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

<ol>
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The authors have nothing to disclose.

El envejecimiento se asocia con aumentos progresivos en la formación de calcificaciones cardiovasculares 1. La estenosis de la válvula aórtica hemodinámicamente significativa afecta al 3% de la población mayor de 65 años 2, y los pacientes con estenosis de la válvula aórtica, incluso moderada (velocidad pico de 3-4 m / s) tienen una supervivencia libre de eventos a los 5 años del 40% menos de 3. En la actualidad, no existen tratamientos eficaces para frenar la progresión de la calcificación de la válvula aórtica, y el reemplazo de la válvula aórtica quirúrgica es el único tratamiento disponible para la estenosis de la válvula aórtica avanzada 4. Estudios dirigidos a obtener una comprensión más profunda de los mecanismos que contribuyen a la iniciación y progresión de la calcificación de la válvula aórtica son un primer paso clave en el camino hacia métodos farmacológicos y no quirúrgicos para tratar la estenosis de la válvula aórtica 5, 6. Genéticoratones alterados-ly han jugado un papel importante en el desarrollo de nuestra comprensión de los mecanismos que contribuyen a una variedad de enfermedades y ahora están llegando a la vanguardia de los estudios sobre los mecanismos destinados a comprender la biología de la estenosis de la válvula aórtica 6, 7, 8. A diferencia de otras enfermedades cardiovasculares tales como la aterosclerosis y la insuficiencia cardíaca, donde protocolos estándar para evaluar la función vascular y ventricular son en su mayor parte bien establecidos-hay retos únicos asociados con el fenotipo in vivo de la función de válvula de corazón en ratones. Aunque recientes han proporcionado discusiones exhaustivas sobre las ventajas y desventajas de numerosas imágenes y modalidades invasivas utilizadas para evaluar la función de la válvula de roedores 9, 10, 11, hasta la fecha, no tenemos conocimiento de una publicación que proporciona una comprehensive, paso a paso protocolo para la función de la válvula del corazón fenotipificación en ratones. El propósito de este manuscrito es describir los métodos y protocolos al fenotipo función de válvula de corazón en ratones. Todos los métodos y procedimientos han sido aprobados por el Comité Institucional de Cuidado y Uso de Animales de la Clínica Mayo. Los componentes clave de este protocolo incluyen la profundidad de la anestesia, la evaluación de la función cardíaca, así como la evaluación de la función de las válvulas cardíacas. Esperamos que este informe no sólo servirá para guiar a los investigadores interesados ​​en seguir la investigación en el campo de la enfermedad de las válvulas del corazón, sino que también se iniciará un diálogo nacional e internacional relacionada con el protocolo de estandarización para asegurar la reproducibilidad y validez de datos en este campo de rápido crecimiento. Es importante destacar que el uso de imágenes exitosa a los sistemas de ultrasonido de alta resolución requiere un conocimiento práctico de los principios de la ecografía (y la terminología de uso común en la ecografía), una comprensión fundamental de la principles de la fisiología cardiaca, y la experiencia significativa con la ecografía para permitir una evaluación precisa y eficiente en el tiempo de la función cardiaca en los roedores.

<table border="0" cellpadding="0" cellspacing="0" width="1906">
	<tbody>
		<tr height="34">
			<td height="34" style="height:34px;width:693px;">High resolution ultrasound machine</td>
			<td style="width:453px;">VisualSonics, Fujifilm</td>
			<td style="width:293px;">Vevo 2100&#160;</td>
			<td style="width:467px;"></td>
		</tr>
		<tr height="68">
			<td height="68" style="height:68px;width:693px;">Isoflurane diffuser (capable of delivering 1 % to 1.5 % isoflurane mixed with 1 L/min 100% O2</td>
			<td>VisualSonics, Fujifilm</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="68">
			<td height="68" style="height:68px;width:693px;">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 height="34">
			<td height="34" style="height:34px;width:693px;">Animal platform</td>
			<td>VisualSonics, Fujifilm</td>
			<td>11503</td>
			<td></td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:693px;">Advanced physiological monitoring unit</td>
			<td>VisualSonics, Fujifilm</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:693px;">Isoflurane</td>
			<td>Terrell</td>
			<td>NDC 66794-019-10</td>
			<td></td>
		</tr>
		<tr height="74">
			<td height="74" style="height:74px;width:693px;">Nose cone and tubing connected to isoflurane diffuser and 100% O2</td>
			<td>Custom Engineered in-house</td>
			<td>--</td>
			<td></td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:693px;">Hair razor</td>
			<td>Andis Super AGR+ vet pack clipper</td>
			<td>AD65340</td>
			<td></td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:693px;">Ultrasound gel</td>
			<td>Parker Laboratories</td>
			<td>REF 01-08</td>
			<td></td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:693px;">Electrode gel&#160;</td>
			<td>Parker Laboratories</td>
			<td>REF 15-25</td>
			<td></td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:693px;">Adhesive tapes</td>
			<td>Fisher Laboratories</td>
			<td>1590120B</td>
			<td></td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:693px;">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.

Los ejemplos de las imágenes que se obtienen de forma rutinaria a partir de imágenes de ultrasonido cardiaco animales se incluyen en este manuscrito. Se proporciona una ilustración de la colocación del transductor en el pecho del animal para dar al lector una comprensión clara de que el transductor se coloca para obtener las imágenes como se describe. Una fotografía del laboratorio de ultrasonido puesta a punto también se incluye para destacar la importancia de los equipos adecuados, en particular el transductor de ultrasonidos para ser utilizado y el método de anestesia. El / B-modo 2D, modo M y Doppler color y muestras de las válvulas normales y anormales, ventrículos derecho e izquierdo, y la raíz aórtica están debidamente etiquetados. Aunque formación de imágenes de la velocidad de deformación no se realiza de forma rutinaria, también se incluye un ejemplo. La regurgitación mitral se caracteriza por una velocidad alta, por lo general no laminar el flujo de sangre (coloración mosaico) a través de la válvula durante la systole (Figura 5). La presencia de un patrón de flujo Doppler tales mosaico de colores desde el ventrículo izquierdo a la aurícula izquierda a través de la MV, que se producen después de que el complejo QRS en el trazado de un ECG, permite un diagnóstico inequívoco de la RM. Cuando esto ocurre en la ausencia de regurgitación de la válvula aórtica y / o disfunción del ventrículo izquierdo, esto puede ser caracterizado como aislado prolapso de la válvula mitral. Si hay dilatación significativa del ventrículo izquierdo (debido a la inducida experimentalmente insuficiencia cardíaca o excesiva profundidad de la anestesia), esto puede ser caracterizado como la regurgitación mitral isquémica (o regurgitación secundaria a la disfunción cardíaca). Una pantalla de Doppler espectral de la onda de impulsos se puede utilizar para confirmar la presencia y el momento de un chorro regurgitante del flujo sanguíneo. Una válvula aórtica normal tiene tres cúspides delgada, flexible que abren y cierran de manera adecuada durante cada ciclo cardiaco. Aórtica separación cúspide de válvula se mide en 2D guiadaModo M de la válvula aórtica en el eje largo. Calibradores electrónicos se utilizan para medir desde el borde delantero de la cúspide aórtica derecho a la vanguardia de la cúspide aórtica izquierda (Figura 3). la distancia de separación de cúspide de la válvula aórtica en ratones normales es de 0,9 a 1,3 mm. Doppler color muestra un flujo laminar a través de la válvula y en la raíz aórtica durante la sístole. El flujo turbulento se puede apreciar en condiciones de aumento del flujo, tales como en la insuficiencia de la válvula aórtica, o aumento de la presión, como en estenosis de la válvula aórtica. Esto se demuestra como la coloración del mosaico en el tracto de salida. Incluso pequeñas cantidades de regurgitación de la válvula aórtica puede resultar en un aumento significativo en la velocidad transvalvular pico debido a la función cardíaca hiperdinámica y el aumento de la precarga del ventrículo izquierdo. la velocidad pico aórtica en rangos normales de ratones de 0,90 m / s a ​​1,50 m / s. la velocidad de la válvula aórtica pico de&gt; 5 m / s se ha registrado en ratones con estenosis de la válvula aórtica severa. <p class="jove_content" fo:keep-together.within-page = &quot;1&quot;&gt; la onda pulsada trazados Doppler espectral puede también ser utilizado para proporcionar un índice de la hemodinámica de la arteria pulmonar 12 (Figura 8). Pulmonar tiempo de aceleración de la arteria es el intervalo de tiempo desde el inicio del flujo arterial sistólica pulmonar a la velocidad de flujo máximo. Justo el tiempo de eyección ventricular es el intervalo entre el inicio de eyección del ventrículo derecho hasta el punto en el que haya una interrupción de flujo sistólica de la arteria pulmonar sistólica. La combinación de un tiempo de aceleración de la arteria pulmonar acortada con una disminución en la proporción de tiempo de aceleración de la arteria pulmonar a tiempo de eyección del ventrículo derecho sugiere la presencia de hipertensión arterial pulmonar (que puede ser confirmado mediante medidas invasivas o directos de la arteria pulmonar o la presión del ventrículo derecho) . <img alt="Figura 1" src="/files/ftp_upload/54110/54110fig1.jpg" /> Figura 1: Animal CLaboratorio de Ultrasonido ardiac. El laboratorio está equipado con el aparato de ultrasonido de pequeños animales-dedicado con alta frecuencia (30 MHz y 40 MHz) transductores (MS 400 y MS 550D), difusor isoflurano, la plataforma de animal, la temperatura y el monitor de frecuencia cardiaca, 1% a 1,5% de isoflurano mezclado con 1 L / min 100% de O 2, cono de la nariz y el tubo conectado al difusor isoflurano y 100% O 2, la maquinilla de afeitar el pelo, gel de ultrasonido, gel del electrodo, cintas adhesivas, y toallas de papel. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig1large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> <img alt="Figura 2" src="/files/ftp_upload/54110/54110fig2.jpg" /> Figura 2: Posiciones básicas del transductor. Ventana (A) paraesternal. El cabezal del transductor se coloca en el borde paraesternal izquierda, con el marcador de índice de imagen del transductor dirigida en sentido caudal. from esta posición, se puede obtener la vista de eje largo del ventrículo izquierdo, la válvula aórtica, y la raíz aórtica y el eje corto de la válvula pulmonar. (B) En la ventana paraesternal, el cabezal del transductor se hace girar en sentido antihorario, con la muesca dirigida posteriormente. Desde esta posición, se puede obtener la vista de eje corto del ventrículo izquierdo y la válvula aórtica y la vista de eje largo de la válvula pulmonar. (C) Apical ventana. El cabezal del transductor está colocado en el ápice del corazón. Desde esta posición, se puede obtener la vista de eje largo del ventrículo derecho e izquierdo y válvulas mitral y tricúspide. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig2large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> <img alt="figura 3" src="/files/ftp_upload/54110/54110fig3.jpg" /> Figura 3: Evaluación de la función de la válvula aórtica en una normaal ratón contra la función de la válvula aórtica en un ratón con la enfermedad de la válvula aórtica calcificada. Imagen (A) 2D de una válvula aórtica normal en el eje largo. Tenga en cuenta que la válvula aórtica se abre así durante la sístole. (B) Imagen en modo M que representa la función valvular aórtica normal (apariencia similar a una caja). Tenga en cuenta que la distancia de cúspide de separación se mide a 1,12 mm. Pantalla (C) Doppler espectral de la velocidad de pico a través de la válvula aórtica normal se meaured a 1,3 m / s. Imagen (D) 2D de una válvula aórtica calcificada en el eje largo de un receptor de lipoproteína de baja densidad deficiente (LDLR - / -) y sólo B100 (apoB 100/100) de ratón apolipoproteína alimentados con la dieta occidental. Las cúspides se espesan y se han incrementado ecogenicidad, lo que resulta en la apertura restringida durante la sístole. (E) Una imagen en modo M que representa la misma válvula aórtica estenótica muestra una medición de distancia de cúspide de separación de 0,7 mm. (F &lt;/ Strong&gt;) La pantalla Doppler espectral de pico de velocidad a través de la válvula aórtica estenótica meaured a 4,6 m / s. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig3large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> <img alt="Figura 4" src="/files/ftp_upload/54110/54110fig4.jpg" /> Figura 4: M-modo de una válvula mitral normal. Desde la ventana apical, se obtiene una vista de eje largo de la válvula mitral. La línea de modo M de interrogación se aplica a través del velo mitral. Mientras que el espesor de los velos mitrales teóricamente se puede medir con calibradores electrónicos, esto puede ser extremadamente difícil dada la delgada, mal ecogénico, y rápidamente valvas de la válvula mitral normal de movimiento. Las flechas señalan el modo M de la valva de la válvula mitral durante la sístole. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig4large.jpg" target="_blank">Por favor cli</a>ck aquí para ver una versión más grande de esta figura. <img alt="Figura 5" src="/files/ftp_upload/54110/54110fig5.jpg" /> Figura 5: La evidencia de una regurgitación de la válvula mitral Jet utilizando Doppler color de imágenes. Desde la ventana paraesternal, se obtiene una vista de eje largo modificada de la válvula mitral. Doppler color muestra un chorro de interrogación mosaico de color en la válvula mitral durante la sístole (resaltado por una flecha). <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig5large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> <img alt="Figura 6" src="/files/ftp_upload/54110/54110fig6.jpg" /> Figura 6: Vista del eje largo de la arteria pulmonar principal y sus ramas principales. El eje largo de la arteria pulmonar principal (MPA) y derecha (RPA) y las ramas izquierda (API) puede ser obtenida de la paraste<html> rno ventana. El tracto de salida del ventrículo derecho (TSVD), la válvula pulmonar (PV), y la aorta (AO) se ven en parte. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig6large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> <img alt="Figura 7" src="/files/ftp_upload/54110/54110fig7.jpg" /> Figura 7: El modo M imagen que representa una válvula pulmonar normal. Desde la ventana paraesternal, ambos puntos de vista de corto y largo del eje de la válvula pulmonar se pueden obtener. La línea de modo M de interrogación se aplica a través de la válvula pulmonar. La distancia pulmonar válvula de cúspide de separación (flechas) se puede medir a partir de este punto de vista. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig7large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> 10fig8.jpg &quot;/&gt; Figura 8: de onda Doppler pulsado interrogatorio de flujo a través de la válvula pulmonar. El tiempo de aceleración de la arteria pulmonar (PCTA) es el intervalo de tiempo desde el inicio del flujo arterial sistólica pulmonar a la velocidad de flujo máximo. tiempo de eyección ventricular derecha (RVet) es el intervalo entre el inicio de la eyección ventricular derecha al punto en el que haya una interrupción de flujo. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig8large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> <img alt="Figura 9" src="/files/ftp_upload/54110/54110fig9.jpg" /> Figura 9: El modo M Imagen que representa un eje menor del ventrículo izquierdo. Desde la ventana paraesternal, el eje corto del ventrículo izquierdo se obtiene mediante la rotación del cabezal del transductor hacia la izquierda de manera que los puntos de las marcas de índice de la imagen posterior o dorsalmente. El M-modlínea e de la interrogación se aplica a través del ventrículo izquierdo a nivel de los músculos papilares. dimensión del ventrículo izquierdo de fin de diástole (DDFVI), diámetro telesistólico del ventrículo izquierdo (DSVI), y la pared anterior (AW) y espesores de pared posterior (PW) se pueden medir fácilmente. Tenga cuidado de no incluir el músculo papilar (*) en las mediciones. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig9large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> <img alt="Figura 10" src="/files/ftp_upload/54110/54110fig10.jpg" /> Figura 10: Evaluación del color y Doppler de onda pulsada Doppler espectral Visualización de flujo de entrada de la válvula mitral. (A) Imagen que muestra una evaluación Doppler color de flujo de entrada de la válvula mitral en el eje largo apical. Tenga en cuenta que la imagen en color Doppler 2D puede ser una herramienta crítica para guiding la posición del volumen de muestra adecuado para la adquisición de los trazados Doppler de onda pulsada (representado en el panel B). (B) Representación espectral de flujo de entrada de la válvula mitral mediante Doppler de onda pulsada. La evaluación Doppler de onda pulsada del flujo de sangre a través de la válvula mitral (en la vista apical del eje largo) se realiza para evaluar la función diastólica del ventrículo izquierdo. El volumen de muestra se coloca en la punta de las valvas de la válvula mitral. El tiempo de isovolumétrica de relajación (TRIV), tiempo de contracción isovolumétrica (IVCT), tiempo de eyección ventricular izquierda (LVET), y la velocidad de flujo de entrada mitral pico (E) pueden todos obtenerse a partir de la visualización espectral de velocidades Doppler de onda pulsada a través de la válvula mitral. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig10large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> <img alt="Figura 11" src="/files/ftp_upload/54110/54110fig11.jpg" /> Fifigura 11: Doppler tisular del anillo mitral septal. Desde la ventana apical, se obtiene una vista de eje largo de la válvula mitral. El volumen de muestra Doppler de tejido se coloca en la región septal del anillo mitral. La relación entre la velocidad de flujo de entrada mitral pico (E variable en la Figura 10B) y la velocidad pico mitral tejido del anillo (e &#39;, denotado por flechas blancas) se utiliza para evaluar la función ventricular izquierda diastólica (comúnmente referido como E / E&#39;). <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig11large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</a> <img alt="Figura 12" src="/files/ftp_upload/54110/54110fig12.jpg" /> Figura 12: Evaluación de la tensión y la velocidad de deformación del miocardio ventricular izquierda. Hay paquetes de software de análisis especializados disponibles en el mercado, y las variables de deformación y velocidad de deformación pueden serobtenido como medidas de los primeros cambios o subclínicas en intrínsecas propiedades contráctiles del miocardio. Los ejemplos mostrados anteriormente representan cepa radial y velocidad de deformación en planos de imágenes adquiridas comúnmente en ratones. Tenga en cuenta que estos planos de imagen (y la subsiguiente forma de los trazados de deformación) pueden diferir de las imágenes en los seres humanos, que se adquieren con frecuencia en el eje largo apical o de 4 cámaras. <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/54110/54110fig12large.jpg" target="_blank">Haga clic aquí para ver una versión más grande de esta figura.</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.

Enfoques y Protocolos para la Integral ecocardiográficos Caracterización fenotípica en las valvulopatías en ratones

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

15.8K vistas.  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

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery (SALLI) MRI in Rats

Small animal magnetic resonance imaging is an important tool to study cardiac function and changes in myocardial tissue. The high heart rates of small animals (200 to 600 beats/min) have previously limited the role of CMR imaging. Small animal Look-Locker inversion recovery (SALLI) is a T1 mapping sequence for small animals to overcome this problem 1. T1 maps provide quantitative information about tissue alterations and contrast agent kinetics. It is also possible to detect diffuse myocardial processes such as interstitial fibrosis or edema 1-6. Furthermore, from a single set of image data, it is possible to examine heart function and myocardial scarring by generating cine and inversion recovery-prepared late gadolinium enhancement-type MR images 1.
	The presented video shows step-by-step the procedures to perform small animal CMR imaging. Here it is presented with a healthy Sprague-Dawley rat, however naturally it can be extended to different cardiac small animal models.

Assessment of Cardiac Function and Myocardial Morphology Using Small Animal Look-locker Inversion Recovery SALLI MRI in Rats

Contrast enhanced cardiac magnetic resonance (CMR) imaging allows comprehensive in vivo assessment of the heart in small animal models of cardiovascular disease. Here we detail the procedures to perform CMR imaging, reconstruction and analysis using Small Animal Lock-Locker Inversion Recovery (SALLI) in rats.

Evaluación de la función cardiaca y la morfología de miocardio utilizando pequeñas Look-locker Recuperación Inversion Animal (SALLI) MRI en ratas

Small animal magnetic resonance  imaging is an important tool to study cardiac function and changes in myocardial  tissue. The high heart rates of small ...

Investigación

Medicina

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.

Evaluación ecocardiográfica de la parte derecha del corazón en ratones

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

Implantation of Total Artificial Heart in Congenital Heart Disease

In patients with end-stage heart failure (HF), a total artificial heart (TAH) may be implanted as a bridge to cardiac transplant. However, in congenital heart disease (CHD), the malformed heart presents a challenge to TAH implantation. 
In the case presented here, a 17 year-old patient with congenital transposition of the great arteries (CCTGA) experienced progressively worsening HF due to his congenital condition. He was hospitalized multiple times and received an implantable cardioverter defibrillator (ICD). However, his condition soon deteriorated to end-stage HF with multisystem organ failure. 
Due to the patient's grave clinical condition and the presence of complex cardiac lesions, the decision was made to proceed with a TAH. The abnormal arrangement of the patient's ventricles and great arteries required modifications to the TAH during implantation.
With the TAH in place, the patient was able to return home and regain strength and physical well-being while awaiting a donor heart. He was successfully bridged to heart transplantation 5 months after receiving the device. This report highlights the TAH is feasible even in patients with structurally abnormal hearts, with technical modification.

This is a case report of a patient with congenitally corrected transposition of the great arteries (CCTGA) who received a total artificial heart (TAH) as a bridge to heart transplant. The TAH was successfully implanted with modifications to accommodate the patient's congenitally malformed heart.

El implante de corazón artificial total en las cardiopatías congénitas

In patients with end-stage heart failure (HF), a total artificial heart (TAH) may be implanted as a bridge to cardiac transplant. However, in congenital ...

Establishment and Characterization of UTI and CAUTI in a Mouse Model

Urinary tract infections (UTI) are highly prevalent, a significant cause of morbidity and are increasingly resistant to treatment with antibiotics. Females are disproportionately afflicted by UTI: 50% of all women will have a UTI in their lifetime. Additionally, 20-40% of these women who have an initial UTI will suffer a recurrence with some suffering frequent recurrences with serious deterioration in the quality of life, pain and discomfort, disruption of daily activities, increased healthcare costs, and few treatment options other than long-term antibiotic prophylaxis. Uropathogenic Escherichia coli (UPEC) is the primary causative agent of community acquired UTI. Catheter-associated UTI (CAUTI) is the most common hospital acquired infection accounting for a million occurrences in the US annually and dramatic healthcare costs. While UPEC is also the primary cause of CAUTI, other causative agents are of increased significance including Enterococcus faecalis. Here we utilize two well-established mouse models that recapitulate many of the clinical characteristics of these human diseases. For UTI, a C3H/HeN model recapitulates many of the features of UPEC virulence observed in humans including host responses, IBC formation and filamentation. For CAUTI, a model using C57BL/6 mice, which retain catheter bladder implants, has been shown to be susceptible to E. faecalis bladder infection. These representative models are being used to gain striking new insights into the pathogenesis of UTI disease, which is leading to the development of novel therapeutics and management or prevention strategies.

The ability to model urinary tract infections (UTI) is crucial in order to be able to understand bacterial pathogenesis and spawn the development of novel therapeutics. This work&#8217;s goal is to demonstrate mouse models of experimental UTI and catheter associated UTI that recapitulate and predict findings seen in humans.

Establecimiento y caracterización de IU y ITUAC en un modelo de ratón

Urinary tract infections (UTI) are highly prevalent, a significant cause of morbidity and are increasingly resistant to treatment with antibiotics. ...

Heterotopic Cervical Heart Transplantation in Mice

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies this model by avoiding challenging suture anastomoses of small vessels thereby reducing warm ischemia time. In comparison to abdominal graft implantation the cervical model is less invasive and the implanted graft is easily accessible for further follow-up examinations. Anastomoses are performed by pulling the ascending aorta of the graft over the cuff with the recipient&#8217;s common carotid artery and by pulling the main pulmonary artery over the cuff with the external jugular vein. Selection of appropriate cuff size and complete mobilization of the vessels are important for successful revascularization. Ischemia-reperfusion (I/R) injury can be minimized by perfusing the graft with a cardioplegic solution and by hypothermia. In this article, we provide technical details for a simplified and improved cuff technique, which should allow surgeons with basic microsurgical skills to perform the procedure with a high success rate.

The cuff technique shortens and facilitates the model of heterotopic cervical heart transplantation in mice by avoiding technically challenging suture anastomoses of small vessels. The technical details shown in this video paper should allow researches to establish this model in their laboratories.

Heterotópico cervical trasplante cardiaco en ratones

The heterotopic cervical heart transplantation in mice is a valuable tool in transplant and cardiovascular research. The cuff technique greatly simplifies ...

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.

Examen ecocardiográfico e histológico de la morfología cardiaca en el ratón

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.

Medición ecocardiográfica de parámetros diastólicos ventriculares derecho en ratón

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

Generation of Human 3D Lung Tissue Cultures (3D-LTCs) for Disease Modeling

Translation of novel discoveries to human disease is limited by the availability of human tissue-based models of disease. Precision-cut lung slices (PCLS) used as 3D lung tissue cultures (3D-LTCs) represent an elegant and biologically highly relevant 3D cell culture model, which highly resemble in situ tissue due to their complexity, biomechanics and molecular composition. Tissue slicing is widely applied in various animal models. 3D-LTCs derived from human PCLS can be used to analyze responses to novel drugs, which might further help to better understand the mechanisms and functional effects of drugs in human tissue. The preparation of PCLS from surgically resected lung tissue samples of patients, who experienced lung lobectomy, increases the accessibility of diseased and peritumoral tissue. Here, we describe a detailed protocol for the generation of human PCLS from surgically resected soft-elastic patient lung tissue. Agarose was introduced into the bronchoalveolar space of the resectates, thus preserving lung structure and increasing the tissue&#39;s stiffness, which is crucial for subsequent slicing. 500 &#181;m thick slices were prepared from the tissue block with a vibratome. Biopsy punches taken from PCLS ensure comparable tissue sample sizes and further increase the amount of tissue samples. The generated lung tissue cultures can be applied in a variety of studies in human lung biology, including the pathophysiology and mechanisms of different diseases, such as fibrotic processes at its best at (sub-)cellular levels. The highest benefit of the 3D-LTC ex vivo model is its close representation of the in situ human lung in respect of 3D tissue architecture, cell type diversity and lung anatomy as well as the potential for assessment of tissue from individual patients, which is relevant to further develop novel strategies for precision medicine.

Generation of Human 3D Lung Tissue Cultures 3D-LTCs for Disease Modeling

Here, we present a protocol for the preparation of agarose-filled human precision-cut lung slices from resected patient tissue that are suitable for generating 3D lung tissue cultures to model human lung diseases in biological and biomedical studies.

Generación de cultivos de tejido de pulmón humano 3D (3D-LTCs) para el modelado de la enfermedad

Translation of novel discoveries to human disease is limited by the availability of human tissue-based models of disease. Precision-cut lung slices (PCLS) ...

Assessment and Characterization of Hyaloid Vessels in Mice

In the eye, the embryonic hyaloid vessels nourish the developing lens and retina and regress when the retinal vessels develop. Persistent or failed regression of hyaloid vessels can be seen in diseases such as persistent hyperplastic primary vitreous (PHPV), leading to an obstructed light path and impaired visual function. Understanding the mechanisms underlying the hyaloid vessel regression may lead to new molecular insights into the vascular regression process and potential new ways to manage diseases with persistent hyaloid vessels. Here we describe the procedures for imaging hyaloid in live mice with optical coherence tomography (OCT) and fundus fluorescein angiography (FFA) and a detailed technical protocol of isolating and flat-mounting hyaloid ex vivo for quantitative analysis. Low-density lipoprotein receptor-related protein 5 (LRP5) knockout mice were used as an experimental model of persistent hyaloid vessels, to illustrate the techniques. Together, these techniques may facilitate a thorough assessment of hyaloid vessels as an experimental model of vascular regression and studies on the mechanism of persistent hyaloid vessels.

This protocol describes both in vivo and ex vivo methods to fully visualize and characterize hyaloid vessels, a model of vascular regression in mouse eyes, using optical coherence tomography and fundus fluorescein angiography for the live imaging and ex vivo isolation and subsequent flat mount of hyaloid for quantitative analysis.

Evaluación y caracterización de recipientes hialoides en ratones

In the eye, the embryonic hyaloid vessels nourish the developing lens and retina and regress when the retinal vessels develop. Persistent or failed ...

Transthoracic Echocardiographic Examination in the Rabbit Model

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to that of humans and that of other large animal models such as dogs and pigs. However, the rabbit model has the additional advantage of lower maintenance costs compared to other large animal models. The longitudinal evaluation of cardiac function using echocardiography, when appropriately implemented, is a useful methodology for preclinical assessment of novel therapies for heart failure with reduced ejection fraction (e.g. cardiac regeneration). The correct use of this non-invasive tool requires the implementation of a standardized examination protocol following international guidelines. Here we describe, step by step, a detailed protocol supervised by veterinary cardiologists for performing echocardiography in the rabbit model, and demonstrate how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

Here we describe, step by step, a detailed protocol for performing echocardiography in the rabbit model. We show how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

Examen Ecocardiográfico Transtorácico en el Modelo Conejo

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to ...