Name: Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice (Video) | JoVE Journal
Uploaded: 2017-02-14T12:30:00.000+00:00
Duration: 12 min 12 s
Description: Learn comprehensive echocardiographic methods for phenotyping valvular heart disease in mice. This standardized protocol enables detection of dysfunction across multiple heart valves and improves cardiac research reproducibility.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

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

<ol><li>Ngo, D. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Determinants+of+occurrence+of+aortic+sclerosis+in+an+aging+population%5BTitle%5D)+AND+%22JACC+Cardiovasc+Imaging%22%5BJournal%5D)">Determinants of occurrence of aortic sclerosis in an aging population.</a> JACC Cardiovasc Imaging. 2, 919-927 (2009).</li>
<li>Nkomo, V. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Epidemiology+and+prevention+of+valvular+heart+diseases+and+infective+endocarditis+in+Africa%5BTitle%5D)+AND+%22Heart%22%5BJournal%5D)">Epidemiology and prevention of valvular heart diseases and infective endocarditis in Africa.</a> Heart. 93, 1510-1519 (2007).</li>
<li>Amato, M. C., Moffa, P. J., Werner, K. E., Ramires, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Treatment+decision+in+asymptomatic+aortic+valve+stenosis%3A+role+of+exercise+testing%5BTitle%5D)+AND+%22Heart%22%5BJournal%5D)">Treatment decision in asymptomatic aortic valve stenosis: role of exercise testing.</a> Heart. 86, 381-386 (2001).</li>
<li>Bonow, R. O., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Focused+update+incorporated+into+the+ACC%2FAHA+2006+guidelines+for+the+management+of+patients+with+valvular+heart+disease%3A+a+report+of+the+American+College+of+Cardiology%2FAmerican+Heart+Association+Task+Force+on+Practice+Guidelines+%28Writing+Committee+to+Revise+the+1998+Guidelines+for+the+Management+of+Patients+With+Valvular+Heart+Disease%29%3A+endorsed+by+the+Society+of+Cardiovascular+Anesthesiologists%2C+Society+for+Cardiovascular+Angiography+and+Interventions%2C+and+Society+of+Thoracic+Surgeons%5BTitle%5D)+AND+%22Circulation%22%5BJournal%5D)">Focused update incorporated into the ACC/AHA 2006 guidelines for the management of patients with valvular heart disease: a report of the American College of Cardiology/American Heart Association Task Force on Practice Guidelines (Writing Committee to Revise the 1998 Guidelines for the Management of Patients With Valvular Heart Disease): endorsed by the Society of Cardiovascular Anesthesiologists, Society for Cardiovascular Angiography and Interventions, and Society of Thoracic Surgeons.</a> Circulation. 118, e523-e661 (2008).</li>
<li>Yutzey, K. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Calcific+aortic+valve+disease%3A+a+consensus+summary+from+the+Alliance+of+Investigators+on+Calcific+Aortic+Valve+Disease%5BTitle%5D)+AND+%22Arterioscler+Thromb+Vasc+Biol%22%5BJournal%5D)">Calcific aortic valve disease: a consensus summary from the Alliance of Investigators on Calcific Aortic Valve Disease.</a> Arterioscler Thromb Vasc Biol. 34, 2387-2393 (2014).</li>
<li>Rajamannan, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Calcific+aortic+valve+disease%3A+cellular+origins+of+valve+calcification%5BTitle%5D)+AND+%22Arterioscler+Thromb+Vasc+Biol%22%5BJournal%5D)">Calcific aortic valve disease: cellular origins of valve calcification.</a> Arterioscler Thromb Vasc Biol. 31, 2777-2778 (2011).</li>
<li>Weiss, R. M., Miller, J. D., Heistad, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Fibrocalcific+aortic+valve+disease%3A+opportunity+to+understand+disease+mechanisms+using+mouse+models%5BTitle%5D)+AND+%22Circ+Res%22%5BJournal%5D)">Fibrocalcific aortic valve disease: opportunity to understand disease mechanisms using mouse models.</a> Circ Res. 113, 209-222 (2013).</li>
<li>Sider, K. L., Blaser, M. C., Simmons, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Animal+models+of+calcific+aortic+valve+disease%5BTitle%5D)+AND+%22Int+J+Inflam%22%5BJournal%5D)">Animal models of calcific aortic valve disease.</a> Int J Inflam. 2011, 364310(2011).</li>
<li>Miller, J. D., Weiss, R. M., Heistad, D. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Calcific+aortic+valve+stenosis%3A+methods%2C+models%2C+and+mechanisms%5BTitle%5D)+AND+%22Circ+Res%22%5BJournal%5D)">Calcific aortic valve stenosis: methods, models, and mechanisms.</a> Circ Res. 108, 1392-1412 (2011).</li>
<li>Ram, R., Mickelsen, D. M., Theodoropoulos, C., Blaxall, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((New+approaches+in+small+animal+echocardiography%3A+imaging+the+sounds+of+silence%5BTitle%5D)+AND+%22Am+J+Physiol+Heart+Circ+Physiol%22%5BJournal%5D)">New approaches in small animal echocardiography: imaging the sounds of silence.</a> Am J Physiol Heart Circ Physiol. 301, H1765-H1780 (2011).</li>
<li>Moran, A. M., Keane, J. F., Colan, S. D. Influence of pressure and volume load on growth of aortic annulus and left ventricle in patients with critical aortic stenosis. J Am Coll Cardiol. 37, 471a(2001).</li>
<li>Thibault, H. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Noninvasive+assessment+of+murine+pulmonary+arterial+pressure%3A+validation+and+application+to+models+of+pulmonary+hypertension%5BTitle%5D)+AND+%22Circ+Cardiovasc+Imaging%22%5BJournal%5D)">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>Baumgartner, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Echocardiographic+assessment+of+valve+stenosis%3A+EAE%2FASE+recommendations+for+clinical+practice%5BTitle%5D)+AND+%22J+Am+Soc+Echocardiogr%22%5BJournal%5D)">Echocardiographic assessment of valve stenosis: EAE/ASE recommendations for clinical practice.</a> J Am Soc Echocardiogr. 22, quiz 101-102 1-23 (2009).</li>
<li>Lang, R. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Recommendations+for+cardiac+chamber+quantification+by+echocardiography+in+adults%3A+an+update+from+the+American+Society+of+Echocardiography+and+the+European+Association+of+Cardiovascular+Imaging%5BTitle%5D)+AND+%22Eur+Heart+J+Cardiovasc+Imaging%22%5BJournal%5D)">Recommendations for cardiac chamber quantification by echocardiography in adults: an update from the American Society of Echocardiography and the European Association of Cardiovascular Imaging.</a> Eur Heart J Cardiovasc Imaging. 16, 233-270 (2015).</li>
<li>Devereux, R. B., Reichek, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Echocardiographic+determination+of+left+ventricular+mass+in+man.+Anatomic+validation+of+the+method%5BTitle%5D)+AND+%22Circulation%22%5BJournal%5D)">Echocardiographic determination of left ventricular mass in man. Anatomic validation of the method.</a> Circulation. 55, 613-618 (1977).</li>
<li>Ommen, S. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Clinical+utility+of+Doppler+echocardiography+and+tissue+Doppler+imaging+in+the+estimation+of+left+ventricular+filling+pressures%3A+A+comparative+simultaneous+Doppler-catheterization+study%5BTitle%5D)+AND+%22Circulation%22%5BJournal%5D)">Clinical utility of Doppler echocardiography and tissue Doppler imaging in the estimation of left ventricular filling pressures: A comparative simultaneous Doppler-catheterization study.</a> Circulation. 102, 1788-1794 (2000).</li>
<li>Tei, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((New+index+of+combined+systolic+and+diastolic+myocardial+performance%3A+a+simple+and+reproducible+measure+of+cardiac+function--a+study+in+normals+and+dilated+cardiomyopathy%5BTitle%5D)+AND+%22J+Cardiol%22%5BJournal%5D)">New index of combined systolic and diastolic myocardial performance: a simple and reproducible measure of cardiac function--a study in normals and dilated cardiomyopathy.</a> J Cardiol. 26, 357-366 (1995).</li>
<li>Koshizuka, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Longitudinal+strain+impairment+as+a+marker+of+the+progression+of+heart+failure+with+preserved+ejection+fraction+in+a+rat+model%5BTitle%5D)+AND+%22J+Am+Soc+Echocardiogr%22%5BJournal%5D)">Longitudinal strain impairment as a marker of the progression of heart failure with preserved ejection fraction in a rat model.</a> J Am Soc Echocardiogr. 26, 316-323 (2013).</li>
<li>Ishizu, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Left+ventricular+strain+and+transmural+distribution+of+structural+remodeling+in+hypertensive+heart+disease%5BTitle%5D)+AND+%22Hypertension%22%5BJournal%5D)">Left ventricular strain and transmural distribution of structural remodeling in hypertensive heart disease.</a> Hypertension. 63, 500-506 (2014).</li>
<li>Yamada, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Induced+pluripotent+stem+cell+intervention+rescues+ventricular+wall+motion+disparity%2C+achieving+biological+cardiac+resynchronization+post-infarction%5BTitle%5D)+AND+%22J+Physiol%22%5BJournal%5D)">Induced pluripotent stem cell intervention rescues ventricular wall motion disparity, achieving biological cardiac resynchronization post-infarction.</a> J Physiol. 591, 4335-4349 (2013).</li>
<li>Andrews, T. G., Lindsey, M. L., Lange, R. A., Aune, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cardiac+Assessment+in+Pediatric+Mice%3A+Strain+Analysis+as+a+Diagnostic+Measurement%5BTitle%5D)+AND+%22Echocardiography%22%5BJournal%5D)">Cardiac Assessment in Pediatric Mice: Strain Analysis as a Diagnostic Measurement.</a> Echocardiography. 31, 375-384 (2014).</li>
<li>Ferferieva, V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Assessment+of+strain+and+strain+rate+by+two-dimensional+speckle+tracking+in+mice%3A+comparison+with+tissue+Doppler+echocardiography+and+conductance+catheter+measurements%5BTitle%5D)+AND+%22Eur+Heart+J+Cardiovasc+Imaging%22%5BJournal%5D)">Assessment of strain and strain rate by two-dimensional speckle tracking in mice: comparison with tissue Doppler echocardiography and conductance catheter measurements.</a> Eur Heart J Cardiovasc Imaging. 14, 765-773 (2013).</li>
<li>Fine, N. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Left+and+right+ventricular+strain+and+strain+rate+measurement+in+normal+adults+using+velocity+vector+imaging%3A+an+assessment+of+reference+values+and+intersystem+agreement%5BTitle%5D)+AND+%22Int+J+Cardiovasc+Imaging%22%5BJournal%5D)">Left and right ventricular strain and strain rate measurement in normal adults using velocity vector imaging: an assessment of reference values and intersystem agreement.</a> Int J Cardiovasc Imaging. 29, 571-580 (2013).</li>
<li>Pernot, M., Fujikura, K., Fung-Kee-Fung, S. D., Konofagou, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((ECG-gated%2C+mechanical+and+electromechanical+wave+imaging+of+cardiovascular+tissues+in+vivo%5BTitle%5D)+AND+%22Ultrasound+Med+Biol%22%5BJournal%5D)">ECG-gated, mechanical and electromechanical wave imaging of cardiovascular tissues in vivo.</a> Ultrasound Med Biol. 33, 1075-1085 (2007).</li>
<li>Liu, J. H., Jeng, G. S., Wu, T. K., Li, P. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((ECG+triggering+and+gating+for+ultrasonic+small+animal+imaging%5BTitle%5D)+AND+%22IEEE+Trans+Ultrason+Ferroelectr+Freq+Control%22%5BJournal%5D)">ECG triggering and gating for ultrasonic small animal imaging.</a> IEEE Trans Ultrason Ferroelectr Freq Control. 53, 1590-1596 (2006).</li>
<li>Monin, J. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Low-gradient+aortic+stenosis%3A+operative+risk+stratification+and+predictors+for+long-term+outcome%3A+a+multicenter+study+using+dobutamine+stress+hemodynamics%5BTitle%5D)+AND+%22Circulation%22%5BJournal%5D)">Low-gradient aortic stenosis: operative risk stratification and predictors for long-term outcome: a multicenter study using dobutamine stress hemodynamics.</a> Circulation. , 319-324 (2003).</li>
</ol>

The authors have nothing to disclose.

Induction of anesthesia
Proper induction and maintenance of anesthesia is critical for the accurate assessment of changes in heart valve and cardiac function in mice. Given the rapid induction of anesthesia elicited by isoflurane and the relatively long wash-out time of this anesthetic following deep anesthesia, we do not use a stand-alone anesthesia chamber for induction. Instead, as noted in detail above, animals are guided directly to the anesthesia cone, which allows for rapi...

Aging is associated with progressive increases in cardiovascular calcification1. Hemodynamically significant aortic valve stenosis affects 3% of the population over the age of 652, and patients with even moderate aortic valve stenosis (peak velocity of 3-4 m/s) have a 5 year event-free survival of less than 40%3. Presently, there are no effective treatments to slow the progression of aortic valve calcification, and surgical aortic valve replacement is the only available treatment for advanced aortic valve stenosis4.
Studies aimed at gaining a deeper understanding of the mechanisms that contribute to the initiation and progression of aortic valve calcification are a key first step in moving towards pharmacological and non-surgical methods to manage aortic valve stenosis5,6. Genetically-altered mice have played a major role in developing our understanding of the mechanisms that contribute to a variety of diseases and are now coming to the forefront of mechanistic studies aimed at understanding the biology of aortic valve stenosis6,7,8. Unlike other cardiovascular diseases such as atherosclerosis and heart failure-where standard protocols for evaluating vascular and ventricular function are for the most part well-established-there are unique challenges associated with in vivo phenotyping of heart valve function in mice. While recent reviews have provided thorough discussions regarding the advantages and disadvantages to numerous imaging and invasive modalities used to assess valve function in rodents9,10,11, to date, we are not aware of a publication that provides a comprehensive, step-by-step protocol for phenotyping heart valve function in mice.
The purpose of this manuscript is to describe the methods and protocols to phenotype heart valve function in mice. All methods and procedures have been approved by the Mayo Clinic Institutional Animal Care and Use Committee. Key components of this protocol include the depth of anesthesia, the evaluation of cardiac function, and the evaluation of heart valve function. We hope this report will not only serve to guide investigators interested in pursuing research in the field of heart valve disease, but will also start a national and international dialogue related to protocol standardization to ensure data reproducibility and validity in this rapidly-growing field. Importantly, successful imaging using high-resolution ultrasound systems requires a working knowledge of the principles of sonography (and terminology commonly used in sonography), an understanding of the fundamental principles of cardiac physiology, and significant experience with sonography to allow for accurate and time-efficient assessment of cardiac function in rodents.

<table><tbody><tr><td>High resolution ultrasound machine</td><td>VisualSonics, Fujifilm</td><td>Vevo 2100&amp;nbsp;</td><td></td></tr><tr><td>Isoflurane diffuser (capable of delivering 1 % to 1.5 % isoflurane mixed with 1 L/min 100% O&lt;sub&gt;2&lt;/sub&gt;</td><td>VisualSonics, Fujifilm</td><td>N/A</td><td></td></tr><tr><td>Transducers for small mice (550D) or larger mice (400)</td><td>MicroScan, VisualSonics, Fujifilm</td><td>MS 550D, MS 400</td><td></td></tr><tr><td>Animal platform</td><td>VisualSonics, Fujifilm</td><td>11503</td><td></td></tr><tr><td>Advanced physiological monitoring unit</td><td>VisualSonics, Fujifilm</td><td>N/A</td><td></td></tr><tr><td>Isoflurane</td><td>Terrell</td><td>NDC 66794-019-10</td><td></td></tr><tr><td>Nose cone and tubing connected to isoflurane diffuser and 100% O&lt;sub&gt;2&lt;/sub&gt;</td><td>Custom Engineered in-house</td><td>--</td><td></td></tr><tr><td>Hair razor</td><td>Andis Super AGR+ vet pack clipper</td><td>AD65340</td><td></td></tr><tr><td>Ultrasound gel</td><td>Parker Laboratories</td><td>REF 01-08</td><td></td></tr><tr><td>Electrode gel&amp;nbsp;</td><td>Parker Laboratories</td><td>REF 15-25</td><td></td></tr><tr><td>Adhesive tapes</td><td>Fisher Laboratories</td><td>1590120B</td><td></td></tr><tr><td>Paper towels</td><td></td><td></td><td></td></tr></tbody></table>

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

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.

Examples of images that are routinely obtained from animal cardiac ultrasound imaging are included in this manuscript. An illustration of transducer placement on the animal's chest is provided to give the reader a clear understanding of where the transducer is positioned to obtain the images as described. A photograph of the ultrasound laboratory set-up is also included to emphasize the importance of the proper equipment, particularly the ultrasound transducer to be used and the method of...

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 ...

echocardiographic-approaches-protocols-for-comprehensive-phenotypic

Research

JoVE Journal

Medicine

Transthoracic echocardiography offers a noninvasive method for the evaluation of cardiac function in mice. A combination of ultrasound and Doppler imaging modalities can be used to obtain dimensional measurements of the heart and intracardiac blood flow, which together provide an assessment of cardiac systolic and diastolic performance.

Transthoracic Echocardiography in Mice

This video demonstrates use of a rail-mounted high-frequency ultrasound probe to perform echocardiography on an anesthetized mouse.  The methods describe both conventional two-dimensional and M-mode measurements of cardiac function in addition to newer, more powerful tools such as color Doppler, strain analysis, as well as general and targeted contrast imaging.

Murine Echocardiography and Ultrasound Imaging

This video describes a protocol to perform echocardiography on a laboratory mouse.

Mouse Echocardiography: A Non-invasive Technique to Assess Cardiac Morphology

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.

Echocardiographic Assessment of the Right Heart in Mice

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.

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

The following protocol describes the methodology for the acquisition and analysis of echocardiographic images used to obtain the Left Atrial Volume (LAV), Aorta (Ao) diameter, and Pulmonary Artery (PA) diameter in mice. This technique is a non-invasive, non-terminal procedure that allows assessment of the cardiopulmonary function.

Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery

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.

Echocardiographic and Histological Examination of Cardiac Morphology in the 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 interchangeably.

Echocardiographic Measurement of Right Ventricular Diastolic Parameters in Mouse

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

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

This article details murine congenital heart disease (CHD) diagnostic methods using fetal echocardiography, necropsy, and Episcopic fluorescence image capture (EFIC) using Episcopic confocal microscopy (ECM) followed by three-dimensional (3D) reconstruction.

A Pipeline to Characterize Structural Heart Defects in the Fetal Mouse

Learn comprehensive echocardiographic methods for phenotyping valvular heart disease in mice. This standardized protocol enables detection of dysfunction across multiple heart valves and improves cardiac research reproducibility.

Echocardiographic Approaches and Protocols for Comprehensive Phenotypic Characterization of Valvular Heart Disease in Mice

Explore More Videos