Name: assessment of right ventricular structure and function in mouse model of pulmonary artery constriction by transthoracic echocardiography
Uploaded: 2014-02-03T14:45:00
Description: Watch this Scientific Journal Video about Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography Video at JoVE.com

We thank Fred Roberts and Chris White for exemplary technical support. We thank Brigham Women&#8217;s Hospital Cardiovascular Physiology Core for providing with the instrumentation and the funds for this work. This work was supported in part by NHLBI grants HL093148, HL086967, and HL 088533(RL), K99HL107642 and the Ellison Foundation (SC).

Surgical Procedure
<ol>
	<li>Obtain 8 week-old male C57BL/6 mice and acclimate for one week before any experimental procedures are performed.</li>
	<li>Prior to imaging, pulmonary artery occlusion is performed as described previously14&#160;in accordance with AVMA guidelines and approved IACUC protocols. 
	Echocardiographic Images Acquisition and Measurements 
	All abbreviations used are summarized in Table 1.</li>
</ol>
1. Parasternal Long Axi...

<ol>
	<li>Anavekar, N. 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=Usefulness+of+right+ventricular+fractional+area+change+to+predict+death,+heart+failure,+and+stroke+following+myocardial+infarction+(from+the+VALIANT+ECHO+Study).">Usefulness of right ventricular fractional area change to predict death, heart failure, and stroke following myocardial infarction (from the VALIANT ECHO Study).</a> Am. J. Cardiol. 101, 607-612 (2008).</li><li>Berger, R. M., Cromme-Dijkhuis, A. H., Witsenburg, M., Hess, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tricuspid+valve+regurgitation+as+a+complication+of+pulmonary+balloon+valvuloplasty+or+transcatheter+closure+of+patent+ductus+arteriosus+in+children+&lt;+or+=+4+years+of+age.">Tricuspid valve regurgitation as a complication of pulmonary balloon valvuloplasty or transcatheter closure of patent ductus arteriosus in children &lt; or = 4 years of age.</a> Am. J. Cardiol. 72, 976-977 (1993).</li><li>Marwick, T. H., Raman, S. V., Carrio, I., Bax, J. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+developments+in+heart+failure+imaging.">Recent developments in heart failure imaging.</a> JACC Cardiovasc. Imaging. 3, 429-439 (2010).</li><li>Souders, C. A., Borg, T. K., Banerjee, I., Baudino, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pressure+overload+induces+early+morphological+changes+in+the+heart.">Pressure overload induces early morphological changes in the heart.</a> Am. J. Pathol. 181, 1226-1235 (2012).</li><li>Karas, M. G., Kizer, J. R. <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+the+right+ventricle+and+associated+hemodynamics.">Echocardiographic assessment of the right ventricle and associated hemodynamics.</a> Prog. Cardiovasc. Dis. 55, 144-160 (2012).</li><li>Lindqvist, P., Calcutteea, A., Henein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Echocardiography+in+the+assessment+of+right+heart+function.">Echocardiography in the assessment of right heart function.</a> Eur. J. Echocardiogr. 9, 225-234 (2008).</li><li>Rudski, L. G., 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=Guidelines+for+the+echocardiographic+assessment+of+the+right+heart+in+adults:+a+report+from+the+American+Society+of+Echocardiography+endorsed+by+the+European+Association+of+Echocardiography,+a+registered+branch+of+the+European+Society+of+Cardiology,+and+the+Canadian+Society+of+Echocardiography.">Guidelines for the echocardiographic assessment of the right heart in adults: a report from the American Society of Echocardiography endorsed by the European Association of Echocardiography, a registered branch of the European Society of Cardiology, and the Canadian Society of Echocardiography.</a> J. Am. Soc. Echocardiogr. 23, 685-713 (2010).</li><li>Scherrer-Crosbie, M., Thibault, H. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Echocardiography+in+translational+research:+of+mice+and+men.">Echocardiography in translational research: of mice and men.</a> J. Am. Soc. Echocardiogr. 21, 1083-1092 (2008).</li><li>Thibault, H. B., 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+assessment+of+murine+pulmonary+arterial+pressure:+validation+and+application+to+models+of+pulmonary+hypertension.">Noninvasive assessment of murine pulmonary arterial pressure: validation and application to models of pulmonary hypertension.</a> Circ. Cardiovasc. Imaging. 3, 157-163 (2010).</li><li>Polak, J. F., Holman, B. L., Wynne, J., Right Colucci, W. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ventricular+ejection+fraction:+an+indicator+of+increased+mortality+in+patients+with+congestive+heart+failure+associated+with+coronary+artery+disease.">ventricular ejection fraction: an indicator of increased mortality in patients with congestive heart failure associated with coronary artery disease.</a> J. Am. Coll. Cardiol. 2, 217-224 (1983).</li><li>Tanaka, N., 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=Transthoracic+echocardiography+in+models+of+cardiac+disease+in+the+mouse.">Transthoracic echocardiography in models of cardiac disease in the mouse.</a> Circulation. 94, 1109-1117 (1996).</li><li>Benza, R., Biederman, R., Murali, S., Gupta, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+cardiac+magnetic+resonance+imaging+in+the+management+of+patients+with+pulmonary+arterial+hypertension.">Role of cardiac magnetic resonance imaging in the management of patients with pulmonary arterial hypertension.</a> J. Am. Coll. Cardiol. 52, 1683-1692 (2008).</li><li>Lang, R. 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=Recommendations+for+chamber+quantification.">Recommendations for chamber quantification.</a> Eur. J. Echocardiogr. 7, 79-108 (2006).</li><li>Tarnavski, O., McMullen, J. R., Schinke, M., Nie, Q., Kong, S., Izumo, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+cardiac+surgery:+comprehensive+techniques+for+the+generation+of+mouse+models+of+human+diseases+and+their+application+for+genomic+studies.">Mouse cardiac surgery: comprehensive techniques for the generation of mouse models of human diseases and their application for genomic studies.</a> Physiol. Genomics. 16, 349-360 (2004).</li><li>Schulz-Menger, , 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=Standardized+image+interpretation+and+post+processing+in+cardiovascular+magnetic+resonance:+Society+for+Cardiovascular+Magnetic+Resonance+(SCMR)+Board+of+Trustees+Task+Force+on+Standardized+Post+Processing.">Standardized image interpretation and post processing in cardiovascular magnetic resonance: Society for Cardiovascular Magnetic Resonance (SCMR) Board of Trustees Task Force on Standardized Post Processing.</a> J. Cardiovasc. Magn. Reson. 15, 35 (2013).</li><li>Williams, R., 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+ultrasonic+measurement+of+regional+and+local+pulse-wave+velocity+in+mice.">Noninvasive ultrasonic measurement of regional and local pulse-wave velocity in mice.</a> Ultrasound Med. Biol. 33, 1368-1375 (2007).</li><li>Senechal, 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+simple+Doppler+echocardiography+method+to+evaluate+pulmonary+capillary+wedge+pressure+in+patients+with+atrial+fibrillation.">A simple Doppler echocardiography method to evaluate pulmonary capillary wedge pressure in patients with atrial fibrillation.</a> Echocardiography. 25, 57-63 (2008).</li><li>Frea, 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=Echocardiographic+evaluation+of+right+ventricular+stroke+work+index+in+advanced+heart+failure:+a+new+index.">Echocardiographic evaluation of right ventricular stroke work index in advanced heart failure: a new index.</a> J. Card. Fail. 18, 886-893 (2012).</li><li>Pokreisz, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pressure+overload-induced+right+ventricular+dysfunction+and+remodelling+in+experimental+pulmonary+hypertension:+the+right+heart+revisited.">Pressure overload-induced right ventricular dysfunction and remodelling in experimental pulmonary hypertension: the right heart revisited.</a> Eur. Heart J. Suppl. , H75-H84 (2007).</li><li>Bauer, 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=Echocardiographic+speckle-tracking+based+strain+imaging+for+rapid+cardiovascular+phenotyping+in+mice.">Echocardiographic speckle-tracking based strain imaging for rapid cardiovascular phenotyping in mice.</a> Circ. Res. 108, 908-916 (2011).</li></ol>

We demonstrate that TTE provides a sensitive and reproducible methodology for routine assessment of RV structure and function in mice. Before the advent of TTE, studies of the RV largely focused on RVSP measurement via right heart catheterization, a terminal and invasive procedure6,9,11,17.
Prior reports have described a variety of techniques for performing right heart measurements3,4,11,17-19. However, the majority of previous studies reported RV size and structural data...

Historically, prognostic assessment of heart failure has focused on the LV, which is easy to image via echocardiography. Numerous studies on LV structure and function using echocardiography have led to the establishment of normal values for LV structure and function1,5,6. Measurements of LV size and systolic function obtained from two-dimensional and Color Doppler images are of great importance as they allow visual delineation of compartments and geometry in great detail for the LV7. M-Mode is often used for measuring LV dimensions and fractional shortening (FS) in mice. Inter-observer and intra-observer variability are low for diameter measurements using this mode, but wall thickness measurements tend to be quite variable7. Pulsed Doppler with color (PW or Color Doppler) has been used to evaluate valvular regurgitation8,9.
Similar to LV, the RV plays an important role and is a significant predictor of morbidity and mortality in patients afflicted with cardiopulmonary disease1,7,10. However, echocardiographic evaluation of RV is inherently challenging due to its complex shape5,11 and its retrosternal position that blocks the ultrasound waves8,9. RV is a crescent shaped structure wrapping around the LV and has a complex anatomy with thin walls that are accustomed to low pressure and resistance to pulmonary vasculature6. To overcome elevated vascular resistance (PVR), the RV first increases in size and undergoes hypertrophies. In chronic diseases like pulmonary hypertension or pulmonary vascular disease, RV undergoes progressive dilatation, eventually resulting in the deterioration of systolic and diastolic function4,5,10.
Echocardiography plays an important role in the screening and diagnosis of PAH despite some limitations present in its clinical diagnostic capability. The main advantage of TTE lies in that it is noninvasive and that it can be performed on lightly sedated, or even conscious animals9. TTE also provides a reasonable estimate of PA pressures, as well as an ongoing assessment of changes in RV structure and function12,13. Due to technical advances in TTE, which include the development of high-frequency mechanical probes, allowing axial resolution of approximately 50 &#956;m at a depth of 5-12 mm, high frame rates (greater than 300 frame/sec), and high sampling rates, echocardiography is a choice tool for imaging the rapidly contracting small sized mouse heart8,11.
Longitudinal monitoring of RV function using multiple views, including 2-dimensional (2D) short and long axis, M-mode and Doppler acoustic windows provide complementary information of RV anatomy and function. Collectively, this methodology permits complete longitudinal assessment of RV hemodynamics in physiology and pathological setting4,7.
Herein, we provide a detailed step-by-step methodology of using noninvasive TTE to characterize RV anatomical and functional changes secondary to PAC in mice.

<table border="0" cellpadding="0" cellspacing="0" width="609">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="61">
			<td height="61" style="height:61px;width:157px;">High Frequency Ultrasound</td>
			<td style="width:108px;">FUJIFILM VisualSonics, Inc.</td>
			<td style="width:96px;">Vevo 2100</td>
			<td style="width:248px;"></td>
		</tr>
		<tr height="60">
			<td height="60" style="height:60px;width:157px;">High-frequency Mechanical Transducer</td>
			<td style="width:108px;">FUJIFILM VisualSonics, Inc.</td>
			<td style="width:96px;">MS250, MS550D, MS400</td>
			<td style="width:248px;"></td>
		</tr>
		<tr height="67">
			<td height="67" style="height:67px;width:157px;">Millar Mikro Pressure Catheter</td>
			<td style="width:108px;">Millar</td>
			<td style="width:96px;">SPR-1000</td>
			<td style="width:248px;"></td>
		</tr>
	</tbody>
</table>

assessment of right ventricular structure and function in mouse model of pulmonary artery constriction by transthoracic echocardiography

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, the RV is significantly affected in pulmonary diseases such as pulmonary artery hypertension (PAH). In addition, the RV is remarkably sensitive to cardiac pathologies, including left ventricular (LV) dysfunction, valvular disease or RV infarction4. To understand the role of RV in the pathogenesis of cardiac diseases, a reliable and noninvasive method to access the RV structurally and functionally is essential.
A noninvasive trans-thoracic echocardiography (TTE) based methodology was established and validated for monitoring dynamic changes in RV structure and function in adult mice. To impose RV stress, we employed a surgical model of pulmonary artery constriction (PAC) and measured the RV response over a 7-day period using a high-frequency ultrasound microimaging system. Sham operated mice were used as controls. Images were acquired in lightly anesthetized mice at baseline (before surgery), day 0 (immediately post-surgery), day 3, and day 7 (post-surgery). Data was analyzed offline using software.
Several acoustic windows (B, M, and Color Doppler modes), which can be consistently obtained in mice, allowed for reliable and reproducible measurement of RV structure (including RV wall thickness, end-diastolic&#160;and end-systolic dimensions), and function (fractional area change, fractional shortening, PA peak velocity, and peak pressure gradient) in normal mice and following PAC.
Using this method, the pressure-gradient resulting from PAC was accurately measured in real-time using Color Doppler mode and was comparable to direct pressure measurements performed with a Millar high-fidelity microtip catheter. Taken together, these data demonstrate that RV measurements obtained from various complimentary views using echocardiography are reliable, reproducible and can provide insights regarding RV structure and function. This method will enable a better understanding of the role of RV cardiac dysfunction.

In this study, baseline echocardiography was performed 48 hr&#160;prior to surgery. Mice were randomized into two groups. Mice received pulmonary artery occlusions (PAC) and sham operations (Sham). Echocardiography was performed at day 0, 3, and 7 following surgical procedure.&#160; The animals were euthanized immediately following the last echocardiography and hearts were harvested for histological assessment.&#160;Catheterization was conducted in subgroup (n=3 and 2 for day 0 and 7, respectively) of PAC mice to measure...

Watch this Scientific Journal Video about Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography Video at JoVE.com

Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography Video (Video) | JoVE

Right ventricle (RV) dysfunction is critical to the pathogenesis of cardiovascular disease, yet limited methodologies are available for its evaluation. Recent advances in ultrasound imaging provide a noninvasive and accurate option for longitudinal RV study. Herein, we detail a step-by-step echocardiographic method using a murine model of RV pressure overload.

Assessment of Right Ventricular Structure and Function in Mouse Model of Pulmonary Artery Constriction by Transthoracic Echocardiography

Emerging clinical data support the notion that RV dysfunction is critical to the pathogenesis of cardiovascular disease and heart failure1-3. Moreover, ...

Research

JoVE Journal

Medicine

Transthoracic Echocardiography in Mice

Transthoracic Echocardiography in Mice Video (Video) | JoVE

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

Echocardiographic Assessment of the Right Heart in Mice

Echocardiographic Assessment of the Right Heart in Mice Video (Video) | JoVE

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

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 Video (Video) | JoVE

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

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 Video (Video) | JoVE

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

Transthoracic Speckle Tracking Echocardiography for the Quantitative Assessment of Left Ventricular Myocardial Deformation

Transthoracic Speckle Tracking Echocardiography for the Quantitative Assessment of Left Ventricular Myocardial Deformation Video (Video) | JoVE

The value of conventional echocardiography is limited by differences in inter-individual image interpretation and therefore largely dependent on the ...

Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery

Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery Video (Video) | JoVE

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

Echocardiographic Measurement of Right Ventricular Diastolic Parameters in Mouse

Echocardiographic Measurement of Right Ventricular Diastolic Parameters in Mouse Video (Video) | JoVE

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

A Pulmonary Trunk Banding Model of Pressure Overload Induced Right Ventricular Hypertrophy and Failure

A Pulmonary Trunk Banding Model of Pressure Overload Induced Right Ventricular Hypertrophy and Failure Video (Video) | JoVE

Right ventricular (RV) failure induced by sustained pressure overload is a major contributor to morbidity and mortality in several cardiopulmonary ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography Video (Video) | JoVE

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

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 Video (Video) | JoVE

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