We thank Gilles Mithieux and Aude Barataud (INSERM U1213, Lyon, France) and Marie Liabeuf and Stephanie Lemarchand-Minde (Animal facility, l'Institut du Thorax, Nantes, France) for their help with the animal care protocol. This work was supported by grants from La r&#233;gion des Pays de la Loire, the Fondation d'Avenir, and the Casden Bank. We would like to thank Catherine Postic, Fadila Benhamed and Michelle Ca&#252;zac from l'institut Cochin for their hospitality and their help during the filming process.

動物とハウジング：8週齢のC57BL / 6雄マウスを入手します。脂肪から35％キロカロリー、25.8パーセントのカゼイン、1.30パーセントのミネラルAIN、1.30パーセントのビタミン、1.70パーセントのリン酸二カルシウム、0.7：10週齢では、水をC57BL / 6マウスの自由なアクセスと高脂肪食（DIOダイエットを与えます8（SG）または14（RYGB）の％炭酸カルシウム2.10％、クエン酸カリウム、0.026％の酒石酸水素コリン、8.9％スクロース、0.384％シスチン、6.5％メチルセルロース、31.7％のラード、3.3％の大豆油、及び16.29％のマルトデキストリン）週肥満手術の前に。体重増加を促進するために、高脂肪食に加えて（w / vの）20％のフルクトースを含むRYGB手術（シャム＆RYGB）水に専用のマウスを与えます。 倫理に関する声明：以下で説明するすべてのプロトコルステップは、動物実験のための倫理委員会のガイドラインに従うの承認番号01953.01下・ド・ラ・ロワールを支払います。 1.一般的な術前の準備<ol><li>手術前に6時間マウスを絶食。ジェルディらの食品は3日、手術の前に手術し、排他的に1日前に与えられました。 5％イソフルラン（0.4 L /分）でチャンバー内で麻酔を誘導し、O 2（二原子酸素、0.4 L /分）。麻酔が有効であることを確認するために、つま先ピンチテストを実行します。皮下注射を介して、鎮痛剤（0.1ミリグラム/キログラムブプレノルフィン）、抗生物質（10mg / kgのmarbofloxacine）、およびプロ動態（1ミリグラム/キログラムメトクロプラミド）を管理します。 </li><li> （実験室でのトラフィックや換気ファンから遠い）の外科的処置のための具体的な作業領域を割り当てます。専用のハード面の消毒剤と手術領域をきれいにしてください。手術中にマウスhomeothermyを維持するために、作業スペース領域に加熱パッドを配置します。加熱パッドの上にきれいな、吸収性アンダーパッドを適用します。 </li><li>電気かみそりを使用して、骨盤に胸骨から腹部を剃ります。滅菌ラップ（ 図1Aおよび2A）でのマウスの腹部をラップし、加熱pに仰臥位でマウスを置きます広告。 </li><li>麻酔下ながら乾燥を防ぐために、目に獣医軟膏を適用します。ノーズコーンにマウスを置き、2％イソフルラン（0.4 L /分）とO 2（0.4 L /分）で麻酔を維持します。 </li><li>手術を開始する前に、無菌試験のガウン、使い捨てスクラブキャップ、フェイスマスク、および滅菌手袋を着用。手術のすべてのステップのために必要とされる滅菌手術器具セットを含むオートクレーブしたパッケージを開きます。滅菌ラップの窓をカットし、2つの連続ポビドンヨードソリューションで皮膚を消毒します。 </li></ol> 2.スリーブ胃切除：手術の手順<ol><li> 中央値開腹（図1B） <ol><li>双眼顕微鏡下とメスで、（腹部の筋肉に損傷を与えない）腹腔を開くには、腹部の中央に胸骨から正中切開を行います。 37℃の生理食塩水に浸した滅菌圧縮で皮膚を保護します。 </li></ol></li><li> 胃の暴露（図1C） <ol><li>静かに湿らせ、綿棒を用いて、胃を動員。湿らせた綿棒やマイクロはさみを使用して、その横に近い結合組織の添付ファイルから胃を解放します。 </li><li>静かに、完全に胃を外部化し、止血コラーゲンは胃の後ろに圧縮配置します。 （ 図1D及びE）を出血未来を避けるために8.0非吸収性縫合糸で胃大弯に沿って、幽門と食道の血管を縫合。注：止血剤の使用は、組織の損傷を引き起こすだけ切除することを目指した組織でそれを使用することを確認することができます。 </li></ol></li><li> 胃の心臓領域の切除（眼底） <ol><li>幽門領域との間の解剖学的ライン存在（コーパス）と胃の心臓領域（眼底）（ 図1F）に胃切開を行います。 </li><li>それぞれの方の2湿らせた綿棒で残留食べ物を削除します胃の側面。マイクロはさみを使用して、眼底および胃（ 図1G）のコーパスとの間の境界に沿って胃を切りました。 </li><li>大出血の場合には2分間圧縮止血コラーゲンを押します。切開（ 図1H）の最後に胃食道接合部から8.0非吸収性縫合糸と胃の開口部を閉じます。 </li></ol></li><li> 胃の幽門部の切除（コーパス） <ol><li>胃（ 図1I）の約80％を除去するように大きな曲率に沿って胃の切除を完了します。 3ミリメートルにSGの幅を標準化。大出血の場合には2分間圧縮止血コラーゲンを押します。 </li><li> 8.0非吸収性縫合糸（ 図1J）と胃の幽門部の開口部を閉じます。静かに縫合糸が漏れ防止であることを保証するために、胃の各側に2湿らせた綿棒を転がし。 </li><li>漏れの場合8.0非吸収性縫合糸で完了します。止血コラーゲン圧縮を削除し、腹腔に胃を返します。縫合糸に対して圧縮止血コラーゲンを残します。 </li></ol></li><li> 腹部閉鎖 <ol><li> 5.0非吸収性縫合糸（ 図1K）と腹壁の筋層を閉じます。 1パーセントにイソフルラン濃度を低減することにより、麻酔を減らします。 5.0非吸収性縫合糸（ 図1L）を用いて皮膚を閉じます。 </li></ol></li></ol> 3.ルーワイ胃バイパス：手術の手順<ol><li> 中央値開腹 <ol><li>双眼顕微鏡下とメスで、（腹部の筋肉に損傷を与えない）腹腔を開くには、腹部の中央に胸骨から正中切開を行います。 37℃の生理食塩水（ 図2B）に浸した滅菌湿布で皮膚を保護します。 </li></ol> &lt;/李&gt; <li> Biliopancreatic手足と消化四肢 <ol><li>腸（ 図2C）を外部化。メジャー8幽門からセンチと5.0非吸収性縫合糸（図2D）と腸の2合字を行います。 </li><li> 2合字（ 図2E）との間の腸をカット。腹部の左上の象限内の2つの端部の近位四肢を置きます。注：これは、消化肢（ 図2F）として使用されます。 </li><li> 6センチメートル四肢近位（ 図2F）以下の消化四肢が直面している両端の遠位四肢を置きます。 注：これは胆汁手足として使用されます。 </li></ol></li><li> Jejuno-空腸造瘻術 <ol><li>マイクロはさみを使用して、四肢近位および腸のループの両方をカットし、同じ長さ（ 図2Gおよび2H）の2 antimesenteric切開を行います。 </li><li> 2 8.0側側吻合を行います非吸収性縫合糸。腹側吻合（ 図2J）に続く背側吻合最初（ 図2I）を実行します。 </li><li>静かに縫合糸が漏れ防止であることを保証するために吻合部の各側に2湿らせた綿棒を転がし。漏れの場合8.0非吸収性縫合糸で完了します。 </li></ol></li><li> 胃空腸造瘻術 <ol><li>静かに湿らせた綿棒を使って胃を動員。湿らせた綿棒やマイクロはさみを使用して、その横に近い結合組織の添付ファイルから胃を解放します。 </li><li>優しく、胃の後ろに再吸収性の止血コラーゲン湿布を配置し、完全に胃を外部化。湾曲したマイクロ鉗子（ 図2Kおよび2L）を使用して、網を通過した5.0非吸収性縫合糸を使用して、幽門結紮を行います。 </li><li>胃、幽門〜1.5 cmであり、遠位四肢の腹側の両方をカット同じ長さ（ 図2M）の2切開部を作成し、マイクロはさみを使用して。 </li><li> 2 8.0非吸収性縫合糸で側側吻合を実行します。背側吻合（ 図2N）で起動し、腹側吻合（ 図2O）を行います。静かに縫合糸が漏れ防止であることを保証するために吻合部の各側に2湿らせ、綿棒を転がし。漏れの場合8.0非吸収性縫合糸で完了します。 </li></ol></li><li> 腹部閉鎖 <ol><li> 5.0非吸収性縫合糸（ 図2P）を使用して腹壁の筋層を閉じます。 1パーセントにイソフルラン濃度を低減することにより、麻酔を減らします。 5.0非吸収性縫合糸を用いて皮膚を閉じます（ 図2Q）.Administer皮下注射を介して、温かい生理食塩水を25mL / kgです。 </li></ol></li></ol> 4.一般的な術後ケア<ol><li>イソフルランを停止マウスが完全に目を覚ましになるまで0.8 L / minのO 2流量を続行します。それは動物によって示されるモータ制御、ケージの周りを移動するために開始し、落下せずに立って歩くことができることを取り戻すまで、無人、マウスを放置しないでください。 </li><li> 5日間30℃の温度条件下でインキュベーターにマウス（ケージごとに1つだけのマウス）を配置します。 </li><li>プロトコルの最後までRYGBマウスにおけるビタミン（800ミリグラム/ 180 mLの水中）の補充;鉄（皮下注射は0.5mg / kg /日）を維持。 </li><li>手術後5日間：ゲルダイエット食品（10％ラード、10％液糖、57％の水高脂肪ゲル食）への無料アクセスを返します。 3日手術後の固形食を再導入。 </li><li>皮下には、メロキシカム（手術後3日目に0日目から1日2回0.1ミリグラム/キログラム、）（1ミリグラム/キログラム、0日目から手術後3日目に）、メトクロプラミド（1ミリグラム/キログラム、0日目からにブプレノルフィンを注入します手術後5日目）、および0日目から3日目AFTEにmarbofloxacine（10mg / kgの、<html> rの手術）。 </li></ol> 5.術後の代謝パラメータの評価<ol><li> 食物摂取量測定 <ol><li>マウスは、手術後1週間回復できるようにします。ハウスマウスケージあたり1。ケージに入れ固体食事の量を測定します。重量24時間後に残っている食べ物。限り、必要に応じてこの手順を繰り返します。 </li></ol></li><li> 経口的ブドウ糖負荷試験 <ol><li>経口ボーラスの前に6時間マウスを絶食。尾の先端から血液滴を収穫し、時間0血糖値を決定するために、グルコメーターに挿入グルコースストリップにそれを適用します。 </li><li> 20％D-グルコース溶液（2グラム/ kg）を経口ボーラスを投与します。 、15、30、60、及び120分後に胃管栄養法の血糖値を測定します。 </li></ol></li><li> ヘモグロビンレベルの分析 <ol><li>血液学システムを使用して、ヘモグロビンレベルをサンプリングするために新鮮な血液の20μLを測定します。 </li></ol></li></ol>&quot;&gt; 6。安楽死<ol><li>腹腔内キシラジン/ケタミン溶液（10/80ミリグラム/キログラム）を注入することにより、マウスを麻酔。頸椎脱臼を行います。心臓が鼓動していないと眼球瞬目反射が失われることを保証することにより、完全な安楽死を確認します。 </li></ol>

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The authors declare no conflicts of interest.

肥満の成長の流行を克服するために、最初の肥満手術の手順は、米国で1960年代に出現しました。それ以来、世界中で毎年行われる手順の数は依然として増加し、今日、これらの技術は、病的肥満6の管理のための最善の治療オプションを表します。開発手順の中では、SGとRYGBは、臨床実習4で使用される2つの最も一般的な方法です。動物モデル、特に齧歯動...

肥満や糖尿病の世界的大流行は、重症度の壊滅的です。世界中で20億以上の成人（人口の30％）が、過体重（BMI&gt; 25キロ/ m 2）で、または肥満（BMI&gt; 30キロ/ m 2）でのいずれかである1。これは、罹患率及び死亡率の増加につながる、このような2型糖尿病、高血圧、脂質異常症などの代謝性合併症と一緒に来ることができます。肥満は全死亡率および癌2の有病率が増加します。何らかの効果的な非侵襲的な治療法がないために、肥満手術は、長期的、持続的な体重減少3、4につながることができます唯一のオプションを表します。異なる外科多くの方法が開発されてきたが、スリーブ胃切除（SG）とルーワイ胃バイパス（RYGB）は、最も一般的に臨床で使用される2つの手順です。 SG手順の間、胃の初期体積の80％です削除;従って、この技術は、満腹感を改善制限手術の一つです。 RYGBは限定的-吸収不良技術の一つです。 RYGBの間に、小さな胃嚢（総胃の容積の1から2パーセント）が作成され、腸は消化と栄養素の吸収を遅延させるY字型、に再配置されます。これら2つの技術がRYGBに見られる高い効率で、大幅な体重の減少と頻繁に関連併存疾患（ 例えば、高血圧症、2型糖尿病、脂質異常症）3の一般的な改善につながります。しかし、肥満手術の多面的な有益な効果の背後にある分子メカニズムは、多くの場合、完全には解明されていません。 、利用可能な多数の遺伝的に改変された株に、マウスモデルは、これらの分子メカニズムを探求するための最も便利な動物モデルです。 しかし、肥満の手順は、小動物モデルとrequirに直接適応することが困難です高い外科手術器用さを電子。 SGが簡単に非常に良好な生存率とげっ歯類で行うことができますが、RYGBは厳しい腸閉塞5匹のマウスで致死的です。異なる修飾RYGB法は、この問題、特にoesojejunostomy 5に対抗するために提案されています。胃切除せずに胃空腸吻合術：ここでは、別の代替案を提示します。この修正されたRYGBは再現ヒトで観察された有益な効果のほとんど（ すなわち、有意な体重減少とグルコースと脂質の恒常性の向上）。 この原稿は、要約したマウスでSGとRYGBの技術的および実験の詳細を議論し、動画の助けを借りて、これらの手順を容易にするためにすることを目指しています。特定のハイライトは、ビタミン、鉄欠乏の低減を可能に術前と術後の医療プロトコルの最適化について説明します。

<table><tbody><tr><td>&lt;strong&gt;Drugs&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>High Fat diet</td><td>DIO diet</td><td>Safe</td><td></td></tr><tr><td>Isoflurane</td><td>Forane</td><td>Baxter</td><td></td></tr><tr><td>Buprenorphin</td><td>Buprecare</td><td>Animalcare</td><td></td></tr><tr><td>Marbofloxacine</td><td>Marbocyl</td><td>Vetoquinol</td><td></td></tr><tr><td>Ammonium iron citrate, vitamins PP-B12</td><td>Fercobsang</td><td>Vetoquinol</td><td></td></tr><tr><td>Vitamins A-D3-E-K-B</td><td>Vita Rongeur</td><td>Virbac</td><td></td></tr><tr><td>NaCl 0.9%</td><td>NaCl 0,9%</td><td></td><td></td></tr><tr><td>Povidone solution</td><td>Betadine Scrub</td><td>Betadine</td><td></td></tr><tr><td>Povidone solution</td><td>Betadine Solution</td><td>Betadine</td><td></td></tr><tr><td>Carboptol 980 NF</td><td>Ocrygel</td><td>TVM</td><td></td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;References&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Sutures&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Prolene&amp;reg;</td><td>8.0, 6,5 mm</td><td>Ethicon</td><td></td></tr><tr><td>Prolene&amp;reg;</td><td>5.0, 13 mm</td><td>Ethicon</td><td></td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;References&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Surgical equipments&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Scissors</td><td></td><td>FST</td><td></td></tr><tr><td>Needle holder</td><td>Olsen-Hegar</td><td>FST</td><td></td></tr><tr><td>Micro scissors</td><td>Vannas</td><td>FST</td><td></td></tr><tr><td>Micro forceps&amp;nbsp;</td><td>Graefe</td><td>FST</td><td></td></tr><tr><td>Micro forceps curved</td><td>Graefe</td><td>FST</td><td></td></tr><tr><td>Curved micro needle holder</td><td>Castroviejo</td><td>FST</td><td></td></tr><tr><td>Hemostatic collagen compress</td><td>Pangen</td><td>Urgo</td><td></td></tr><tr><td>Absorbent underpads</td><td></td><td>VWR</td><td></td></tr><tr><td>&lt;strong&gt;Name&lt;/strong&gt;</td><td>&lt;strong&gt;References&lt;/strong&gt;</td><td>&lt;strong&gt;Company&lt;/strong&gt;</td><td>&lt;strong&gt;Comments&lt;/strong&gt;</td></tr><tr><td>&lt;strong&gt;Specific equipments&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Hematology system&amp;nbsp;</td><td>Hemavet 950FS</td><td>Hemavet</td><td></td></tr><tr><td>Glucose strips and glucometer</td><td>One touch Verio</td><td>Life scan</td><td></td></tr><tr><td>Stereo microscope</td><td>MZ6</td><td>Leica&amp;nbsp;</td><td></td></tr></tbody></table>

techniques of sleeve gastrectomy and modified roux-en-y gastric bypass in mice

肥満は（2003年〜2008年）欧州における女性のための男性のための4〜28％と6.2 36.5％の有病率と、主要な公衆衛生問題です。病的肥満は、しばしば寿命と品質を低下させる、例えば、2型糖尿病、高血圧、及び脂質異常症などの代謝性合併症と関連しています。任意の効果的な非侵襲的治療の非存在下では、肥満手術は、病的肥満の患者（ボディ・マス・インデックス（BMI）&gt; 40 kg /日M 2）のための貴重な治療選択肢が長期につながるで、代謝合併症における体重減少および改善を持続。しかし、肥満手術の有益な効果を維持する基礎となる細胞および分子メカニズムはまだ完全には理解されていません。 、利用可能な多数の遺伝的に改変された株に、マウスモデルは、肥満手術の多面的な有益な効果の背後にある分子メカニズムを探求するための最も便利な動物モデルです。ここでは、最適化されたhealthcを詳述しましたスリーブ胃切除および修正ルーワイ胃バイパス：2、最も広く使用されている肥満症治療手術のためのマウスでの方法と外科的なプロトコルがあります。肥満手術の治療効果の根底にある分子メカニズムを解読すると、新しい治療薬の標的を同定するの約束を提供しています。

一般的な条件 SG手続きの平均手術時間は49.3±1.5分でした。私たちは、胃の約80％のパーセントを表し、胃の62.8±5.0 mgの、削除しました。いいえマウスは、手術中に、または次の7日の間に死亡しませんでした。 Oneマウス（7.1％）があるため胃石によって引き起こされる胃閉塞の11 番目の術後日目に死?...

Watch this Scientific Journal Video about Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice at JoVE.com

Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice

Bariatric surgery is the most efficient way to reduce body weight and the deadly metabolic complications (diabetes, obesity, and dyslipidemia) frequently associated with morbid obesity. Mouse models of bariatric surgery represent a unique asset for deciphering molecular mechanisms behind the beneficial effects of these surgeries on diabetes, hypertension, and dyslipidemia.

スリーブ胃切除およびマウスにおける変更されたルーワイ胃バイパスのテクニック

Obesity is a major public health issue, with a prevalence of 4 to 28% for men and 6.2 to 36.5% for women in Europe (from 2003 to 2008). Morbid obesity is ...

techniques-sleeve-gastrectomy-modified-roux-en-y-gastric-bypass

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

11.9K ビュー.  INSERM, CNRS, UNIV Nantes. Bariatric surgery is the most efficient way to reduce body weight and the deadly metabolic complications (diabetes, obesity, and dyslipidemia) frequently associated with morbid obesity. Mouse models of bariatric surgery represent a unique asset for deciphering molecular mechanisms behind the beneficial effects of these surgeries on diabetes, hypertension, and dyslipidemia.

ビデオ: Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice

Modified Technique for Coronary Artery Ligation in Mice

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI we need better knowledge about pathophysiology of myocardial ischemia. This knowledge may be valuable to define new therapeutic targets for innovative cardiovascular therapies4. Experimental MI model in mice is an increasingly popular small-animal model in preclinical research in which MI is induced by means of permanent or temporary ligation of left coronary artery (LCA)5. In this video, we describe the step-by-step method of how to induce experimental MI in mice.
The animal is first anesthetized with 2% isoflurane. The unconscious mouse is then intubated and connected to a ventilator for artificial ventilation. The left chest is shaved and 1.5 cm incision along mid-axillary line is made in the skin. The left pectoralis major muscle is bluntly dissociated until the ribs are exposed. The muscle layers are pulled aside and fixed with an eyelid-retractor. After these preparations, left thoracotomy is performed between the third and fourth ribs in order to visualize the anterior surface of the heart and left lung. The proximal segment of LCA artery is then ligated with a 7-0 ethilon suture which typically induces an infarct size ~40% of left ventricle. At the end, the chest is closed and the animals receive postoperative analgesia (Temgesic, 0.3 mg/50 ml, ip). The animals are kept in a warm cage until spontaneous recovery.

The surgical procedure used to induce experimental myocardial infarction in mice begins with left thoracotomy between the third and the fourth ribs in order to visualize the anterior surface of the heart and left lung. The left coronary artery is ligated, the chest is closed and the mouse is allowed to recover spontaneously.

マウスにおける冠動脈結紮のために修飾されたテクニック

Myocardial infarction (MI) is one of the most important causes of mortality in humans1-3. In order to improve morbidity and mortality in patients with MI ...

医学

Roux-en-Y Gastric Bypass Operation in Rats

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality benefit is bariatric surgery1,2. Consequently, there has been a steady rise in the number of bariatric operations done worldwide in recent years with the Roux-en-Y gastric bypass (gastric bypass) being the most commonly performed operation3. Against this background, it is important to understand the physiological mechanisms by which gastric bypass induces and maintains body weight loss. These mechanisms are yet not fully understood, but may include reduced hunger and increased satiation4,5, increased energy expenditure6,7, altered preference for food high in fat and sugar8,9, altered salt and water handling of the kidney10 as well as alterations in gut microbiota11. Such changes seen after gastric bypass may at least partly stem from how the surgery alters the hormonal milieu because gastric bypass increases the postprandial release of peptide-YY (PYY) and glucagon-like-peptide-1 (GLP-1), hormones that are released by the gut in the presence of nutrients and that reduce eating12.
During the last two decades numerous studies using rats have been carried out to further investigate physiological changes after gastric bypass. The gastric bypass rat model has proven to be a valuable experimental tool not least as it closely mimics the time profile and magnitude of human weight loss, but also allows researchers to control and manipulate critical anatomic and physiologic factors including the use of appropriate controls. Consequently, there is a wide array of rat gastric bypass models available in the literature reviewed elsewhere in more detail 13-15. The description of the exact surgical technique of these models varies widely and differs e.g. in terms of pouch size, limb lengths, and the preservation of the vagal nerve. If reported, mortality rates seem to range from 0 to 35%15. Furthermore, surgery has been carried out almost exclusively in male rats of different strains and ages. Pre- and postoperative diets also varied significantly.
Technical and experimental variations in published gastric bypass rat models complicate the comparison and identification of potential physiological mechanisms involved in gastric bypass. There is no clear evidence that any of these models is superior, but there is an emerging need for standardization of the procedure to achieve consistent and comparable data. This article therefore aims to summarize and discuss technical and experimental details of our previously validated and published gastric bypass rat model.

Numerous studies using gastric bypass rat models have been recently conducted to uncover the underlying physiological mechanisms of Roux-en-Y gastric bypass operations. This article aims to demonstrate and discuss the technical and experimental details of our published gastric bypass rat model to understand advantages and limitations of this experimental tool.

ラットにおけるルー-en-Y法胃バイパス手術

Currently, the most effective therapy for the treatment of morbid obesity to induce significant and maintained body weight loss with a proven mortality ...

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used techniques require killing of the animals and postmortem recovery of the meal1,2. This approach prevents longitudinal studies to determine changes in gastric emptying with age and progression of disease. The commonly used [13C]-octanoic acid breath test for humans3 has been modified for use in mice4-6 and rats7 and we previously showed that this test is reliable and responsive to changes in gastric emptying in response to drugs and during diabetic disease progression8. In this video presentation the principle and practical implementation of this modified test is explained. As in the previous study, NOD LtJ mice are used, a model of type 1 diabetes9. A proportion of these mice develop the symptoms of gastroparesis, a complication of diabetes characterized by delayed gastric emptying without mechanical obstruction of the stomach10.
This paper demonstrates how to train the mice for testing, how to prepare the test meal and obtain 4 hr gastric emptying data and how to analyze the obtained data. The carbon isotope analyzer used in the present study is suitable for the automatic sampling of the air samples from up to 12 mice at the same time. This technique allows the longitudinal follow-up of gastric emptying from larger groups of mice with diabetes or other long-standing diseases.

Assessment of Gastric Emptying in Non-obese Diabetic Mice Using a [13C]-octanoic Acid Breath Test

Determination of gastric emptying with a non-invasive [13C]-octanoic acid breath test for tracking gastroparesis in female NOD LtJ mice.

[を使用した非肥満糖尿病マウスにおける胃排出能の評価<sup&gt; 13</sup&gt; C] - オクタン酸呼気テスト

Gastric emptying studies in mice have been limited by the inability to follow gastric emptying changes in the same animal since the most commonly used ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the aorta in mice using the sleeve method was short and easy averaging 20 min, permitting studies of iso/allo grafts. The following article describes the aortic transplantation procedure used in our laboratory. The mice were anesthetized with a mixture of 1.5% volume isoflurane and 100% oxygen through a face mask. At this point, the segment of the aorta between the renal arteries and its bifurcation was separated from the vena cava, freely prepared and clampedat the proximal and distal segments with a single silk suture. Prior to the removal of the aorta, a saline solution containing heparin was injected into the inferior vena cava. Then the aorta was cut between the clamps and a saline heparin solution was used to flush the lumen. The sleeve technique with monofilament sutures was used in order to transplant the abdominal aorta in the orthotopic position.

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

大動脈弁移植 - 教則ビデオのマウスモデルでスリーブを用いた

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

Sleeve Gastrectomy in Mice using Surgical Clips

The number of people who are overweight and obese is continually increasing both in the adult and adolescent populations. This coincides with the increased universal phenomenon of type 2 diabetes (T2D) and other metabolic problems. Bariatric surgery, such as SG, is currently one of the most effective and commonly used long-term treatment for obesity and T2D, but the association between them is not completely explored yet. The mechanisms underlying the outcomes seen after bariatric surgery in humans can be investigated based on preclinical animal studies. The SG reduces body weight, glucose levels and many metabolic parameters, and is easy to perform with a low incidence of complications. The goal of this work is to provide a simple method and an uncomplicated preclinical model of bariatric surgery in animals for researchers.

The prevalence of diabetes and obesity is continuously increasing worldwide. The mechanisms between diabetes, obesity and their associated mortality and co-morbidities need to be further investigated. Here, we present a protocol for sleeve gastrectomy (SG) in animals as an uncomplicated preclinical model of bariatric surgery.

外科用クリップを用いたマウスにおけるスリーブ胃切れ術

The number of people who are overweight and obese is continually increasing both in the adult and adolescent populations. This coincides with the ...

A Protocol for Roux-en-Y Gastric Bypass in Rats using Linear Staplers

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and metabolic changes are not well understood. Multiple factors are thought to play a role, including reduced caloric intake, decreased nutrient absorption, increased satiety, the release of satiety-promoting hormones, shifts in bile acid metabolism, and alterations in the gut microbiota.
The rat RYGB model presents an ideal framework to study these mechanisms. Prior work on mouse models have had high mortality rates, ranging from 17 to 52%, limiting their adoption. Rat models demonstrate more physiologic reserve to surgical stimulus and are technically easier to adopt as they allow for the use of surgical staplers. One challenge with surgical staplers, however, is that they often leave a large gastric pouch which is not representative of RYGB in humans. 
In this protocol, we present a RYGB protocol in rats that result in a small gastric pouch using surgical staplers. Utilizing two stapler fires which remove the forestomach of the rat, we obtain a smaller gastric pouch similar to that following a typical human RYGB. Surgical stapling also results in better hemostasis than sharp division. Additionally, the forestomach of the rat does not contain any glands and its removal should not alter the physiology of RYGB.
Weight loss and metabolic changes in the RYGB cohort were significant compared to the sham cohort, with significantly lower glucose tolerance at 14 weeks. Furthermore, this protocol has an excellent survival of 88.9% after RYGB. The skills described in this protocol can be acquired without previous microsurgical experience. Once mastered, this procedure will provide a reproducible tool for studying the mechanisms and effects of RYGB.

Roux-en-Y gastric bypass (RYGB) is performed to treat obesity and diabetes. However, the mechanisms underlying RYGB's efficacy are not fully understood, and studies are limited by technical difficulty leading to high mortality in animal models. This article provides instructions on how to perform RYGB in rats with high success rates.

リニア・ステープラーを用いたラットにおけるルー・アン・Y胃バイパスのプロトコル

Roux-en-Y gastric bypass (RYGB) is commonly performed for the treatment of severe obesity and type 2 diabetes. However, the mechanism of weight loss and ...

Single-Anastomosis Duodeno-Ileal Bypass with Sleeve Gastrectomy Model in Mice

Obesity is a major health issue worldwide. As a response, bariatric surgeries have emerged to treat obesity and its related comorbidities (e.g., diabetes mellitus, dyslipidemia, non-alcoholic steatohepatitis, cardiovascular events, and cancers) through restrictive and malabsorptive mechanisms. Understanding the mechanisms by which these procedures allow such improvements often require their transposition into animals, especially in mice, because of the ease of generating genetically modified animals. Recently, the single-anastomosis duodeno-ileal bypass with sleeve gastrectomy (SADI-S) has emerged as a procedure that uses both restrictive and malabsorptive effects, which is being used as an alternative to gastric bypass in case of major obesity. Thus far, this procedure has been associated with strong metabolic improvements, which has led to a marked increase in its use in daily clinical practice. However, the mechanisms underlying these metabolic effects have been poorly studied as a result of a lack of animal models. In this article, we present a reliable and reproducible model of SADI-S in mice, with a special focus on perioperative management. The description and use of this new rodent model will be helpful for the scientific community to better understand the molecular, metabolic, and structural changes induced by the SADI-S and to better define the surgical indications for clinical practice.

Single-anastomosis duodeno-ileal bypass (SADI-S) is an emerging bariatric procedure with important metabolic effects. In this article, we present a reliable and reproducible model of SADI-S in mice.

マウスにおけるスリーブ状胃切除モデルを用いた単一吻合十二指腸回腸バイパス術

Obesity is a major health issue worldwide. As a response, bariatric surgeries have emerged to treat obesity and its related comorbidities (e.g., diabetes ...

A Murine Model of Vertical Sleeve Gastrectomy

Bariatric surgery, such as vertical sleeve gastrectomy (VSG), is a surgery of the gastrointestinal tract that is performed for the purpose of weight loss. Bariatric surgery is currently the most effective long-term treatment for obesity. In addition to weight loss, bariatric surgery produces additional health benefits such as remission of type 2 diabetes, remission of hypertension, and decreased risk of developing certain types of cancer. The mechanisms beyond weight loss for these benefits remain incompletely defined. Therefore, animal models of bariatric surgery are being developed and validated to identify the mechanisms leading to these benefits, with the goal of improving understanding of gastrointestinal physiology and identifying new therapeutic targets. VSG has become the most commonly performed bariatric procedure in the clinic in the United States because it is highly effective at producing weight loss and metabolic improvement, and is simpler to perform than other bariatric procedures. Therefore, we have developed and validated a murine model of VSG. This murine VSG model recapitulates many of the effects of VSG seen in humans, including improved glucose and blood pressure regulation. The method is based on isolation of the stomach, ligation of gastric vessels, and removal of 70% of the stomach by transecting along the greater curvature of the stomach. We have successfully applied this surgical protocol to various genetically modified mouse lines to define the mechanistic contributors to the benefits of VSG. Furthermore, this murine VSG model has been combined with other surgical techniques, to achieve deeper mechanistic insight. Therefore, this is a simple and versatile model for studying gastrointestinal physiology and the health benefits of bariatric surgery.

The following describes the performance of vertical sleeve gastrectomy in mice. This is a type of weight-loss surgery that involves removal of approximately 70% of the stomach.

垂直方向のスリーブ胃切除術のマウスモデル

Bariatric surgery, such as vertical sleeve gastrectomy (VSG), is a surgery of the gastrointestinal tract that is performed for the purpose of weight loss. ...

生物学

Roux-en-Y Gastric Bypass Procedure: A Bariatric Surgical Procedure for Gastric Resectioning in Murine Model

Source: Ayer, A., et al., <a href="https://jove.unicartagenaproxy.elogim.com/v/54905">Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice.</a> J. Vis. Exp. (2017).
This video describes a bariatric surgery-Roux-en-Y procedure. This involves the cutting of the jejunum- the middle portion of the intestine- to generate two intestinal limbs. The biliary limb is attached to the existing intestinal loop and the alimentary limb is attached to the stomach, creating a gastric bypass. The gastric bypass created here helps to decrease the nutrient uptake and efficiently reduces body weight.

Roux-en-Y胃バイパス術:マウスモデルにおける胃切除術のための肥満外科手術

Source: Ayer, A., et al., Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice. J. Vis. Exp. (2017).
This video describes a ...

JoVE Encyclopedia of Experiments

げっ歯類モデル

成虫

Murine Vertical Sleeve Gastrectomy: A Surgical Procedure to Downsize Stomach in Experimental Mouse Models

Before beginning the procedure, apply veterinary ointment to the eyes of an anesthetized mouse and place the mouse on a heating pad. Remove the hair from the umbilicus to the axilla, gently holding the skin taut to avoid skin lesions, and use cotton tip applicators to sequentially disinfect the exposed skin with povidone-iodine and ethanol.
Wearing sterile gloves and using sterile technique, place sterile surgical drapes on either side of the animal. Cut a small hole in the foil to allow access to the mouse&#39;s abdomen and use a piece of autoclaved aluminum foil as a surgical drape. Make a skin incision from the mid-abdomen to the xiphoid cartilage.
Under a dissecting microscope, use iris scissors to cut through the body wall along the linea alba and use new cotton tip applicators to gently elevate the stomach out of the abdomen. Bluntly dissect the greater omentum from the greater curvature of the stomach.
Then, use 7.0 monofilament absorbable sutures to place two ligatures around the short gastric artery that runs between the fundus of the stomach and the spleen. When both ligatures have been placed, use spring scissors to cut between the two sutures and fully exteriorize the stomach from the abdominal cavity.
Place gauze under the stomach and hydrate the organ with saline. For a vertical sleeve gastrectomy procedure, use a 7.0 monofilament absorbable suture with a taper needle to ligate the prominent branches of the gastric artery and vein, just below the intended line of transection. Place a simple continuous line of 6.0 monofilament absorbable suture through both gastric walls, just below the intended line of transection, to prevent spillage of the gastric contents.
Place thin-tipped hemostats above the suture line and use spring scissors to cut between the suture line and the hemostats. Remove the transected gastric tissue from the sterile surgical field and use fresh cotton tip applicators to clean the blood and digesta from the stomach.
Use a 6.0 monofilament absorbable suture with a taper needle to reinforce the apposition of the gastric walls in a simple discontinuous pattern. Using a 20G gavage needle attached to a 20-milliliter syringe, perform gastric lavage with fresh saline and securely close the stomach with a minimum of 20 knots.
Then, gently press the stomach with a cotton applicator to test for leaks. If no leaks are detected, lavage the stomach with at least 60 more milliliters of fresh saline to remove any infectious particulates.
After the procedure, use a cotton tip applicator to return the stomach under the liver within the abdominal cavity and place an applicator along the dorsal aspect of the abdominal cavity to absorb any excess fluid.
Use a blunt 18G needle to inject lactated Ringer&#39;s solution with or without antibiotics directly into the abdominal cavity, and use 6.0 monofilament absorbable sutures to close the abdominal muscle layer in a simple discontinuous pattern and the skin layer in a simple continuous pattern. Then, place tissue adhesive on the skin and fold the skin over the suture line to protect the suture from disruption by the animal.

スリーブ胃切除およびマウスにおける変更されたルーワイ胃バイパスのテクニック

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