The authors would like to acknowledge the Cincinnati Children&#8217;s Hospital Division of Veterinary Services for their assistance. This manuscript was supported by the National Institutes of Health K12 HD028827. Anna Perl PhD taught the authors this surgical procedure.

注：動物use文：本研究におけるすべての手順を承認し、シンシナティ小児病院での動物実験使用とケア委員会（IACUC）のガイドラインに従って行った。 8週齢のC57BL / 6J雄マウスをジャクソン·ラボラトリーズ（バーハーバー、ME）から入手し、使用前に一週間順応させた。手術まで、動物を、無病原体バリア施設で飼育し、オートクレーブした食餌及びろ過水広告libdiumを与えた。各マウスのケージは、専用の空気と水を供給し、客室は、12時間の昼夜サイクルで維持した。手術からの回復に続いて、マウスは、濾過トップとのケージに維持し、オートクレーブ処理飼料適宜該ギプスを提供し、水のボトルからの水を濾過して設けられた。 楽器の調製<ol><li>ペーパークリップやピンを使用して6スキンリトラクターを作る。ツイストがl、紙·ピンのシャンク上のペーパークリップを伸ばした一端の5センチメートルストレート鋼線を庇ワイヤの端部にフック状の単0.5センチメートル &quot;U&quot;を作る。 </li><li>プラスチック製のラップを使用して、いくつかの15×15四方の手術用ドレープを作る。マウス当たり1ドレッシングを準備します。各ラップの間に紙塔を置く。 </li><li> 12×12インチのコル​​クタイル、ガーゼ、および綿棒のスタックと一緒にすべての手術器具を滅菌する。 </li></ol> 2.マウスの準備<ol><li> 2％イソフルランで麻酔を誘導する。動物を計量。 </li><li>専用の外科準備領域では、電気シェーバーと左胸部と首の領域を剃る。 </li><li>マウスの目に人工涙軟膏の低下を適用します。 </li><li>クロルヘキシジン、イソプロピルアルコールで首と左胸部を除染。さらに2回繰り返します。 </li></ol> 3.マウスオロ気管挿管および機械的人工換気<ol><li>予め温めておいた手術領域における非滅菌外科技術者の場所でマウス仰臥位を持っている。</li><li>足のピンチに応答がないことを文書化することにより、麻酔の深さを確認してください。 </li><li>手を洗うと手術衣装、マスク、帽子を着用した後、無菌手術用手袋を着用。 </li><li>ドレープと無菌技術を使用した後、喉頭を露出させ、前方半ば首オーバー1センチメートル垂直切開する。軽く湾曲し、鋸歯状の10cmの鉗子でストラップの筋肉を撤回し、ストレートハサミの先端でストラップの筋肉を広げることによって喉頭と気管を公開。 </li><li>経口半ば気管（ 図1A）に22 G鈍先端の血管カテーテルを挿入し、視覚的に配置します（ 図1B）を確認します。麻酔を維持し、げっ歯類の人工呼吸器を介して1〜3％イソフルランを使用して換気（;毎分200ストークスストロークあたり225μL）。 15cmのH 2 Oの圧力限界を採用</li></ol> 4.マウスの肺<ol><li>マウスの背中を向けた状態で、右側臥位でマウスを置き演算子（左側上）。無菌ドレープのように自己シールプラスチックラップを使用してください。ドレープを切断、4 番目と5 番目の肋間で肋骨に2cmの長さのカット並列を作るために鈍い傾いて湾曲したハサミを使用しています。鈍先端湾曲したハサミを挿入し、根底にある肋骨と肋間筋から離れて皮膚を解剖。 </li><li>正方形の1.5×1.5センチメートル外科ウィンドウ（ 図2A）を露出するために4つのリトラクターで皮膚を撤回。コルクボードにリトラクターを固定します。 </li><li>湾曲した鉗子を用いてリブまで細かく分析し、胸腔を入力するように湾曲した鉗子の1チップを使用しています。 </li><li>丸い先端マイクロはさみを使用して、胸腔を入力するように下刃を使用しています。反対方向にリブを繰り返し間で0.5cmの切開を行います。 </li><li>残りの2つのリトラクターを使用して、前後軸に胸部を開き、コルクボード（ 図2B）に開創を確保。 </li> &lt;LI&gt;左肺をつかみ、左肺動脈と気管支が（ 図3A、B）が露出するまで開胸を通って横方向及び下方に左肺の上部を変位、左手に湾曲した鈍先端が鉗子を使用。 </li><li>手のひらでアプリケーターとパーム（ 図3C）からポインティング曲がった先端の本体と右手にロードされたチタン血管をマイクロアプリケーターを持ち、左の後面の曲率に沿って胸部にアプリケータ先端部をスライドさせ左気管支及び肺動脈（ 図3D）の肺およびクリップ。 </li><li>アプリケーターを取り外しますが後退し左肺を保つ。右手で鈍先端マイクロハサミを握り、クリップに気管支及び肺動脈遠位をカットし、左肺（ 図3E）を削除。 </li><li>リブリトラクターを削除します。 </li><li> incisiに劣る皮膚から1cmを挟まないように湾曲した鈍鉗子を使用してくださいではなく、振動板のレベルを超えて、皮膚を通して、左胸腔（ 図4A、B）への24のG血管カテーテルを挿入します。 </li><li>胸腔を閉じるには、4 番目と5 番目の肋骨の周りに2結節縫合を配置する5-0プロレン縫合糸を使用してください。 </li><li>肌のリトラクターを削除します。切開の長さに沿って皮膚を近似し、肌が閉じ接着する鉗子の二組を使用してください。 </li><li>血管カテーテルに3ミリリットルルアーロックシリンジを接続し、穏やかな吸引を適用し、血管カテーテルを引き出すことにより、残留空気を除去する。 </li><li>首の切開が以前鉗子の二組を使用して閉じた接着。 </li></ol> 5.マウスシャム肺全摘術<ol><li> 「マウス肺」プロトコルに記載されているように左肺を公開します。左胸腔内に空気を可能にするために湾曲した鈍鉗子を用いて胸郭を持ち上げて（ 図5A、B）。 </li><li>左カントーに24のG血管カテーテルを配置左肺を傷つけないように注意しながら、上記のようにracic空洞。 </li><li> 5-0プロレン縫合糸を使用し、肺（ 図5C）を穿刺しないように注意しながら、 第 3 / 第 4に縫合材料の2つの長さと5 番目 / 6 番目の肋骨隙間（ 図5D）を配置。左肺ヘルニアのリスクを軽減するために抱き合わせ前に縫合材料の両方の長さを置きます。 2、中断ステッチ（ 図5E）を作るために縫合材料を結ぶ。 </li><li>胸部切開上の皮膚を接着、血管カテーテルで残留空気を除去し、上記のように首の切開を接着。 </li></ol> 6.蘇生、鎮痛、および回復<ol><li>イソフルランの電源を切り、ブプレノルフィンの0.1 mg / kgを投与し、生理食塩水を皮下の0.5ミリリットル。 </li><li>自発呼吸が再開すると、気管内チューブを取り外します。 </li><li>それが再び歩行になるまでマウスを観察します。通常resumウォーキング気管内チューブを除去した後、数分、ES。 </li><li> O / Nを回復するために27℃のインキュベーター（加湿され、25％の酸素）でマウスを置きます。 注：我々は、手術後の最初の24時間、ケージの床に水で湿らせた固形飼料のいくつかのペレットを配置。 </li><li>手術後3日間、一日二回腹腔内注射によりブプレノルフィンの0.1 mg / kgをを管理します。動物を扱うときに手術部位をオープンしないように注意してください。 </li></ol> 7.マウスの監視<ol><li>手術後1、3、5、および7日目にマウスを秤量する。 </li></ol>

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The authors have nothing to disclose.

我々はこれまでに報告マウスPNXとマウス偽PNX手順のほとんどの詳細な説明を提供してきました。私たちは、手順を学ぶ研究者は、一般的に遭遇する、と​​我々はこれらの落とし穴を軽減するために私たちの研究室が開発したいくつかの技術を概説している共通の落とし穴のいくつかの読者は意識してきた。このモデルを利用する他の研究所は、他の技術の変更を開発したか、別の商品を利?...

肺の主な機能は、生物と大気の酸素と二酸化炭素の交換を提供することである。ヒトでは、先天性および後天条件のホストが損なわれた肺機能をもたらす減少、肺の表面積をも​​たらす。 すなわち、肺-そのような吸入コルチコステロイド、気管支拡張薬、酸素補給、および慢性の機械的換気などの治療のホストが損なわれた肺機能1-3の影響を緩和するために使用されているが、これらの状態のための理想的な治療は、機能的な肺組織の再生を促進する再生。 哺乳動物の組織再生は十分に立証されている。アフリカのトゲマウスは瘢痕形成4なく、皮膚の大部分を再生することができる。ヒトでの末節骨は、以下の傷害または切断5-7を再生成することができます。以下の肺（PNX）、代償肺の成長が行い、9ラット、マウス8で発生gsは10、およびヒト11。定義により、代償肺の成長は既存の空域の拡大が、関連した微小循環12の拡大にこれらの拡大空域の再隔壁形成だけではないが含まれます。遺伝子発現解析は、このモデルが肺の発達13のシグナル伝達事象の多くを再現することを示した。四週間マウスPNXた後、肺胞の表面積は、偽手術動物14と同等である。本稿では、我々は、マウスPNXおよび偽PNX手順を説明します。

<table border="0" cellpadding="0" cellspacing="0" width="1052">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:263px;">Name of the reagent</td>
			<td style="width:263px;">Company</td>
			<td style="width:263px;">Catalogue number</td>
			<td style="width:263px;">Comments (optional)</td>
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			<td height="101" style="height:101px;width:263px;">6-inch Vascular Clip Applicator</td>
			<td style="width:263px;">Teleflex Medical (WECK)</td>
			<td style="width:263px;">137062</td>
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			<td height="101" style="height:101px;width:263px;">Horizon Small Titanium Red Clip</td>
			<td style="width:263px;">Teleflex Medical (WECK)</td>
			<td style="width:263px;">1201</td>
			<td style="width:263px;"></td>
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			<td height="121" style="height:121px;width:263px;">Narrow Pattern 12cm Curved Forceps</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">11003-12</td>
			<td style="width:263px;"></td>
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			<td height="141" style="height:141px;width:263px;">Curved Serated 10 cm Graefe Forceps&#160;</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">11052-10</td>
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			<td height="21" style="height:21px;width:263px;">Castroviejo Needle Holder</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">12565-14</td>
			<td style="width:263px;"></td>
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			<td height="41" style="height:41px;width:263px;">Straight 9 cm Strabismus Scissors (Blunt Tip)</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">14075-09</td>
			<td style="width:263px;"></td>
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			<td height="41" style="height:41px;width:263px;">Straight 8.5 cm Hardened Fine Scissors</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">14090-09</td>
			<td style="width:263px;"></td>
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			<td height="41" style="height:41px;width:263px;">Straight, Blunt Tip Cohan-Vannas Spring Scissors</td>
			<td style="width:263px;">Fine Science Tools</td>
			<td style="width:263px;">15000-12</td>
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			<td height="21" style="height:21px;width:263px;">Skin Glue</td>
			<td style="width:263px;">Gluture</td>
			<td style="width:263px;">32046</td>
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			<td height="21" style="height:21px;width:263px;">22 ga angiocatheter</td>
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			<td height="21" style="height:21px;width:263px;">24 ga angiocatheter</td>
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			<td style="width:263px;"></td>
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		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:263px;">3 mL luer lock syringe</td>
			<td style="width:263px;"></td>
			<td style="width:263px;"></td>
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			<td height="21" style="height:21px;width:263px;">4 short retractors</td>
			<td style="width:263px;"></td>
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			<td style="width:263px;"></td>
		</tr>
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			<td height="21" style="height:21px;width:263px;">2 long retractors</td>
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			<td height="41" style="height:41px;width:263px;">5-0 Prolene on curved cutting needle</td>
			<td style="width:263px;">Ethicon</td>
			<td style="width:263px;">8698G</td>
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			<td height="21" style="height:21px;width:263px;">0.5 mL syringe on 27 ga needle</td>
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			<td height="21" style="height:21px;width:263px;">Normal Saline</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:263px;">Buprenorphine</td>
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			<td height="21" style="height:21px;width:263px;">Press-n-Seal Wrap</td>
			<td style="width:263px;">Glad Products Company</td>
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			<td height="21" style="height:21px;width:263px;">12 inch X 12 inch cork board stack</td>
			<td style="width:263px;">Office Depot</td>
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			<td height="21" style="height:21px;width:263px;">70% Ethanol</td>
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			<td height="21" style="height:21px;width:263px;">Betadine</td>
			<td style="width:263px;"></td>
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			<td height="21" style="height:21px;width:263px;">Mouse Ventilator</td>
			<td style="width:263px;">Hugo Sachs Elektronnik&#160;</td>
			<td style="width:263px;">Minivent Type 845</td>
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			<td height="21" style="height:21px;width:263px;">Isoflurane Vaporizer</td>
			<td style="width:263px;">OHMEDA</td>
			<td style="width:263px;">Excel 210 SE</td>
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			<td height="21" style="height:21px;width:263px;">artificial tear ointment</td>
			<td style="width:263px;">puralube</td>
			<td style="width:263px;">NDC: 17033-211-38</td>
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</table>

mouse pneumonectomy model of compensatory lung growth

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to disability or death. Injury-based animal models of lung repair and regeneration are limited by injury-specific responses making it difficult to differentiate changes related to the injury response and injury resolution from changes related to lung repair and lung regeneration. However, use of animal models to identify these repair and regeneration signaling pathways is critical to the development of new therapies aimed at improving pulmonary function following lung injury. The mouse pneumonectomy model utilizes compensatory lung growth to isolate those repair and regeneration signals in order to more clearly define mechanisms of alveolar re-septation. Here, we describe our technique for performing mouse pneumonectomy and sham pneumonectomy. This technique may be utilized in conjunction with lineage tracing or other transgenic mouse models to define molecular and cellular mechanism of lung repair and regeneration.

。PNXと偽肺の両方のための100％ - PNXと偽手術マウスの体重のプロットを図6に提供されている我々の手では、生存率は一貫して95です。右肺がこのモデルと予想される時間経過に再成長する方法の詳細については、我々はギブニーら 15及びWang らの原稿を読者に参照してください。14 いくつかの一般的な落とし穴が正常マウ...

Watch this Scientific Journal Video about Mouse Pneumonectomy Model of Compensatory Lung Growth at JoVE.com

Mouse Pneumonectomy Model of Compensatory Lung Growth

Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

代償肺の成長のマウス肺モデル

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to ...

mouse-pneumonectomy-model-of-compensatory-lung-growth

Research

JoVE Journal

Medicine

12.7K ビュー.  Cincinnati Children's Hospital Medical Center. Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

ビデオ: Mouse Pneumonectomy Model of Compensatory Lung Growth

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

マウス肺挿管の簡単な方法

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

医学

Diagnostic Ultrasound Imaging of Mouse Diaphragm Function

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive surgical procedures. Although this is an effective method of assessing in vitro diaphragm activity, it involves non-survival surgery. The application of non-invasive ultrasound imaging as an in vivo procedure is beneficial since it not only reduces the number of animals sacrificed, but is also suitable for monitoring disease progression in live mice. Thus, our ultrasound imaging method may likely assist in the development of novel therapies that alleviate muscle injury induced by various respiratory diseases. Particularly, in clinical diagnoses of obstructive lung diseases, ultrasound imaging has the potential to be used in conjunction with other standard tests to detect the early onset of diaphragm muscle fatigue. In the current protocol, we describe how to accurately evaluate diaphragm contractility in a mouse model using a diagnostic ultrasound imaging technique.

Diagnostic ultrasound imaging has proven to be effective in diagnosing various respiratory diseases in human and animal subjects. We demonstrate a comprehensive ultrasound protocol utilized by Dr. Zuo's lab to analyze diaphragm kinetics specifically in mouse models. This is also a non-invasive research technique which can provide quantitative information on mouse respiratory muscle function.

マウス絞り機能の診断用超音波イメージング

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive ...

A Murine Model of Myocardial Ischemia-reperfusion Injury through Ligation of the Left Anterior Descending Artery

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms at work during MI and developing effective therapeutic approaches requires methodology to reproducibly simulate the clinical incidence and reflect the pathophysiological changes associated with MI. Here, we describe a surgical method to induce MI in mouse models that can be used for short-term ischemia-reperfusion (I/R) injury as well as permanent ligation. The major advantage of this method is to facilitate location of the left anterior descending artery (LAD) to allow for accurate ligation of this artery to induce ischemia in the left ventricle of the mouse heart. Accurate positioning of the ligature on the LAD increases reproducibility of infarct size and thus produces more reliable results. Greater precision in placement of the ligature will improve the standard surgical approaches to simulate MI in mice, thus reducing the number of experimental animals necessary for statistically relevant studies and improving our understanding of the mechanisms producing cardiac dysfunction following MI. This mouse model of MI is also useful for the preclinical testing of treatments targeting myocardial damage following MI.

We introduce a surgical method to induce experimental ischemia/reperfusion (I/R) injury to simulate myocardial infarction (MI) in mouse models that allows for more clarity in positioning of the ligation on the left anterior descending artery (LAD) to increase the reproducibility of MI experiments in mice.

左前下の動脈の結紮を通して心筋虚血再灌流傷害のマウスモデル

Acute or chronic myocardial infarction (MI) are cardiovascular events resulting in high morbidity and mortality. Establishing the pathological mechanisms ...

Implantation of Fibrin Gel on Mouse Lung to Study Lung-specific Angiogenesis

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and repair damage in simple tissues in the orthopedic and periodontal fields. However, attempts to regenerate the structures and functions of more complex three-dimensional (3D) organs such as lungs have not been very successful because the biological processes of organ regeneration have not been well explored. It is becoming clear that angiogenesis, the formation of new blood vessels, plays key roles in organ regeneration. Newly formed vasculatures not only deliver oxygen, nutrients and various cell components that are required for organ regeneration but also provide instructive signals to the regenerating local tissues. Therefore, to successfully regenerate lungs in an adult, it is necessary to recapitulate the lung-specific microenvironments in which angiogenesis drives regeneration of local lung tissues. Although conventional in vivo angiogenesis assays, such as subcutaneous implantation of extracellular matrix (ECM)-rich hydrogels (e.g., fibrin or collagen gels or Matrigel - ECM protein mixture secreted by Engelbreth-Holm-Swarm mouse sarcoma cells), are extensively utilized to explore the general mechanisms of angiogenesis, lung-specific angiogenesis has not been well characterized because methods for orthotopic implantation of biomaterials in the lung have not been well established. The goal of this protocol is to introduce a unique method to implant fibrin gel on the lung surface of living adult mouse, allowing for the successful recapitulation of host lung-derived angiogenesis inside the gel. This approach enables researchers to explore the mechanisms by which the lung-specific microenvironment controls angiogenesis and alveolar regeneration in both normal and pathological conditions. Since implanted biomaterials release and supply physical and chemical signals to adjacent lung tissues, implantation of these biomaterials on diseased lung can potentially normalize the adjacent diseased tissues, enabling researchers to develop new therapeutic approaches for various types of lung diseases.

Recapitulation of the organ-specific microenvironment, which stimulates local angiogenesis, is indispensable for successful regeneration of damaged tissues. This report demonstrates a novel method to implant fibrin gels on the lung surface of living mouse in order to explore how the lung-specific microenvironment modulates angiogenesis and alveolar regeneration in adult mouse.

肺特異的血管形成を研究するためにマウスの肺にフィブリンゲルの移植

Recent significant advances in stem cell research and bioengineering techniques have made great progress in utilizing biomaterials to regenerate and ...

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. As this technology expands and improves, the ability to potentially evaluate and improve the quality of substandard lungs prior to transplant is a critical need. In order to more rigorously evaluate these approaches, a reproducible animal model needs to be established that would allow for testing of improved techniques and management of the donated lungs as well as to the lung-transplant recipient. In addition, an EVLP animal model of associated pathologies, e.g., ventilation induced lung injury (VILI), would provide a novel method to evaluate treatments for these pathologies. Here, we describe the development of a rat EVLP lung program and refinements to this method that allow for a reproducible model for future expansion. We also describe the application of this EVLP system to model VILI in rat lungs. The goal is to provide the research community with key information and &#8220;pearls of wisdom&#8221;/techniques that arose from trial and error and are critical to establishing an EVLP system that is robust and reproducible.

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

孤立の方法<em&gt;エクスビボ</emラットモデルにおける&gt;肺灌流：ラットEVLPプログラムの開発の教訓

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

Real-time X-ray Imaging of Lung Fluid Volumes in Neonatal Mouse Lung

At birth, the lung undergoes a profound phenotypic switch from secretion to absorption, which allows for adaptation to breathing independently. Promoting and sustaining this phenotype is critically important in normal alveolar growth and gas exchange throughout life. Several in vitro studies have characterized the role of key regulatory proteins, signaling molecules, and steroid hormones that can influence the rate of lung fluid clearance. However, in vivo examinations must be performed to evaluate whether these regulatory factors play important physiological roles in regulating perinatal lung liquid absorption. As such, the utilization of real time X-ray imaging to determine perinatal lung fluid clearance, or pulmonary edema, represents a technological advancement in the field. Herein, we explain and illustrate an approach to assess the rate of alveolar lung fluid clearance and alveolar flooding in C57BL/6 mice at post natal day 10 using X-ray imaging and analysis. Successful implementation of this protocol requires prior approval from institutional animal care and use committees (IACUC), an in vivo small animal X-ray imaging system, and compatible molecular imaging software.

We present a protocol to assess the rate of alveolar fluid clearance or pulmonary edema in neonatal mouse lung using X-ray imaging technology.

新生児マウスの肺における肺流体体積のリアルタイムX線イメージング

At birth, the lung undergoes a profound phenotypic switch from secretion to absorption, which allows for adaptation to breathing independently. Promoting ...

Apical Resection Mouse Model to Study Early Mammalian Heart Regeneration

Cardiovascular disease plagues the whole world due to intensive lifestyle changes. Heart regeneration holds great promise for repairing and restoring cardiomyocytes lost due to injury and disease. In contrast to the robust cardiac regeneration of certain lower vertebrates, adult mammalian hearts typically show minimal capacity for heart regeneration and repair. However, recent studies have sparked considerable scientific interest with the finding that, between postnatal day 1 to 7 (P1 to P7), the neonatal mouse heart retains significant regenerative potential after apical resection (i.e., surgical amputation and exposure of left ventricular apex). One major controversy over this finding might be due to the diverse surgery-related procedures used in efforts to replicate or expand upon this important finding. These instructions dynamically present the materials and methodology for apical resection in a mouse model. The salient steps of this rodent survival surgery involve hypothermia anesthesia, thoracotomy, surgical amputation of heart ventricular apex, and suture and recovery of mice. The approach described could expand the application of the apical resection mouse model for cardiovascular research.

A step-by-step video protocol of apical resection is demonstrated in this study. Apical resection is a recently highlighted surgical approach in mammalian heart regeneration research. This study may promote the application of apical resection as a standard methodology in research into the mechanism underlying heart regeneration.

アピカル切除マウスモデルは、初期の哺乳類の心臓の再生を研究するために

Cardiovascular disease plagues the whole world due to intensive lifestyle changes. Heart regeneration holds great promise for repairing and restoring ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

最大急性非致死肺損傷における急性肺損傷の結果のオープン気管切開胃酸吸引マウスモデル

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...

Integrated Compensatory Responses in a Human Model of Hemorrhage

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of hemorrhaging patients is difficult because current clinical tools provide measures of vital signs that remain stable during the early stages of bleeding due to compensatory mechanisms. Consequently, there is a need to understand and measure the total integration of mechanisms that compensate for reduced circulating blood volume and how they change during ongoing progressive hemorrhage. The body&#39;s reserve to compensate for reduced circulating blood volume is called the &#39;compensatory reserve&#39;. The compensatory reserve can be accurately evaluated with real-time measurements of changes in the features of the arterial waveform measured with the use of a high-powered computer. Lower Body Negative Pressure (LBNP) has been shown to simulate many of the physiological responses in humans associated with hemorrhage, and is used to study the compensatory response to hemorrhage. The purpose of this study is to demonstrate how compensatory reserve is assessed during progressive reductions in central blood volume with LBNP as a simulation of hemorrhage.

The purpose of this protocol is to demonstrate the techniques for measuring compensatory responses to reduced central blood volume using lower body negative pressure as a noninvasive experimental model of human hemorrhage which can be used to quantify the total integration of compensatory mechanisms to blood volume deficit in humans.

出血の人体モデルで統合代償応答

Hemorrhage is the leading cause of trauma-related deaths, partly because early diagnosis of the severity of blood loss is difficult. Assessment of ...

Echocardiographic Measurement of Right Ventricular Diastolic Parameters in Mouse

Diastolic dysfunction is a prominent feature of right ventricular (RV) remodeling associated with conditions of pressure overload. However, the RV diastolic function is rarely quantified in experimental studies. This might be due to technical difficulties in the visualization of the RV in the apical four-chamber view in rodents. Here we describe two positions facilitating the visualization of the apical four-chamber view in mice to assess the RV diastolic function.
The apical four-chamber view is enabled by tilting the mouse fixation platform to the left and caudally (LeCa) or to the right and cranially (RiCr). Both positions provide images of comparable quality. The results of the RV diastolic function obtained from two positions are not significantly different. Both positions are comparably easy to perform. This protocol can be incorporated into published protocols and enables detailed investigations of the RV function.

Here we describe and compare two positions for obtaining the apical four-chamber view in mice. These positions enable the quantification of the right ventricular function, provide comparable results, and can be used interchangeably.