この作品は、OU一般財源G110および生物医学研究およびOSU-HRS基金013000の優良研究基金の助成金によってサポートされています。著者はこの原稿を作成する際に、彼女の援助のためローレン·チェンに感謝したいと思います。

動物を対象とするすべての手順は、オハイオ州立大学施設内動物管理使用委員会（IACUC）の規制やガイドラインに従っておよびコンプライアンスに承認され、完了した。 1。マウス麻酔<ol><li>外科タオルに包まれた加熱された等温パッドでクリーンな手順テーブルを設定します。加熱パッドは、動物への潜在的な応力を低減しつつ、動物の中核温度を安定化させるために30°Cと34°Cの間に維持されるべきである。 </li><li>次のパラメータを使用して麻酔導入室でマウスを配置します。酸素流量は1.5リットル/分、3.5％に設定さイソフルラン気化器にセット。完全な鎮静は1〜2分以内に行われるべきである。導入室が利用できない場合、ベルジャーをイソフルランと直接動物の接触を避けるために底部に配置金網で使用することができる。 </li><li>すぐに誘導室からマウスを削除するには、一度（マウスは自発的な運動機能を失ったときに達成される）、完全に麻酔をかける。麻酔の維持のために動物にノーズコーンを適用します。酸素流量は約0.5 L /分に低減されるべきであるとイソフルラン気化器は、1.5から2.5パーセントの範囲に設定されるべきである。 <ol><li>目の乾き8を減らすために角膜に直接眼軟膏を少量を適用します。また、麻酔中、マウスは粘膜がピンク色のまま必要があり、呼吸が安定した表示され、ペダル引っ込め反射がないことを維持する必要があります。 </li></ol></li></ol> 2。超音波診断イメージング手続きの準備<ol><li>このような外科テープなどの取り外し可能な接着剤を用いて加熱された手順テーブルの上でマウスの各脚を抑制。 </li><li>電気かみそりを使用して、胸腔まで腹部と半分の方法間の腹側の体表面の毛を削除します。さらに再除毛クリームを塗るかみそりでカットされていない残りの髪​​を移動します。 2〜3分後に湿ったガーゼパッドでクリームを拭き取ってください。 </li><li>水で湿らせたガーゼパッドを使用して余分な毛を削除し、70％アルコールや消毒薬と同等で剃毛地域を清掃してください。超音波プローブは、絞り機能を可視化するためにこの領域に適用される。局所用鎮痛剤は、毛髪除去に起因する軽度の皮膚刺激を経験した動物に提供することができる。 </li></ol> 3。超音波診断イメージング·プロトコル<ol><li>超音波装置の電源をオンにして、最適な解像度を得るために、パーセンテージで機器に出力電力を必要に応じて調整してください。 </li><li>マウス横隔膜の収縮を適切に可視化を可能にし、B（明るさ）モード、M（モーション）モード、またはその両方イメージング前に、次のいずれかに超音波装置を設定します。 </li><li>マウスの上腹部に超音波ゲルを少量を適用し、胸腔に向かってゲルをマッサージ。 </li><li>超を配置心に向けたこの分野でのトランスデューサと上向きの角度に聞こえる。画像の最適化された解像度が達成されるまで、プローブを調整します。注意：このプロトコルのために、マイクロコンベックスアレイまたはリニアフェイズドアレイトランスデューサは、小さなフットプリントと優れた距離分解能9に使用するのに理想的なプローブである。周波数帯域にわたって調整されると、これらの実験のために6.5から12メガヘルツの範囲を利用することができる必要がある。 </li><li>一時的にダイヤフラムの画像を保存し、選択された収縮を表示するには、フリーズボタンを押してください。 </li><li>横隔膜遠足など呼吸数の後の測定が可能になるシネ·ループなどの記録を保存します。注意：画像のフレームは将来の分析9にコンピュータ内にまたは外付けハードドライブに保存することができます。 <ol><li>正確に超音波ソフトウェアの一部である電子ノギスを使用して収縮緩和から横隔膜の動きの深さを測定します。 </li><li>変換MPEGファイルにシネループファイルおよび記録期間横隔膜収縮の回数をカウントすることによって、呼吸速度を決定する。あるいは、分（呼吸数）当たりの収縮数をMモード画像から数えてもよい。 </li></ol></li></ol> 4。ポスト麻酔動物の回復<ol><li>マウスが完全に1時間以内に麻酔から回復する必要があります。それは胸骨横臥位を維持するのに十分な意識を取り戻したまでは無人の動物を放置しないでください。 </li></ol>

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	<li>Snyder, C. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complementarity+of+ultrasound+and+fluorescence+imaging+in+an+orthotopic+mouse+model+of+pancreatic+cancer.">Complementarity of ultrasound and fluorescence imaging in an orthotopic mouse model of pancreatic cancer.</a> BMC Cancer. 9, 106 (2009).</li><li>Franchi, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+assessment+of+cardiac+function+in+a+mouse+model+of+renovascular+hypertension.">Non-invasive assessment of cardiac function in a mouse model of renovascular hypertension.</a> Hypertension Research: Official Journal of the Japanese Society of Hypertension. , (2013).</li><li>Coatney, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+imaging:+principles+and+applications+in+rodent+research.">Ultrasound imaging: principles and applications in rodent research.</a> ILAR Journal / National Research Council, Institute of Laboratory Animal Resources. 42, 233-247 (2001).</li><li>Morenz, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+air+trapping+in+chronic+obstructive+pulmonary+disease+by+low+frequency+ultrasound.">Detection of air trapping in chronic obstructive pulmonary disease by low frequency ultrasound.</a> BMC Pulmonary Medicine. 12, 8 (2012).</li><li>Gilliam, L. A., Moylan, J. S., Ann Callahan, L., Sumandea, M. P., Reid, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doxorubicin+causes+diaphragm+weakness+in+murine+models+of+cancer+chemotherapy.">Doxorubicin causes diaphragm weakness in murine models of cancer chemotherapy.</a> Muscle &amp; Nerve. 43, 94-102 (2011).</li><li>Ferreira, L. F., Campbell, K. S., Reid, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effectiveness+of+sulfur-containing+antioxidants+in+delaying+skeletal+muscle+fatigue.">Effectiveness of sulfur-containing antioxidants in delaying skeletal muscle fatigue.</a> Medicine and Science in Sports and Exercise. 43, 1025-1031 (2011).</li><li>Zuo, L., Clanton, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reactive+oxygen+species+formation+in+the+transition+to+hypoxia+in+skeletal+muscle.">Reactive oxygen species formation in the transition to hypoxia in skeletal muscle.</a> American Journal of Physiology. Cell Physiology. 289, 207-216 (2005).</li><li>Helms, M. N., Torres-Gonzalez, E., Goodson, P., Rojas, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+tracheal+instillation+of+solutes+into+mouse+lung.">Direct tracheal instillation of solutes into mouse lung.</a> J. Vis. Exp. , (2010).</li><li>Hedrick, W. R., Hykes, D. L., Starchman, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Ultrasound Physics and Instrumentation. , 445 (2005).</li><li>von Sarnowski, B., Khaw, A. V., Kessler, C., Schminke, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+a+microconvex+array+transducer+for+the+ultrasonographic+examination+of+the+intrathoracic+segments+of+the+supraaortic+arteries.">Evaluation of a microconvex array transducer for the ultrasonographic examination of the intrathoracic segments of the supraaortic arteries.</a> Journal of Neuroimaging: Official Journal of the American Society of Neuroimaging. 20, 246-250 (2010).</li><li>Stocksley, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Abdominal Ultrasound. , 7-8 (2001).</li><li>Kremkau, F. W., Taylor, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Artifacts+in+ultrasound+imaging.">Artifacts in ultrasound imaging.</a> Journal of Ultrasound in Medicine: Official Journal of the American Institute of Ultrasound in Medicine. 5, 227-237 (1986).</li><li>Kremkau, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnostic+Ultrasound:+Principles+and+Instruments.">Diagnostic Ultrasound: Principles and Instruments.</a> Saunders Elsevier. , 521 (2006).</li><li>Laing, F. C., Kurtz, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+importance+of+ultrasonic+side-lobe+artifacts.">The importance of ultrasonic side-lobe artifacts.</a> Radiology. 145, 763-768 (1982).</li><li>Abu-Zidan, F. M., Hefny, A. F., Corr, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+ultrasound+physics.">Clinical ultrasound physics.</a> Journal of Emergencies, Trauma, and Shock. 4, 501-503 (2011).</li><li>Gargani, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+ultrasound:+a+new+tool+for+the+cardiologist.">Lung ultrasound: a new tool for the cardiologist.</a> Cardiovascular Ultrasound. 9, 6 (2011).</li><li>Sanders, R. C., Winter, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Clinical Sonography A Practical Guide. , 632 (2007).</li></ol>

著者は、彼らが競合する経済的利益を持っていないことを宣言します。

現在の実験プロトコルは、非侵襲性のインビボのアプローチを介して、マウスモデルにおける絞り活性に特異的な診断用超音波イメージング技術を開発する。麻酔器の設定は、個々のマウスを麻酔に異なって応答することができるので、わずかに各動物について調整してもよい近似値である。不適切な麻酔の投与を防止するためには、定期的に、心拍数、呼吸数、および体温を含むマ?...

近年、超音波診断技術は、腎血管性高血圧および膵臓癌1,2のマウスモデルに適用されている。しかしながら、これらの技術が広く齧歯類呼吸筋機能アッセイにおいて使用されていない。したがって、我々はマウスにおけるダイヤフラムの移動の長手インビボ評価のための貴重なツールとして、超音波診断法を開発した。 診断用超音波イメージングにはいくつかの利点がある。例えば、それは、非侵襲的に安全、ポータブルであり、比較的低コストで3リアルタイム測定を可能にする。特に、特定の低周波数超音波装置は、空気トラッピング、重度の気流の制限〜4の軽度の慢性閉塞性肺疾患（COPD）の臨床的特徴を検出することができた。従って、診断用超音波イメージングは​​、リアルタイム監視のための容易にアクセス可能で再現可能なスクリーニング方法として機能することができる<html>呼吸器疾患の。 診断用超音波イメージング技術は、しばしば、大型動物またはヒト対象に適用される。しかし、小規模なテーマについて、超音波を実施する課題への可能性が高いマウスモデルでの超音波イメージング研究、限られた数がありました。現在のプロトコルは、マウスに絞り機能を測定するための新たな手順の概要を説明します。絞り機能に関するいくつかのげっ歯類の研究が行われているが、また、結果のほとんどは安楽死させ、動物5-7から直接、筋肉片を単離することによって生成した。対照的に、ダイヤフラム活性を分析するためのインビボ診断超音波イメージング法を用いて、実験のために屠殺した動物の数を減少させるであろう。また、振動板の収縮性の強化に焦点を当てた長期治療を正確に動物を犠牲にすることなく、げっ歯類モデルにおける超音波によって評価してもよい。 ntent」は&gt;我々の研究室では、可視化、ならびに、 生体内で絞り機能の理解を助ける動物に侵襲的な方法を回避し、治療薬の開発を補助する超音波装置を用いて、マウスの横隔膜活性を分析するための有効な方法を開発した呼吸機能障害の治療。

<table><tbody><tr><td>Veterinary digital ultrasonic diagnostic imaging system</td><td>Edan</td><td>DUS 3 VET</td><td>Ultrasound parameters include: frequency of 6.5 MHz, Depth of 29 mm. Note: An equivalent ultrasound machine may be used for this protocol</td></tr><tr><td>Micro-convex array transducer</td><td>Edan</td><td>C611</td><td>Or equivalent</td></tr><tr><td>GE Logiq i hand-carried unit (HCU)&amp;nbsp;</td><td>GE Healthcare</td><td>GE Logiq i hand-carried unit (HCU)</td><td>Or equivalent</td></tr><tr><td>GE 12 MHz linear array probe</td><td>GE Healthcare</td><td>12L-RS</td><td>Or equivalent</td></tr><tr><td>Veterinary anesthetic vaporizer</td><td>Webster Veterinary</td><td>Serial #: W422021</td><td>Isoflurane was exclusively used with this vaporizer (or equivalent). A custom made induction chamber for anesthesia was assembled for initial anesthesia. Maintenance anesthesia was performed using a nose cone</td></tr><tr><td>Isothesia (Isoflurane, USP)</td><td>Butler Schein</td><td>29405 250ML PVL</td><td>Or equivalent</td></tr><tr><td>Enviro-pure anesthesia absorbing canister</td><td>Surgivet Smiths Medical PM, Inc.</td><td>Part #: 32373B10</td><td>Or equivalent</td></tr><tr><td>Ultrasound transmission gel</td><td>HM Sonic</td><td>N/A</td><td>Or equivalent&amp;nbsp;</td></tr><tr><td>Puralube vet ointment</td><td>Puralube</td><td>NDC 17033-211-38</td><td>Or equivalent</td></tr><tr><td>Deltaphase isothermal pad</td><td>Braintree Scientific Inc.</td><td>39DP</td><td>Or equivalent</td></tr><tr><td>Hair remover</td><td>Nair</td><td>N/A</td><td>Or equivalent</td></tr><tr><td>Electric razor</td><td>Remington</td><td>HC-5015</td><td>Or equivalent</td></tr><tr><td>Surgical tape</td><td>3M Micropore</td><td>1530-1</td><td>Or equivalent</td></tr><tr><td>Gauze sponges</td><td>Dynarex</td><td>3262</td><td>Or equivalent</td></tr></tbody></table>

diagnostic ultrasound imaging of mouse diaphragm function

齧歯類呼吸骨格筋の機能解析、特にダイヤフラムは、一般に侵襲性外科手術手順を使用して筋肉ストリップを単離することによって行われる。これは、インビトロ絞り活性を評価する有効な方法であるが、非生存手術を伴う。 この生体内における方法のような非侵襲性の超音波イメージングのアプリケーションは、屠殺動物の数を減少させるのみならずので有益であるが、生きたマウスにおける疾患の進行をモニターするのに適している。したがって、我々の超音波イメージング方法は、おそらく、種々の呼吸器疾患によって誘発される筋肉損傷を軽減する新規な治療法の開発を支援することができる。特に、閉塞性肺疾患の臨床診断において、超音波撮像は、横隔膜筋疲労の早期発症を検出するための他の標準的試験と併せて使用する可能性を有する。現在のプロトコルでは、我々は正確にダイヤフラムcontractiを評価する方法について説明診断用超音波イメージング技術を用いてマウスモデルでのリティ。

マウスの横隔膜の典型的な超音波画像を、 図1Aに示されている。マウスの横隔膜の最大垂直変位を記録した。この距離は、正確に超音波ソフトウェア。 表1に、三つの異なるマウスからの横隔膜収縮のこれらの距離測定値の一部である電子カリパスを用いて収縮弛緩ダイアフラムの動きの深さを測定することにより算出した。 MPEGファイルにシネループファイルを変?...

Watch this Scientific Journal Video about Diagnostic Ultrasound Imaging of Mouse Diaphragm Function at JoVE.com

Diagnostic Ultrasound Imaging of Mouse Diaphragm Function

診断用超音波イメージングは​​、ヒトおよび動物対象の様々な呼吸器疾患の診断に有効であることが証明されている。我々は、特異的マウスモデルにおいて振動板動態を分析するためにZuoの博士の研究室によって利用される総合的な超音波プロトコルを示す。これはまた、マウス呼吸筋機能に関する定量的情報を提供することができる非侵襲的な研究技術である。

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 ...

diagnostic-ultrasound-imaging-of-mouse-diaphragm-function

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

16.0K ビュー.  The Ohio State University College of Medicine. 診断用超音波イメージングは​​、ヒトおよび動物対象の様々な呼吸器疾患の診断に有効であることが証明されている。我々は、特異的マウスモデルにおいて振動板動態を分析するためにZuoの博士の研究室によって利用される総合的な超音波プロトコルを示す。これはまた、マウス呼吸筋機能に関する定量的情報を提供することができる非侵襲的な研究?...

ビデオ: Diagnostic Ultrasound Imaging of Mouse Diaphragm Function

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  ...

Murine Fetal Echocardiography

Transgenic mice displaying abnormalities in cardiac development and function represent a powerful tool for the understanding the molecular mechanisms underlying both normal cardiovascular function and the pathophysiological basis of human cardiovascular disease. Fetal and perinatal death is a common feature when studying genetic alterations affecting cardiac development 1-3. In order to study the role of genetic or pharmacologic alterations in the early development of cardiac function, ultrasound imaging of the live fetus has become an important tool for early recognition of abnormalities and longitudinal follow-up. Noninvasive ultrasound imaging is an ideal method for detecting and studying congenital malformations and the impact on cardiac function prior to death 4. It allows early recognition of abnormalities in the living fetus and the progression of disease can be followed in utero with longitudinal studies 5,6. Until recently, imaging of fetal mouse hearts frequently involved invasive methods. The fetus had to be sacrificed to perform magnetic resonance microscopy and electron microscopy or surgically delivered for transillumination microscopy. An application of high-frequency probes with conventional 2-D and pulsed-wave Doppler imaging has been shown to provide measurements of cardiac contraction and heart rates during embryonic development with databases of normal developmental changes now available 6-10. M-mode imaging further provides important functional data, although, the proper imaging planes are often difficult to obtain. High-frequency ultrasound imaging of the fetus has improved 2-D resolution and can provide excellent information on the early development of cardiac structures 11.

Fetal and perinatal death is a common feature when studying genetic alterations affecting cardiac development. High-frequency ultrasound imaging has improved 2-D resolution and can provide excellent information on early cardiac development and is an ideal method to detect the impact on cardiac structure and function prior to death.

ネズミの胎児心エコー検査

Transgenic mice displaying abnormalities in  cardiac development and function represent a powerful tool for the understanding  the molecular mechanisms ...

Detection of Lung Tumor Progression in Mice by Ultrasound Imaging

With ~1.6 million victims per year, lung cancer contributes tremendously to the worldwide burden of cancer. Lung cancer is partly driven by genetic alterations in oncogenes such as the KRAS oncogene, which constitutes ~25% of lung cancer cases. The difficulty in therapeutically targeting KRAS-driven lung cancer partly stems from having poor models that can mimic the progression of the disease in the lab. We describe a method that permits the relative quantification of primary KRAS lung tumors in a Cre-inducible LSL-KRAS G12D mouse model via ultrasound imaging. This method relies on brightness (B)-mode acquisition of the lung parenchyma. Tumors that are initially formed in this model are visualized as B-lines and can be quantified by counting the number of B-lines present in the acquired images. These would represent the relative tumor number formed on the surface of the mouse lung. As the formed tumors develop with time, they are perceived as deep clefts within the lung parenchyma. Since the circumference of the formed tumor is well-defined, calculating the relative tumor volume is achieved by measuring the length and width of the tumor and applying them in the formula used for tumor caliper measurements. Ultrasound imaging is a non-invasive, fast and user-friendly technique that is often used for tumor quantifications in mice. Although artifacts may appear when obtaining ultrasound images, it has been shown that this imaging technique is more advantageous for tumor quantifications in mice compared to other imaging techniques such as computed tomography (CT) imaging and bioluminescence imaging (BLI). Researchers can investigate novel therapeutic targets using this technique by comparing lung tumor initiation and progression between different groups of mice.

This protocol describes the steps taken to induce KRAS lung tumors in mice as well as the quantification of formed tumors by ultrasound imaging. Small tumors are visualized in early timepoints as B-lines. At later timepoints, relative tumor volume measurements are achieved by the measurement tool in the ultrasound software.

超音波画像によるマウスにおける肺腫瘍の進行の検出

With ~1.6 million victims per year, lung cancer contributes tremendously to the worldwide burden of cancer. Lung cancer is partly driven by genetic ...

がん研究

Non-Invasive Ultrasound Assessment of Endometrial Cancer Progression in Pax8-Directed Deletion of the Tumor Suppressors Arid1a and Pten in Mice

Uterine cancers can be studied in mice due to the ease of handling and genetic manipulation in these models. However, these studies are often limited to assessing pathology post-mortem in animals euthanized at multiple time points in different cohorts, which increases the number of mice needed for a study. Imaging mice in longitudinal studies can track the progression of disease in individual animals, reducing the number of mice needed. Advances in ultrasound technology have allowed for the detection of micrometer-level changes in tissues. Ultrasound has been used to study follicle maturation in ovaries and xenograft growth but has not been applied to morphological changes in the mouse uterus. This protocol examines the juxtaposition of pathology with in vivo imaging comparisons in an induced endometrial cancer mouse model. The features observed by ultrasound were consistent with the degree of change seen by gross pathology and histology. Ultrasound was found to be highly predictive of the observed pathology, supporting the incorporation of ultrasonography into longitudinal studies of uterine diseases such as cancer in mice.

Non-Invasive Ultrasound Assessment of Endometrial Cancer Progression in Pax8-Directed Deletion of the Tumor Suppressors Arid1a and Pten in Mice

This protocol describes a method for monitoring the progression of morphological changes over time in the uterus in an inducible mouse model of endometrial cancer using ultrasound imaging with correlation to gross and histological changes.

マウスにおける腫瘍抑制因子 Arid1a および Pten のPax8指向性欠失における子宮内膜癌進行の非侵襲的超音波評価

Uterine cancers can be studied in mice due to the ease of handling and genetic manipulation in these models. However, these studies are often limited to ...

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 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 ...

横隔膜の機能障害の評価と治療のモニタリングにおける超音波

Measuring Diaphragm Thickness and Function Using Point-of-Care Ultrasound

The diaphragm is the main component of the respiratory muscle pump. Diaphragm dysfunction can cause dyspnea and exercise intolerance, and predisposes affected individuals to respiratory failure. In mechanically ventilated patients, the diaphragm is susceptible to atrophy and dysfunction through disuse and other mechanisms. This contributes to failure to wean and poor long-term clinical outcomes. Point-of-care ultrasound provides a valid and reproducible method for evaluating diaphragm thickness and contractile activity (thickening fraction during inspiration) that can be readily employed by clinicians and researchers alike. This article presents best practices for measuring diaphragm thickness and quantifying diaphragm thickening during tidal breathing or maximal inspiration. Once mastered, this technique can be used to diagnose and prognosticate diaphragm dysfunction, and guide and monitor response to treatment over time in both healthy individuals and acute or chronically ill patients.

著者スポットライト:横隔膜機能と患者の転帰に対する人工呼吸器の影響を解明する 

Diaphragm thickness and function can be assessed in healthy individuals and critically ill patients using point-of-care ultrasound. This technique offers an accurate, reproducible, feasible, and well-tolerated method for evaluating diaphragm structure and function.

ポイントオブケア超音波を用いたダイアフラムの厚さと機能の測定

The diaphragm is the main component of the respiratory muscle pump. Diaphragm dysfunction can cause dyspnea and exercise intolerance, and predisposes ...

マウス絞り機能の診断用超音波イメージング

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マウスにおける経胸壁心エコー

マウス心エコー検査と超音波イメージング

ネズミの胎児心エコー検査

超音波画像によるマウスにおける肺腫瘍の進行の検出

マウスにおける腫瘍抑制因子 Arid1a および Pten のPax8指向性欠失における子宮内膜癌進行の非侵襲的超音波評価

左心房、大動脈、肺動脈のマウス心エコー

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

ポイントオブケア超音波を用いたダイアフラムの厚さと機能の測定

ポイントオブケア超音波を用いたダイアフラムの厚さと機能の測定

ポイントオブケア超音波を用いたダイアフラムの厚さと機能の測定