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&#39;-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&#8242;-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 ...

echocardiographic-histological-examination-cardiac-morphology

Research

JoVE Journal

Medicine

21.2K 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

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

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

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

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

הערכה של תפקוד לב ומורפולוגיה שריר הלב באמצעות שחזור קטן בעלי חיים תראו-הלבשה היפוך (SALLI) בדיקת MRI בחולדות

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

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

היבטים טכניים של מורסת Aortocaval מאוס

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

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 ...

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled microscopic lesions (microlesions) that are remarkable due to the absence of infiltrating immune cells. Due to their release of cardiotoxic products, S. pneumoniae within microlesions are thought to contribute to the heart failure that is frequently observed during fulminate invasive pneumococcal disease in adults. Herein is demonstrated a protocol for experimental mouse infection that leads to reproducible cardiac microlesion formation within 30 hr. Instruction is provided on microlesion identification in hematoxylin &#38; eosin stained heart sections and the morphological distinctions between early and late microlesions are highlighted. Instruction is provided on a protocol for verification of S. pneumoniae within microlesions using antibodies against pneumococcal capsular polysaccharide and immunofluorescent microscopy. Last, a protocol for antibiotic intervention that rescues infected mice and for the detection and assessment of scar formation in the hearts of convalescent mice is provided. Together, these protocols will facilitate the investigation of the molecular mechanisms underlying pneumococcal cardiac invasion, cardiomyocyte death, cardiac remodeling as a result of exposure to S. pneumoniae, and the immune response to the pneumococci in the heart.

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

Streptococcus pneumoniae forms discrete non-purulent microscopic lesions in the heart. Outlined is the protocol for a murine model of cardiac microlesion formation. Instruction is provided on microlesion visualization using microscopy, discrimination between early and late microlesions, and methods to detect cardiac remodeling in hearts of convalescent animals.

ויזואליזציה של<em&gt; Streptococcus pneumoniae</em&gt; בתוך Microlesions הלב וRemodeling לב לאחר

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled ...

Do's and Don'ts in the Preparation of Muscle Cryosections for Histological Analysis

Histological evaluation of muscle biopsies has served as an indispensable tool in the understanding of the development and progression of pathology of neuromuscular disorders. However, in order to do so, proper care needs to be taken when excising and preserving tissues to achieve optimal staining. One method of tissue preservation involves fixing tissues in formaldehyde and then embedding them with paraffin wax. This method preserves morphology well and allows for long-term storage at RT but is cumbersome and requires handling of toxic chemicals. Further, formaldehyde fixation results in antigen cross-linking, which necessitates antigen retrieval protocols for effective immunostaining. On the contrary, frozen sectioning does not require fixation and thus retains biological antigen conformation. This method also provides a distinct advantage in quick turn around time, making it especially useful in situations needing quick histological evaluation like intraoperative surgical biopsies. Here we describe the most effective method of preparing muscle biopsies for visualization with different histological and immunological stains.

Here we demonstrate the most efficient methods for freezing, embedding, cryosectioning, and staining of muscle biopsies to avoid freezing artifacts.

עשה ואל תעשה בהכנת השריר cryosections לניתוח היסטולוגית

Histological evaluation of muscle biopsies has served as an indispensable tool in the understanding of the development and progression of pathology of ...

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 ...

Dissection of the Mouse Pancreas for Histological Analysis and Metabolic Profiling

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at the 12 codon (Ptf1acre/+;LSL-KrasG12D/+) mouse strain as a model of human pancreatic cancer. The goal of our current studies is to identify novel metabolic biomarkers of pancreatic cancer progression. We have performed metabolic profiling of urine, feces, blood, and pancreas tissue extracts, as well as histological analyses of the pancreas to stage the cancer progression. The mouse pancreas is not a well-defined solid organ like in humans, but rather is a diffusely distributed soft tissue that is not easily identified by individuals unfamiliar with mouse internal anatomy or by individuals that have little or no experience performing mouse organ dissections. The purpose of this article is to provide a detailed step-wise visual demonstration to guide novices in the removal of the mouse pancreas by dissection. This article should be especially valuable to students and investigators new to research that requires harvesting of the mouse pancreas by dissection for metabolic profiling or histological analyses.

This video article provides a detailed demonstration of the procedures required to successfully remove the pancreas from a mouse by dissection for histological analysis and metabolic profiling.

דיסקציה של הלבלב העכבר עבור ניתוח היסטולוגית והגדרת פרופיל מטבולי

We have been investigating the pancreas specific transcription factor, 1a cre-recombinase; lox-stop-lox- Kristen rat sarcoma, glycine to aspartic acid at ...

Cardiac Magnetic Resonance Imaging at 7 Tesla

CMR at an ultra-high field (magnetic field strength B0&#160;&#8805; 7 Tesla) benefits from the signal-to-noise ratio (SNR) advantage inherent at higher magnetic field strengths and potentially provides improved signal contrast and spatial resolution. While promising results have been achieved, ultra-high field CMR is challenging due to energy deposition constraints and physical phenomena such as transmission field non-uniformities and magnetic field inhomogeneities. In addition, the magneto-hydrodynamic effect renders the synchronization of the data acquisition with the cardiac motion difficult. The challenges are currently addressed by explorations into novel magnetic resonance technology. If all impediments can be overcome, ultra-high field CMR may generate new opportunities for functional CMR, myocardial tissue characterization, microstructure imaging or metabolic imaging. Recognizing this potential, we show that multi-channel radio frequency (RF) coil technology tailored for CMR at 7 Tesla together with higher order B0 shimming and a backup signal for cardiac triggering facilitates high fidelity functional CMR. With the proposed setup, cardiac chamber quantification can be accomplished in examination times similar to those achieved at lower field strengths. To share this experience and to support the dissemination of this expertise, this work describes our setup and protocol tailored for functional CMR at 7 Tesla.

The sensitivity gain inherent to ultrahigh field magnetic resonance holds promise for high spatial resolution imaging of the heart. Here, we describe a protocol customized for functional cardiovascular magnetic resonance (CMR) at 7 Tesla using an advanced multi-channel radio-frequency coil, magnetic field shimming and a triggering concept.

דימות תהודה מגנטית לב בטסלה 7

CMR at an ultra-high field (magnetic field strength B0&#160;&#8805; 7 Tesla) benefits from the signal-to-noise ratio (SNR) advantage inherent at higher ...

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 ...

Transthoracic Echocardiographic Examination in the Rabbit Model

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

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

בחינת החזה הקרדיוגרפית במודל הארנב

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