Name: echocardiographic approaches and protocols for comprehensive phenotypic characterization of valvular heart disease in mice
Uploaded: 2017-02-14T12:30:00.000+00:00
Duration: 12 min 12 s
Description: Watch this Scientific Journal Video about Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice at JoVE.com

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

<ol>
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The authors have nothing to disclose.

Yaşlanma Kardiyovasküler kalsifikasyon 1 ilerici artış ile ilişkilidir. Hemodinamik olarak önemli aort darlığı 65 2 yaşın üzerindeki nüfusun% 3 etkiler ve hatta ılımlı aort darlığı (3-4 m / s pik hız) olan hastaların% 40&#39;tan az bir 5 yıllık olaysız sağkalım 3.. Halen, orada aort kapak kalsifikasyonu ilerlemesini yavaşlatmak için etkili tedaviler ve cerrahi aort kapak replasmanı ileri aort darlığı 4 için kullanılabilir tedavidir. Başlatılması ve aort kapak kalsifikasyonu ilerlemesine katkıda mekanizmaların daha derin bir anlayış kazanıyor amaçlayan çalışmalar aort kapak darlığı 5, 6 yönetmek için farmakolojik ve cerrahi olmayan yöntemlerle doğru hareket önemli bir ilk adımdır. Genetikly-değiştirilmiş fareler çeşitli hastalıklara katkıda bulunmak ve şimdi aort darlığı 6, 7, 8 biyolojisi anlamaya yönelik mekanik çalışmalar ön plana geliyor mekanizmaların anlayışımızı geliştirilmesinde önemli bir rol oynamıştır. Gibi diğer kardiyovasküler hastalıklar aksine ateroskleroz ve kalp yetmezliği-damar ve ventrikül fonksiyonunu değerlendirmek için standart protokoller köklü-orada farelerde kalp kapak fonksiyonunun in vivo feno ilişkilendirilmiş benzersiz zorluklar çoğunlukla vardır. Son yorumları sayıda görüntüleme ve kemirgenler 9, 10, 11 valf fonksiyonunu değerlendirmek için kullanılan invaziv modalitelere avantajları ve dezavantajları konusunda kapsamlı tartışmalar sağlanan olsa da, bugüne kadar, biz bir COMPRE sağlayan bir yayının farkında değildirmlam, adım-adım farelerde fenotiplendirme kalp kapak fonksiyonu için protokol. Bu yazının amacı, farelerde kalp kapak işlevi fenotipe yöntem ve protokolleri tanımlamaktır. Tüm yöntemler ve prosedürler Mayo Kliniği Kurumsal Hayvan Bakım ve Kullanım Kurulu tarafından onaylanmıştır. Bu protokolün temel bileşenleri anestezi derinliği, kalp fonksiyonunun değerlendirilmesi ve kalp kapak fonksiyonlarının değerlendirilmesini içerir. Biz bu raporun sadece kalp kapak hastalığı alanında araştırma peşinde ilgilenen araştırmacılar rehberlik hizmet etmeyecek, aynı zamanda bu hızla büyüyen alandaki veri tekrarlanabilirlik ve geçerliliğini sağlamak için protokol standardizasyon ile ilgili ulusal ve uluslararası diyalog başlatmak umuyoruz. Önemli olan, yüksek çözünürlüklü ultrason sistemlerini kullanarak başarılı görüntüleme (yaygın sonografi kullanılan ve terminoloji) sonografi ilkelerinin bir çalışma bilgi, temel Princip bir anlayış gerektirirkalp fizyolojisi ve sonografi ile önemli bir deneyime les kemirgenlerde kardiyak fonksiyonların doğru ve zaman verimli değerlendirilmesi için izin vermek.

<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.

rutin hayvan kalp ultrason görüntülemede elde edilen görüntü örnekleri Bu yazıda yer almaktadır. Hayvanın göğüs dönüştürücü yerleştirme bir illüstrasyon okuyucu dönüştürücü açıklandığı gibi görüntüler elde etmek için konumlandırılmış nerede net bir anlayış vermek için sağlanmıştır. ultrason laboratuvar set-up bir fotoğraf da uygun ekipman, özellikle kullanılacak ultrason dönüştürücü ve anestezi yönteminin önemini vurgulamak dahildir. 2D / B-mod, M-mode ve normal ve anormal vanalar, sağ ve sol ventriküller ve aort kökü renk ve Doppler görüntüler düzgün etiketli. Strain-rate görüntüleme rutin olmasa da, bir örnek de dahildir. Mitral yetersizliği s sırasında vananın karşısında yüksek, genellikle olmayan laminer kan akış hızı (mozaik boyama) ile karakterize edilirystole (Şekil 5). EKG izleme QRS kompleksinin sonra meydana gelen MV karşısında sol atrium, sol karıncıktan böyle bir mozaik renkli Doppler akım paterni varlığı, MR kesin bir tanı için izin verir. Bu aort kapak yetersizliği ve / veya sol ventrikül disfonksiyonu yokluğunda oluştuğunda, bu izole mitral kapak prolapsusu olarak karakterize edilebilir. sol ventrikül önemli dilatasyon (nedeniyle kalp yetmezliği veya anestezi aşırı derinliğini deneysel kaynaklı için) varsa, bu iskemik mitral yetersizliği (veya yetersizliği, kardiyak disfonksiyona sekonder) olarak karakterize edilebilir. Bir nabızlı spektral Doppler görüntüleme, kan akışının bir kaçak jetinin varlığını ve zamanlamasını teyit etmek için kullanılabilir. Normal aort kapağı açın ve her kardiyak döngüsü sırasında yeterince yakın üç ince, esnek dişçiklidir. Aort kapak doruk ayırma 2D-güdümlü ölçülüruzun eksen görünümünde aort kapağının M-mod. Elektronik kaliperler sol aort zirve ön kenarına doğru aortik doruk (Şekil 3) ön kenarının ölçmek için kullanılır. Normal farelerde aort kapak doruk-ayırma mesafesi 0.9 1.3 mm&#39;dir. Renkli Doppler sistol sırasında aort kökü valfi genelinde ve içine laminar akışını gösterir. Türbülanslı akış gibi aort kapak yetersizliği gibi arttığı akışının şartlarında takdir ya da aort darlığı gibi basınç artırılabilir. Bu çıkış yolunda mozaik boyama olarak gösterilmiştir. aort kapak yetersizliğinin bile küçük miktarlarda nedeniyle hiperdinamik kalp fonksiyonu için zirve transvalvüler hızında önemli artışlar neden olabilir ve ventrikül ön artmış sol. 1.50 m / s 0.90 m / s normal fareler aralıklarında Tepe aort hız. &gt; 5 m Tepe aort kapak hızı / s ciddi aort kapak darlığı olan farelerde kaydedilmiştir. 12 (Şekil 8) bir dizin sağlamak için kullanılabilir. Pulmoner arter hızlanma süresi pik akım hızına sistolik pulmoner arter akımının başlangıcından zaman aralığıdır. Sağ ventrikül ejeksiyon zamanı sistolik pulmoner arter sistolik akışının kesilmesi var olduğu noktaya kadar sağ ventrikül ejeksiyon başlangıcı arasındaki aralığıdır. sağ ventrikül ejeksiyon zamanına pulmoner arter hızlanma süresi oranında bir azalma ile kısaltılmış pulmoner arter hızlanma zamanı kombinasyonu (pulmoner arter veya sağ ventrikül basıncının invaziv ya da doğrudan önlemler kullanılarak teyit edilebilir) Pulmoner arteriyel hipertansiyon varlığını düşündürmektedir . <img alt="Şekil 1" src="/files/ftp_upload/54110/54110fig1.jpg" /> Şekil 1: Hayvan Cardiac Ultrason Laboratuvarı. Laboratuvar yüksek frekansta (30 MHz ve 40 MHz) dönüştürücüler (MS 400 ve MS 550D), izofluran difüzör, hayvan platformu, sıcaklık ve kalp atış hızı monitörü,% 1 ila 1.5% izofluran ile küçük hayvan adanmış ultrason makinesi ile donatılmıştır 1 L / dk% 100 O 2, burun konisi ve izofluran difüzör ve% 100 O 2, saç jilet, ultrason jeli, elektrot jeli, yapışkan bantlar ve kağıt havlu bağlı boru ile karıştırılır. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig1large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> <img alt="şekil 2" src="/files/ftp_upload/54110/54110fig2.jpg" /> Şekil 2: Temel Dönüştürücü Pozisyonlar. (A) Parasternal penceresi. dönüştürücü kafa kaudal yönettiği transdüser görüntü indeksi işaretleyici ile sol parasternal sınırında yer alıyor. FrBu pozisyon om, sol ventrikül, aort kapak ve aort kökü ve pulmoner kapağın kısa eksen görünümü uzun eksen görünümü elde edilebilir. Parasternal pencereden (B), dönüştürücü kafa posterior yönettiği çentik ile, saat yönünün tersine döndürülür. Bu pozisyonda, sol ventrikül ve aort kapağına ve pulmoner vana uzun eksen görünümü kısa eksen görünümü elde edilebilir. (C) Apikal penceresi. dönüştürücü kafa kalbin tepesinde konumlandırılmış. Bu pozisyondan, sağ ve sol ventriküller mitral ve triküspit kapaklarda uzun ekseni görünümü elde edilebilir. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig2large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> <img alt="Şekil 3," src="/files/ftp_upload/54110/54110fig3.jpg" /> Şekil 3: Norm içinde Aort Kapak Fonksiyon DeğerlendirilmesiKalsifik Aort Kapak Hastalığı ile Fare Aort Kapak Fonksiyon karşı el Fare. Uzun eksen görünümünde normal aort kapağının (A) 2D görüntü. Aort kapak sistol sırasında iyi açılır unutmayın. (B) M-mod görüntü normal aort kapak fonksiyonunu (kutu gibi bir görünüm) tasvir. doruk-ayırma mesafesi 1.12 mm ölçülen unutmayın. Normal aort kapağı boyunca zirve hızı (C) spektral Doppler görüntüleme 1,3 m / s meaured edildi. Düşük dansiteli lipoprotein reseptörü uzun eksen görünümünde kalsifiye aort kapağının (D) 2D görüntü eksik (LDLR - / -) ve batı diyet ile beslenen apolipoprotein B100 sadece (apo B 100/100) fare. tüberkülü, kalınlaşmış ve sistol sırasında kısıtlı açılış sonuçlanır ekojenitenin artmıştır. Aynı stenotik aort kapak gösteren (E) Bir M-mod görüntü 0,7 mm doruk ayırma mesafe ölçümü gösterir. (F &lt;) / Strong&gt; stenotik aort kapağı boyunca zirve hızının spektral Doppler görüntüleme 4,6 m / s meaured edildi. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig3large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> <img alt="Şekil 4," src="/files/ftp_upload/54110/54110fig4.jpg" /> Şekil 4: Normal Mitral Kapak M-mode. apikal pencereden, mitral kapak uzun eksen görünümü elde edilir. sorgulama M-mod hat mitral kapak broşür karşısında uygulanır. mitral yaprakçık kalınlığı teorik elektronik kaliperler kullanılarak ölçülebilir, ancak bu normal bir mitral kapak broşürler hareketli hızla ince, zayıf ekojenik verilen son derece zorlu olabilir ve. oklar sistolde mitral kapak broşürün M-mod işaret etmektedir. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig4large.jpg" target="_blank">cli Lütfen</a>Bu rakamın büyük halini görmek için buraya ck. <img alt="Şekil 5," src="/files/ftp_upload/54110/54110fig5.jpg" /> Şekil 5: Renkli Doppler Görüntüleme kullanarak Mitral Kapak Regurjitan Jet kanıtı. parasternal pencere, mitral kapak değiştirilmiş uzun eksen görünümü elde edilir. Renkli Doppler sorgulama (bir okla vurgulanan) sistol sırasında mitral kapak bir mozaik renkli jet gösterir. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig5large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> <img alt="Şekil 6," src="/files/ftp_upload/54110/54110fig6.jpg" /> Şekil 6: Uzun eksen Ana Pulmoner Arter Görünüm ve Binbaşı Şubeler. Ana pulmoner arter uzun eksen görünümü (MPA) ve sağ (ECP) ve sol (LPA) dalları paraste elde edilebilir<html> rnal pencere. Sağ ventrikül çıkış yolu (RVOT), pulmoner kapak (PV) ve aort (AO) kısmen görülmektedir. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig6large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> <img alt="Şekil 7," src="/files/ftp_upload/54110/54110fig7.jpg" /> Şekil 7: Normal Pulmonik Vana tasvir M-mod görüntü. parasternal pencere, pulmoner vananın kısa ve uzun eksen görünümleri elde edilebilir. sorgulama M-mod hat pulmoner kapaktaki uygulanır. pulmoner kapak doruk ayırma (oklar) mesafe bu görünümden ölçülebilir. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig7large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> 10fig8.jpg &quot;/&gt; Şekil 8: Pulmonik Vana karşısında Akış Pulsed-dalga Doppler Sorgulama. pulmoner arter hızlanma süresi (Paat) pik akım hızına sistolik pulmoner arter akımının başlangıcından zaman aralığıdır. Sağ ventrikül ejeksiyon zamanı (RVET) akış kesilmesi var olduğu noktaya kadar sağ ventrikül ejeksiyon başlangıcı arasındaki aralığıdır. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig8large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> <img alt="Şekil 9," src="/files/ftp_upload/54110/54110fig9.jpg" /> Şekil 9: Sol Ventrikül Kısa eksenli Görünüm tasvir M-mod görüntü. parasternal pencere, sol ventrikül kısa eksen görünümü yönünün tersine dönüştürücü kafa döndürülerek elde edildiği şekilde posterior veya dorsal görüntü indeks işaretleyici noktaları. M-modsorgulama e hattı papiller kas seviyesinde sol ventrikül boyunca uygulanır. Sol ventrikül diyastol sonu çapı (SVDSÇ), sol ventrikül sistol sonu boyut (SVSSÇ) ve ön duvar (AW) ve arka duvar (PW) kalınlıkları kolaylıkla ölçülebilir. Herhangi bir ölçümlerde papiller kas (*) dahil vermemek için dikkatli olun. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig9large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> <img alt="Şekil 10," src="/files/ftp_upload/54110/54110fig10.jpg" /> Şekil 10: Renkli Doppler Değerlendirme ve Mitral Kapak Girişi Pulsed-dalga Doppler spektral görüntü. (A) apikal uzun eksen görünümünde mitral kapak girişi renkli doppler değerlendirme gösteren resim. 2B renkli Doppler görüntü g için kritik bir araç olabilir unutmayın(Panel B tasvir) pulsed Doppler traselerin edinimi için uygun numune hacmi pozisyonunu uiding. (B) darbeli dalga Doppler ile mitral kapak girişi spektral ekran. (Apikal uzun eksen görünümünde) mitral kapak genelinde kan akışının darbeli dalga Doppler değerlendirmesi, sol ventrikül diyastolik fonksiyonu değerlendirmek için yapılır. numune hacmi mitral kapaktaki ucunda yer almaktadır. izovolumik gevşeme zamanı (IVRT), izovolümik kasılma zamanı (İVKZ), sol ventrikül ejeksiyon zamanı (LVET), ve tepe mitral hızı (E) tüm mitral kapak genelinde darbeli dalga Doppler hızları spektral ekran elde edilebilir. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig10large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> <img alt="Şekil 11," src="/files/ftp_upload/54110/54110fig11.jpg" /> Fişekil 11: Septal Mitral anulus Doku Doppler Görüntüleme. apikal pencereden, mitral kapak uzun eksen görünümü elde edilir. Doku Doppler örnek hacim mitral annulus septal bölgede konumlandırılmış. zirve mitral akım hızı (Şekil 10B değişken E) ve (e zirve mitral anulus doku hızı arasındaki oran &#39;(beyaz oklarla gösterilen) sol ventrikül diyastolik fonksiyonlarını değerlendirmek için kullanılır yaygın e / e olarak anılacaktır&#39;). <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig11large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</a> <img alt="Şekil 12," src="/files/ftp_upload/54110/54110fig12.jpg" /> Şekil 12: Sol ventrikülün Strain ve Strain Oranı değerlendirilmesi. Orada mevcut uzman analiz yazılım paketleri ticari, ve strain ve strain rate değişkenleri olabiliriçsel miyokard kasılma özelliklerinin erken veya sub-klinik değişiklikler tedbirler olarak elde edilmiştir. Yukarıda gösterilen örnekler farelerde yaygın edinilmiş görüntüleme düzlemlerde radyal gerginlik ve gerilme oranı betimliyor. Bu görüntüleme uçakları (ve gerilme traselerin sonraki şekli) sık sık apikal uzun eksen ya da 4 odacıklı görünümünde elde edilen insanlarda görüntüleri, farklı olabileceğini unutmayın. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig12large.jpg" target="_blank">Bu rakamın büyük halini görmek için lütfen buraya tıklayınız.</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.

Farelerde Kalp Kapak Hastalıkları Kapsamlı Fenotipik Karakterizasyonu için ekokardiyografik Yaklaşımlar ve Protokoller

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

Video: 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.

Fareler de Transtorasik Ekokardiyografi

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

Tıp

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.

Murin Ekokardiyografi ve Ultrasonik Görüntüleme

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.

Farelerde Sağ Kalp Ekokardiyografik Değerlendirilmesi

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.

Fetal Faredeki Yapısal Kalp Kusurlarını Karakterize Etmek İçin Bir Boru Hattı

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 ...

Gelişim Biyolojisi

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.

Ekokardiyografik Imaging tarafından Enine Aort Konstrüksiyon Mouse Modeli Kardiyak Morfolojik Değerlendirilmesi ve Fonksiyonel Değişiklikler

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.

Sol Atrium, Aort ve Pulmoner Arter Murin Ekokardiyografi

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.

Kardiyak Morfoloji Ekokardiyografik ve histolojik muayene fare

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.

Fareyle sağ ventriküler diyastolik parametrelerin ekokardiyografik ölçümü

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.

Yenidoğan Farelerde Sol Ventrikül Yapısının, Fonksiyonunun ve Koroner Akımın Ekokardiyografik Karakterizasyonu

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