Name: echocardiographic and histological examination of cardiac morphology in the mouse
Uploaded: 2017-10-26T12:30:00.000+00:00
Duration: 10 min 22 s
Description: Watch this Scientific Journal Video about Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse at JoVE.com

העבודה נתמך על ידי הממשלה הצרפתית (סוכנות המחקר הלאומית, ANR) דרך התוכנית "השקעות עבור העתיד" LABEX SIGNALIFE (הפניה ANR-11-LABX-0028-01) ויוצקים על ידי מענקים ק' וו ד מ האגודה la Recherche sur le סרטן, Fondation דה פראנס, לתכנן Inserm סרטן. ד B. נ א קיבלו מלגות את Fondation pour la Médicale רשרש ומן העיר ניס, בהתאמה. Echocardiograph את המתמר היו מתבקשים המסופקים על ידי פיליפס. אנו מודים Borderie א, ס Destree, מ Cutajar-Bossert, א Landouar, א Martres, Biancardini א ו ס מ וגנר לסיוע טכני מיומן שלהם.

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המחברים אין לחשוף.

שיטות שונות פותחו כדי להעריך את מבנה הלב ותפקודו בעכברים, כולל אקוקרדיוגרפיה חדות משופרת MRI, מיקרו CT, סריקת PET. בשל העלות האפקטיביות שלה פשטות, אקו הוא הטכניקה הנפוצה ביותר לניתוח פונקציונלי עכברים11. באופן כללי, בגלל גודלו הקטן של הלב ואת תדירות גבוהה של קצב הלב בעכברים, מתמרים ?...

מגוון רחב של מודלים מהונדס גנטית עכבר זמינים, מספר מחקרים בעכברים גבוהה1,2. אקוקרדיוגרפיה MRI הגישות הנפוצות עבור אפיון פנוטיפי מורפולוגיה של מודלים אלה עכבר3ותפקוד הלב בקרב אנשי עסקים ותיירים כאחד. המטרה של פרוטוקול הציג היא לנתח את תפקוד הלב ואת מורפולוגיה בעכברים בוגרים. הוא משלב echocardiographic, היסטולוגית, מדידות immunohistochemical. בדיקה echocardiographic נעשה שימוש נרחב עכברים4,5,6,7,8,9,10,11, 12. . Pachon et al. 11 מזוהה 205 מחקרים שפורסמו בזרימת הדם, זרימת הדם מחקר, כתב העת האמריקאי לפיזיולוגיה - פיזיולוגיה הדם והלבומחקר לב וכלי דם בין 2012 ו 2015 זה שימוש בבדיקה echocardiographic בבעלי חיים.
אקוקרדיוגרפיה משמש לזיהוי פנוטיפים לב עכברים מהונדסים5,6,13,14,15,16, 17 , 18 , 19 , 20 , 21 , 22, כמו גם וניתוח תפקוד הלב אצל היפרטרופיה הנוצרות על-ידי עומס יתר כרונית, איסכמיה בשריר הלב, ומודלים של שריר הלב בעכברים (נבדקה12). ציוד אקוקרדיוגרפיה משופר מאפשר המדד הסטנדרטי של שמאל-חדרית (LV) בהתכווצות והתרחבות של מידות, הדמיה דופלר רקמות, ניגודיות שריר הלב echography את ההערכה של הפונקציה אזוריים LV ושמורת כלילית 12. באופן אידיאלי, יש לבצע בדיקה echocardiographic בעכברים בהכרה כדי למנוע את ההשפעות השליליות של הרדמה על הפונקציה כויץ, שליטה רפלקס אוטונומי, ולדרג את הלב11. עם זאת, גישה זו הוא מוגבל על ידי הדרישה לאמן את החיות; קשיים בשמירה על טמפרטורת הגוף יציב; תנועת חפצים; לחץ; תדרים לב גבוהה מאוד; את הדרישה לפחות שני חוקרים כדי לבצע את הניסוי, במיוחד אם מספר גדול של בעלי חיים נמצאים תחת חקירה. מעניין, מחקר שנערך לאחרונה דיווח אין הבדלים בפרמטרים echocardiographic מיומנים ובעלי לא מאומן19. אנו מבצעים מדידות echocardiographic בעכברים מורדם. פרוטוקולי הרדמה שונות יידונו להלן.
למרות אקוקרדיוגרפיה ברזולוציה תקנית (&#62; 10 מגה-הרץ) היא מספקת מידה LV סיסטולי, דיאסטולי מידות, תפקוד הלב בעכברים מבוגרים, השיטה היא מוגבלת בתיאור שלה התופעות המבנית הבסיסית. לפיכך, אנו משלבים את המידות ויוו עם היסטולוגית וניתוחים immunohistological כדי למדוד, לדוגמה, צפיפות cardiomyocyte בקוטר של כלי השיט. חקירות היסטולוגית ו immunohistological אחרים, כגון הקביעה של התפשטות, בחינת אפופטוזיס, מדידות גודל לאוטם, קביעה של פיברוזיס, ועל חופש הביטוי סמן ספציפיים, ניתן גם לבצע את אותו סוג של עובד הרקמה אך אינם הנושא של פרוטוקול זה. השילוב של ויוו echocardiographic בדיקה היסטולוגית ניתוחים מספק תובנות נוספות המשמשות כבסיס שינויים מבניים. בצעד נוסף, אנחנו יכולים להשלים מדידות אלה עם חקירות מולקולרית ומבניים אולטרה. ניתוחים היסטולוגית לא רק להשלים את הבדיקה echocardiographic, אלא גם להפוך חיוניות כאשר הרזולוציה של אקוקרדיוגרפיה אינה מספיקה. זהו המצב, מודלים של עכברים מהונדסים כי הם מתחלקים קטלני23,24.

<table><tbody><tr><td>Wheat germ agglutinin (WGA) conjugated tetramethylrhodamine</td><td>Life Technologies, Molecular Probes</td><td>W849</td><td></td></tr><tr><td>Biotinylated Goat Anti-Rabbit IgG Antibody</td><td>Vectorlabs</td><td>BA-1000</td><td></td></tr><tr><td>Avidin/Biotin Blocking Kit</td><td>Vectorlabs</td><td>SP-2001</td><td></td></tr><tr><td>VECTASTAIN Elite ABC HRP Kit (Peroxidase, Standard)</td><td>Vectorlabs</td><td>PK-6100</td><td></td></tr><tr><td>VECTASHIELD Antifade Mounting Medium with DAPI</td><td>Vectorlabs</td><td>H-1200</td><td></td></tr><tr><td>SIGMAFAST 3,3&#039;-Diaminobenzidine tablets</td><td>Sigma</td><td>D4168</td><td></td></tr><tr><td>Hydrogen peroxide solution</td><td>Sigma</td><td>H1009</td><td></td></tr><tr><td>Anti-Pecam-1 (CD31) antibody</td><td>Abcam</td><td>ab28364</td><td></td></tr><tr><td>Ultrasound transmission gel, Gel Aquasonic 100</td><td>Parker</td><td></td><td></td></tr><tr><td>Linear ultrasound probe, L15-7io</td><td>Philips Healthcare</td><td></td><td></td></tr><tr><td>Echocardiograph, IE33 xMATRIX</td><td>Philips Healthcare</td><td></td><td></td></tr><tr><td>Microscope, Leica DMi8</td><td>Leica</td><td></td><td></td></tr><tr><td>Fluorescence Filterset DAPI</td><td>Leica</td><td>11525304</td><td></td></tr><tr><td>Filterset TxR</td><td>Leica</td><td>11525310</td><td></td></tr><tr><td>Digital Camera, SPOT RT3 Color Slider</td><td>Spot Imaging</td><td></td><td></td></tr><tr><td>Imaging Software, SPOT 5.2 Advanced and Basic Software</td><td>Spot Imaging</td><td></td><td></td></tr><tr><td>Imaging Computer</td><td>Dell</td><td></td><td></td></tr><tr><td>Fine Scissors</td><td>Fine Science Tools</td><td>14028-10</td><td></td></tr><tr><td>Large Scissors</td><td>Fine Science Tools</td><td>14501-14</td><td></td></tr><tr><td>Scalpel blades</td><td>Fine Science Tools</td><td>10023-00</td><td></td></tr><tr><td>Graefe Forceps</td><td>Fine Science Tools</td><td>11650-10</td><td></td></tr><tr><td>Rodent shaver</td><td>Harvard Apparatus</td><td>34-0243</td><td></td></tr><tr><td>cassettes for paraffin embedding</td><td>Sakura</td><td>4155F</td><td></td></tr><tr><td>neutral buffered Formalin</td><td>Sakura</td><td>8727</td><td></td></tr><tr><td>Xylene</td><td>Sakura</td><td>8733</td><td></td></tr><tr><td>Paraffine TEK III</td><td>Sakura</td><td>4511</td><td></td></tr><tr><td>automated embedding apparatus, Tissue-Tek VIP</td><td>Sakura</td><td>6032</td><td></td></tr><tr><td>paraffin-embedding station Tissue-Tek TEC 5</td><td>Sakura</td><td>5229</td><td></td></tr><tr><td>microtome blades,Accu-Edge S35</td><td>Sakura</td><td>4685</td><td></td></tr><tr><td>microscopy slides, Tissue-Tek</td><td>Sakura</td><td>9533</td><td></td></tr><tr><td>cover slips, Tissue-Tek</td><td>Sakura</td><td>9582</td><td></td></tr><tr><td>Mounting medium Tissue-Tek</td><td>Sakura</td><td>1408</td><td></td></tr><tr><td>slide boxes</td><td>Sakura</td><td>3958</td><td></td></tr><tr><td>eosine solution</td><td>Sakura</td><td>8703</td><td></td></tr><tr><td>hematoxyline solution</td><td>Sakura</td><td>8711</td><td></td></tr><tr><td>microtome, RM2125RT</td><td>Leica</td><td>720-1880 (VWR)</td><td></td></tr><tr><td>water bath, Leica HI1210</td><td>Leica</td><td>720-0113(VWR)</td><td></td></tr><tr><td>Ethanol</td><td>VWR</td><td>ACRO444220050</td><td></td></tr><tr><td>15 ml tubes</td><td>VWR</td><td>734-0451</td><td></td></tr><tr><td>staining glass dish</td><td>VWR</td><td>MARI4220004</td><td></td></tr><tr><td>staining jars</td><td>VWR</td><td>MARI4200005</td><td></td></tr><tr><td>Incubator</td><td>Binder</td><td>9010-0012</td><td></td></tr><tr><td>DAB and urea hydrogen peroxide tablets, SIGMAFAST 3,3&amp;prime;-Diaminobenzidine tablets</td><td>Sigma</td><td>D4293</td><td></td></tr><tr><td>PBS (10X)</td><td>Thermo Fisher Scientific</td><td>70011044</td><td></td></tr></tbody></table>

echocardiographic and histological examination of cardiac morphology in the mouse

מספר גדל והולך של מודלים מהונדס גנטית העכבר הפך זמין בשנים האחרונות. יתר על כן, מספר מחקרים שבוצעו בעכברים הוא גבוה. אפיון פנוטיפי של מודלים אלה העכבר גם דורש הבחינה של תפקוד הלב ואת מורפולוגיה. אקוקרדיוגרפיה דימות תהודה מגנטית (MRI) גישות נפוצות כדי לאפיין מורפולוגיה בעכברים ותפקוד הלב בקרב אנשי עסקים ותיירים כאחד. ציוד Echocardiographic ו- MRI המתמחה בשימוש מכרסמים קטנים הוא יקר ודורש שטח ייעודי. פרוטוקול זה מתאר מידות הלב בעכברים באמצעות מערכת echocardiographic קליניים גשש כלי דם אנושי 15 מגה-הרץ. המדידות מבוצעות על עכברים בוגרים מורדם. רצפי תמונות לפחות שלושה מוקלט, נותחו עבור כל בעל חיים במצב M בתצוגת ציר קצר מתחת. לאחר מכן מבוצעת בדיקה היסטולוגית הלב, קטרים cardiomyocyte נקבעים ב hematoxylin-agglutinin eosin - או נבט חיטה (WGA)-פרפין סעיפים מוכתמים. צפיפות כלי הוא נחוש morphometrically אחרי immunostaining Pecam-1. הפרוטוקול עבר בהצלחה שימושית ללימודים תרופתי ושונה גנטי מודלים בעלי חיים בתנאים בסיסית, כמו גם לאחר ניסיוני אוטם שריר הלב על ידי מצדו קבוע של השמאלי הקדמי היורד עורקים ( LAD). מניסיוננו, חקירה echocardiographic מוגבל לבעלי מרדימים, הוא ריאלי בעכברים בוגרים במשקל של פחות 25 גרם.

איור 1, הקלטות echocardiographic נציג מדגימים את התועלת אקוקרדיוגרפיה כדי לזהות פנוטיפים הלב בעכברים מהונדסים. ניתן בקלות לזהות את ההבדל בין עכבר עם תפקוד הלב תקין (איור 1 א') חיה עם החדר השמאלי מורחבים, LV מופחתת פונקציה (איור 1B). <strong ...

Watch this Scientific Journal Video about Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse at JoVE.com

Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

בדיקה echocardiographic משמש לעתים קרובות בעכברים. מכשירי אולטרסאונד ברזולוציה גבוהה יקר פותחו למטרה זו. פרוטוקול זה מתאר הליך echocardiographic סבירים בשילוב עם morphometric היסטולוגית ניתוחים כדי לקבוע מורפולוגיה לב.

בבדיקה היסטולוגית ו echocardiographic של מורפולוגיה לב העכבר

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

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Research

JoVE Journal

Medicine

21.3K Views.  CNRS, INSERM, iBV. בדיקה echocardiographic משמש לעתים קרובות בעכברים. מכשירי אולטרסאונד ברזולוציה גבוהה יקר פותחו למטרה זו. פרוטוקול זה מתאר הליך echocardiographic סבירים בשילוב עם morphometric היסטולוגית ניתוחים כדי לקבוע מורפולוגיה לב.

Video: Echocardiographic and Histological Examination of Cardiac Morphology in the Mouse

Transthoracic Echocardiography in Mice

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene expression. Transgenic and gene targeting methods can be used to generate mice with altered cardiac size and function,1-3 and as a result, in vivo techniques are needed to evaluate their cardiac phenotype. Transthoracic echocardiography, pulse wave Doppler (PWD), and tissue Doppler imaging (TDI) can be used to provide dimensional measurements of the mouse heart and to quantify the degree of cardiac systolic and diastolic performance. Two-dimensional imaging is used to detect abnormal anatomy or movements of the left ventricle, whereas M-mode echo is used for quantification of cardiac dimensions and contractility.4,5 In addition, PWD is used to quantify localized velocity of turbulent flow,6 whereas TDI is used to measure the velocity of myocardial motion.7 Thus, transthoracic echocardiography offers a comprehensive method for the noninvasive evaluation of cardiac function in mice.

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

אקו Transthoracic ב עכברים

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

Murine Echocardiography and Ultrasound Imaging

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1 Echocardiography provides a powerful, non-invasive tool to serially assess cardiac morphometry and function in a living animal.2 However, using this technique on mice poses unique challenges owing to the small size and rapid heart rate of these animals.3 Until recently, few ultrasound systems were capable of performing quality echocardiography on mice, and those generally lacked the image resolution and frame rate necessary to obtain truly quantitative measurements. Newly released systems such as the VisualSonics Vevo2100 provide new tools for researchers to carefully and non-invasively investigate cardiac function in mice. This system generates high resolution images and provides analysis capabilities similar to those used with human patients. Although color Doppler has been available for over 30 years in humans, this valuable technology has only recently been possible in rodent ultrasound.4,5 Color Doppler has broad applications for echocardiography, including the ability to quickly assess flow directionality in vessels and through valves, and to rapidly identify valve regurgitation. Strain analysis is a critical advance that is utilized to quantitatively measure regional myocardial function.6 This technique has the potential to detect changes in pathology, or resolution of pathology, earlier than conventional techniques. Coupled with the addition of three-dimensional image reconstruction, volumetric assessment of whole-organs is possible, including visualization and assessment of cardiac and vascular structures. Murine-compatible contrast imaging can also allow for volumetric measurements and tissue perfusion assessment.

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

אקו Murine והדמיה אולטראסאונד

Rodent models of cardiac pathophysiology represent a valuable research tool to investigate mechanism of disease as well as test new therapeutics.1  ...

Echocardiographic Assessment of the Right Heart in Mice

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential therapies. Given the expense and time involved in creating animal models of disease, it is critical that researchers have tools to accurately assess phenotypic expression of disease. Right ventricular dysfunction is the major manifestation of pulmonary hypertension. Echocardiography is the mainstay of the noninvasive assessment of right ventricular function in rodent models and has the advantage of clear translation to humans in whom the same tool is used. Published echocardiography protocols in murine models of PAH are lacking.
In this article, we describe a protocol for assessing RV and pulmonary vascular function in a mouse model of PAH with a dominant negative BMPRII mutation; however, this protocol is applicable to any diseases affecting the pulmonary vasculature or right heart. We provide a detailed description of animal preparation, image acquisition and hemodynamic calculation of stroke volume, cardiac output and an estimate of pulmonary artery pressure.

This article provides a protocol for the echocardiographic assessment of right ventricular size and pulmonary hypertension in mice. Applications include phenotype determination and serial assessment in transgenic and toxin-induced mouse models of cardiomyopathy and pulmonary vascular disease.

אקו הערכה של הלב הימני בעכברים

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

Analysis of Congenital Heart Defects in Mouse Embryos Using Qualitative and Quantitative Histological Methods

Congenital heart defects (CHD) are the most common type of birth defect in humans, affecting up to 1% of all live births. However, the underlying causes for CHD are still poorly understood. The developing mouse constitutes a valuable model for the study of CHD, because cardiac developmental programs between mice and humans are highly conserved. The protocol describes in detail how to produce mouse embryos of the desired gestational stage, methods to isolate and preserve the heart for downstream processing, quantitative methods to identify common types of CHD by histology (e.g., ventricular septal defects, atrial septal defects, patent ductus arteriosus), and quantitative histomorphometry methods to measure common muscular compaction phenotypes. These methods articulate all the steps involved in sample preparation, collection, and analysis, allowing scientists to correctly and reproducibly measure CHD.

In this protocol, we describe procedures to qualitatively and quantitatively analyze developmental phenotypes in mice associated with congenital heart defects.

ניתוח של פגמים לב מולדים בעוברי עכבר באמצעות שיטות היסטולוגית איכותית וכמותית

Congenital heart defects (CHD) are the most common type of birth defect in humans, affecting up to 1% of all live births. However, the underlying causes ...

Developmental Biology

A Pipeline to Characterize Structural Heart Defects in the Fetal Mouse

Congenital heart diseases (CHDs) are major causes of infant death in the United States. In the 1980s and earlier, most patients with moderate or severe CHD died before adulthood, with the maximum mortality during the first week of life. Remarkable advances in surgical techniques, diagnostic approaches, and medical management have led to marked improvements in outcomes. To address the critical research needs of understanding congenital heart defects, murine models have provided an ideal research platform, as they have very similar heart anatomy to humans and short gestation rates. The combination of genetic engineering with high-throughput phenotyping tools has allowed for the replication and diagnosis of structural heart defects to further elucidate the molecular pathways behind CHDs. The use of noninvasive fetal echocardiography to screen the cardiac phenotypes in mouse models coupled with the high fidelity of Episcopic fluorescence image capture (EFIC) using Episcopic confocal microscopy (ECM) histopathology with three-dimensional (3D) reconstructions enables a detailed view into the anatomy of various congenital heart defects. This protocol outlines a complete workflow of these methods to obtain an accurate diagnosis of murine congenital heart defects. Applying this phenotyping protocol to model organisms will allow for accurate CHD diagnosis, yielding insights into the mechanisms of CHD. Identifying the underlying mechanisms of CHD provide opportunities for potential therapies and interventions.

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

צינור לאפיון מומי לב מבניים בעכבר העוברי

Congenital heart diseases (CHDs) are major causes of infant death in the United States. In the 1980s and earlier, most patients with moderate or severe ...

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

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable noninvasive method is essential to assess real-time cardiac morphological and functional changes in animal models of heart disease. Transthoracic echocardiography represents an important tool for noninvasive assessment of cardiac structure and function2. Here we used a high-resolution ultrasound imaging system to monitor myocardial remodeling and heart failure progression over time in a mouse model of TAC. B-mode, M-mode, and Doppler imaging were used to precisely assess cardiac hypertrophy, ventricular dilatation, and functional deterioration in mice following TAC. Color and pulse wave (PW) Doppler imaging was used to noninvasively measure pressure gradient across the aortic constriction created by TAC and to assess transmitral blood flow in mice. Thus transthoracic echocardiographic imaging provides comprehensive noninvasive measurements of cardiac dimensions and function in mouse models of heart disease.

The goal of this protocol is to noninvasively assess cardiac structural and functional changes in a mouse model of heart disease created by transverse aortic constriction, using B- and M-mode echocardiography and color/pulse wave Doppler imaging.

שינויים הערכה לב מורפולוגי תפקודי עכבר דגם של מועקה הרוחבית אב עורקים על ידי הדמית אקו

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

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.

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

גישות פרוטוקולים אקו לאפיון פנוטיפי מקיף של מסתמית מחלות לב בעכברים

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 ...

Murine Echocardiography of Left Atrium, Aorta, and Pulmonary Artery

The present methodology teaches the investigator how to measure and use the LAV as a surrogate of chronic elevations in Left Ventricular diastolic pressure through echocardiography, as well as to obtain measurements of the Aorta and PA diameter in mice.
Mice older than 10 d of age can be analyzed using the present technique. The technique is composed of 3 main steps: set-up, image acquisition, and image analysis. The set-up step consists of getting the mouse anesthetized with 1% isoflurane, shaving it, and taping it in a supine position to a heated EKG board where the image acquisition will take place. The image acquisition step consists of learning to identify the cardiac structures and obtaining all the required images with its correspondent probe and axis in order to be able to calculate volumes and diameters. The image analysis step consists of measuring the previously acquired images with the aid of computer software.
Advantages of the proposed technique include a fast (15 min) procedure that would allow the researcher to evaluate interventions in a non-invasive, non-terminal approach and therefore follow the same mouse over time; each mouse can be used as its own control. This fact plus having the same operator perform all the acquisition and analysis for the entire experiment minimizes the limitation of operator-dependency. The present methodology is useful for mouse researchers in cardiovascular and pulmonary medicine.

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

אקו Murine של השמאל אטריום, אבי העורקים, ו עורק הריאה

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

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.

המדידה האקוקרדיוגרפית של פרמטרים של הימני החדרית בעכבר

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

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

Echocardiography is a non-invasive procedure that enables the evaluation of structural and functional parameters in animal models of cardiovascular disease and is used to assess the impact of potential treatments in preclinical studies. Echocardiographic studies are usually conducted in young adult mice (i.e., 4-6 weeks of age). The evaluation of early neonatal cardiovascular function is not usually performed because of the small size of the mouse pups and the associated technical difficulties. One of the most important challenges is that the short length of the pups&#39; limbs prevents them from reaching the electrodes in the echocardiography platform. Body temperature is the other challenge, as pups are very susceptible to changes in temperature. Therefore, it is important to establish a practical guide for performing echocardiographic studies in small mouse pups to help researchers detect early pathological changes and study the progression of cardiovascular disease over time. The current work describes a protocol for performing echocardiography in mouse pups at the early age of 7 days old. The echocardiographic characterization of cardiac morphology, function, and coronary flow in neonatal mice is also described.

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

אפיון אקוקרדיוגרפי של מבנה החדר השמאלי, תפקודו וזרימה כלילית בעכברים ילודים

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