Questo lavoro è sostenuto da finanziamenti del Fondo UO generale G110 e ricerca di eccellenza Fondo della ricerca biomedica e OSU-HRS Fondo 013.000. Gli autori desiderano ringraziare Lauren Chen per la sua assistenza nella preparazione di questo manoscritto.

Tutte le procedure che coinvolgono soggetti animali sono stati approvati e completati in conformità e il rispetto degli Institutional Animal Care ed uso commissione Ohio State University (IACUC) norme e linee guida. 1. Topo Anestesia <ol><li> Istituire un tavolo procedura pulita con un pad isotermica riscaldata avvolta in un asciugamano chirurgico. Il rilievo di riscaldamento deve essere mantenuta tra 30 ° C e 34 ° C per stabilizzare la temperatura interna dell&#39;animale, riducendo la probabilità di stress per l&#39;animale. </li><li> Posizionare il mouse in una camera di induzione di anestesia con i seguenti parametri: flusso di ossigeno impostato a 1,5 L / min e vaporizzatore isoflurano impostato a 3,5%. Sedazione completa dovrebbe avvenire entro 1-2 min. Se una camera di induzione non è disponibile, una campana di vetro può essere utilizzato con una rete metallica posizionata nella parte inferiore per evitare il contatto diretto con l&#39;animale isoflurano. </li><li> Rimuovere immediatamente il mouse dalla camera di induzione voltaè completamente anestetizzato (raggiunta quando il mouse perde la funzione motoria volontaria). Applicare un naso-cono all&#39;animale per il mantenimento dell&#39;anestesia. Il flusso di ossigeno deve essere diminuito di circa 0,5 L / min e il vaporizzatore isoflurano dovrebbe essere impostato in un range da 1,5 a 2,5%. <ol><li> Applicare una piccola quantità di pomata oftalmica direttamente alle cornee per ridurre la secchezza degli occhi 8. Inoltre, durante l&#39;anestesia, il mouse dovrebbe mantenere l&#39;assenza del riflesso ritiro pedale, le mucose dovrebbero rimanere di colore rosa, e la respirazione dovrebbe apparire costante. </li></ol></li></ol> 2. Preparazione per Diagnostic Ultrasound Imaging procedura <ol><li> Trattenere ogni tappa del mouse sulla tabella procedura riscaldata con un adesivo rimovibile, come nastro chirurgico. </li><li> Usando un rasoio elettrico, rimuovere i capelli sulla superficie ventrale del corpo tra il modo addome e mezza la cavità toracica. Applicare crema depilatoria per ulteriori respostare i capelli rimanenti non tagliata dal rasoio. Pulire la crema con una garza umida dopo 2-3 minuti. </li><li> Rimuovere i peli in eccesso utilizzando una garza imbevuta di acqua e pulire la regione rasata con il 70% di alcool o equivalente antisettico. La sonda ad ultrasuoni viene applicato a questa zona per visualizzare la funzione diaframma. Un analgesico topico può essere fornito agli animali che avvertono lievi irritazioni cutanee a causa di depilazione. </li></ol> 3. Diagnostic Ultrasound Imaging Protocol <ol><li> Accendere il dispositivo ad ultrasuoni e regolare la potenza di uscita (se necessario) sull&#39;apparecchio per percentuale avere risoluzione ottimale. </li><li> Impostare la macchina ad ultrasuoni a uno B (luminosità)-mode, M (moto)-mode, o entrambi prima di imaging, che consente la corretta visualizzazione della contrazione diaframma mouse. </li><li> Applicare una piccola quantità di gel ultrasuoni sull&#39;addome superiore del mouse e massaggiare il gel verso la cavità toracica. </li><li> Posizionare l&#39;ultrasuono trasduttore in questa zona e l&#39;angolo verso l&#39;alto verso il cuore. Regolare la sonda fino a raggiungere una risoluzione ottimizzata dell&#39;immagine. Nota: per questo protocollo, una matrice di micro-convessa o lineare phased array di trasduttori è una sonda ideale da usare grazie al minimo ingombro e un&#39;eccellente risoluzione assiale 9; richiede la frequenza da regolare attraverso la larghezza di banda e per questi esperimenti una gamma di 6.5-12 MHz può essere utilizzata. </li><li> Premere il pulsante FREEZE per salvare temporaneamente le immagini diaframma e visualizzare le contrazioni selezionati. </li><li> Salvare la registrazione come cine loop, che permette di successive misure di escursione diaframmatica, così come il tasso di respirazione. Nota: fotogrammi di immagini possono essere salvate nella memoria del computer o su un disco rigido esterno per le analisi future 9. <ol><li> Proprio misurare la profondità del movimento del diaframma dal relax alla contrazione utilizzando calibri elettronici che fanno parte del software ultrasuoni. </li><li> Convertire ilfile di cine loop in un file MPEG e determinare il tasso di respirazione contando il numero di contrazioni diaframmatiche durante il periodo di registrazione. In alternativa, il numero di contrazioni al minuto (frequenza respiratoria) può essere conteggiato dall&#39;immagine M-mode. </li></ol></li></ol> 4. Post anestesia recupero degli animali <ol><li> Il mouse dovrebbe recuperare completamente dall&#39;anestesia entro 1 ora. Non lasciare l&#39;animale incustodito fino a quando non ha ripreso conoscenza sufficiente per mantenere decubito sternale. </li></ol>

<ol>
	<li>Snyder, C. 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=Complementarity+of+ultrasound+and+fluorescence+imaging+in+an+orthotopic+mouse+model+of+pancreatic+cancer.">Complementarity of ultrasound and fluorescence imaging in an orthotopic mouse model of pancreatic cancer.</a> BMC Cancer. 9, 106 (2009).</li><li>Franchi, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+assessment+of+cardiac+function+in+a+mouse+model+of+renovascular+hypertension.">Non-invasive assessment of cardiac function in a mouse model of renovascular hypertension.</a> Hypertension Research: Official Journal of the Japanese Society of Hypertension. , (2013).</li><li>Coatney, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ultrasound+imaging:+principles+and+applications+in+rodent+research.">Ultrasound imaging: principles and applications in rodent research.</a> ILAR Journal / National Research Council, Institute of Laboratory Animal Resources. 42, 233-247 (2001).</li><li>Morenz, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+air+trapping+in+chronic+obstructive+pulmonary+disease+by+low+frequency+ultrasound.">Detection of air trapping in chronic obstructive pulmonary disease by low frequency ultrasound.</a> BMC Pulmonary Medicine. 12, 8 (2012).</li><li>Gilliam, L. A., Moylan, J. S., Ann Callahan, L., Sumandea, M. P., Reid, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Doxorubicin+causes+diaphragm+weakness+in+murine+models+of+cancer+chemotherapy.">Doxorubicin causes diaphragm weakness in murine models of cancer chemotherapy.</a> Muscle &amp; Nerve. 43, 94-102 (2011).</li><li>Ferreira, L. F., Campbell, K. S., Reid, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effectiveness+of+sulfur-containing+antioxidants+in+delaying+skeletal+muscle+fatigue.">Effectiveness of sulfur-containing antioxidants in delaying skeletal muscle fatigue.</a> Medicine and Science in Sports and Exercise. 43, 1025-1031 (2011).</li><li>Zuo, L., Clanton, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reactive+oxygen+species+formation+in+the+transition+to+hypoxia+in+skeletal+muscle.">Reactive oxygen species formation in the transition to hypoxia in skeletal muscle.</a> American Journal of Physiology. Cell Physiology. 289, 207-216 (2005).</li><li>Helms, M. N., Torres-Gonzalez, E., Goodson, P., Rojas, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+tracheal+instillation+of+solutes+into+mouse+lung.">Direct tracheal instillation of solutes into mouse lung.</a> J. Vis. Exp. , (2010).</li><li>Hedrick, W. R., Hykes, D. L., Starchman, D. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Ultrasound Physics and Instrumentation. , 445 (2005).</li><li>von Sarnowski, B., Khaw, A. V., Kessler, C., Schminke, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+a+microconvex+array+transducer+for+the+ultrasonographic+examination+of+the+intrathoracic+segments+of+the+supraaortic+arteries.">Evaluation of a microconvex array transducer for the ultrasonographic examination of the intrathoracic segments of the supraaortic arteries.</a> Journal of Neuroimaging: Official Journal of the American Society of Neuroimaging. 20, 246-250 (2010).</li><li>Stocksley, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Abdominal Ultrasound. , 7-8 (2001).</li><li>Kremkau, F. W., Taylor, K. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Artifacts+in+ultrasound+imaging.">Artifacts in ultrasound imaging.</a> Journal of Ultrasound in Medicine: Official Journal of the American Institute of Ultrasound in Medicine. 5, 227-237 (1986).</li><li>Kremkau, F. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diagnostic+Ultrasound:+Principles+and+Instruments.">Diagnostic Ultrasound: Principles and Instruments.</a> Saunders Elsevier. , 521 (2006).</li><li>Laing, F. C., Kurtz, A. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+importance+of+ultrasonic+side-lobe+artifacts.">The importance of ultrasonic side-lobe artifacts.</a> Radiology. 145, 763-768 (1982).</li><li>Abu-Zidan, F. M., Hefny, A. F., Corr, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+ultrasound+physics.">Clinical ultrasound physics.</a> Journal of Emergencies, Trauma, and Shock. 4, 501-503 (2011).</li><li>Gargani, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+ultrasound:+a+new+tool+for+the+cardiologist.">Lung ultrasound: a new tool for the cardiologist.</a> Cardiovascular Ultrasound. 9, 6 (2011).</li><li>Sanders, R. C., Winter, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Clinical Sonography A Practical Guide. , 632 (2007).</li></ol>

Gli autori dichiarano di non avere interessi finanziari in competizione.

Il protocollo sperimentale attuale sviluppa diagnostici tecniche di imaging ad ultrasuoni specifica dell&#39;attività diaframma in un modello murino mediante un approccio non invasivo vivo. Le impostazioni di apparecchi per anestesia sono valori approssimati, che possono essere leggermente adattate per ciascun animale in quanto topi individuo può rispondere in modo diverso ad anestesia. Per evitare somministrazione anestesia improprio, è importante controllare regolarmente i segni vitali del topo co...

Recentemente, tecniche di imaging ad ultrasuoni di diagnostica sono state applicate a modelli murini di ipertensione renovascolare e pancreas 1,2 cancro. Tuttavia, queste tecniche non sono stati ampiamente utilizzati in roditori respiratoria saggio di funzione muscolare. Pertanto, abbiamo sviluppato un metodo di imaging ad ultrasuoni di diagnostica come un valido strumento per le valutazioni in vivo longitudinali di mobilità del diaframma nei topi. Ci sono diversi vantaggi per diagnostica ecografica. Per esempio, è invasivo, sicuro, portatile, e consente misurazioni in tempo reale ad un costo relativamente basso 3. In particolare, alcuni dispositivi ad ultrasuoni a bassa frequenza sono stati in grado di rilevare air trapping, una caratteristica clinica della malattia polmonare ostruttiva cronica (BPCO) con lieve a grave limitazione al flusso aereo 4. Così, diagnostica ecografica può servire come un metodo di screening facilmente accessibile e riproducibile per il monitoraggio in tempo reale<html> di disturbi respiratori. Tecniche di imaging ad ultrasuoni di diagnostica sono spesso applicati agli animali più grandi o soggetti umani. Tuttavia, ci sono stati un numero limitato di studi di imaging ad ultrasuoni su modelli murini, che è probabilmente dovuto alle sfide di eseguire ultrasuoni su soggetti di piccole dimensioni. Il protocollo attuale delinea una nuova procedura per misurare la funzione diaframma nel topo. Inoltre, anche se ci sono stati diversi studi sui roditori sulla funzione del diaframma, la maggior parte dei risultati sono stati ottenuti isolando fasce muscolari direttamente dall&#39;animale eutanasia 5-7. Al contrario, utilizzando un metodo di imaging diagnostico ad ultrasuoni in vivo per analizzare l&#39;attività diaframma diminuirebbe il numero di animali sacrificati per la sperimentazione. Inoltre, i trattamenti a lungo termine incentrati sul rafforzamento della membrana contrattilità possono essere valutati con precisione con gli ultrasuoni in modelli di roditori senza sacrificare animali. S copi &quot;&gt; Nel nostro laboratorio, abbiamo sviluppato un metodo efficace per la visualizzazione e l&#39;analisi dell&#39;attività diaframma mouse usando una macchina ad ultrasuoni, che aiuta la comprensione della funzione di diaframma in vivo, evita metodi invasivi agli animali, e facilita lo sviluppo di terapeutica trattamenti per la disfunzione respiratoria.

<table><tbody><tr><td>Veterinary digital ultrasonic diagnostic imaging system</td><td>Edan</td><td>DUS 3 VET</td><td>Ultrasound parameters include: frequency of 6.5 MHz, Depth of 29 mm. Note: An equivalent ultrasound machine may be used for this protocol</td></tr><tr><td>Micro-convex array transducer</td><td>Edan</td><td>C611</td><td>Or equivalent</td></tr><tr><td>GE Logiq i hand-carried unit (HCU)&amp;nbsp;</td><td>GE Healthcare</td><td>GE Logiq i hand-carried unit (HCU)</td><td>Or equivalent</td></tr><tr><td>GE 12 MHz linear array probe</td><td>GE Healthcare</td><td>12L-RS</td><td>Or equivalent</td></tr><tr><td>Veterinary anesthetic vaporizer</td><td>Webster Veterinary</td><td>Serial #: W422021</td><td>Isoflurane was exclusively used with this vaporizer (or equivalent). A custom made induction chamber for anesthesia was assembled for initial anesthesia. Maintenance anesthesia was performed using a nose cone</td></tr><tr><td>Isothesia (Isoflurane, USP)</td><td>Butler Schein</td><td>29405 250ML PVL</td><td>Or equivalent</td></tr><tr><td>Enviro-pure anesthesia absorbing canister</td><td>Surgivet Smiths Medical PM, Inc.</td><td>Part #: 32373B10</td><td>Or equivalent</td></tr><tr><td>Ultrasound transmission gel</td><td>HM Sonic</td><td>N/A</td><td>Or equivalent&amp;nbsp;</td></tr><tr><td>Puralube vet ointment</td><td>Puralube</td><td>NDC 17033-211-38</td><td>Or equivalent</td></tr><tr><td>Deltaphase isothermal pad</td><td>Braintree Scientific Inc.</td><td>39DP</td><td>Or equivalent</td></tr><tr><td>Hair remover</td><td>Nair</td><td>N/A</td><td>Or equivalent</td></tr><tr><td>Electric razor</td><td>Remington</td><td>HC-5015</td><td>Or equivalent</td></tr><tr><td>Surgical tape</td><td>3M Micropore</td><td>1530-1</td><td>Or equivalent</td></tr><tr><td>Gauze sponges</td><td>Dynarex</td><td>3262</td><td>Or equivalent</td></tr></tbody></table>

diagnostic ultrasound imaging of mouse diaphragm function

Analisi Funzione di roditore muscoli scheletrici respiratorio, in particolare il diaframma, viene comunemente eseguita isolando strisce muscolari utilizzando procedure chirurgiche invasive. Anche se questo è un metodo efficace di valutazione in vitro dell&#39;attività diaframma, esso comporta un intervento chirurgico non-sopravvivenza. L&#39;applicazione di non invasiva ecografia come una procedura in vivo è utile in quanto non solo riduce il numero di animali sacrificati, ma è adatto anche per monitorare la progressione della malattia in topi vivi. Così, il nostro metodo di imaging ad ultrasuoni può probabilmente contribuire allo sviluppo di nuove terapie che alleviano lesione muscolare indotto da varie malattie respiratorie. In particolare, nella diagnosi clinica delle malattie polmonari ostruttive, ecografia ha il potenziale di essere utilizzato in combinazione con altri test standard per rilevare l&#39;insorgenza precoce di affaticamento muscolare diaframma. Nel protocollo attuale, si descrive come valutare con precisione diaframma contractilità in un modello murino utilizzando una tecnica diagnostica ecografica.

Una tipica immagine ecografica di un diaframma mouse viene mostrato nella Figura 1A. Il diaframma topo spostamento verticale massima è stata registrata. Questa distanza è stata calcolata misurando con precisione la profondità di movimento del diaframma da relax contrazione utilizzando calibri elettronici che fanno parte del software ultrasuoni. Tabella 1 visualizza queste misurazioni della distanza di contrazioni diaframmatici da tre diversi topi. Dopo la conversione del file cine lo...

Watch this Scientific Journal Video about Diagnostic Ultrasound Imaging of Mouse Diaphragm Function at JoVE.com

Diagnostic Ultrasound Imaging of Mouse Diaphragm Function

Diagnostica ecografica ha dimostrato di essere efficace nella diagnosi delle diverse malattie respiratorie in soggetti umani e animali. Dimostriamo un protocollo ecografia completa utilizzata da laboratorio del Dr. Zuo per analizzare la cinetica del diaframma in particolare in modelli murini. Questa è anche una tecnica di ricerca non invasiva che può fornire informazioni quantitative sulla funzione dei muscoli respiratori mouse.

Diagnostic Ultrasound Imaging di funzione mouse Diaframma

Function analysis of rodent respiratory skeletal muscles, particularly the diaphragm, is commonly performed by isolating muscle strips using invasive ...

diagnostic-ultrasound-imaging-of-mouse-diaphragm-function

Research

JoVE Journal

Medicine

16.0K Views.  The Ohio State University College of Medicine.  Diagnostica ecografica ha dimostrato di essere efficace nella diagnosi delle diverse malattie respiratorie in soggetti umani e animali. Dimostriamo un protocollo ecografia completa utilizzata da laboratorio del Dr. Zuo per analizzare la cinetica del diaframma in particolare in modelli murini. Questa è anche una tecnica di ricerca non invasiva che può fornire informazioni quantitative sulla funzione dei muscoli respiratori mouse. 

Video: Diagnostic Ultrasound Imaging of Mouse Diaphragm Function

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.

L&#39;ecocardiografia transtoracica in topi

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

Ricerca

Medicina

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.

L&#39;ecocardiografia murini e imaging a ultrasuoni

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

Murine Fetal Echocardiography

Transgenic mice displaying abnormalities in cardiac development and function represent a powerful tool for the understanding the molecular mechanisms underlying both normal cardiovascular function and the pathophysiological basis of human cardiovascular disease. Fetal and perinatal death is a common feature when studying genetic alterations affecting cardiac development 1-3. In order to study the role of genetic or pharmacologic alterations in the early development of cardiac function, ultrasound imaging of the live fetus has become an important tool for early recognition of abnormalities and longitudinal follow-up. Noninvasive ultrasound imaging is an ideal method for detecting and studying congenital malformations and the impact on cardiac function prior to death 4. It allows early recognition of abnormalities in the living fetus and the progression of disease can be followed in utero with longitudinal studies 5,6. Until recently, imaging of fetal mouse hearts frequently involved invasive methods. The fetus had to be sacrificed to perform magnetic resonance microscopy and electron microscopy or surgically delivered for transillumination microscopy. An application of high-frequency probes with conventional 2-D and pulsed-wave Doppler imaging has been shown to provide measurements of cardiac contraction and heart rates during embryonic development with databases of normal developmental changes now available 6-10. M-mode imaging further provides important functional data, although, the proper imaging planes are often difficult to obtain. High-frequency ultrasound imaging of the fetus has improved 2-D resolution and can provide excellent information on the early development of cardiac structures 11.

Fetal and perinatal death is a common feature when studying genetic alterations affecting cardiac development. High-frequency ultrasound imaging has improved 2-D resolution and can provide excellent information on early cardiac development and is an ideal method to detect the impact on cardiac structure and function prior to death.

Murine ecocardiografia fetale

Transgenic mice displaying abnormalities in  cardiac development and function represent a powerful tool for the understanding  the molecular mechanisms ...

Detection of Lung Tumor Progression in Mice by Ultrasound Imaging

With ~1.6 million victims per year, lung cancer contributes tremendously to the worldwide burden of cancer. Lung cancer is partly driven by genetic alterations in oncogenes such as the KRAS oncogene, which constitutes ~25% of lung cancer cases. The difficulty in therapeutically targeting KRAS-driven lung cancer partly stems from having poor models that can mimic the progression of the disease in the lab. We describe a method that permits the relative quantification of primary KRAS lung tumors in a Cre-inducible LSL-KRAS G12D mouse model via ultrasound imaging. This method relies on brightness (B)-mode acquisition of the lung parenchyma. Tumors that are initially formed in this model are visualized as B-lines and can be quantified by counting the number of B-lines present in the acquired images. These would represent the relative tumor number formed on the surface of the mouse lung. As the formed tumors develop with time, they are perceived as deep clefts within the lung parenchyma. Since the circumference of the formed tumor is well-defined, calculating the relative tumor volume is achieved by measuring the length and width of the tumor and applying them in the formula used for tumor caliper measurements. Ultrasound imaging is a non-invasive, fast and user-friendly technique that is often used for tumor quantifications in mice. Although artifacts may appear when obtaining ultrasound images, it has been shown that this imaging technique is more advantageous for tumor quantifications in mice compared to other imaging techniques such as computed tomography (CT) imaging and bioluminescence imaging (BLI). Researchers can investigate novel therapeutic targets using this technique by comparing lung tumor initiation and progression between different groups of mice.

This protocol describes the steps taken to induce KRAS lung tumors in mice as well as the quantification of formed tumors by ultrasound imaging. Small tumors are visualized in early timepoints as B-lines. At later timepoints, relative tumor volume measurements are achieved by the measurement tool in the ultrasound software.

Rilevamento della progressione del tumore del polmone nei topi mediante Ultrasound Imaging

With ~1.6 million victims per year, lung cancer contributes tremendously to the worldwide burden of cancer. Lung cancer is partly driven by genetic ...

Ricerca sul cancro

Non-Invasive Ultrasound Assessment of Endometrial Cancer Progression in Pax8-Directed Deletion of the Tumor Suppressors Arid1a and Pten in Mice

Uterine cancers can be studied in mice due to the ease of handling and genetic manipulation in these models. However, these studies are often limited to assessing pathology post-mortem in animals euthanized at multiple time points in different cohorts, which increases the number of mice needed for a study. Imaging mice in longitudinal studies can track the progression of disease in individual animals, reducing the number of mice needed. Advances in ultrasound technology have allowed for the detection of micrometer-level changes in tissues. Ultrasound has been used to study follicle maturation in ovaries and xenograft growth but has not been applied to morphological changes in the mouse uterus. This protocol examines the juxtaposition of pathology with in vivo imaging comparisons in an induced endometrial cancer mouse model. The features observed by ultrasound were consistent with the degree of change seen by gross pathology and histology. Ultrasound was found to be highly predictive of the observed pathology, supporting the incorporation of ultrasonography into longitudinal studies of uterine diseases such as cancer in mice.

Non-Invasive Ultrasound Assessment of Endometrial Cancer Progression in Pax8-Directed Deletion of the Tumor Suppressors Arid1a and Pten in Mice

This protocol describes a method for monitoring the progression of morphological changes over time in the uterus in an inducible mouse model of endometrial cancer using ultrasound imaging with correlation to gross and histological changes.

Valutazione ecografica non invasiva della progressione del cancro dell'endometrio nella delezione diretta da Pax8 dei soppressori tumorali Arid1a e Pten nei topi

Uterine cancers can be studied in mice due to the ease of handling and genetic manipulation in these models. However, these studies are often limited to ...

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.

Murine ecocardiografia di Sinistra Atrium, Aorta, e l&#39;arteria polmonare

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.

Misurazione ecocardiografica dei parametri diastolici ventricolari a destra nel mouse

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

Ecografia nella valutazione della disfunzione del diaframma e nel monitoraggio del trattamento

Measuring Diaphragm Thickness and Function Using Point-of-Care Ultrasound

The diaphragm is the main component of the respiratory muscle pump. Diaphragm dysfunction can cause dyspnea and exercise intolerance, and predisposes affected individuals to respiratory failure. In mechanically ventilated patients, the diaphragm is susceptible to atrophy and dysfunction through disuse and other mechanisms. This contributes to failure to wean and poor long-term clinical outcomes. Point-of-care ultrasound provides a valid and reproducible method for evaluating diaphragm thickness and contractile activity (thickening fraction during inspiration) that can be readily employed by clinicians and researchers alike. This article presents best practices for measuring diaphragm thickness and quantifying diaphragm thickening during tidal breathing or maximal inspiration. Once mastered, this technique can be used to diagnose and prognosticate diaphragm dysfunction, and guide and monitor response to treatment over time in both healthy individuals and acute or chronically ill patients.

Autore in primo piano: Svelare l'impatto della ventilazione meccanica sulla funzione del diaframma e sugli esiti dei pazienti 

Diaphragm thickness and function can be assessed in healthy individuals and critically ill patients using point-of-care ultrasound. This technique offers an accurate, reproducible, feasible, and well-tolerated method for evaluating diaphragm structure and function.

Misurazione dello spessore e della funzione del diaframma mediante ecografia point-of-care

The diaphragm is the main component of the respiratory muscle pump. Diaphragm dysfunction can cause dyspnea and exercise intolerance, and predisposes ...

Diagnostic Ultrasound Imaging di funzione mouse Diaframma

Riepilogo

Esplora Altri Video

Capitoli in questo video

L'ecocardiografia transtoracica in topi

L'ecocardiografia murini e imaging a ultrasuoni

Murine ecocardiografia fetale

Rilevamento della progressione del tumore del polmone nei topi mediante Ultrasound Imaging

Valutazione ecografica non invasiva della progressione del cancro dell'endometrio nella delezione diretta da Pax8 dei soppressori tumorali Arid1a e Pten nei topi

Murine ecocardiografia di Sinistra Atrium, Aorta, e l'arteria polmonare

Misurazione ecocardiografica dei parametri diastolici ventricolari a destra nel mouse

Misurazione dello spessore e della funzione del diaframma mediante ecografia point-of-care

Misurazione dello spessore e della funzione del diaframma mediante ecografia point-of-care

Misurazione dello spessore e della funzione del diaframma mediante ecografia point-of-care