1. Equipment Preparation&#160;
<ol>
	<li>Examine the ultrasound transducer for defects. Depending on the equipment used, this step may be unnecessary.
	<ol>
		<li>If an air bubble is observed, remove the screw located on the right side of the transducer head, and add sterile water through the hole with a 26 G needle. Air bubbles inside the transducer head are common. They will impede the acquisition of quality images.</li>
		<li>Check the membrane covering the probe for leaks or holes. Replace if necessary.</li>
	</ol>...

<ol>
	<li>Urboniene, D., Haber, I., Fang, Y. H., Thenappan, T., Archer, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Validation+of+high-resolution+echocardiography+and+magnetic+resonance+imaging+vs.+high-fidelity+catheterization+in+experimental+pulmonary+hypertension.">Validation of high-resolution echocardiography and magnetic resonance imaging vs. high-fidelity catheterization in experimental pulmonary hypertension.</a> Am. J. Physiol. Lung Cell Mol. Physiol. 299, 401-412 (2010).</li><li>Rottman, J. N., Ni, G., Brown, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Echocardiographic+evaluation+of+ventricular+function+in+mice.">Echocardiographic evaluation of ventricular function in mice.</a> Echocardiography. 24, 83-89 (2007).</li><li>West, J., 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=Pulmonary+hypertension+in+transgenic+mice+expressing+a+dominant-negative+BMPRII+gene+in+smooth+muscle.">Pulmonary hypertension in transgenic mice expressing a dominant-negative BMPRII gene in smooth muscle.</a> Circ. Res. 94, 1109-1114 (2004).</li><li>Ghofrani, H. A., Seeger, W., Grimminger, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imatinib+for+the+treatment+of+pulmonary+arterial+hypertension.">Imatinib for the treatment of pulmonary arterial hypertension.</a> N. Engl. J. Med. 353, 1412-1413 (2005).</li><li>Gomberg-Maitland, M., 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=A+dosing/cross-development+study+of+the+multikinase+inhibitor+sorafenib+in+patients+with+pulmonary+arterial+hypertension.">A dosing/cross-development study of the multikinase inhibitor sorafenib in patients with pulmonary arterial hypertension.</a> Clin. Pharmacol. Ther. 87, 303-310 (2010).</li><li>Brittain, E., 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=Right+ventricular+plasticity+and+functional+imaging.">Right ventricular plasticity and functional imaging.</a> Pulm. Circ. 2, 309-326 (2012).</li><li>Yang, X. P., 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=Echocardiographic+assessment+of+cardiac+function+in+conscious+and+anesthetized+mice.">Echocardiographic assessment of cardiac function in conscious and anesthetized mice.</a> Am. J. Physiol. 277, 1967-1974 (1999).</li><li>Suehiro, 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=Assessment+of+segmental+wall+motion+abnormalities+using+contrast+two-dimensional+echocardiography+in+awake+mice.">Assessment of segmental wall motion abnormalities using contrast two-dimensional echocardiography in awake mice.</a> Am. J. Physiol. Heart Circ Physiol. 280, 1729-1735 (2001).</li><li>Jones, J. E., 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=Serial+noninvasive+assessment+of+progressive+pulmonary+hypertension+in+a+rat+model.">Serial noninvasive assessment of progressive pulmonary hypertension in a rat model.</a> Am. J. Physiol. Heart Circ. Physiol. 283, 364-371 (2002).</li><li>Devaraj, A., 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+pulmonary+hypertension+with+multidetector+CT+and+echocardiography+alone+and+in+combination.">Detection of pulmonary hypertension with multidetector CT and echocardiography alone and in combination.</a> Radiology. 254, 609-616 (2010).</li><li>Kitabatake, A., 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=Noninvasive+evaluation+of+pulmonary+hypertension+by+a+pulsed+Doppler+technique.">Noninvasive evaluation of pulmonary hypertension by a pulsed Doppler technique.</a> Circulation. 68, 302-309 (1983).</li><li>Yared, 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=Pulmonary+artery+acceleration+time+provides+an+accurate+estimate+of+systolic+pulmonary+arterial+pressure+during+transthoracic+echocardiography.">Pulmonary artery acceleration time provides an accurate estimate of systolic pulmonary arterial pressure during transthoracic echocardiography.</a> J. Am. Soc. Echocardiogr. 24, 687-692 (2011).</li><li>Cheung, M. C., 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=Body+surface+area+prediction+in+normal,+hypermuscular,+and+obese+mice.">Body surface area prediction in normal, hypermuscular, and obese mice.</a> J. Surg. Res. 153, 326-331 (2009).</li><li>Patten, R. D., Hall-Porter, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+animal+models+of+heart+failure:+development+of+novel+therapies,+past+and+present.">Small animal models of heart failure: development of novel therapies, past and present.</a> Circ. Heart Fail. 2, 138-144 (2009).</li><li>Baumgartner, H., 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=Echocardiographic+assessment+of+valve+stenosis:+EAE/ASE+recommendations+for+clinical+practice.">Echocardiographic assessment of valve stenosis: EAE/ASE recommendations for clinical practice.</a> J. Am. Soc. Echocardiogr. 22, 1-23 (2009).</li><li>Arkles, J. 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=Shape+of+the+right+ventricular+Doppler+envelope+predicts+hemodynamics+and+right+heart+function+in+pulmonary+hypertension.">Shape of the right ventricular Doppler envelope predicts hemodynamics and right heart function in pulmonary hypertension.</a> Am. J. Respir. Crit. Care Med. 183, 268-276 (2011).</li><li>Wiesmann, 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=Analysis+of+right+ventricular+function+in+healthy+mice+and+a+murine+model+of+heart+failure+by+in+vivo+MRI.">Analysis of right ventricular function in healthy mice and a murine model of heart failure by in vivo MRI.</a> Am. J. Physiol. Heart Circ. Physiol. 283, 1065-1071 (2002).</li><li>West, J., 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=A+potential+role+for+Insulin+resistance+in+experimental+pulmonary+hypertension.">A potential role for Insulin resistance in experimental pulmonary hypertension.</a> Eur. Respir. J. , (2012).</li><li>Johnson, J. A., 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=Cytoskeletal+defects+in+Bmpr2-associated+pulmonary+arterial+hypertension.">Cytoskeletal defects in Bmpr2-associated pulmonary arterial hypertension.</a> Am. J. Physiol. Lung Cell Mol. Physiol. 302, L474-L484 (2012).</li><li>Johnson, J. A., West, J., Maynard, K. B., Hemnes, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ACE2+improves+right+ventricular+function+in+a+pressure+overload+model.">ACE2 improves right ventricular function in a pressure overload model.</a> PLoS One. 6, e20828 (2011).</li></ol>

Mouse models of disease, either transgenic or toxin-related, require phenotypic validation that the model actually recapitulates the human disease it is intended to emulate. This validation can often be accomplished by the presence or absence of a particular feature, for example development of a tumor. However, models that result in hemodynamic abnormalities such as aortic constriction models of left ventricular hypertrophy or our transgenic model of PAH are more difficult to validate. These models require either termina...

Elevated pulmonary pressure and right ventricular (RV) dysfunction are the hallmarks of pulmonary vascular disease in animal models and human patients with pulmonary arterial hypertension (PAH). Transgenic and toxic (e.g. monocrotaline or hypoxia) models of 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.
Echocardiography is the mainstay of the noninvasive assessment of ventricular function in rodent models1,2. Echocardiography has the advantage of clear translation to humans in whom the same tool is used. In addition, some genetic models exhibit incomplete penetrance3; the ability to noninvasively identify affected animals saves valuable time and resources. Noninvasive assessment of disease severity without sacrificing an animal also allows researchers to serially study the effects of investigative therapies. This is especially important given the rapidity with which translational therapies can progress to human trials4,5.
In humans, echocardiographic assessment of RV size and pulmonary hypertension is particularly challenging due to the retrosternal position and irregular shape of the RV6. Rodent models have the added challenges of small size and extremely rapid heart rates (300-700 beat/min). Recent advances including higher frame rates and smaller transducers have improved image quality and even allowed conscious imaging in some experimental protocols, though most rodent imaging is done under anesthesia7,8. Excellent experimental protocols of echocardiography in rat models of PAH have been described and validated against both MRI and invasive hemodynamics1,9. However, 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 and a model of isolated RV afterload after pulmonary artery banding; however, this protocol is applicable to any diseases affecting the pulmonary vasculature or right heart. We will describe animal preparation and detailed assessment of RV size and function as well as main pulmonary artery (PA) size. We also demonstrate the techniques and calculations needed to estimate stroke volume and cardiac output. Technical limitations preclude accurate Doppler estimates of pulmonary pressure, but we have applied a well-validated human surrogate, pulmonary artery acceleration time, to estimate PA pressure.

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			<td height="55" style="height:55px;width:251px;">Vevo 770 High Resolution Micro-Ultrasound System</td>
			<td style="width:164px;">Visualsonics Inc.</td>
			<td style="width:188px;"></td>
			<td style="width:247px;">get more info at www.visualsonics.com/products</td>
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			<td height="66" style="height:66px;width:251px;">RMV (Real-Time MicroVisualization) 704B 40 mH Scanhead w/ Encapsulated Transducer</td>
			<td style="width:164px;">Visualsonics Inc.</td>
			<td style="width:188px;"></td>
			<td style="width:247px;">get more info at www.visualsonics.com/products</td>
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			<td height="69" style="height:69px;width:251px;">Vevo Integrated Rail System including the Physioogical Monitoring System</td>
			<td style="width:164px;">Visualsonics Inc.</td>
			<td style="width:188px;"></td>
			<td style="width:247px;">get more info at www.visualsonics.com/products</td>
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			<td height="62" style="height:62px;width:251px;">Computer Monitor set up for use with the Vevo770</td>
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			<td style="width:247px;">get more info at www.visualsonics.com/products</td>
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			<td height="86" style="height:86px;width:251px;">VetEquip Portable Tabletop Anesthesia Machine with an Isoflurane Vaporizer</td>
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			<td height="62" style="height:62px;width:251px;">Activated Charcoal Waste Gas Containers</td>
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			<td height="62" style="height:62px;width:251px;">Puralube Eye Ointment</td>
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			<td height="63" style="height:63px;width:251px;">Ecogel 100 Ultrasound Gel</td>
			<td style="width:164px;">EcoMed Pharmaceuticals</td>
			<td style="width:188px;">30GB</td>
			<td>get more info at ecomed.com</td>
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			<td height="59" style="height:59px;width:251px;">3M Transpore Tape</td>
			<td style="width:164px;">Fisher Scientific</td>
			<td style="width:188px;">1527-0</td>
			<td>get more info at fishersci.com</td>
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			<td height="61" style="height:61px;width:251px;">Sterile H2O</td>
			<td style="width:164px;">DDI H2O from faucet and then autoclave</td>
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			<td height="59" style="height:59px;width:251px;">6 in Cotton Tipped Applicators</td>
			<td style="width:164px;">Fisher Scientific</td>
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			<td>get more info at fishersci.com</td>
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			<td height="64" style="height:64px;width:251px;">2 in x 2 in Gauze Sponges</td>
			<td style="width:164px;">Fisher Scientific</td>
			<td style="width:188px;"></td>
			<td>get more info at fishersci.com</td>
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</table>

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.

The principal goals of this protocol are to quantify RV size and function, and to understand the degree to which the pulmonary vasculature is diseased. Appropriate preparation of both the mouse and echocardiography equipment is essential to obtaining accurate and reproducible results. Mice should have their chest depilated and limbs secured to the imaging platform with tape. Anesthesia, in this case isoflurane, is administered via nose cone. The transducer should be checked for defects, particularly air bubbles, which ca...

Watch this Scientific Journal Video about Echocardiographic Assessment of the Right Heart in Mice at JoVE.com

Echocardiographic Assessment of the Right Heart in Mice

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.

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

echocardiographic-assessment-of-the-right-heart-in-mice

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22.6K Views.  Vanderbilt University Medical Center. 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.

Video: Echocardiographic Assessment of the Right Heart in Mice

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...

hemodynamic characterization of rodent models of pulmonary arterial hypertension

Hemodynamic Characterization of Rodent Models of Pulmonary Arterial Hypertension

Pulmonary arterial hypertension (PAH) is a disease of pulmonary arterioles that leads to their obliteration and the development of right ventricular failure. Rodent models of PAH are critical in understanding the pathophysiology of PAH. Here we demonstrate hemodynamic characterization, with right heart catheterization and echocardiography, in the mouse and...

echocardiographic measurement of right ventricular diastolic parameters in mouse

Echocardiographic Measurement of Right Ventricular Diastolic Parameters in Mouse

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...

shunt surgery, right heart catheterization, and vascular morphometry in a rat model for flow-induced pulmonary arterial hypertension

Shunt Surgery, Right Heart Catheterization, and Vascular Morphometry in a Rat Model for Flow-induced Pulmonary Arterial Hypertension

This protocol describes a surgical procedure to create a model for flow-induced pulmonary arterial hypertension (PAH) in rats and the procedures to analyze the principle hemodynamic and histological end-points in this...

comprehensive echocardiographic assessment of right ventricle function in a rat model of pulmonary arterial hypertension

Comprehensive Echocardiographic Assessment of Right Ventricle Function in a Rat Model of Pulmonary Arterial Hypertension

The present protocol describes the echocardiographic characterization of right ventricular morphology and function in a rat model of pulmonary arterial...

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

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.

echocardiographic assessment of cardiac anatomy and function in adult rats

Echocardiographic Assessment of Cardiac Anatomy and Function in Adult Rats

A non-invasive protocol for transthoracic echocardiography assessment of cardiac anatomy and function for adult rats is presented in the current study. The heart valves, all four cardiac chambers and the ascending aorta, aortic arch and descending aorta are studied in...

invasive hemodynamic assessment for the right ventricular system and hypoxia-induced pulmonary arterial hypertension in mice

Invasive Hemodynamic Assessment for the Right Ventricular System and Hypoxia-Induced Pulmonary Arterial Hypertension in Mice

Here, we present a protocol to perform an invasive hemodynamic assessment of the right ventricle and pulmonary artery in mice using an open-chest surgery...

echocardiographic and histological examination of cardiac morphology in the mouse

Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

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...

morphological and functional assessment of the right ventricle using 3d echocardiography

Morphological and Functional Assessment of the Right Ventricle Using 3D Echocardiography

Here, we provide a step-by-step acquisition and analysis protocol for the 3D volumetric assessment of the right ventricle, mainly focusing on the practical aspects that maximize the feasibility of this...

generation and characterization of right ventricular myocardial infarction induced by permanent ligation of the right coronary artery in mice

Generation and Characterization of Right Ventricular Myocardial Infarction Induced by Permanent Ligation of the Right Coronary Artery in Mice

There are several differences between the right and left ventricles. However, the pathophysiology of right ventricular infarction (RVI) has not been clarified. In the present protocol, a reproducible method for RVI mouse model generation is introduced, which may provide a means to explain the mechanism of RVI.

three-dimensional echocardiographic method for the visualization and assessment of specific parameters of the pulmonary veins

Three-Dimensional Echocardiographic Method for the Visualization and Assessment of Specific Parameters of the Pulmonary Veins

The dimensions of the pulmonary veins (PV) are important parameters when planning pulmonary vein isolation. 2D transoesophageal echocardiography can only provide limited data about the PVs; however, 3D echocardiography can evaluate relevant diameters and areas of the PVs, as well as their spatial relationship to surrounding...

echocardiographic characterization of left ventricular structure, function, and coronary flow in neonate mice

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

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

transthoracic echocardiographic examination in the rabbit model

Transthoracic Echocardiographic Examination in the Rabbit Model

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...

chronic thromboembolic pulmonary hypertension and assessment of right ventricular function in the piglet

Chronic Thromboembolic Pulmonary Hypertension and Assessment of Right Ventricular Function in the Piglet

Chronic Thromboembolic Pulmonary Hypertension (CTEPH) and Right Ventricular (RV) dysfunction were induced in piglets by progressive obstruction of the pulmonary arteries. Consequences were remarkably similar to those observed in CTEPH patients. This animal model would be a very useful tool for pathophysiology and therapeutic experiments on CTEPH and RV...

assessment of cardiac morphological and functional changes in mouse model of transverse aortic constriction by echocardiographic imaging

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

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...

induction of right ventricular failure by pulmonary artery constriction and evaluation of right ventricular function in mice

Induction of Right Ventricular Failure by Pulmonary Artery Constriction and Evaluation of Right Ventricular Function in Mice

Here, we provide a useful approach for studying the mechanism of right ventricular failure. A more convenient and efficient approach to pulmonary artery constriction is established using surgical instruments made inhouse. In addition, methods to evaluate the quality of this approach by echocardiography and catheterization are...

a novel right ventricular volume and pressure loaded piglet heart model for the study of tricuspid valve function.

A Novel Right Ventricular Volume and Pressure Loaded Piglet Heart Model for the Study of Tricuspid Valve Function.

A novel recovery piglet heart model with combined pressure and volume overload on the right ventricle is described for the study of tricuspid valve...

heterotopic heart transplantation in mice

Heterotopic Heart Transplantation in Mice

The mouse heterotopic heart transplantation model has been proven by many investigators to be an important method for studying mechanisms of rejection and immune response.  However, the techniques involved are still challenging.  By modifying standard techniques we have had success with more than 1000...

induction and phenotyping of acute right heart failure in a large animal model of chronic thromboembolic pulmonary hypertension

Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension

We present a protocol to induce and phenotype an acute right heart failure in a large animal model with chronic pulmonary hypertension. This model can be used to test therapeutic interventions, to develop right heart metrics&#160;or to improve the understanding of acute right heart failure...

Magnetic Resonance Derived Myocardial Strain Assessment Using Feature Tracking

Purpose: An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that strain is a more sensitive and earlier marker for contractile dysfunction than the frequently used parameter EF. Current technologies for CMR are time consuming and difficult to implement in clinical practice. Feature tracking is a technology that can lead to more automization and robustness of quantitative analysis of medical images with less time consumption than comparable methods.
Methods: An automatic or manual input in a single phase serves as an initialization from which the system starts to track the displacement of individual patterns representing anatomical structures over time. The specialty of this method is that the images do not need to be manipulated in any way beforehand like e.g. tagging of CMR images.
Results: The method is very well suited for tracking muscular tissue and with this allowing quantitative elaboration of myocardium and also blood flow.
Conclusions: This new method offers a robust and time saving procedure to quantify myocardial tissue and blood with displacement, velocity and deformation parameters on regular sequences of CMR imaging. It therefore can be implemented in clinical practice.

An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that strain is a more sensitive and earlier marker for contractile dysfunction than the frequently used parameter EF.

Purpose: An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that ...

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. Pulmonary Hypertension (PH) was induced in 3-week-old piglets by a progressive obstruction of the pulmonary vascular bed. A ligation of the left Pulmonary Artery (PA) was performed first through a mini-thoracotomy. Second, weekly embolizations of the right lower pulmonary lobe were done under fluoroscopic guidance with n-butyl-2-cyanoacrylate during 5 weeks. Mean Pulmonary Arterial Pressure (mPAP) measured by ritght heart catheterism, increased progressively, as well as Right Atrial pressure and Pulmonary Vascular Resistances (PVR) after 5 weeks compared to sham animals. Right Ventricular (RV) structural and functional remodeling were assessed by transthoracic echocardiography (RV diameters, RV wall thickness, RV systolic function). RV elastance and RV-pulmonary coupling were assessed by Pressure-Volume Loops (PVL) analysis with conductance method. Histologic study of the lung and the right ventricle were also performed. Molecular analyses on RV fresh tissues could be performed through repeated transcutaneous endomyocardial biopsies. Pulmonary microvascular disease in obstructed and unobstructed territories was studied from lung biopsies using molecular analyses and pathology. Furthermore, the reliability and the reproducibility was associated with a range of PH severity in animals. Most aspects of the human CTEPH disease were reproduced in this model, which allows new perspectives for the understanding of the underlying mechanisms (mitochondria, inflammation) and new therapeutic approaches (targeted, cellular or gene therapies) of the overloaded right ventricle but also pulmonary microvascular disease.

Chronic Thromboembolic Pulmonary Hypertension (CTEPH) and Right Ventricular (RV) dysfunction were induced in piglets by progressive obstruction of the pulmonary arteries. Consequences were remarkably similar to those observed in CTEPH patients. This animal model would be a very useful tool for pathophysiology and therapeutic experiments on CTEPH and RV failure.

An original piglet model of Chronic Thromboembolic Pulmonary Hypertension (CTEPH) associated with chronic Right Ventricular (RV) dysfunction is described. ...

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.

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

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.

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

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.

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

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.

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

Harvest of Endothelial Cells from the Balloon Tips of Swan-Ganz Catheters after Right Heart Catheterization

A variety of pathologies lead to pulmonary hypertension (PH), which is defined as a mean pulmonary artery pressure exceeding 25 mmHg at rest. To further diagnose and manage PH, patients undergo repeated right heart catheterizations (RHC) wherein a Swan-Ganz catheter is advanced into a branch of the pulmonary artery and a balloon is inflated to wedge the catheter tip. This article illustrates a protocol whereby pulmonary artery endothelial cells (PAECs) may be harvested from the balloon tips of Swan-Ganz catheters after RHC, and purified with an anti- CD146 affinity column technique to purify putative PAECs. These cells might be used to provide an in situ snapshot of the biological state of the pulmonary vasculature endothelium to complement hemodynamic measurements obtained during RHC. Harvested and purified PAECs may be used for either cell culture or for subsequent analytical assays such as flow cytometery.

This protocol describes the removal and purification of pulmonary artery endothelial cells (PAECs) that adhere to the balloon tips of Swan-Ganz catheters while they are wedged in the pulmonary artery during right heart catheterization and remain adherent to the deflated balloon following removal from the patient's body.

A variety of pathologies lead to pulmonary hypertension (PH), which is defined as a mean pulmonary artery pressure exceeding 25 mmHg at rest. To further ...

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.

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

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.

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

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. However, the evaluation of right ventricular pressure (RVP) and pulmonary artery pressure (PAP) remains technically challenging in mice. RVP and PAP are more difficult to measure than left ventricular pressure because of the anatomical differences between the left and right heart systems. In this paper, we describe a stable right heart hemodynamic measurement method and its validation using healthy and PAH mice. This method is based on open-chest surgery and mechanical ventilation support. It is a complicated procedure compared to closed chest procedures. While a well-trained surgeon is required for this surgery, the advantage of this procedure is that it can generate both RVP and PAP parameters at the same time, so it is a preferable procedure for the evaluation of PAH models.

Here, we present a protocol to perform an invasive hemodynamic assessment of the right ventricle and pulmonary artery in mice using an open-chest surgery approach.

Pulmonary arterial hypertension (PAH) is a chronic and severe cardiopulmonary disorder. Mice are a popular animal model used to mimic this disease. ...