Name: technique of minimally invasive transverse aortic constriction in mice for induction of left ventricular hypertrophy
Uploaded: 2017-09-25T12:30:00.000+00:00
Duration: 8 min 34 s
Description: Watch this Scientific Journal Video about Technique of Minimally Invasive Transverse Aortic Constriction in Mice for Induction of Left Ventricular Hypertrophy at JoVE.com

この仕事にしたスイスの心血管系財団の助成金 (N ° 32016) 支えられ

男性 c57bl/6 j マウス (10 週間、25 ~ 30 g、n = 60)、このプロトコルで使用されます。動物は、農学部と高等教育と研究のフランスの省によって策定されたガイドラインに準拠して人道的なケアを受けるし、1986 年 11 月 24 日の欧州共同体理事会指令に従ってすべてのプロシージャを実行 (86/609/EEC) およびフランスの法律。プロトコルによって承認された、&#34; 地域倫理委員会動物実験 CREMEAS &#34; (#2016092816207606).
 1 です手術に備えて <ol><li>12/12 h ダーク/ライト サイクル (詳細資料の表を参照) のための食糧との標準的なケージで飼養施設到着後 1 週間のマウスを維持し、水の利用可能な 広告。自由.</li>
	<li>操作の日にマウス個々 のケージ、麻酔導入前に数分動物に追加のストレスを避けるために。手術の前日すべて手術器具を滅菌します</li>。
	<li>腹腔内ベンゼン ケタミン (51.4 mg/kg) およびキシラジン (3.3 mg/kg) 食塩水で希釈し混合物の単回投与を注入 (0.9 %nacl).</li>
	<li>つま先リトリート反射の不在によって麻酔の深さを確認してください</li>。
	<li>首と動物の胸を市販のカミソリで剃るし、剃毛エリア 70% アルコールで消毒します</li>。
	<li>動物を配置するには、臥位きれいなコルク作業パッドと粘着テープで足を修正します</li>。
</ol> 2。手術 <ol><li>無菌手技が手順全体で使用されます。自発的に呼吸で動物、胸骨 ( 図 1) を公開するために中間の胸にスープラ胸骨切痕から 11 刃ナイフで縦正中頸部切開 10 mm 以上を実行します</li>。
	<li>Crile 木 4/0 モノフィラメント ポリプロピレン宿泊縫合針ホルダーを渡すことによって、甲状腺を撤回し、作業パッドをテープします</li>。
	<li>気管を明らかにするマイクロ手術用鉗子でにべもなくあらかじめ気管の筋肉を区切ります</li>。
	<li>優しく滑らかな先端スライド閉じた顎を持つマイクロ手術用鉗子の湾曲した気管上、胸骨の後ろにします</li>。
	<li>によって慎重に滑らかな先端カーブタイプ顕微鏡鉗子の顎の開閉を実施済み気管の筋肉の下と、胸膜を離れて移動する胸骨の後ろに鈍的切離</li>。
	<li>滑らかな先端ストレート マイクロ手術鉗子で右スープラ鎖骨の筋肉を持ち、動物の優しく胸を引き出します</li>。
	<li>は、胸骨下骨ニッパーの下の顎をスライドさせ、3-4 mm の上部の部分胸骨切開 ( 図 2) を実行します。左に向かって少しミニ胸骨の下の部分を直接します</li>。
	<li>は、マイクロ手術針ホルダーを使用してミニ胸骨の両側に第 2 肋間スペースを 7/0 モノフィラメント ポリプロピレン宿泊縫合から内部に外を通過します。肋間と内部胸部血管または胸膜への損傷を避けるために肋骨胸骨角の近くに滞在します</li>。
	<li>両側 7/0 モノフィラメント宿泊ポリプロピレン縫合糸を使用して胸骨端を広げるし、粘着テープで作業台に固定します</li>。
	<li>ゆっくりの移動は、さておきあらかじめ気管筋、縦隔脂肪と低倍率 (2-3 X) 下大動脈弓を可視化する顕微鏡下に鉗子胸腺が滑らかな先端を使用して ( 図 3)。触れたり気胸の開発を防ぐために壁側胸膜を損傷しないよう注意します</li>。
	<li>は、鉗子 ( 図 4 A) を結ぶことによって大動脈のアーチの下の軟部組織を公開し、その顎を優しく 。優しく軟部組織の中の顎の開閉により 2 番目の抱き合わせ鉗子で大動脈のアーチの下の軟部組織にトンネルを準備します</li>。
	<li>は、大動脈弓の下で左の手にて結紮援助 ( 図 4 B) の目に通して 6/0 絹糸を結紮のセグメントを渡すし、鉗子の起源の間右手で開催を結ぶことによってそれを取得右の腕、左総頚動脈 ( 図 5).</li>
	<li>は 27 ゲージの針を 5 mm の長さにカット、Crile 針ホルダーとそれらを押すことによって両端を鈍らせます。滑らかな先端ストレート マイクロ手術鉗子で大動脈弓 ( 図 6) の横にある鈍い 27 ゲージ針を置き、針と右腕と左総頸動脈の間に大動脈の周りにぴったりと縫合糸を結ぶ( 図 7) 2 つの抱き合わせ鉗子を用いた動脈。ぴったりと縫合糸を結ぶ、4 つ追加ノットに続いて初期の二重結び目を実行します。すべての結び目がフラットであることを確認してください</li>。
	<li>, 退院後すぐにでもやさしく 0.4 mm 径の狭小化を達成するために針と削除再現横 65-70% 大動脈狭窄</li>。
	<li>は、胸骨端とあらかじめ気管の筋肉、大動脈弓周囲軟部組織の止血を確認します。吸収性止血ガーゼを入れてどこに血の滲みが観察されます。7/0 モノフィラメントポリプロピレン滞在胸骨端を広めるために使用される縫合糸削除します</li>。
	<li>は左第 2 肋間スペースの中に外からマイクロ手術針ホルダーに簡単な 6/0 モノフィラメント ポリプロピレン縫合を通過し、内部から右側第 2 肋間スペースの外に、です。肋間と内部胸部血管または胸膜への損傷を避けるために肋骨胸骨角の近くに滞在します</li>。
	<li>エッジをもたらす一緒に胸骨 Crile 木 6/0 モノフィラメント ポリプロピレン縫合糸を結ぶことによって針ホルダー</li>。
	<li>針ホルダー Crile 木と 1 つの層で縫合を実行 5/0 モノフィラメントポリプロピレンと皮膚を閉じます</li>。
	<li>同一の偽の手順を実行大動脈周囲縫合糸を結ぶことがなく、収縮操作します</li>。
</ol> 3。手術後の回復 <ol><li>動物を非常に密接に監視します個々 のケージにマウスを移動し、腹臥位。</li>。
	<li>許可まで完全に温暖化の光の下で回復するマウス覚醒 (麻酔を誘導した後 1 h 未満).</li>
	<li>術後鎮痛に対するブプレノルフィン 0.1 mg/kg を腹腔内注入します示されるように最初の 3 日のブプレノルフィンすべての 8 h の 0.1 mg/kg の皮下注射を繰り返します。</li>。
	<li>標準ケージ (檻あたり最大 3 マウス) とケージあたり最低 2 マウス操作マウスを配置します</li>。
</ol> 4。心の収穫 <ol><li>分析の日、ケタミンの溶液でマウスを安楽死させる 300 mg/kg とキシラジン 20 mg/kg 腹腔内投与による生理食塩水中で</li>。
	<li>下大静脈から血液をまず収穫し、同じラインを生理食塩水で 2.6 mm EDTA 溶液 5 mL を注入します</li>。
	<li>心を収穫、心房を削除および重量の心臓 (心房なし左と右心室).</li>
	<li>は右の ventr から左側を分離します。残りの左心室部分に隔壁を持つ icle。両方の組織のサンプルの重量を量るし、液体窒素で凍結します</li>。
</ol>

<ol>
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著者はある利益相反を開示します。

このプロトコルの目的は、マウスにおける低侵襲の横方向の大動脈狭窄に対する手技の順を追って図を提示することです。マウスで横方向の大動脈狭窄の詳細な技術的な説明は、他の著者2,8によって報告されています。ただし、これらの調査官は、手術を行う次の挿管や動物の換気。挿管換気の追加手順の使用が増加の複雑さと全体のプロシージ?...

過去年にわたって心不全の研究で実施されている実行可能な動物モデル1。小動物モデル心不全の大動物モデルと比較して、多数の潜在的な利点があります。住宅とメンテナンス コストの削減の横にある小さな動物モデルはより少なく複雑な設備を必要な2のための多くの研究者にアクセス可能です。
マウスモデル心不全ラットのモデルと同じ利点の多くを提供しています。さらに減少住宅コスト3、マウス モデルの恩恵を関連するトランスジェニック、ノックアウト (KO) の系統の可用性。携帯型固有の誘導 KO や遺伝子組換え戦略の可能性はマウスに心不全の病態を研究し、新規化学療法3を識別しようとする非常に貴重なツールを確認します。
マウスの中の心不全モデルは現在4を使用、最初のロックマン5で記述された横大動脈狭窄 (TAC) は圧力の過負荷による左心室の肥大 (LVH)1を生成する推奨モデル,3。 このモデルの最大の利点は LVH2の階層を許可する能力、左室が TAC に対応改造が異なるマウス系統間で変数。特に、c57bl/6 マウスは他系統4,6,7では発生可能性があります TAC 後急速な左心室拡張を開発します。
TAC の原因で高血圧症の突然の発症を約 50% 量の増加 LV 2 週間以内に急速に変調 LVH4の開発を目指して、薬理学的または分子の介入のアクティビティを確認することができます。TAC によって重篤な高血圧症の急性の誘導は進歩的な左室肥大を再現して丁度と大動脈弁狭窄症または高血圧症の臨床で観察された改造します。それにもかかわらず、多くの研究者がこのモデルを使用して、識別し、心不全4における新たな治療標的を変更します。
マウスでタックを実行すると、LVH とその後心不全2を誘導するために使用される他の技術のために必要なより大きい外科の専門知識が必要です。ほとんどの著者は、挿、換気、動物2,8, この手順より厳しい、時間がかかるし、動物の手術の負担を追加してこの手順を実行します。いくつか捜査官だけは、手術9,10,11を簡単に参照を研究で低侵襲の TAC を使用しています。
このプロトコルの目的は、ステップバイ ステップを説明するプロシージャの重要な段階を強調表示、マウスにおける低侵襲横大動脈狭窄の簡易で使いやすいテクニックです。これらのキーの手順を実行このテクニックを簡単に実行できます 1 つ。

<table><tbody><tr><td>Surgical microscope</td><td>Olympus</td><td>SZX2-TR30</td><td></td></tr><tr><td>Razor</td><td>Rowenta</td><td>Nomad TN3650FO</td><td></td></tr><tr><td>&lt;strong&gt;Sutures:&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Polypropylene 7/0</td><td>Ethicon</td><td>BV-1X</td><td></td></tr><tr><td>Polypropylene 6/0</td><td>BBraun</td><td>C0862061</td><td></td></tr><tr><td>Silk 6/0 ligature&amp;nbsp;</td><td>FST</td><td>18020-60</td><td></td></tr><tr><td>Polypropylene 4/0</td><td>Ethicon</td><td>8683</td><td></td></tr><tr><td>Polypropylene 5/0</td><td>Ethicon</td><td>Z303</td><td></td></tr><tr><td>&lt;strong&gt;Drugs:&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Ketamin</td><td>Merial</td><td>Imalg&amp;egrave;ne 1000, LBM154AD</td><td></td></tr><tr><td>Xylazine</td><td>Bayer</td><td>Rompun 2%, KP09PPC</td><td></td></tr><tr><td>Buprenorphine</td><td>Ceva</td><td>Vetergesic, 072013</td><td></td></tr><tr><td>&lt;strong&gt;Instruments:&amp;nbsp;&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Bone nippers</td><td>Fine Surgical Tools</td><td>16101-10</td><td></td></tr><tr><td>Ligation aid</td><td>Fine Surgical Tools</td><td>18062-12</td><td></td></tr><tr><td>Tying forceps</td><td>Fine Surgical Tools</td><td>18026-10</td><td></td></tr><tr><td>Needle holder Crile-Wood</td><td>Fine Surgical Tools</td><td>12003-15</td><td></td></tr><tr><td>Microsurgery forceps&amp;nbsp;</td><td>Fine Surgical Tools</td><td>11003-12</td><td></td></tr><tr><td>Microsurgery forceps&amp;nbsp;</td><td>Fine Surgical Tools</td><td>11002-12</td><td></td></tr><tr><td>Tissue forceps</td><td>Fine Surgical Tools</td><td>11021-12</td><td></td></tr><tr><td>Microsurgery needle holder</td><td>Fine Surgical Tools</td><td>12076-12</td><td></td></tr><tr><td>Microsurgery scissors</td><td>Fine Surgical Tools</td><td>91501-09</td><td></td></tr><tr><td>Mayo scissors</td><td>Fine Surgical Tools</td><td>14511-15</td><td></td></tr><tr><td>11-blade knife</td><td>Fine Surgical Tools</td><td>10011-00</td><td></td></tr><tr><td>&lt;strong&gt;RNA extraction and qPCR:&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>TriReagent</td><td>Euromedex</td><td>TR-118-200</td><td></td></tr><tr><td>Rneasy Mini kit</td><td>Qiagen</td><td>74704</td><td></td></tr><tr><td>Qubit Fluorimetric RNA assay</td><td>Fisher Scientific</td><td>10034622</td><td></td></tr><tr><td>RNA 6000 Nano kit</td><td>Agilent</td><td>5067-1511</td><td></td></tr><tr><td>High Capacity cDNA kit</td><td>Fisher Scientific</td><td>10400745</td><td></td></tr><tr><td>Taqman Master Mix</td><td>Fisher Scientific</td><td>10157154</td><td></td></tr><tr><td>Taqman BNP primers</td><td>Fisher Scientific</td><td>Mm01255770_g1</td><td></td></tr><tr><td>Taqman ANP primers</td><td>Fisher Scientific</td><td>Mm01255747_g1&amp;nbsp;</td><td></td></tr><tr><td>Taqman ACE primers</td><td>Fisher Scientific</td><td>Mm00802048_m1</td><td></td></tr><tr><td>Taqman Col1a1 primers&amp;nbsp;</td><td>Fisher Scientific</td><td>Mm00801666_g1</td><td></td></tr><tr><td>Taqman TGFb primers</td><td>Fisher Scientific</td><td>Mm01178820_m1</td><td></td></tr><tr><td>Taqman Gapdh primers</td><td>Fisher Scientific</td><td>Mm99999915_g1</td><td></td></tr><tr><td>ABIPrism&amp;nbsp; Thermocycler</td><td>Applied Biosystems</td><td>7000</td><td></td></tr><tr><td>&lt;strong&gt;Software:&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>GraphPad Prism</td><td>GraphPad</td><td>Prism 7</td><td></td></tr><tr><td>&lt;strong&gt;Animal food&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Complete diet for adult rats/mice</td><td>Safe</td><td>UB220610R</td><td></td></tr></tbody></table>

technique of minimally invasive transverse aortic constriction in mice for induction of left ventricular hypertrophy

マウスの横の大動脈狭窄 (TAC) は、心不全、心室の肥大 (LVH) とその進行を左オーバー ロードによる圧力の実験調査のため最もよく使用される手技のひとつです。報告された調査の大半は、この手順を実行して、挿管と厳しい、時間のかかるレンダリングし、動物に外科的負担に追加動物の換気。このプロトコルの目的は、挿管することがなく低侵襲の TAC の簡略化手法とマウスの換気を記述することです。技術の重要なステップは、死亡率は低い、LVH の誘導で高効率を達成するために強調されます。
男性の c57bl/6 マウス (10 週齢, 25-30 g, n = 60) ケタミン ・ キシラジンの混合物の腹腔内注射の麻酔をかけられました。自発的に呼吸動物 3-4 mm 上部部分の胸骨を次 6/0 絹糸結紮援助の目に通してのセグメントは大動脈弓の下を通過し、鈍化 27 ゲージ針を結んだ。動物の偽手術を受けた同じ手術の準備なく大動脈狭窄。LVH を誘導の手順の有効性は、心/体重比の大幅な増加によって立証されています。術後後 3、7、14、28 日でこの比率が得られた (n = 6-10 各グループと各時点)。私たちの技術を使用して、LVH は 28 日目から 7 日目から偽の動物と比較して TAC で観察されます。手術と後半 (28 日間) 死亡率がともに 1.7% で非常に低い。
結論としては、マウスにおける低侵襲の TAC の本のコスト効果の高い手法は非常に低い手術と術後の死亡率を運ぶ、LVH を誘導する効率が高い。それは手術の手順を簡素化し、動物の負担を軽減します。このプロトコルで説明する重要な手順に従うことによって簡単に実行できます。

手術と後半の生存 手術の生存率は非常に高かった (TAC と sham 動物) シリーズ全体の 98.3% (59 のうち 60)。のみ手術死はかんな偽操作のマウスで出血合併症が原因でした。28 日の観察期間中に術後生存率もすばらしい, 98.3% (59 のうち 58) だった後半のみ術後死は心臓由来の可能性がある日 (D) 16、TAC マウスで発生しました。
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Watch this Scientific Journal Video about Technique of Minimally Invasive Transverse Aortic Constriction in Mice for Induction of Left Ventricular Hypertrophy at JoVE.com

Technique of Minimally Invasive Transverse Aortic Constriction in Mice for Induction of Left Ventricular Hypertrophy

このプロトコルの目的は、段階的に説明するマウスにおける低侵襲横大動脈狭窄 (TAC) の技術。挿管や一般的に使用される標準的な手順に必要な換気の除去、TAC の低侵襲手術を簡素化し、動物の負担を軽減します。

左室肥大の誘導マウスにおける低侵襲の横大動脈狭窄のテクニック

Transverse aortic constriction (TAC) in mice is one of the most commonly used surgical techniques for experimental investigation of pressure ...

technique-minimally-invasive-transverse-aortic-constriction-mice-for

Research

JoVE Journal

Medicine

24.2K ビュー.  University of Zurich. このプロトコルの目的は、段階的に説明するマウスにおける低侵襲横大動脈狭窄 (TAC) の技術。挿管や一般的に使用される標準的な手順に必要な換気の除去、TAC の低侵襲手術を簡素化し、動物の負担を軽減します。

ビデオ: Technique of Minimally Invasive Transverse Aortic Constriction in Mice for Induction of Left Ventricular Hypertrophy

Ascending Aortic Constriction in Rats for Creation of Pressure Overload Cardiac Hypertrophy Model

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart failure. Here, we describe a detailed surgical procedure for creating pressure overload and cardiac hypertrophy in rats by constriction of the ascending aorta using a small metallic clip. After anesthesia, the trachea is intubated by inserting a cannula through a half way incision made between two cartilage rings of trachea. Then a skin incision is made at the level of the second intercostal space on the left chest wall and muscle layers are cleared to locate the ascending portion of aorta. The ascending aorta is constricted to 50&#8211;60% of its original diameter by application of a small sized titanium clip. Following aortic constriction, the second and third ribs are approximated with prolene sutures. The tracheal cannula is removed once spontaneous breathing was re-established. The animal is allowed to recover on the heating pad by gradually lowering anesthesia. The intensity of pressure overload created by constriction of the ascending aorta is determined by recording the pressure gradient using trans-thoracic two dimensional Doppler-echocardiography. Overall this protocol is useful to study the remodeling events and contractile properties of the heart during the gradual onset and progression from compensated cardiac hypertrophy to heart failure stage.

We describe a stepwise procedure for creating pressure overload and left ventricular hypertrophy in Wistar rats by constriction of the ascending aorta using a small metallic clip. This model is extensively used for studying remodeling changes during cardiac hypertrophy and for identifying and evaluating strategies for regression of such changes.

圧負荷心肥大モデルの作成のためのラットの大動脈狭窄昇順

Ascending aortic constriction is the most common and successful surgical model for creating pressure overload induced cardiac hypertrophy and heart ...

医学

Transverse Aortic Constriction in Mice

Transverse aortic constriction (TAC) in the mouse is a commonly used experimental model for pressure overload-induced cardiac hypertrophy and heart failure.1 TAC initially leads to compensated hypertrophy of the heart, which often is associated with a temporary enhancement of cardiac contractility. Over time, however, the response to the chronic hemodynamic overload becomes maladaptive, resulting in cardiac dilatation and heart failure.2 The murine TAC model was first validated by Rockman et al.1, and has since been extensively used as a valuable tool to mimic human cardiovascular diseases and elucidate fundamental signaling processes involved in the cardiac hypertrophic response and heart failure development. When compared to other experimental models of heart failure, such as complete occlusion of the left anterior descending (LAD) coronary artery, TAC provides a more reproducible model of cardiac hypertrophy and a more gradual time course in the development of heart failure. Here, we describe a step-by-step procedure to perform surgical TAC in mice. To determine the level of pressure overload produced by the aortic ligation, a high frequency Doppler probe is used to measure the ratio between blood flow velocities in the right and left carotid arteries.3, 4 With surgical survival rates of 80-90%, transverse aortic banding is an effective technique of inducing left ventricular hypertrophy and heart failure in mice.

Transverse aortic constriction (TAC) in the mouse is a commonly used experimental model to study mechanisms underlying cardiac hypertrophy and heart failure development. Here, we describe procedures to constrict the aorta to create a reproducible degree of cardiac hypertrophy in mice.

マウスの横大動脈狭窄

Transverse aortic constriction (TAC) in the mouse is a commonly used experimental model for pressure overload-induced cardiac hypertrophy and heart ...

生物学

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable noninvasive method is essential to assess real-time cardiac morphological and functional changes in animal models of heart disease. Transthoracic echocardiography represents an important tool for noninvasive assessment of cardiac structure and function2. Here we used a high-resolution ultrasound imaging system to monitor myocardial remodeling and heart failure progression over time in a mouse model of TAC. B-mode, M-mode, and Doppler imaging were used to precisely assess cardiac hypertrophy, ventricular dilatation, and functional deterioration in mice following TAC. Color and pulse wave (PW) Doppler imaging was used to noninvasively measure pressure gradient across the aortic constriction created by TAC and to assess transmitral blood flow in mice. Thus transthoracic echocardiographic imaging provides comprehensive noninvasive measurements of cardiac dimensions and function in mouse models of heart disease.

The goal of this protocol is to noninvasively assess cardiac structural and functional changes in a mouse model of heart disease created by transverse aortic constriction, using B- and M-mode echocardiography and color/pulse wave Doppler imaging.

心臓形態学的および心エコーイメージングによって横大動脈狭窄のマウスモデルにおける機能の変更の評価

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

Minimally Invasive Transverse Aortic Constriction in Mice

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard open-chest transverse aortic constriction (TAC). Although transverse aortic constriction is a highly functional method for the induction of high pressure in the left ventricle, it is a more difficult and lengthy procedure due to its use of artificial ventilation with tracheal intubation. TAC is oftentimes also less survivable, as the newer method, MTAC, neither requires the cutting of the ribs and intercostal muscles nor tracheal intubation with a ventilation setup. In MTAC, as opposed to a thoracotomy to access to the chest cavity, the aortic arch is reached through a midline incision in the anterior neck. The thyroid is pulled back to reveal the sternal notch. The sternum is subsequently cut down to the second rib level, and the aortic arch is reached simply by separating the connective tissues and thymus. From there, a suture can be wrapped around the arch and tied with a spacer, and then the sternal cut and skin can be closed. MTAC is a much faster and less invasive way to induce left ventricular hypertension and enables the possibility for high-throughput studies. The success of the constriction can be verified using high-frequency trans-thoracic echocardiography, particularly color Doppler and pulsed-wave Doppler, to determine the flow velocities of the aortic arch and left and right carotid arteries, the dimension of the blood vessels, and the left ventricular function and morphology. A successful constriction will also trigger significant histopathological changes, such as cardiac muscle cell hypertrophy with interstitial and perivascular fibrosis. Here, the procedure of MTAC is described, demonstrating how the resulting flow changes in the carotid arteries can be examined with echocardiography, gross morphology, and histopathological changes in the heart.

Minimally invasive transverse aortic constriction (MTAC) conserves the essentials of regular transverse aortic constriction (TAC) while eliminating the use of a ventilator with tracheal intubation. It proves to be a highly desirable method for high-throughput studies on left ventricular overload, particularly in translational studies.

マウスにおける低侵襲横大動脈狭窄

Minimally invasive transverse aortic constriction (MTAC) is a more desirable method for the constriction of the transverse aorta in mice than standard ...

A Closed-chest Model to Induce Transverse Aortic Constriction in Mice

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to perform a partial thoracotomy to visualize the transverse aortic arch. However, the surgical trauma caused by the thoracotomy in open-chest models changes the respiratory physiology as the ribs are dissected and left unattached after chest closure. To prevent this, we established a minimally invasive, closed chest approach via lateral thoracotomy. Herein we approach the aortic arch via the 2nd intercostal space without entering the chest cavities, leaving the mouse with a less traumatic injury to recover from. We perform this operation using standard laboratory settings for open chest TAC procedures with equal survival rates. Apart from maintaining physiological breathing patterns due to the closed chest approach, the mice seem to benefit by showing rapid recovery, as the less invasive technique appears to facilitate a fast healing process and to reduce immune response after trauma.

Here, we present a protocol of Transverse Aortic constriction (TAC) via a lateral thoracotomy. This technique is a minimally invasive, closed chest surgical procedure aiming to simulate pressure overload and heart failure in mice utilizing standard TAC laboratory settings.

マウスで横方向の大動脈狭窄を誘導するために閉じて胸モデル

Research on cardiac hypertrophy and heart failure is frequently based on pressure overload mouse models induced by TAC. The standard procedure is to ...

Studying Left Ventricular Reverse Remodeling by Aortic Debanding in Rodents

To better understand the left ventricular (LV) reverse remodeling (RR), we describe a rodent model wherein, after aortic banding-induced LV remodeling, mice undergo RR upon removal of the aortic constriction. In this paper, we describe a step-by-step procedure to perform a minimally invasive surgical aortic debanding in mice. Echocardiography was subsequently used to assess the degree of cardiac hypertrophy and dysfunction during LV remodeling and RR and to determine the best timing for aortic debanding. At the end of the protocol, terminal hemodynamic evaluation of the cardiac function was conducted, and samples were collected for histological studies. We showed that debanding is associated with surgical survival rates of 70-80%. Moreover, two weeks after debanding, the significant reduction of ventricular afterload triggers the regression of ventricular hypertrophy (~20%) and fibrosis (~26%), recovery of diastolic dysfunction as assessed by the normalization of left ventricular filling and end-diastolic pressures (E/e&#39;&#160;and LVEDP). Aortic debanding is a useful experimental model to study LV RR in rodents. The extent of myocardial recovery is variable between subjects, therefore, mimicking the diversity of RR that occurs in the clinical context, such as aortic valve replacement. We conclude that the aortic banding/debanding model represents a valuable tool to unravel novel insights into the mechanisms of RR, namely the regression of cardiac hypertrophy and the recovery of diastolic dysfunction.

Here we describe a step-by-step protocol of surgical aorta debanding in the well-established mice model of aortic-constriction. This procedure not only allows studying the mechanisms underlying the left ventricular reverse remodeling and regression of hypertrophy but also to test novel therapeutic options that might accelerate myocardial recovery.

げっ歯類における大動脈デベンディングによる左心室逆改造の研究

To better understand the left ventricular (LV) reverse remodeling (RR), we describe a rodent model wherein, after aortic banding-induced LV remodeling, ...

Operating Transverse Aortic Constriction with Absorbable Suture to Obtain Transient Myocardial Hypertrophy

Based on twice transverse aortic constrictions (TACs) in mice, it is proved that myocardial hypertrophic preconditioning (MHP) could attenuate cardiomyocyte hypertrophy and slow down progression to heart failure. For novices, however, the MHP model is usually quite difficult to establish because of the technical obstacles in ventilator operation, opening the chest repeatedly, and bleeding caused by debanding. To facilitate this model, to increase the surgical success rate and to reduce the incidence of bleeding, we switched to absorbable sutures for the first TAC combing with a ventilator-free technique. Using a 2-week absorbable suture, we demonstrated that this procedure could cause significant myocardial hypertrophy in 2 weeks; and 4 weeks after surgery, myocardial hypertrophy was almost completely regressed to the baseline. Using this protocol, the operators could master the MHP model easily with a lower operation mortality.

This protocol presents an improved method to obtain transient myocardial hypertrophy with absorbable suture, simulating left ventricular hypertrophy decrease after removing pressure overload. It could be valuable for the studies on myocardial hypertrophic preconditioning.

過渡性心筋肥大を得るために吸収性縫合を用いて横方向の大動脈収縮を操作する

Based on twice transverse aortic constrictions (TACs) in mice, it is proved that myocardial hypertrophic preconditioning (MHP) could attenuate ...

A Modified Technique for Transverse Aortic Constriction in Mice

Transverse aortic constriction (TAC) is a frequently used surgery in research regarding heart failure and cardiac hypertrophy based on the formation of pressure overload in mouse models. The main challenge of this procedure is to clearly visualize the transverse aortic arch and precisely band the target vessel. Classical approaches perform a partial thoracotomy to expose the transverse aortic arch. However, it is an open-chest model that causes a rather large surgical trauma and requires a ventilator during the surgery. To prevent unnecessary trauma and simplify the operating procedure, the aortic arch is approached via the proximal proportion of the sternum, reaching and binding the target vessel using a small self-made retractor that contains a snare. This procedure can be conducted without entering the pleura cavity and does not need a ventilator or microsurgical operation, which leaves the mice with physiological breathing patterns, simplifies the procedure, and significantly reduces operation time. Due to the less invasive approach and less operation time, mice can undergo fewer stress reactions and recover rapidly.

The present protocol describes a modified and simplified technique with a minimally invasive transverse aortic constriction (TAC) procedure using a self-made retractor. This procedure can be conducted without a ventilator or microscope and introduces pressure overload, eventually leading to cardiac hypertrophy or heart failure.

マウスにおける横行大動脈狭窄の修正技術

Transverse aortic constriction (TAC) is a frequently used surgery in research regarding heart failure and cardiac hypertrophy based on the formation of ...

Transverse Aortic Arch Banding in Mouse Model: A Minimally Invasive Technique in Mice for Induction of High Pressure in the Left Ventricle

Source: Zaw, A. M. et al. <a href="https://www.jove.com/video/55293" target="_blank">Minimally Invasive Transverse Aortic Constriction in Mice</a>. J. Vis. Exp. (2017)
In this video, we demonstrate an experimental protocol for minimally invasive transverse aortic constriction in mice to examine pressure overload-induced heart failure.

マウスモデルにおける横方向大動脈弓バンディング:左心室における高圧誘導のためのマウスにおける低侵襲技術

Source: Zaw, A. M. et al. Minimally Invasive Transverse Aortic Constriction in Mice. J. Vis. Exp. (2017)
In this video, we demonstrate an experimental ...

JoVE Encyclopedia of Experiments

げっ歯類モデル

成虫

Surgical Aortic Debanding: A Procedure to Study Left Ventricular Reverse Remodeling in Murine Model of Aortic Constriction

Source: Goncalves-Rodrigues, P. et al. <a href="https://www.jove.com/video/60036" target="_blank">Studying Left Ventricular Reverse Remodeling by Aortic Debanding in Rodents</a>. J. Vis. Exp. (2021)
This video demonstrates the technique of aortic debanding, which can lead to reverse remodeling to limit the progression towards heart failure in a murine model. Post debanding, a significant reduction of ventricular afterload triggers regression of ventricular hypertrophy and fibrosis, causing a recovery in cardiac function.