著者らは、初期の草稿の校正のためのローリー·フォン·Melchner、ベルン大学病院、脳神経外科、ベルン、スイス、校正のための原稿を編集しPaskusエレミヤ、ボストン小児病院、マサチューセッツ州ボストンに感謝します。私たちは、ダニエル·メトラー、DVM、マックス·ミュラー、DVM、ダニエルZalokar、およびOlgica Beslac、実験外科研究所、臨床研究科、ベルン大学、ベルン、スイスからの動物の世話、麻酔および手術支援の熟練した管理を感謝しています。私たちは、生理学的パラメータのリアルタイムデータの監視と事後処理のために、マイケルLensch、ヘッドリサーチナース、集中治療医学科、ベルン大学病院やベルン大学、ベルン、スイスに感謝します。私たちは、その優れた研究室の技術的および手術支援のため、エディンNevzati、カール·室井、そしてサロメエルハルトに感謝します。 この作品はIntensiv省によってサポートされていましたEケア医学、ベルン大学病院やベルン、ベルン、スイス、臨床研究科、ベルン、ベルン、スイスの大学の大学、Kantonsspitalアーラウ、アーラウ、スイス研究基金。私たちは、 図1および2のためのリプリント許可、エルゼビアに感謝します。

2.5計量三ヶ月齢の雌のニュージーランドウサギ - 3.5キロは、この手順のために使用された。この研究は、実験動物の管理と使用のための健康ガイドラインの国立研究所に従ってベルン、スイス（承認＃105/13）のカントンの動物管理委員会の承認を得て実施した。すべての外科的処置はベルン、スイスのベルン大学病院での臨床研究科の実験的手術研究所で、無菌条件下で行った。獣医麻酔科医は手術中におよび回復を通して動物を監視した。 1動物の準備、位置決めおよび鎖骨下動脈のカニューレ挿入<ol><li>有害に対するウサギの反応をチェックすることによって、麻酔の制御深さケタミンの筋肉内注射（30mg / kgの;ケタラール、50 mg / ml）をウサギに全身麻酔を誘導し、キシラジン（Xylapan 20 mg / mlのkgを6ミリグラム/） stimulatイオン（ 例えば、つま先のピンチ）。肯定的な応答の場合には1.7を参照してください。 </li><li>両眼の下蓋をプルダウンして、乾燥やさらなる刺激を防止するために、まぶたに軟膏を少量を適用します。 </li><li> 、20 Gbutterflyのvenflon（20 G血管カテーテル）と横方向の耳静脈にカテーテルを挿入粘着テープで固定し、0.9％塩化ナトリウム（500ミリリットル/ 24時間）およびケタミン（40ミリグラム/ kg /時間）を含む重力バッグに接続する持続静脈内（IV）麻酔用/キシラジン（4 / kg /時間）。追加の鎮痛薬を15分毎に静脈（フェンタニル、1マイクログラム/ kg）を管理します。注意：CPPの低下と関連している揮発性のガス麻酔薬、CBFを増加し、酸素が ​​脳代謝率を低下させるの回避11静脈麻酔薬が脳研究する最上重要であり、CBF、脳血管収縮、12を保存することにより、脳神経外科麻酔のためのより理想的な特性を提供する。血管攣縮。加えて、死亡率がSPONTで増加するもののaneously動物呼吸、それはより良い急性SAHのヒトの状況を模倣することがあります。 </li><li>呼気終末二酸化炭素（ETCO 2）監視を可能にする呼吸マスクを介して- （2L /分1）酸素吸入を行う。 </li><li>ウサギの腹側の三角形の配置で3チャネル心電図（ECG）.Place 3皮下電極をインストールします。具体的には、両方の手足に分布下腹部に二つの電極（鎖骨下動脈カニューレ挿入のための無菌剃らフィールドまでの距離で）右midthoracic領域上に一方の電極を配置します。 </li><li>呼吸速度に追従して、手術中の麻酔深度を15分毎に監視し、心拍数（HR）は、ECG信号から監視し、侵害刺激に対する反応。 </li><li>侵害刺激に対する肯定応答（つま先ピンチ）の場合は、ケタミンボーラスによって麻酔深度を適応させる（6ミリグラム/ kg）を静脈内およびキシラジンボーラス（0.05ミリグラム/ kg）を静脈内および/またはフェンタニル（1 MCでさらに鎮痛ボーラスグラム/ kg）を静脈内</li><li>ダウンヘッド20°に傾けると鎖骨下動脈が露出される側に対側に少し回転、ボディ温暖化プレート上の仰臥位でウサギを修正しました。 </li><li>眼軟膏を適用し、鎖骨の中央3分の周りの右胸筋の上、およびfrontal-、parietal-と後頭部頭蓋骨、首の上に、右大腿動脈の上の毛を剃ることにより、手術の領域を用意。 </li><li>防腐剤、広域スペクトル、 例えば 、ポビドンヨードで3分間の皮膚を消毒。 </li><li>無菌シートウサギをカバー。無菌条件下でさらなるすべての手順を実行し、頻繁に局所的に血管の操作や地元の感染によって動脈血管攣縮を防ぐために、HClおよび抗生物質溶液（硫酸ネオマイシン5 mg / ml）をパパベリン4％を適用します。 </li><li>局所麻酔薬（リドカイン1％の最大6ミリグラム/ kg）で胸筋潜入。胸骨皮膚切開を行い、準備を胸筋。顕微鏡を使用して、鎖骨下動脈を解剖し、露出端の周り近位および遠位合字（4-0 polyfilament縫合糸）で固定します。カテーテルを確保し、遠位に容器を連結する場所に近い端の制御に1合字にしてください。 </li><li>湾曲したmicroscissorで動脈を切開することによって鎖骨下動脈の壁に動脈切開を行い、小血管内三方活栓に逆行鎖骨下動脈にカニューレを挿入。動脈の近位部の動脈ねじれや曲がりを防ぎ、滑りや大量出血を回避するために、遠位合字に向かってダブルノット結紮によってカテーテルを固定します。 </li></ol> 2血圧や動脈血ガスモニタリング<ol><li>動脈血ガス（ABG）用の血管内カニューレは、pHが、PACO 2、のPaO 2、重炭酸塩、塩基過剰、およびSO 2、ⅱ）侵襲動脈血を分析）をiに3方コックを接続します圧力測定装置、およびiii）シャント装置。 </li><li> ABGステータスの血液サンプルを（パコ2のPaO 2）を収集し、連続的にアナログアナログ出力インタフェースを介して、標準的な心血管及び呼吸パラメータ（血圧、心拍、ECG、呼吸数および呼吸終期CO 2）と転送データを監視デジタル·コンバータ/データロガーとストア。 注：圧力が前と各セッションの後に心臓レベルでゼロにされ、シリーズが始まる前に、アナログ/デジタル変換器とデータロギングシステムの圧力校正は一回行われます。 </li></ol> 3ベースラインデジタルサブトラクション血管造影<ol><li>血管造影を校正するために、両方の下顎角にわたって外部サイジング装置（小球）を配置します。 注：これは、ベースラインの事後測定値の正確な比較を可能にし、血管径をフォローアップ。 </li><li>逆行内aでデジタルサブトラクション血管造影（DSA）を実行非イオン性イオパミドールのrterialボーラス注入カニューレを挿入し動脈を通る（0.6ミリリットル/ kgで、2秒、5ミリリットル/秒）、後者の閉塞を防止するために、生理食塩水ボーラス注射の直後にカニューレをフラッシュ。 </li><li> 5°、左前方の斜めの位置に迅速なシーケンシャル血管造影記録を使用して、椎骨システムの画像（14秒7イメージ）を取得します。 </li><li>局所麻酔薬（リドカイン1％、最大で6ミリグラム/ kg）で右総大腿動脈の周りの領域を潜入。小さな鼠径皮膚切開を行います。可視化のための顕微鏡を用いて、総大腿動脈を解剖し、露出端の周りの近位および遠位合字（4-0 polyfilament縫合）で固定します。 </li><li>動脈切開後、5-Fシースと大腿動脈にカニューレを挿入。生理食塩水でシースのサイドポートをフラッシュします。 </li><li> X線透視下でシースを通して腕頭動脈内に5-Fカテーテルを進める。目にガイドワイヤを進めるその後、ロードマップを作成します。電子椎骨システム。ステップ3.2で説明したように、DSAのための非イオン性イオパミドール（0.6ミリリットル/ kgで、2秒での5ミリリットル/秒）のボーラスを注入。 </li></ol>腹臥位に4回転<ol><li>ベースラインDSAに続いて、仰臥位から​​腹臥位にウサギを再配置する。動脈内のカテーテルの位置を操作したり、再配置しないように注意してください。 </li><li>ダウン頭を重視、30°の角度でヘッドホルダーヘッドを置きます。 </li></ol> 5。大槽穿刺<ol><li> 1分ごとに、ポビドンヨード3回で頭と首の上の皮膚を消毒し、滅菌シーツ手術領域をカバーしています。 </li><li>事前の皮膚切開や筋肉のずれなく経皮大槽内に22 Gのx 40ミリメートル小児脊髄アクセス針を挿入します。 </li><li>動物が完全に骨の外後頭隆起に沿って針を滑り落ちる前に、つま先のピンチ応答の欠如を確実にすることによって麻酔していることを確認してくださいギャップが検出されるまで;それ以上の針をプッシュしないでください。 </li><li>少数分間20°の角度で下方に傾斜したウサギの頭部に脳脊髄液の自発的な滴下を観察することによって、針の正確な位置を確認してください。 </li></ol>頭蓋内圧と脳血流のモニタリング6。インストール<ol><li>正中線皮膚や帽状切開した後、小さな手術用リトラクターを挿入します。 </li><li>嗅球とneuromonitoringデバイスを配置するための二国間の前頭場合の上、外側頭蓋骨のランドマーク（ 図1）9、 すなわちに従って頭蓋骨の正面部分の高速マイクロドリルを使用して、3ラウンドの骨切り術（直径2mm）を作る必要に応じて。次のようにバーホールの配置の座標を決定するために、ミリメートルスケールの定規を使用します。midpupillaryラインの頭蓋内圧（ICP）の監視、正中矢状線から1〜2ミリメートル;実質内レーザードップラープローブ4〜5ミリメートルの前方およびブレグマの側方（ 図1）。 </li><li>硬膜を視覚化し、細心の止血を行います。そのタンポナーデ作用により骨止血用の骨ろうを使用し、硬膜のバイポーラ凝固を使用してローカル止血を行う。 </li><li>実質内頭蓋内圧（ICP）2ミリメートルの深さまで右嗅球にヒントを監視し、キャリブレーションを配置します。 </li><li>外部クランプ·システムを使用して2レーザードップラーフローメトリ細い針プローブを置き、左右の前頭半球の両方が脳脊髄液との干渉を避けるために、正中線で、脳室系、 すなわちに横に対応したバーホールに挿入してください。 2.5mmの深さまで針プローブを配置します。 </li><li> neuromonitoringプローブを配置した後、頭蓋の流体密封を維持するために骨ろうの厚いプラグのすべてのバリ穴をシール。 </li><li>平均動脈血圧のベースラインパラメータを測定（MAP）、ICPおよびマルチパラメータモニタと4チャンネルレーザードップラー組織血灌流モニターを使用して脳血流量（CBF）。 </li></ol> 7。シャント誘導<ol><li>血液で満たされた圧力モニタリング管を介して以前にカテーテルを挿入鎖骨下動脈に、大槽内、脊髄アクセス針を接続します。血圧測定のための採血ポートに三方活栓を使用する。 NOTE：SAHの重症度は、血液の量に依存し、ほぼ脳収穫5,11時のくも膜下凝血塊の広大によって推定することができる。 </li><li>継続的SAH後少なくとも20分まで前に6分〜1Hzのサンプリングレートで、MAP、HR、心電図、呼吸数や呼吸終期CO 2を監視します。 </li><li>動物が完全に鎖骨下動脈と大槽圧力gradieによるSAHを誘導するために間にシャント接続を開く前に足指のピンチ応答の欠如を確実にすることによって麻酔していることを確認してくださいNT。 NOTE：（ たとえば 、ICPの所望のレベル）に制御SAHは、任意の時点でシャントを閉じることによって達成することができる。 </li><li>約15分の期間中に定常状態の値を記録する。 </li><li> ICPはそのピークに達した後、ICPは、ベースライン値に近い定常状態に戻るまで、所定の位置に脊椎アクセス針を保持します。 ICP高原以上10秒間維持されるか、またはICPは自然に減少した場合、シャントを閉じます。された場合</li><li> 、（連続的な出血カテーテル操作が高い罹患率と死亡率と関連し、感染率が増加しているため、鎖骨下カテーテルを含む）すべてのカテーテルを取り外し、骨ろうでバー​​ホールを差し込み、CBF細針プローブおよびICPプローブを削除した厳格な創傷洗浄を行う硫酸ネオマイシンと皮膚を縫合。 </li></ol> 8。術後管理<ol><li>手順は約2時間持続する。原因ケタミンおよびキシラジン、アニマの回復時間の半減期に約1時間 - lはかなり短いです。動物は、回復時の加熱ランプの下で保持されます。追加の流体が供給されない。この最初の術後の回復期の間、ブプレノルフィン0.02ミリグラム/ kg皮下24時間毎に8時間適用されます。 </li><li>次の72時間にわたって効果的な鎮痛のための動物の毛を剃り、首領域において12.5μgの/時間を解放する経皮フェンタニルマトリックスパッチを適用します。 </li><li>それは胸骨横臥位を維持するのに十分な意識を取り戻したまでは無人の動物を放置しないでください。 </li><li>完全に回復するまで、他の動物の会社で手術を受けた動物を返さないでください。 </li></ol> 9。3日目でDCVSを評価するために、デジタルサブトラクション血管造影をフォロー<ol><li>前述したように、ステップ1.1〜3.6を実行してください。 </li><li>ナトリウムthiopenthalの動脈内ボーラス注射（40 mgの/ kg）を（ペントタール、Ospedalia AG、Hünenberg、スイス）により動物を安楽死させる。例組織学およびimmunohistでochemistry必要とされる100センチのH 2 Oの灌流圧でRTで心臓内灌流固定を行う</li></ol>

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W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Platelet-derived+growth+factor-induced+severe+and+chronic+vasoconstriction+of+cerebral+arteries:+proposed+growth+factor+explanation+of+cerebral+vasospasm.">Platelet-derived growth factor-induced severe and chronic vasoconstriction of cerebral arteries: proposed growth factor explanation of cerebral vasospasm.</a> Neurosurgery. 66, 728-735 (2010).</li><li>Laslo, A. M., Eastwood, J. D., Chen, F. X., Lee, T. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+CT+perfusion+imaging+in+subarachnoid+hemorrhage-related+vasospasm.">Dynamic CT perfusion imaging in subarachnoid hemorrhage-related vasospasm.</a> AJNR. American journal of neuroradiology. 27, 624-631 (2006).</li><li>Shao, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+tetramethylpyrazine+on+nitric+oxide/cGMP+signaling+after+cerebral+vasospasm+in+rabbits.">Effects of tetramethylpyrazine on nitric oxide/cGMP signaling after cerebral vasospasm in rabbits.</a> Brain research. 1361, 67-75 (2010).</li><li>Bederson, J. B., Germano, I. M., Guarino, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cortical+blood+flow+and+cerebral+perfusion+pressure+in+a+new+noncraniotomy+model+of+subarachnoid+hemorrhage+in+the+rat.">Cortical blood flow and cerebral perfusion pressure in a new noncraniotomy model of subarachnoid hemorrhage in the rat.</a> Stroke; a journal of cerebral circulation. 26, 1086-1091 (1995).</li><li>Veelken, J. A., Laing, R. J., Jakubowski, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Sheffield+model+of+subarachnoid+hemorrhage+in+rats.">The Sheffield model of subarachnoid hemorrhage in rats.</a> Stroke; a journal of cerebral circulation. 26, 1279-1283 (1995).</li><li>Zakhartchenko, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell-mediated+transgenesis+in+rabbits:+chimeric+and+nuclear+transfer+animals.">Cell-mediated transgenesis in rabbits: chimeric and nuclear transfer animals.</a> Biology of reproduction. 84, 229-237 (2011).</li><li>Capecchi, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+targeting+in+mice:+functional+analysis+of+the+mammalian+genome+for+the+twenty-first+century.">Gene targeting in mice: functional analysis of the mammalian genome for the twenty-first century.</a> Nature reviews. Genetics. 6, 507-512 (2005).</li><li>Flisikowska, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+immunoglobulin+gene+disruption+and+targeted+replacement+in+rabbit+using+zinc+finger+nucleases.">Efficient immunoglobulin gene disruption and targeted replacement in rabbit using zinc finger nucleases.</a> PloS one. 6, e21045 (2011).</li><li>Nakajima, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+aging+on+cerebral+vasospasm+after+subarachnoid+hemorrhage+in+rabbits.">Effects of aging on cerebral vasospasm after subarachnoid hemorrhage in rabbits.</a> Stroke. 32, 620-628 (2001).</li></ol>

None. The authors have no financial or commercial interest in any of the drugs, materials, or equipment used. No specific funding was received for this work. The authors are solely responsible for the design and conduct of the presented study and report no conflict of interest concerning the materials and methods used in this study or the findings specified in the paper. They confirm the adherence of ethical standards. The study was performed in accordance with the National Institutes of Health guidelines for the care and use of experimental animals and with the approval of the Animal Care Committee of the Canton of Bern, Switzerland (approval #109/07 and #107/09).

シャントモデルは、急性SAH 3,8,10後にヒトにおいて観察されたものと同様の病理を生成する。それはEBIは、悪化させるの維持、さらにはDCVS 12をトリガし、そのようなこのモデルはSAH後のEBIとDCVS相互作用を含む初期および後期DCVSフェーズ、の両方を調査するのに役立つ可能性があることが示唆されている。具体的には、DSA 13、コンピュータ断層撮影血管造影14、</sup...

くも膜下出血（SAH）は、頻繁に永続的な神経障害や死につながる1、神経病理学的な条件を脅かす最も生命の一つです。過去の研究では、SAH 2に関連する神経障害の主要な病因として、遅延脳血管攣縮（DCVS）に焦点を当てている。しかし、血管痙攣の治療後にSAHに罹患している患者の一般的に不良な臨床転帰は、SAH 3後早期脳損傷（EBI）の影響を含めるための研究の焦点の拡大をもたらした。 SAH後の不良な臨床転帰に貢献するEBIとDCVS両方の重要性の理解は、より効果的な治療戦略の開発に不可欠である。 これまで、大槽へのシングルとダブルの自己血注入はDCVS 2-6の研究のためのSAH誘導のための標準的な方法であった。一般的に、以前の研究で使用されているが、このモデルは最も可能性の高いEBI 7誘発性SAHに関連した神経病理学主要な変更点を再現していません。対照的に、血管穿孔を部分的EBI 7の症状を模倣する重度の急性病態生理学的変化を生じることが知られている。 このレポートでは、SAHの新規ウサギモデルが記載された技術では、標準的な大槽モデルは動脈系を接続することにより適合されている。それによって、SAH誘発性病理8-10のより正確な特性評価が可能、EBIとDCVS両方の調査を可能にするように設計について説明します体外シャントを経由して鎖骨下動脈と大槽の。血流がそれによってウサギの生理機能にリンクされ、動脈血および頭蓋内圧との間の圧力勾配によって駆動される。脳内の圧力（ICP）は、拡張期血圧が等しく、シャントシステム内の血液が凝固する際の出血が停止します。ホスト＆＃を利用し8217;の生理機能を確実EBIとDCVS両方の表現型3,8-10を生産SAHのより一貫性のあるモデルにつながる、審査官依存のSAH誘導を低減します。

<table border="0" cellpadding="0" cellspacing="0" width="905">
	<tbody>
		<tr height="34">
			<td height="34" style="height:35px;width:279px;">Name</td>
			<td style="width:236px;">Company</td>
			<td style="width:223px;">Catalog Number</td>
			<td style="width:167px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:279px;">Equipment</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">operation microscope</td>
			<td style="width:236px;">Zeiss, Jena, Germany</td>
			<td colspan="2">Zeiss, OPMI-MD surgical microscope</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:279px;">surgical equipment</td>
			<td style="width:236px;">B. Braun, Germany</td>
			<td style="width:223px;">forceps medical n&#176;5, vessel sciccors 8cm, microclip 4mm</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">respirator</td>
			<td style="width:236px;">Hugo Sachs</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">hair clipper</td>
			<td style="width:236px;">3M Surgical Clipper&#160;</td>
			<td style="width:223px;">&#160;Starter Kit 9667A</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">body warm plate</td>
			<td style="width:236px;">FHC</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:279px;">blood gas analyzer</td>
			<td style="width:236px;">Radiometer, Copenhagen, Denmark</td>
			<td style="width:223px;">ABL 725</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:279px;">cardiac monitoring</td>
			<td style="width:236px;">Camino Multi-Parameter Monitor, Integra, Plainsboro, NJ, US</td>
			<td style="width:223px;">AP-05</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;">software analysis</td>
			<td style="width:236px;">BIOPAC Systems, Inc., Goleta, CA, USA</td>
			<td style="width:223px;">Biopac MP100 and acqKnowledge software,version 3.8.1</td>
			<td></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;">software analysis</td>
			<td style="width:236px;">ImagePro Discovery, MediaCybernetics, Silver Spring, MD, USA</td>
			<td style="width:223px;">image-Pro Plus version&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">angiography apparatus</td>
			<td style="width:236px;">DFP 2000 A-Toshiba</td>
			<td style="width:223px;">MIIXR0001EAA</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">ICP monitor</td>
			<td style="width:236px;">Camino Laboratories, San Diego, CA, USA</td>
			<td style="width:223px;">ICP monitor, Model 110-4B</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">blood flow monitor</td>
			<td style="width:236px;">Oxford Optronix Ltd., Oxford, UK</td>
			<td style="width:223px;">CAL KIT microsphere solution</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">laser-Doppler flowmetry fine needle probes&#160;</td>
			<td style="width:236px;">Oxford Optronix Ltd., Oxford, UK</td>
			<td style="width:223px;">MNP110XP, 0.48 mm diameter</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">pressure tube</td>
			<td>B. Braun, Germay</td>
			<td style="width:223px;">PE 1.0 mm &#215; 2.0 mm</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">anesthesia monitor</td>
			<td>GE Medical Systems, Switzerland&#160;</td>
			<td style="width:223px;">Datex S5 Monitor</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:279px;">Material</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">20 G vascular catheter</td>
			<td style="width:236px;">Smiths Medical</td>
			<td style="width:223px;">Jelco i.v. catheter, REF 4057</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">5.5F three-lumen central venous catheter&#160;</td>
			<td style="width:236px;">Connectors, Tagelswangen, Switzerland</td>
			<td>silicone catheter STH-C040</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">22Gx40mm needle&#160;</td>
			<td style="width:236px;">Emergo Group Inc., Netherlands</td>
			<td style="width:223px;"></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">high-speed microdrill</td>
			<td style="width:236px;">Stryker, Solothurn, Switzerland</td>
			<td>5400-15&#160;</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">bone wax</td>
			<td style="width:236px;">Ethicon, Johnson &#38; Johnson,NJ, USA</td>
			<td style="width:223px;">ETHW31G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">bipolar forceps</td>
			<td style="width:236px;">Aesculap, Inc., PA, US</td>
			<td>US349SP&#160;</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Ketamin</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Xylazine</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Buprenorphine</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fentanyl</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">transdermal fentanyl matrix patches&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Lidocaine 1%&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">4% papaverin HCl&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Neomycin sulfate&#160;</td>
			<td style="width:236px;">Research Organics Inc., OH, USA</td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Povidone-iodine&#160;</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">0.9% sodium chloride</td>
			<td style="width:236px;"></td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">Iopamidol&#160;</td>
			<td style="width:236px;">Abott Laboratories, IL, USA</td>
			<td style="width:223px;"></td>
			<td>Any generic product</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">3-0 resorbable suture</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td>VCP824G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">5-0 non absorbable suture</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td>8618G</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:279px;">4-0 polyfilament sutures</td>
			<td style="width:236px;">Ethicon Inc., USA</td>
			<td style="width:223px;">VCP284G</td>
			<td></td>
		</tr>
	</tbody>
</table>

the rabbit blood-shunt model for the study of acute and late sequelae of subarachnoid hemorrhage: technical aspects

初期の脳損傷および遅延脳血管攣縮の両方が、くも膜下出血（SAH）後に不利な結果に貢献しています。両方の条件をシミュレートし、再現可能かつ制御可能な動物モデルは、現在、まれである。したがって、新しいモデルはSAHに起因するヒトの病態生理学的状態を模倣するために必要とされる。 このレポートでは、脳内の圧力（ICP）の制御を可能にウサギ血液シャントSAHモデルの技術的なニュアンスを説明しています。体外シャントは、動脈系と閉じた頭蓋内、検者に依存しないSAHを可能にくも膜下腔、の間に配置される。ステップバイステップの手続きの手順や必要な機器が記載されているだけでなく、技術的な検討事項最小限の死亡率および罹患率を持つモデルを生成する。この堅牢シンプルで一貫性のICP-制御SAHのウサギモデルの成功の外科的作成に必要な重要な詳細が説明されている。

<html> SAHのウサギ血液シャントモデルは、このレポートに記載されて負傷した後、早ければ24時間として、海馬（ 図2A、B）、基底皮質（ 図2A、B）及び脳血管系（ 図2C）でEBIを生成し、特性を示している血液分布（ 図2D）8。また、モデルはSAH誘導後3日目（ 図3）10 日にDCVS重度の度に中程度をトリガします?...

Watch this Scientific Journal Video about The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects at JoVE.com

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

くも膜下出血の急性および後期後遺症の研究のためのウサギの血液シャントモデル：技術的側面

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

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13.5K ビュー.  University and Bern University Hospital (Inselspital). The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures — subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

ビデオ: The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Microsurgical Venous Pouch Arterial-Bifurcation Aneurysms in the Rabbit Model: Technical Aspects

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials have shown disappointing long-term results with unacceptable high rates of aneurysm recanalization and delayed aneurysm rupture.2 To overcome these problems, animal experimental studies are crucial for the development of better endovascular devices.3-5
Several animal models in rats, rabbits, canines and swine are available.6-8 Comparisons of the different animal models showed the superiority of the rabbit model with regard to hemodynamics and comparability of the coagulation system and cost-effectiveness.9-11 
The venous pouch arterial bifurcation model in rabbits is formed by a venous pouch sutured into an artificially created true bifurcation of both common carotid arteries (CCA). The main advantage of this model are true bifurcational hemodynamics.12 The major drawbacks are the sofar high microsurgical technical demands and high morbidity and mortality rates of up to 50%.13 These limitations have resulted in less frequent use of this aneurysm model in the recent years. These shortcomings could be overcome with improved surgical procedures and modified peri- and postoperative analgetic management and anticoagulation.14-16 Our techniques reported in this paper demonstrate this optimized technique for microsurgical creation of arterial bifurcation aneurysms.

An optimized technique for the microsurgical creation of arterial bifurcation aneurysms mimicking bifurcation human cerebral aneurysms is described. A venous pouch is sutured into an artificially created true bifurcation of both common carotid arteries. Facilitated microsurgical techniques and aggressive postoperative anticoagulation and analgesia lead to minimized morbidity rates and high aneurysm patency rates.

ウサギモデルにおける顕微静脈ポーチ動脈 - 分岐瘤：技術的側面

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials ...

医学

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

PTSDの、ニューラル血液学的および行動的相関関係を明らかにするための動物モデルにおけるバイオマーカー

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

NADH Fluorescence Imaging of Isolated Biventricular Working Rabbit Hearts

Since its inception by Langendorff1, the isolated perfused heart remains a prominent tool for studying cardiac physiology2. However, it is not well-suited for studies of cardiac metabolism, which require the heart to perform work within the context of physiologic preload and afterload pressures. Neely introduced modifications to the Langendorff technique to establish appropriate left ventricular (LV) preload and afterload pressures3. The model is known as the isolated LV working heart model and has been used extensively to study LV performance and metabolism4-6. This model, however, does not provide a properly loaded right ventricle (RV). Demmy et al. first reported a biventricular model as a modification of the LV working heart model7, 8. They found that stroke volume, cardiac output, and pressure development improved in hearts converted from working LV mode to biventricular working mode8. A properly loaded RV also diminishes abnormal pressure gradients across the septum to improve septal function. Biventricular working hearts have been shown to maintain aortic output, pulmonary flow, mean aortic pressure, heart rate, and myocardial ATP levels for up to 3 hours8.
When studying the metabolic effects of myocardial injury, such as ischemia, it is often necessary to identify the location of the affected tissue. This can be done by imaging the fluorescence of NADH (the reduced form of nicotinamide adenine dinucleotide)9-11, a coenzyme found in large quantities in the mitochondria. NADH fluorescence (fNADH) displays a near linearly inverse relationship with local oxygen concentration12 and provides a measure of mitochondrial redox state13. fNADH imaging during hypoxic and ischemic conditions has been used as a dye-free method to identify hypoxic regions14, 15 and to monitor the progression of hypoxic conditions over time10.
The objective of the method is to monitor the mitochondrial redox state of biventricular working hearts during protocols that alter the rate of myocyte metabolism or induce hypoxia or create a combination of the two. Hearts from New Zealand white rabbits were connected to a biventricular working heart system (Hugo Sachs Elektronik) and perfused with modified Krebs-Henseleit solution16 at 37 &deg;C. Aortic, LV, pulmonary artery, and left &amp; right atrial pressures were recorded. Electrical activity was measured using a monophasic action potential electrode. To image fNADH, light from a mercury lamp was filtered (350&plusmn;25 nm) and used to illuminate the epicardium. Emitted light was filtered (460&plusmn;20 nm) and imaged using a CCD camera. Changes in the epicardial fNADH of biventricular working hearts during different pacing rates are presented. The combination of the heart model and fNADH imaging provides a new and valuable experimental tool for studying acute cardiac pathologies within the context of realistic physiological conditions.

The objective is to monitor the mitochondrial redox state of isolated hearts within the context of physiologic preload and afterload pressures. A biventricular working rabbit heart model is presented. High spatiotemporal resolution fluorescence imaging of NADH is used to monitor the mitochondrial redox state of epicardial tissue.

隔離された両心室ワーキングウサギ心臓のNADH蛍光イメージング

Since its inception by Langendorff1, the isolated perfused heart remains a prominent tool for studying cardiac physiology2. However, it is not well-suited ...

A Low Mortality Rat Model to Assess Delayed Cerebral Vasospasm After Experimental Subarachnoid Hemorrhage

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) in rats, in order to identify the initiating events, pathophysiological changes and potential targets for treatment.
Methods: Twenty-eight male Sprague-Dawley rats (250 - 300 g) were arbitrarily assigned to one of two groups - SAH or saline control. Rat subarachnoid hemorrhage in the SAH group (n=15) was induced by double injection of autologous blood, 48 hr apart, into the cisterna magna. Similarly, normal saline (n=13) was injected into the cisterna magna of the saline control group. Rats were sacrificed on day five after the second blood injection and the brains were preserved for histological analysis. The degree of vasospasm was measured using sections of the basilar artery, by measuring the internal luminal cross sectional area using NIH Image-J software. The significance was tested using Tukey/Kramer's statistical analysis.
Results: After analysis of histological sections, basilar artery luminal cross sectional area were smaller in the SAH than in the saline group, consistent with cerebral vasospasm in the former group. In the SAH group, basilar artery internal area (.056 &mu;m &plusmn; 3) were significantly smaller from vasospasm five days after the second blood injection (seven days after the initial blood injection), compared to the saline control group with internal area (.069 &plusmn; 3; p=0.004). There were no mortalities from cerebral vasospasm.
Conclusion: The rat double SAH model induces a mild, survivable, basilar artery vasospasm that can be used to study the pathophysiological mechanisms of cerebral vasospasm in a small animal model. A low and acceptable mortality rate is a significant criterion to be satisfied for an ideal SAH animal model so that the mechanisms of vasospasm can be elucidated 7, 8. Further modifications of the model can be made to adjust for increased severity of vasospasm and neurological exams.

Aneurysmal subarachnoid hemorrhage (SAH) is bleeding that occurs into the subarachnoid space when an aneurysm ruptures. While the morbidity and mortality from this event has been on a decline due to improved treatment approaches, the risk of vasospasm after subarachnoid hemorrhage continues to be the same as it was several years ago. The importance of establishing a comprehensive and reproducible animal model to identify initiating events of cerebral vasospasm has been the focus of research since the first use of rats in an experimental vasospasm model in 1979 by Barry et al. Early work in rats demonstrated that a single injection of autologous blood into the cisterna magna led to acute (within minutes) but not delayed cerebral vasospasm 3, 6, 14. Here we characterize a low mortality SAH rat model that results in reproducible delayed vasospasm.

実験的くも膜下出血後の脳血管攣縮遅延を評価するための低死亡率のラットモデル

Objective: To characterize and establish a reproducible model that demonstrates delayed cerebral vasospasm after aneurysmal subarachnoid hemorrhage (SAH) ...

Human Neuroendocrine Tumor Cell Lines as a Three-Dimensional Model for the Study of Human Neuroendocrine Tumor Therapy

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human malignancies.1 Other than surgery for the minority of patients who present with localized disease, there is little or no survival benefit of systemic therapy. Therefore, there is a great need to better understand the biology of NETs, and in particular define new therapeutic targets for patients with nonresectable or metastatic neuroendocrine tumors. 3D cell culture is becoming a popular method for drug screening due to its relevance in modeling the in vivo tumor tissue organization and microenvironment.2,3 The 3D multicellular spheroids could provide valuable information in a more timely and less expensive manner than directly proceeding from 2D cell culture experiments to animal (murine) models.
To facilitate the discovery of new therapeutics for NET patients, we have developed an in vitro 3D multicellular spheroids model using the human NET cell lines. The NET cells are plated in a non-adhesive agarose-coated 24-well plate and incubated under physiological conditions (5% CO2, 37 &deg;C) with a very slow agitation for 16-24 hr after plating. The cells form multicellular spheroids starting on the 3rd or 4th day. The spheroids become more spherical by the 6th day, at which point the drug treatments are initiated. The efficacy of the drug treatments on the NET spheroids is monitored based on the morphology, shape and size of the spheroids with a phase-contrast light microscope. The size of the spheroids is estimated automatically using a custom-developed MATLAB program based on an active contour algorithm. Further, we demonstrate a simple method to process the HistoGel embedding on these 3D spheroids, allowing the use of standard histological and immunohistochemical techniques. 
This is the first report on generating 3D spheroids using NET cell lines to examine the effect of therapeutic drugs. We have also performed histology on these 3D spheroids, and displayed an example of a single drug's effect on growth and proliferation of the NET spheroids. Our results support that the NET spheroids are valuable for further studies of NET biology and drug development.

We present a simple agarose overlay platform to grow 3D multicellular spheroids using neuroendocrine cancer cell lines. This method provides a very convenient way to examine the effect of therapeutic drugs on the neuroendocrine tumor cells. It could also help us establish human neuroendocrine tumor spheroids for cancer therapy.

人間の神経内分泌腫瘍療法の研究のための三次元モデルとしての人間の神経内分泌腫瘍細胞株

Neuroendocrine tumors (NETs) are rare tumors, with an incidence of two per 100,&#x200A;000 individuals per year, and they account for 0.5% of all human ...

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

再狭窄の研究のための頸動脈内ステント留置のマウスモデル

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

マウスAortocaval瘻の技術的な側面

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

くも膜下出血のマウスモデル

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to atherosclerotic plaque development or the rupturing of established plaques, interrupts normal blood flow leading to various morbidities such as myocardial infarction and stroke. In the clinic endovascular procedures such as angioplasty are commonly performed to reopen the lumen. However, these treatments inevitably damage the vessel wall as well as the vascular endothelium, triggering an excessive healing response and the development of a neointimal plaque that extends into the lumen causing vessel restenosis (re-narrowing). Restenosis remains a major cause of failure of endovascular treatments for atherosclerosis. Thus, preclinical animal models of restenosis are vitally important for investigating the pathophysiological mechanisms as well as translational approaches to vascular interventions. Among several murine experimental models, femoral artery wire injury is widely accepted as the most suitable for studies of post-angioplasty restenosis because it closely resembles the angioplasty procedure that injures both endothelium and vessel wall. However, many researchers have difficulty utilizing this model due to its high degree of technical difficulty. This is primarily because a metal wire needs to be inserted into the femoral artery, which is approximately three times thinner than the wire, to generate sufficient injury to induce prominent neointima. Here, we describe the essential surgical details to effectively overcome the major technical difficulties of this model. By following the presented procedures, performing the mouse femoral artery wire injury becomes easier. Once familiarized, the whole procedure can be completed within 20 min.

The mouse femoral artery wire injury model of restenosis is technically challenging. In this protocol we show the key technical details essential for successfully performing wire injury to induce consistent&#160;neointima for studies of restenosis.

動脈再狭窄のマウスモデル：大腿ワイヤー傷害の技術的な側面

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

ワイヤー図法を使用してマウスの腸間膜動脈の等尺性収縮力測定

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...