾燥を防ぐために、両眼に眼軟膏を適用します。 </li><li>素早く低酸素システムにマウスを転送し、30分間、0.5 L /分の流速で7.5％のO 2中の2％イソフルランでフェイスマスクで鼻と口を置きます。 <ol><li>低酸素時には、37.5±0.5℃の直腸温度を制御する加熱ランプと温度コントローラを使用しています。 80〜120呼吸/分で呼吸速度を監視します。低酸素時に37℃以上の体温を維持することは、一貫性の脳梗塞を作成することが重要です。低呼吸数は、通常20分の低酸素症の後に発生します。フェイスマスクを外し、呼吸数は40に下回る場合、これは1〜2分を取り、30分間の低酸素症の期間にカウントされません通常の空気の供給を可能にします。 </li></ol></li><li>低酸素症の後、転送手術台のマウスとはRCCAから2本の縫合糸を解放します。組織接着剤を使用して傷を閉じて、ケージにマウスを返します。 2つのライブノットの両方が予期せず、低酸素後にリリースされた場合、動物を除外します。 </li><li>低酸素および麻酔から回復するために5〜10分間マウスを監視します。ケージに濡れた食品を置き、動物飼育施設に戻します。 注：THI後24時間で重度の旋回動作に軽度を示す動物は脳梗塞と相関しています。発作症状を持つほとんどの動物は、Th1 24時間後の時点前に死亡します。 </li></ol> 3.レーザースペックルコントラストイメージング注意：これはTHIモデルの本質的な手順ではないが、二次元のレーザースペックルコントラスト画像化システム16は、中または一時低酸素性虚血後の脳の血流（CBF）の変化を特徴づけるために使用することができます。 STの直後に、THIで、レコードをCB​​Fの変化を文書化することEP 2.6。あるいは、THI傷害後のCBFの回復を比較するために、これらの手順は、ステップ2.10以下を行うことができます。 <ol><li>腹臥位で麻酔したマウスを置き、頭蓋骨露出したが、未開封で、頭皮に約1cm長の正中切開を行います。 </li><li>製造業者のプロトコルに従って、血流画像化装置の下で両方の大脳半球にCBFを監視し、CCAO手術（ステップ2.6）の直後に脳血流の記録を開始。 50分間続けます。 </li><li> 16色のパレットで任意の単位でCBF画像を表示して、製造業者の指示（ 図2）は、次のMoorFLPIソフトウェアを使用してリアルタイムで選択した領域を分析します。 </li><li> CBF画像を記録した後、組織接着剤で頭皮を閉じて、ケージに動物を返します。 </li></ol> 4. tPAの管理<ol><li> 220〜300（溶媒または10mg / kgの組換えtPAを尾静脈で動物を注入＆＃956; 1ミリグラム/ M TPA）のL 0.5、1、またはtCCAoプラス低酸素症後4時間で（ 図4）。 </li></ol>いくつかの異なるオプションで5脳の損傷検出注：THI後、脳試料を採取1でマウスを安楽死させる、4または24時間にします。 <ol><li> in vivoでの塩化2,3,5-トリフェニルテトラゾリウム（TTC）が記載以前のようなTHI傷害後24時間目の方法で梗塞体積の定量化を行う。17 <ol><li>腹腔内transcardial灌流前に1.4 Mマンニトール溶液（〜0.1ミリリットル/ g体重）を30分で動物を注入します。 PBSでTranscardial散らすマウスを2％TTCを10mlました。 </li><li> 10分後に手術器具を用いた動物の脳を削除し、ビブラトームで、1mmの厚さに一晩固定し、セクションの4％パラホルムアルデヒド中に配置します。 </li><li>デジタル顕微鏡および定量によって4区分脳のスライドの一連のスナップImageJソフトウェアを使用して、損傷していない、反対側の半球の面積に対する梗塞領域（右側の白い部分）の割合としての梗塞体積をIFY。 </li></ol></li><li>あるいは、THI傷害後の1時間後に免疫蛍光法により血栓形成を行います。 <ol><li> OCT化合物および部分クライオスタットを用いて、12μmの厚さで脳内の固定脳をフリーズします。 </li><li>蛍光顕微鏡で蛍光を観察するために：（200 1）ヤギ抗ウサギアレクサFluro 488色素により以下：（100 1）ウサギ抗フィブリノゲン抗体を用いた脳のスライドをインキュベートします。 </li></ol></li><li>あるいは、THI傷害の後4時間で100μlの2％エバンスブルー色素の尾静脈注射により血管閉塞を行います。 <ol><li>マウスを安楽死させるとすぐにエバンスブルー注射後、4％パラホルムアルデヒド中に脳を除去するために頭を切りました。注：これは、両方の前足の青色とエバンスブルー循環のための5-10分を取り、後肢。 </li><li> Secti100ミクロンの厚さで固定された脳にスライディングミクロトームを用いて、蛍光顕微鏡で680 nmの発光フィルターを用いて蛍光を観察します。 </li></ol></li></ol>

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脳卒中は、人口の高齢化との任意の社会の高まる重要性の主要な健康問題です。世界的には、ストロークが全死亡18の11.1％に相当する2010年の推定590万致命的なイベント、との死因の第2位です。脳卒中は障害調整生存年数（DALYs）の第三の主要な原因は、1990年19で5位から上昇し、2010年に世界的に失われたもである。これらの疫学データは、急性のより有効な治療法（虚血性）...

血栓溶解および再疎通は、臨床診療1急性虚血性脳卒中の最も効果的な治療法です。しかし、前臨床神経保護研究の大部分は、血管閉塞を除去すると脳血流量の急速な回復を生成したtPAの血栓によるない利点のほとんどを示す過渡メカニック閉塞モデル（腔内縫合中大脳動脈閉塞）で行いました。これは、脳卒中モデルの選択は、疑わしい患者2,3に神経保護療法を変換することが困難に、少なくとも部分的に寄与することが示唆されています。したがって、前臨床研究におけるtPAの応答性血栓塞栓性脳卒中モデルを採用したため増加コールがありますが、このようなモデルはまた、技術的な問題（説明を参照）4-7を有しています。ここでは、一方的な一過性低酸素性虚血性（THI）侮辱および静脈内のtPA療法8への応答に基づいて新たな血栓性脳卒中モデルについて説明します。 THIストロークモデルは、1960年9成体ラットを用いた実験のために発明されたLevineの手順（チャンバ内の過渡的低酸素への曝露が続く一方的な総頸動脈の永久結紮）に基づいて開発されました。オリジナルのレバイン手順はあいまいに色あせたため、それが唯一の変数脳の損傷を生じたが、それは1981年10新生児低酸素性虚血性脳症（HIE）のモデルとしてロバート·バンヌッチと彼の同僚によって再導入されたときと同じ侮辱は、げっ歯類の仔で一貫性のある神経病理の原因となった。近年では、いくつかの低酸素室11内の温度を調整することにより、成体マウスの研究者の再適応レヴァイン-バンヌッチモデル。これは、元レヴァインの手順で一貫性のない脳病変は、低酸素室に大人のげっ歯類の体温の変動から生じ得ることがもっともらしいです。この仮説を試験するために、我々は、低酸素ガスを投与することにより、レヴァイン手順を改変しましたフェイスマスクを介して、手術台12の上に、37℃でげっ歯類コア温度を維持します。予想されるように、ストリンジェントな体温調節が大きくHI誘発性脳病変の再現性を増加させました。 HI傷害はまた、凝固、オートファジー、とグレーと白質損傷13をトリガします。他の研究者らはまた、脳卒中後の炎症反応14を調査するためにHIモデルを使用しています。 HIストロークモデルのユニークな特徴、それは密接に血流のうっ滞を含む、血栓形成のウィルヒョウのトライアドに従うことで、内皮損傷（ 例えばによるHI-誘導される酸化ストレスに対する）、および凝固亢進（HI誘発性血小板活性化）（ 図1A）15。このように、HIモデルは、実世界の虚血性脳卒中に関連したいくつかの病態生理学的メカニズムを取り込むことができます。念頭に置いてこの考えでは、我々はさらに、国連の可逆的ライゲーションとHIモデルを洗練しましたilateral総頸動脈（したがって過渡HI傷害を作成する）、およびまたはエダラボンなしのtPA血栓溶解にその応答をテストしました。エダラボンは、すでに発症9の24時間以内の虚血性脳卒中を治療するために、日本で承認され、フリーラジカルスカベンジャーです。我々の実験は、短い30分の過渡HIは、血栓性梗塞をトリガし、その組み合わせのtPA-エダラボン処理が相乗便益8を与えることを示しました。ここでは、詳細な外科的手順および急性虚血性脳卒中の再灌流治療を最適化するために使用することができますTHIモデルの方法論的な考慮事項について説明します。

<table><tbody><tr><td>adult male mice</td><td>Charles River</td><td>C57BL/6&amp;nbsp;</td><td>10-13 weeks old (22-30 g)</td></tr><tr><td>Mobile Laboratory Animal Anesthesia System</td><td>VetEquip</td><td>901807</td><td>anesthesia</td></tr><tr><td>Medical air (Compressed) air tank</td><td>Airgas</td><td>UN1002</td><td>anesthesia</td></tr><tr><td>Isoflurane</td><td>Piramal Healthcare</td><td>NDC 66794-013-25</td><td>anesthesia</td></tr><tr><td>Multi-Station Lab Animal AnesthesiaSystem</td><td>Surgivet</td><td>V703501</td><td>hypoxia system</td></tr><tr><td>7.5% O&lt;sub&gt;2&lt;/sub&gt; balanced by 92.5% N&lt;sub&gt;2&lt;/sub&gt; tank</td><td>Airgas</td><td>UN1956</td><td>hypoxia system</td></tr><tr><td>Temperature Controller with heating lamp&amp;nbsp;</td><td>Cole Parmer&amp;nbsp;</td><td>EW-89000-10</td><td>temperature controllers</td></tr><tr><td>Rectal probe</td><td>Cole Parmer&amp;nbsp;</td><td>NCI-00141PG</td><td>temperature controllers</td></tr><tr><td>Dissecting microscope&amp;nbsp;</td><td>Olympus&amp;nbsp;</td><td>SZ40</td><td>surgical setup</td></tr><tr><td>Heat pump with warming pad</td><td>Gaymar&amp;nbsp;</td><td>TP700</td><td>surgical setup</td></tr><tr><td>Fine curved forceps (serrated)</td><td>FST</td><td>11370-31</td><td>surgical instrument</td></tr><tr><td>Fine curved forceps (smooth)</td><td>FST</td><td>11373-12</td><td>surgical instrument</td></tr><tr><td>micro scissors</td><td>FST</td><td>15000-03</td><td>surgical instrument</td></tr><tr><td>micro needle holders</td><td>FST</td><td>12060-01</td><td>surgical instrument</td></tr><tr><td>Halsted-Mosquito hemostats</td><td>FST</td><td>13008-12</td><td>surgical instrument</td></tr><tr><td>5-0 silk suture&amp;nbsp;</td><td>Harvard Apparatus</td><td>624143</td><td>surgical supplies</td></tr><tr><td>4-0 Nylon monofilament suture</td><td>LOOK</td><td>766B</td><td>surgical supplies</td></tr><tr><td>Tissue glue</td><td>Abbott Laboratories</td><td>NC9855218</td><td>surgical supplies</td></tr><tr><td>Puralube Vet ointment</td><td>Fisher</td><td>NC0138063&amp;nbsp;</td><td>eye dryness prevention&amp;nbsp;</td></tr><tr><td>MoorFLPI-2 blood flow imager</td><td>Moor</td><td>780-nm laser source</td><td>Laser Speckle Contrast Imaging</td></tr><tr><td>Mannitol</td><td>Sigma</td><td>M4125</td><td>&lt;i&gt;in vivo&lt;/i&gt; TTC</td></tr><tr><td>2,3,5-triphenyltetrazolium chloride (TTC)&amp;nbsp;</td><td>Sigma</td><td>T8877</td><td>&lt;i&gt;in vivo&lt;/i&gt; TTC</td></tr><tr><td>Vibratome</td><td>Stoelting</td><td>51425</td><td>brain section for &lt;i&gt;in vivo&lt;/i&gt; TTC&amp;nbsp;</td></tr><tr><td>Digital microscope</td><td>Dino-Lite</td><td>AM2111</td><td>whole-brain imaging</td></tr><tr><td>O.C.T compound</td><td>Sakura Finetek</td><td>4583</td><td></td></tr><tr><td>goat anti-rabbit Alexa Fluro 488</td><td>Invitrogen</td><td>A11008</td><td>Immunohistochemistry</td></tr><tr><td>Cryostat</td><td>Vibratome</td><td>ultrapro 5000</td><td>brain section for IHC</td></tr><tr><td>Evans blue</td><td>Sigma</td><td>E2129</td><td>Detecting vascular perfusion</td></tr><tr><td>Microtome</td><td>Electron Microscopy Sciences</td><td>5000</td><td>brain section for histology</td></tr><tr><td>Avertin (2, 2, 2-Tribromoethanol)</td><td>Sigma</td><td>T48402</td><td>euthanasia</td></tr><tr><td>Fluorescent microscope</td><td>Olympus</td><td>DP73</td><td></td></tr><tr><td>Meloxicam SR</td><td>ZooPharm</td><td></td><td>NSAID analgesia</td></tr></tbody></table>

a thrombotic stroke model based on transient cerebral hypoxia-ischemia

脳卒中の研究は、臨床実践に神経保護療法を変換するときに、多くの挫折に耐えてきました。これとは対照的に、現実世界の治療因子（tPAの血栓溶解）はめったに前臨床ストローク研究を支配する機械的閉塞ベースの実験モデルで利益を、生成しません。ベンチとベッドサイドの間のこの分割は、前臨床ストローク研究でのtPA応答モデルを採用する必要性を示唆しています。この目的のために、簡単かつ反応性のtPA血栓性脳卒中モデルを考案し、ここで説明されています。 37.5±0.5℃で、動物の直腸温度を維持しながら、このモデルは、30分間、成体マウスにおいて、フェイスマスクを介して片側の総頸動脈および7.5％酸素の送達の一過性閉塞から成ります。片側頚動脈または低酸素の可逆的連結各のみ一過性脳血流を抑制しているが、両方の損傷の組合せが持続する再灌流障害、フィブリンおよび血小板沈着、および大infarを引き起こし中大脳動脈が提供する地域におけるCT。重要なことには0.5で、組換えtPAの、尾静脈注射、1、または4時間後THI（10mg / kg）は死亡率および梗塞サイズの時間依存的な減少をもたらしました。この新たな脳卒中モデルが単純であり、実験結果を比較するために実験室を横切って標準化することができます。また、頭蓋骨切除術または予め形成された塞栓を導入することなく、血栓症を誘発します。これらのユニークなメリットを考えると、THIモデルは、前臨床ストローク研究のレパートリーに有用な付加です。

二次元レーザスペックルコントラストイメージング（LSCI）16は、30分の過渡片側頚動脈閉塞（tCCAO）、低酸素状態に30分間露光（7.5％酸素）、および30分片側によって脳血流（CBF）の変化を比較しました低酸素症（THI）の下で連結を頸動脈。この実験は、tCCAOが通常酸素下で迅速に頸動脈閉塞（ 図2AのR）のリリース後85％以上に回復し、ベースライン値の〜50％に頸動脈結紮...

Watch this Scientific Journal Video about A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia at JoVE.com

A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia

Thromboembolic stroke models are vital tools for optimizing the recanalization therapy. Here we report a murine thrombotic stroke model based on transient cerebral hypoxic-ischemic (tHI) insult, which triggers thrombosis and infarction, and responds favorably to tissue plasminogen activator (tPA)-mediated fibrinolysis in a therapeutic window similar to those in stroke patients.

一過性脳低酸素性虚血に基づく血栓性脳卒中モデル

Stroke research has endured many setbacks in translating neuroprotective therapies into clinical practice. In contrast, the real-world therapy (tPA ...

a-thrombotic-stroke-model-based-on-transient-cerebral-hypoxia-ischemia

Research

JoVE Journal

Medicine

14.7K ビュー.  The Emory University School of Medicine. Thromboembolic stroke models are vital tools for optimizing the recanalization therapy. Here we report a murine thrombotic stroke model based on transient cerebral hypoxic-ischemic (tHI) insult, which triggers thrombosis and infarction, and responds favorably to tissue plasminogen activator (tPA)-mediated fibrinolysis in a therapeutic window similar to those in stroke patients. 

ビデオ: A Thrombotic Stroke Model Based On Transient Cerebral Hypoxia-ischemia

The Hypoxic Ischemic Encephalopathy Model of Perinatal Ischemia

Hypoxic-Ischemic Encephalopathy (HIE) is the consequence of systemic asphyxia occurring at birth. Twenty five percent of neonates with HIE develop severe and permanent neuropsychological sequelae, including mental retardation, cerebral palsy, and epilepsy. The outcomes of HIE are devastating and permanent, making it critical to identify and develop therapeutic strategies to reduce brain injury in newborns with HIE. To that end, the neonatal rat model for hypoxic-ischemic brain injury has been developed to model this human condition. The HIE model was first validated by Vannucci et al 1 and has since been extensively used to identify mechanisms of brain injury resulting from perinatal hypoxia-ischemia 2 and to test potential therapeutic interventions 3,4. The HIE model is a two step process and involves the ligation of the left common carotid artery followed by exposure to a hypoxic environment. Cerebral blood flow (CBF) in the hemisphere ipsilateral to the ligated carotid artery does not decrease because of the collateral blood flow via the circle of Willis; however with lower oxygen tension, the CBF in the ipsilateral hemisphere decreases significantly and results in unilateral ischemic injury. The use of 2,3,5-triphenyltetrazolium chloride (TTC) to stain and identify ischemic brain tissue was originally developed for adult models of rodent cerebral ischemia 5, and is used to evaluate the extent of cerebral infarctin at early time points up to 72 hours after the ischemic event 6. In this video, we demonstrate the hypoxic-ischemic injury model in postnatal rat brain and the evaluation of the infarct size using TTC staining.

The postnatal rat model for hypoxic-ischemic brain injury is a well-established model of human neonatal hypoxic ischemic encephalopathy (HIE). In this article, we describe the model of HIE in post-natal rat pups.

周産期虚血低酸素虚血性脳症モデル

Hypoxic-Ischemic Encephalopathy (HIE) is the consequence of systemic asphyxia occurring at birth. Twenty five percent of neonates with HIE develop severe ...

生物学

Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical Lesion Model for Mouse Stroke Studies

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected light-sensitive dye. Following illumination, the dye is activated and produces singlet oxygen that damages components of endothelial cell membranes, with subsequent platelet aggregation and thrombi formation, which eventually determines the interruption of local blood flow. This approach, initially proposed by Rosenblum and El-Sabban in 1977, was later improved by Watson in 1985 in rat brain and set the basis of the current model. Also, the increased availability of transgenic mouse lines further contributed to raise the interest on the photothrombosis model. Briefly, a photosensitive dye (Rose Bengal) is injected intraperitoneally and enters the blood stream. When illuminated by a cold light source, the dye becomes activated and induces endothelial damage with platelet activation and thrombosis, resulting in local blood flow interruption. The light source can be applied on the intact skull with no need of craniotomy, which allows targeting of any cortical area of interest in a reproducible and non-invasive way. The mouse is then sutured and allowed to wake up. The evaluation of ischemic damage can be quickly accomplished by triphenyl-tetrazolium chloride or cresyl violet staining. This technique produces infarction of small size and well-delimited boundaries, which is highly advantageous for precise cell characterization or functional studies. Furthermore, it is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

Photothrombosis is a quick, minimally-invasive technique for inducing small and well-delimited infarction in areas of interest in highly reproducible manner. It is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

Photothrombotic虚：マウス脳卒中研究のための低侵襲かつ再現性の光化学皮質病変モデル

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected ...

医学

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 ...

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 ...

A Fibrin-Enriched and tPA-Sensitive Photothrombotic Stroke Model

An ideal thromboembolic stroke model requires certain properties, including relatively simple surgical procedures with low mortality, a consistent infarction size and location, precipitation of platelet:fibrin intermixed blood clots similar to those in patients, and an adequate sensitivity to fibrinolytic treatment. The rose bengal (RB) dye-based photothrombotic stroke model meets the first two requirements but is highly refractory to tPA-mediated lytic treatment, presumably due to its platelet-rich, but fibrin-poor clot composition. We reason that combination of RB dye (50 mg/kg) and a sub-thrombotic dose of thrombin (80 U/kg) for photoactivation aimed at the proximal branch of middle cerebral artery (MCA) may produce fibrin-enriched and tPA-sensitive clots. Indeed, the thrombin and RB (T+RB)-combined photothrombosis model triggered mixed platelet:fibrin blood clots, as shown by immunostaining and immunoblots, and maintained consistent infarct sizes and locations plus low mortality. Moreover, intravenous injection of tPA (Alteplase, 10 mg/kg) within 2 h post-photoactivation significantly decreased the infarct size in T+RB photothrombosis. Thus, the thrombin-enhanced photothrombotic stroke model may be a useful experimental model to test novel thrombolytic therapies.

Traditional photothrombotic stroke (PTS) models mainly induce dense platelet aggregates of a high resistance to tissue plasminogen activator (tPA)-lytic treatment. Here a modified murine PTS model is introduced by co-injecting thrombin and photosensitive dye for photoactivation. The thrombin-enhanced PTS model produces mixed platelet:fibrin clots and is highly sensitive to tPA-thrombolysis.

フィブリン富化およびtPA感受性光血栓性脳卒中モデル

An ideal thromboembolic stroke model requires certain properties, including relatively simple surgical procedures with low mortality, a consistent ...

神経科学

Modeling Stroke in Mice: Focal Cortical Lesions by Photothrombosis

Stroke is a leading cause of death and acquired adult disability in developed countries. Despite extensive investigation for novel therapeutic strategies, there remain limited therapeutic options for stroke patients. Therefore, more research is needed for pathophysiological pathways such as post-stroke inflammation, angiogenesis, neuronal plasticity, and regeneration. Given the inability of in vitro models to reproduce the complexity of the brain, experimental stroke models are essential for the analysis and subsequent evaluation of novel drug targets for these mechanisms. In addition, detailed standardized models for all procedures are urgently needed to overcome the so-called replication crisis. As an effort within the ImmunoStroke research consortium, a standardized photothrombotic mouse model using an intraperitoneal injection of Rose Bengal and the illumination of the intact skull with a 561 nm laser is described. This model allows the performance of stroke in mice with allocation to any cortical region of the brain without invasive surgery; thus, enabling the study of stroke in various areas of the brain. In this video, the surgical methods of stroke induction in the photothrombotic model along with&#160;histological analysis are demonstrated.

Described here is the photothrombotic stroke model, where a stroke is produced through the intact skull by inducing permanent microvascular occlusion using laser illumination after administration of a photosensitive dye.

マウスにおける脳卒中のモデリング:光血栓症による焦点性皮質病変

Stroke is a leading cause of death and acquired adult disability in developed countries. Despite extensive investigation for novel therapeutic strategies, ...

Modeling Stroke in Mice: Transient Middle Cerebral Artery Occlusion via the External Carotid Artery

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options are limited to a small proportion of stroke patients within the first hours after stroke. Novel therapeutic strategies are being extensively investigated, especially to prolong the therapeutic time window. These current investigations include the study of important pathophysiological pathways after stroke, such as post-stroke inflammation, angiogenesis, neuronal plasticity, and regeneration. Over the last decade, there has been increasing concern about the poor reproducibility of experimental results and scientific findings among independent research groups. To overcome the so-called &#8220;replication crisis&#8221;, detailed standardized models for all procedures are urgently needed. As an effort within the &#8220;ImmunoStroke&#8221; research consortium (https://immunostroke.de/), a standardized mouse model of transient middle cerebral artery occlusion (MCAo) is proposed. This model allows the complete restoration of the blood flow upon removal of the filament, simulating the therapeutic or spontaneous clot lysis that occurs in a large proportion of human strokes. The surgical procedure of this &#8220;filament&#8221; stroke model and tools for its functional analysis are demonstrated in the accompanying video.

Different models of middle cerebral artery occlusion (MCAo) are used in experimental stroke research. Here, an experimental stroke model of transient MCAo via the external carotid artery (ECA) is described, which aims to mimic human stroke, in which the cerebrovascular thrombus is removed due to spontaneous clot lysis or therapy.

マウスにおける脳卒中のモデリング:外頸動脈を介した過渡中脳動脈閉塞

Stroke is the third most common cause of mortality and the leading cause of acquired adult disability in developed countries. To date, therapeutic options ...

中大脳動脈の閉塞による一過性脳虚血のモデル

Transient Middle Cerebral Artery Occlusion Model of Stroke

Stroke stands as a major cause of death or chronic disability globally. Nevertheless, existing optimal treatments are limited to reperfusion therapies during the acute phase of ischemic stroke. To gain insights into stroke physiopathology and develop innovative therapeutic approaches, in vivo rodent models of stroke play a fundamental role. The availability of genetically modified animals has particularly propelled the use of mice as experimental stroke models.
In stroke patients, occlusion of the middle cerebral artery (MCA) is a common occurrence. Consequently, the most prevalent experimental model involves intraluminal occlusion of the MCA, a minimally invasive technique that doesn't require craniectomy. This procedure involves inserting a monofilament through the external carotid artery (ECA) and advancing it through the internal carotid artery (ICA) until it reaches the branching point of the MCA. After a 45 min arterial occlusion, the monofilament is removed to allow reperfusion. Throughout the process, cerebral blood flow is monitored to confirm the reduction during occlusion and subsequent recovery upon reperfusion. Neurological and tissue outcomes are evaluated using behavioral tests and magnetic resonance imaging (MRI) studies.

著者スポットライト:中大脳動脈閉塞モデルによる虚血性脳卒中損傷の評価 

This protocol describes the model of transient focal cerebral ischemia in mice through intraluminal occlusion of the middle cerebral artery. Additionally, examples of outcome assessment are shown using magnetic resonance imaging and behavioral tests.

脳卒中の一過性中大脳動脈閉塞モデル

Stroke stands as a major cause of death or chronic disability globally. Nevertheless, existing optimal treatments are limited to reperfusion therapies ...

一過性脳低酸素性虚血に基づく血栓性脳卒中モデル

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