This work was supported by NIH grants HL111121 (JDM) and TR000954 (JDM).

<ol>
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The authors have nothing to disclose.

エージングは、心血管石灰化1の漸進的増加と関連しています。血行動態的に重要な大動脈弁狭窄症は、65 2歳以上の人口の3％に影響を及ぼし、さらには中程度の大動脈弁狭窄（3-4メートル/秒のピーク速度）を有する患者は、40％未満の5年無イベント生存率を持っています3。現在、そこに大動脈弁の石灰化の進行を遅らせるために有効な治療法はなく、外科的大動脈弁置換術は、高度な大動脈弁狭窄4のための唯一の可能な治療です。 大動脈弁石灰化の開始および進行に寄与するメカニズムのより深い理解を得ることを目的とした研究は、大動脈弁狭窄5、6を管理するための薬理学的および非外科的方法に向かって移動中の重要な最初のステップです。遺伝学的なLY-変更されたマウスは、種々の疾患に貢献し、今大動脈弁狭窄6、7、8の生物学を理解することを目的とした機構研究の最前線に来ているメカニズムの理解を開発する上で大きな役割を果たしてきました。アテローム性動脈硬化症や心不全、血管および心室機能を評価するための標準プロトコルは大部分がされているような他の心血管疾患とは異なり、マウスにおける心臓弁の機能のin vivoでの表現型に関連付けられている固有の課題は、十分に確立された-があります。最近のレビューは、多数のイメージングおよびげっ歯類9、10、11における弁の機能を評価するために使用侵襲的モダリティの長所と短所についての徹底的な議論を提供してきたが、今日まで、私たちはCOMPREを提供する出版物を認識していませんhensive、マウスでは心臓弁の機能を表現型のためのステップバイステップのプロトコル。 本稿の目的は、マウスにおける心臓弁の機能を表現型にする方法およびプロトコルを記述することです。すべての方法及び手順は、メイヨークリニック制度動物実験委員会によって承認されています。このプロトコルの主な構成要素は、麻酔の深さ、心機能の評価、および心臓弁の機能の評価を含みます。我々は、この報告書は、心臓弁疾患の分野の研究を追求に興味を持って研究者を案内するのに役立つだけでなく、この急速に成長している分野で、データの再現性と妥当性を確保するためのプロトコル標準化に関連する国内および国際的な対話を開始することを願っています。重要なことには、高解像度の超音波システムを使用して成功したイメージングは​​、基本Principでの理解を加工超音波検査の原理の知識（及び一般的に超音波検査で使用される用語）を必要とレ心臓生理学、およびげっ歯類における心機能の正確かつ時間効率の評価を可能にするために超音波検査での豊富な経験の。

<table><tbody><tr><td>High resolution ultrasound machine</td><td>VisualSonics, Fujifilm</td><td>Vevo 2100&amp;nbsp;</td><td></td></tr><tr><td>Isoflurane diffuser (capable of delivering 1 % to 1.5 % isoflurane mixed with 1 L/min 100% O&lt;sub&gt;2&lt;/sub&gt;</td><td>VisualSonics, Fujifilm</td><td>N/A</td><td></td></tr><tr><td>Transducers for small mice (550D) or larger mice (400)</td><td>MicroScan, VisualSonics, Fujifilm</td><td>MS 550D, MS 400</td><td></td></tr><tr><td>Animal platform</td><td>VisualSonics, Fujifilm</td><td>11503</td><td></td></tr><tr><td>Advanced physiological monitoring unit</td><td>VisualSonics, Fujifilm</td><td>N/A</td><td></td></tr><tr><td>Isoflurane</td><td>Terrell</td><td>NDC 66794-019-10</td><td></td></tr><tr><td>Nose cone and tubing connected to isoflurane diffuser and 100% O&lt;sub&gt;2&lt;/sub&gt;</td><td>Custom Engineered in-house</td><td>--</td><td></td></tr><tr><td>Hair razor</td><td>Andis Super AGR+ vet pack clipper</td><td>AD65340</td><td></td></tr><tr><td>Ultrasound gel</td><td>Parker Laboratories</td><td>REF 01-08</td><td></td></tr><tr><td>Electrode gel&amp;nbsp;</td><td>Parker Laboratories</td><td>REF 15-25</td><td></td></tr><tr><td>Adhesive tapes</td><td>Fisher Laboratories</td><td>1590120B</td><td></td></tr><tr><td>Paper towels</td><td></td><td></td><td></td></tr></tbody></table>

echocardiographic approaches and protocols for comprehensive phenotypic characterization of valvular heart disease in mice

The aim of this manuscript and accompanying video is to provide an overview of the methods and approaches used for imaging heart valve function in rodents, with detailed descriptions of the appropriate methods for anesthesia, the echocardiographic windows used, the imaging planes and probe orientations for image acquisition, the methods for data analysis, and the limitations of emerging technologies for the evaluation of cardiac and valvular function. Importantly, we also highlight several future areas of research in cardiac and heart valve imaging that may be leveraged to gain insights into the pathogenesis of valve disease in preclinical animal models. We propose that using a systematic approach to evaluating cardiac and heart valve function in mice can result in more robust and reproducible data, as well as facilitate the discovery of previously underappreciated phenotypes in genetically-altered and/or physiologically-stressed mice.

日常的に動物の心臓の超音波画像から得られる画像の例は、この原稿に含まれています。動物の胸にトランスデューサ配置の図が読者に変換器が記載されているように画像を取得するように配置されるの明確な理解を与えるために設けられています。超音波実験室セットアップの写真はまた、適切な装置、特に、使用される超音波トランスデューサと麻酔の方法の重要性を強調するために含まれています。 2D / Bモード、Mモード、カラードップラー、正常および異常な弁のディスプレイ、右心室および左心室と大動脈根が適切に標識されています。ひずみ速度イメージングが日常的に行われていないが、例も含まれています。 僧帽弁逆流は、S中に弁を横切って高い、通常、非層状血流速度（モザイク着色）することを特徴としますystole（ 図5）。 ECGトレース中のQRS群の後に発生するMV全体の左心房、左心室から、このようなモザイクカラードップラーフローパターンの存在は、MRの明確な診断を可能にします。これは、大動脈弁逆流および/または左心室機能障害の不存在下で発生した場合、これは、単離された僧帽弁逸脱として特徴付けることができます。左心室（これは実験的に誘導する心不全または麻酔の過剰な深さ）の著しい拡張がある場合、これは、虚血性僧帽弁逆流（または心機能不全に続発逆流）として特徴付けることができます。パルス波スペクトルドップラ表示は、血流の逆流ジェットの存在およびタイミングを確認するために使用することができます。 通常の大動脈弁が開き、各心周期の間に十分に閉じて3薄く、柔軟な尖を持っています。大動脈弁尖の分離は、2D-案内さで測定されます長軸ビューで大動脈弁のMモード。電子キャリパーは、左大動脈尖の先端に右大動脈尖（ 図3）の先端から測定するのに使用されています。正常マウスにおける大動脈弁尖分離距離は、0.9〜1.3 mmです。カラードップラーでは収縮期バルブ前後と大動脈根への層流を示しています。乱流は、大動脈弁逆流と同様に、増加した流量の条件で認識、または大動脈弁狭窄症のように、圧力を増加させることができます。これは、流出路内のモザイク着色として示されています。大動脈弁逆流の少量でも起因亢進、心機能にピークtransvalvular速度の大幅な増加をもたらし、左心室前負荷を増加することができます。正常マウスにおけるピーク大動脈速度1.50メートル/秒0.90メートル/秒の範囲です。 &gt; 5メートル/秒のピーク大動脈弁の速度は、重度の大動脈弁狭窄症を有するマウスで記録されています。 12（ 図8）の指標を提供するために使用することができます。肺動脈加速時間は、ピーク流速の収縮期肺動脈流の開始からの時間間隔です。右心室駆出時間は、収縮期肺動脈収縮期流の停止がある点まで右心室駆出の開始との間の間隔です。右心室駆出時間に肺動脈加速時間の割合の減少に短縮肺動脈加速時間の組み合わせは、（肺動脈または右心室圧の侵襲的または直接的尺度を用いて確認することができる）、肺動脈高血圧症の存在を示唆しています。 <img alt="図1" src="/files/ftp_upload/54110/54110fig1.jpg" /> 図1：動物C超音波研究所ardiac。実験室は、高周波（30 MHzおよび40 MHz）のトランスデューサ（MS 400およびMS 550D）、イソフルランディフューザー、動物プラットフォーム、温度および心拍数モニター、1％〜1.5％イソフルランと小動物専用の超音波装置が装備されていますイソフルランディフューザ100％O 2、毛かみそり、超音波ゲル、電極ゲル、粘着テープ、紙タオルに接続された1 L /分、100％O 2、ノーズコーンとチューブと混合しました。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig1large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図2" src="/files/ftp_upload/54110/54110fig2.jpg" /> 図2：基本的なトランスデューサ体位。 （A）胸骨傍ウィンドウ。トランスデューサヘッドは、尾側指向トランスデューサの画像インデックスマーカーと、左胸骨傍の境界に位置しています。金曜日OMこの位置、左心室、大動脈弁、及び大動脈根の長軸図及び肺動脈弁の短軸像を得ることができます。胸骨傍ウィンドウ（B）は 、トランスデューサヘッドは、後方に向けられたノッチと、反時計方向に回転させます。この位置から、左心室と大動脈弁及び肺動脈弁の長軸ビューの短軸像を得ることができます。 （C）アピカルウィンドウ。トランスデューサヘッドは、心臓の頂点に位置しています。この位置から、右心室および左心室と僧帽弁と三尖弁の長軸像を得ることができます。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig2large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図3" src="/files/ftp_upload/54110/54110fig3.jpg" /> 図3：規範で大動脈弁機能の評価石灰大動脈弁疾患を持つマウスにおける大動脈弁の機能対アルマウス。 （A）長軸ビューで正常な大動脈弁の2D画像。大動脈弁が収縮期にも開くことに注意してください。 （B）Mモード画像は、通常の大動脈弁機能（箱状の外観）を示します。カスプ分離距離は1.12ミリメートルで測定されることに注意してください。通常の大動脈弁全体のピーク速度の（C）スペクトルドップラー表示は、1.3メートル/ sでmeauredました。 （D）欠乏、低密度リポタンパク質受容体（LDLR - / - ）から長軸ビューにおける石灰化大動脈弁の2D画像と西洋食を与えたアポリポタンパク質B100のみ（アポB 100/100）マウス。尖は肥厚しており、収縮期に制限された開口部に生じるエコー輝度を、増加しています。 （E）は、同じ狭窄大動脈弁を示すMモード画像は、0.7mmでカスプ分離距離の測定を示します。 （F &lt;狭窄大動脈弁全体のピーク速度の/ strong&gt;の）スペクトルドップラ表示は4.6メートル/秒でmeauredました。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig3large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図4" src="/files/ftp_upload/54110/54110fig4.jpg" /> 図4：通常の僧帽弁のMモード。頂端ウィンドウから、僧帽弁の長軸像が得られます。質問のMモード線は、僧帽弁尖に印加されます。僧帽弁のリーフレットの厚さは、理論的に電子ノギスを用いて測定することができるが、これは薄く、不十分なエコー発生与えられた非常に挑戦すること、及び急速に正常な僧帽弁のリーフレットを移動することができます。矢印は、収縮期における僧帽弁尖のMモードを指します。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig4large.jpg" target="_blank">CLIください</a>この図の拡大版を表示するには、こちらのCK。 <img alt="図5" src="/files/ftp_upload/54110/54110fig5.jpg" /> 図5：カラードップラーイメージングを使用して、僧帽弁逆流ジェットの証拠。胸骨傍ウィンドウから、僧帽弁の変形長軸像が得られます。カラードップラー問い合わせは（矢印で強調）収縮期の僧帽弁でモザイク色のジェットを示しています。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig5large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図6" src="/files/ftp_upload/54110/54110fig6.jpg" /> 図6：主肺動脈とその主要な枝の長軸ビュー。主肺動脈の長軸ビュー（MPA）および右（RPA）及び左（LPA）ブランチがparasteから得ることができ<html> rnalウィンドウ。右心室流出路（RVOT）、肺動脈弁（PV）、および大動脈（AO）が部分的に見られます。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig6large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図7" src="/files/ftp_upload/54110/54110fig7.jpg" /> 図7：通常の肺動脈弁を描いたMモード画像。胸骨ウィンドウから、肺動脈弁の両方の短期および長期軸像を得ることができます。質問のMモード線は肺動脈弁に印加されます。肺動脈弁尖分離（矢印）の距離は、この図から測定することができます。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig7large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> 10fig8.jpg &quot;/&gt; 図8：肺動脈弁を横切る流れのパルス波ドップラー尋問。肺動脈の加速時間（PAAT）は、ピーク流速の収縮期肺動脈流の開始からの時間間隔です。右心室駆出時間（RVET）は流れの停止がある点まで右心室駆出の開始との間の間隔です。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig8large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図9" src="/files/ftp_upload/54110/54110fig9.jpg" /> 図9：左心室の短軸ビューを描いたMモード画像。胸骨傍ウィンドウから、左心室の短軸像をするよう後方にまたは背反時計画像インデックスマーカ点をトランスデューサヘッドを回転させることによって得られます。 M-MOD質問のe線は、乳頭筋のレベルで左心室に印加されます。左心室拡張終期寸法（LVEDD）、左心室収縮終期寸法（LVESD）、及び前壁（AW）および後壁（PW）の厚さを容易に測定することができます。いずれの測定で乳頭筋（*）を含めるしないように注意してください。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig9large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図10" src="/files/ftp_upload/54110/54110fig10.jpg" /> 図10：カラードップラー評価と僧帽弁流入のパルス波ドップラースペクトル表示。 （A）頂端長軸図で僧帽弁流入のカラードップラー評価を示す画像。 2Dカラードプラ画像はグラムのための重要なツールであることに注意してください（パネルBに示されている）、パルス波ドップラー追跡の取得のための適切なサンプルボリューム位置をuiding。 （B）、パルス波ドップラーを使用して、僧帽弁流入のスペクトル表示。 （頂端長軸ビュー）僧帽弁を横切って血流のパルス波ドップラー評価は、左心室拡張期機能を評価するために行われます。サンプル体積は、僧帽弁尖の先端に配置されています。等容性弛緩時間（IVRT）、等容性収縮時間（IVCT）、左心室駆出時間（LVET）、ピーク僧帽弁流入速度（E）は、すべての僧帽弁の両端のパルス波ドップラー速度のスペクトル表示から導出することができます。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig10large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図11" src="/files/ftp_upload/54110/54110fig11.jpg" /> Fiのグレ11：中隔僧帽弁輪の組織ドップラーイメージング。頂端ウィンドウから、僧帽弁の長軸像が得られます。組織ドプラサンプルボリュームは、僧帽弁輪の中隔領域に配置されています。ピーク僧帽弁流入速度（図10（b）中の変数E）及び（eピーク僧帽弁輪の組織速度の比」（白い矢印で示されるが、）、左心室拡張期機能を評価するために使用される一般的E / Eと呼びます」）。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig11large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a> <img alt="図12" src="/files/ftp_upload/54110/54110fig12.jpg" /> 図12：左心室心筋のひずみとひずみ速度の評価。そこに特殊な解析ソフトウェアパッケージが市販されており、歪み及び歪み速度変数とすることができます内因性の心筋の収縮特性の初期またはサブ臨床変化の尺度として得られます。上に示した例は、マウスにおいて、一般的に取得された撮像面における半径方向の歪み及び歪み速度を示します。これらの撮像面（や歪みトレーシングのその後の形状）が頻繁に頂端長軸または4室ビューで取得されたヒトでの画像とは異なることに注意してください。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig12large.jpg" target="_blank">この図の拡大版をご覧になるにはこちらをクリックしてください。</a>

Watch this Scientific Journal Video about Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice at JoVE.com

Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice

This protocol provides a detailed description of the echocardiographic approach for comprehensive phenotyping of heart and heart valve function in mice.

マウスの弁膜症の包括的表現型の特徴のための心エコーアプローチとプロトコル

The aim of this manuscript and accompanying video is to provide an overview of the methods and approaches used for imaging heart valve function in ...

echocardiographic-approaches-protocols-for-comprehensive-phenotypic

Nanyang Technological University

Research

JoVE Journal

Medicine

15.9K ビュー.  Mayo Clinic. This protocol provides a detailed description of the echocardiographic approach for comprehensive phenotyping of heart and heart valve function in mice. 

ビデオ: Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice

Transthoracic Echocardiography in Mice

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene expression. Transgenic and gene targeting methods can be used to generate mice with altered cardiac size and function,1-3 and as a result, in vivo techniques are needed to evaluate their cardiac phenotype. Transthoracic echocardiography, pulse wave Doppler (PWD), and tissue Doppler imaging (TDI) can be used to provide dimensional measurements of the mouse heart and to quantify the degree of cardiac systolic and diastolic performance. Two-dimensional imaging is used to detect abnormal anatomy or movements of the left ventricle, whereas M-mode echo is used for quantification of cardiac dimensions and contractility.4,5 In addition, PWD is used to quantify localized velocity of turbulent flow,6 whereas TDI is used to measure the velocity of myocardial motion.7 Thus, transthoracic echocardiography offers a comprehensive method for the noninvasive evaluation of cardiac function in mice.

Transthoracic echocardiography offers a noninvasive method for the evaluation of cardiac function in mice. A combination of ultrasound and Doppler imaging modalities can be used to obtain dimensional measurements of the heart and intracardiac blood flow, which together provide an assessment of cardiac systolic and diastolic performance.

マウスにおける経胸壁心エコー

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene ...

医学

Murine Echocardiography and Ultrasound Imaging

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1 Echocardiography provides a powerful, non-invasive tool to serially assess cardiac morphometry and function in a living animal.2 However, using this technique on mice poses unique challenges owing to the small size and rapid heart rate of these animals.3 Until recently, few ultrasound systems were capable of performing quality echocardiography on mice, and those generally lacked the image resolution and frame rate necessary to obtain truly quantitative measurements. Newly released systems such as the VisualSonics Vevo2100 provide new tools for researchers to carefully and non-invasively investigate cardiac function in mice. This system generates high resolution images and provides analysis capabilities similar to those used with human patients. Although color Doppler has been available for over 30 years in humans, this valuable technology has only recently been possible in rodent ultrasound.4,5 Color Doppler has broad applications for echocardiography, including the ability to quickly assess flow directionality in vessels and through valves, and to rapidly identify valve regurgitation. Strain analysis is a critical advance that is utilized to quantitatively measure regional myocardial function.6 This technique has the potential to detect changes in pathology, or resolution of pathology, earlier than conventional techniques. Coupled with the addition of three-dimensional image reconstruction, volumetric assessment of whole-organs is possible, including visualization and assessment of cardiac and vascular structures. Murine-compatible contrast imaging can also allow for volumetric measurements and tissue perfusion assessment.

This video demonstrates use of a rail-mounted high-frequency ultrasound probe to perform echocardiography on an anesthetized mouse. The methods describe both conventional two-dimensional and M-mode measurements of cardiac function in addition to newer, more powerful tools such as color Doppler, strain analysis, as well as general and targeted contrast imaging.

マウス心エコー検査と超音波イメージング

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1  ...

Echocardiographic Assessment of the Right Heart in Mice

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential therapies. Given the expense and time involved in creating animal models of disease, it is critical that researchers have tools to accurately assess phenotypic expression of disease. Right ventricular dysfunction is the major manifestation of pulmonary hypertension. Echocardiography is the mainstay of the noninvasive assessment of right ventricular function in rodent models and has the advantage of clear translation to humans in whom the same tool is used. Published echocardiography protocols in murine models of PAH are lacking.
In this article, we describe a protocol for assessing RV and pulmonary vascular function in a mouse model of PAH with a dominant negative BMPRII mutation; however, this protocol is applicable to any diseases affecting the pulmonary vasculature or right heart. We provide a detailed description of animal preparation, image acquisition and hemodynamic calculation of stroke volume, cardiac output and an estimate of pulmonary artery pressure.

This article provides a protocol for the echocardiographic assessment of right ventricular size and pulmonary hypertension in mice. Applications include phenotype determination and serial assessment in transgenic and toxin-induced mouse models of cardiomyopathy and pulmonary vascular disease.

マウスの右心の心エコー評価

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

A Pipeline to Characterize Structural Heart Defects in the Fetal Mouse

Congenital heart diseases (CHDs) are major causes of infant death in the United States. In the 1980s and earlier, most patients with moderate or severe CHD died before adulthood, with the maximum mortality during the first week of life. Remarkable advances in surgical techniques, diagnostic approaches, and medical management have led to marked improvements in outcomes. To address the critical research needs of understanding congenital heart defects, murine models have provided an ideal research platform, as they have very similar heart anatomy to humans and short gestation rates. The combination of genetic engineering with high-throughput phenotyping tools has allowed for the replication and diagnosis of structural heart defects to further elucidate the molecular pathways behind CHDs. The use of noninvasive fetal echocardiography to screen the cardiac phenotypes in mouse models coupled with the high fidelity of Episcopic fluorescence image capture (EFIC) using Episcopic confocal microscopy (ECM) histopathology with three-dimensional (3D) reconstructions enables a detailed view into the anatomy of various congenital heart defects. This protocol outlines a complete workflow of these methods to obtain an accurate diagnosis of murine congenital heart defects. Applying this phenotyping protocol to model organisms will allow for accurate CHD diagnosis, yielding insights into the mechanisms of CHD. Identifying the underlying mechanisms of CHD provide opportunities for potential therapies and interventions.

This article details murine congenital heart disease (CHD) diagnostic methods using fetal echocardiography, necropsy, and Episcopic fluorescence image capture (EFIC) using Episcopic confocal microscopy (ECM) followed by three-dimensional (3D) reconstruction.

マウス胎児の構造的心臓欠損を特徴付けるパイプライン

Congenital heart diseases (CHDs) are major causes of infant death in the United States. In the 1980s and earlier, most patients with moderate or severe ...

発生生物学

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 ...

Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery

The present methodology teaches the investigator how to measure and use the LAV as a surrogate of chronic elevations in Left Ventricular diastolic pressure through echocardiography, as well as to obtain measurements of the Aorta and PA diameter in mice.
Mice older than 10 d of age can be analyzed using the present technique. The technique is composed of 3 main steps: set-up, image acquisition, and image analysis. The set-up step consists of getting the mouse anesthetized with 1% isoflurane, shaving it, and taping it in a supine position to a heated EKG board where the image acquisition will take place. The image acquisition step consists of learning to identify the cardiac structures and obtaining all the required images with its correspondent probe and axis in order to be able to calculate volumes and diameters. The image analysis step consists of measuring the previously acquired images with the aid of computer software.
Advantages of the proposed technique include a fast (15 min) procedure that would allow the researcher to evaluate interventions in a non-invasive, non-terminal approach and therefore follow the same mouse over time; each mouse can be used as its own control. This fact plus having the same operator perform all the acquisition and analysis for the entire experiment minimizes the limitation of operator-dependency. The present methodology is useful for mouse researchers in cardiovascular and pulmonary medicine.

The following protocol describes the methodology for the acquisition and analysis of echocardiographic images used to obtain the Left Atrial Volume (LAV), Aorta (Ao) diameter, and Pulmonary Artery (PA) diameter in mice. This technique is a non-invasive, non-terminal procedure that allows assessment of the cardiopulmonary function.

左心房、大動脈、肺動脈のマウス心エコー

The present methodology teaches the investigator how to measure and use the LAV as a surrogate of chronic elevations in Left Ventricular diastolic ...

Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed in mice is high. Phenotypic characterization of these mouse models also requires the examination of cardiac function and morphology. Echocardiography and magnetic resonance imaging (MRI) are commonly used approaches to characterize cardiac function and morphology in mice. Echocardiographic and MRI equipment specialized for use in small rodents is&#160;expensive and requires a dedicated space. This protocol describes cardiac measurements in mice using a clinical echocardiographic system with a 15 MHz human vascular probe. Measurements are performed on anesthetized adult mice. At least three image sequences are recorded and analyzed for each animal in M-mode in the parasternal short-axis view. Afterwards, cardiac histological examination is performed, and cardiomyocyte diameters are determined on hematoxylin-eosin- or wheat germ agglutinin (WGA)-stained paraffin sections. Vessel density is determined morphometrically after Pecam-1 immunostaining. The protocol has been applied successfully to pharmacological studies and different genetic animal models under baseline conditions, as well as after experimental myocardial infarction by the permanent ligation of the left anterior descending coronary artery (LAD). In our experience, echocardiographic investigation is limited to anesthetized animals and is feasible in adult mice weighing at least 25 g.

Echocardiographic examination is frequently used in mice. Expensive high-resolution ultrasound devices have been developed for this purpose. This protocol describes an affordable echocardiographic procedure combined with histological morphometric analyses to determine cardiac morphology.

マウスの心臓形態の心エコー検査・組織学的検討

An increasing number of genetically modified mouse models has become available in recent years. Moreover, the number of pharmacological studies performed ...

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.

マウスにおける右心室拡張期パラメータの心エコー測定

Diastolic dysfunction is a prominent feature of right ventricular (RV) remodeling associated with conditions of pressure overload. However, the RV ...

Revealing Subtle Changes in Cardiac Function using Transthoracic Dobutamine Stress Echocardiography in Mice

Left ventricular (LV) dysfunction paves the final pathway for a multitude of cardiac disorders. With the non-invasive high-frequency transthoracic dobutamine stress echocardiography in humans, a reductionist investigation approach to unmask subtle changes in cardiac function has become possible. Here, we provide a protocol for using this technique in mice to facilitate expanded analysis of LV architecture and function in physiology and pathology enabling the observation of alterations in models of cardiac disease hidden in unstressed hearts. This investigation can be performed in one and the same animal and allows both, basal and pharmacologically stress-induced measurements. We outline detailed criteria for appropriate anesthesia, imaging-based LV analysis, consideration of intra- and interobserver variability, and obtaining positive inotrope response that can be attained in mice after intraperitoneal injection of dobutamine under near physiological conditions. To recapitulate the characteristics of human physiology and disease in small animal models, we highlight critical pitfalls in evaluation, e.g., a pronounced Bowditch effect in mice. To further meet translational objectives, we compare stress-induced effects in humans and mice. When used in translational studies, attention must be paid to physiological differences between mice and human. Experimental rigor dictates that some parameters assessed in patients can only be used with caution due to restrictions in spatial and temporal resolution in mouse models.

Left ventricular dysfunction constitutes the final common pathway for a host of cardiac disorders. We here provide a detailed protocol of transthoracic dobutamine stress echocardiography approach for comprehensive evaluation of the left ventricular function of mouse models of cardiac disease as well as cardiac phenotyping.

Left ventricular (LV) dysfunction paves the final pathway for a multitude of cardiac disorders. With the non-invasive high-frequency transthoracic ...

Echocardiographic Characterization of Left Ventricular Structure, Function, and Coronary Flow in Neonate Mice

Echocardiography is a non-invasive procedure that enables the evaluation of structural and functional parameters in animal models of cardiovascular disease and is used to assess the impact of potential treatments in preclinical studies. Echocardiographic studies are usually conducted in young adult mice (i.e., 4-6 weeks of age). The evaluation of early neonatal cardiovascular function is not usually performed because of the small size of the mouse pups and the associated technical difficulties. One of the most important challenges is that the short length of the pups&#39; limbs prevents them from reaching the electrodes in the echocardiography platform. Body temperature is the other challenge, as pups are very susceptible to changes in temperature. Therefore, it is important to establish a practical guide for performing echocardiographic studies in small mouse pups to help researchers detect early pathological changes and study the progression of cardiovascular disease over time. The current work describes a protocol for performing echocardiography in mouse pups at the early age of 7 days old. The echocardiographic characterization of cardiac morphology, function, and coronary flow in neonatal mice is also described.

The present protocol describes the echocardiographic assessment of left ventricular morphology, function, and coronary blood flow in 7-day old neonate mice.

新生児マウスにおける左心室構造、機能、冠動脈流の心エコー特性評価

Echocardiography is a non-invasive procedure that enables the evaluation of structural and functional parameters in animal models of cardiovascular ...