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Tutte le procedure sugli animali sono state eseguite con l&#39;approvazione della Cura e uso Comitato Animal Istituzionale dell&#39;Università di Cincinnati e in conformità con la guida per la cura e l&#39;uso di animali da laboratorio dal National Institutes of Health (NIH Publication No. 85-23, Revised 1996). 1. Preparare il mouse <ol><li> Euthanize esponendo l&#39;animale ad una dose di anestetico sovraterapeutiche, isoflurano inalato a effetto. Verificare l&#39;eutanasia primaria via pizzico punta come uno stimolo nocivo. Metodo secondario di eutanasia, il taglio del diaframma, si verificherà in un passaggio imminente. I metodi alternativi possono essere usati eutanasia concesso si ottiene. </li><li> Sanitizie la superficie esterna del mouse spruzzando il pelo lungo la regione addominale, dove verranno effettuati i tagli iniziali, con il 70% di etanolo al punto di umidità in modo che il pelo è bagnato con etanolo ed eventuali peli residui / secco non immettere la regione. </li><li> Disporre il mouse su una surbordo chirurgico in posizione supina, con appendici superiori e inferiori esteso verso l&#39;esterno. </li><li> Appendici sicuro alla scheda con nastro chirurgico. </li></ol> 2. Isolamento del Cuore e Aorta <ol><li> Dopo un adeguato posizionamento del mouse, utilizzare pinze per localizzare e isolare pelle addominale appena inferiore al processo xifoideo dello sterno. </li><li> Creare tensione sollevando la pelle verso l&#39;alto e usare le forbici per rimuovere la pelle superficiale, esponendo la parte superiore del peritoneo e porzione inferiore della cavità toracica. </li><li> Inserire la cavità peritoneale sollevando il processo xifoideo e facendo incisioni laterali appena inferiori al processo lungo i margini sottocostali. </li><li> Sezionare up nella cavità toracica, passando attraverso il diaframma, assicurandosi di non lacerare il cuore o qualsiasi vasi sanguigni principali. </li><li> Estendere le incisioni laterali in passo 2.3 cranialmente per rimuovere la porzione anteriore della gabbia toracica. Senecessario, liberare il cuore dalla parete toracica anteriore utilizzando smussa. </li><li> Pulire la cavità toracica di sangue estraneo e fluido, utilizzando garza sterile per assorbire il materiale. Una volta che la zona è più visibile, rimuovere i polmoni per esporre meglio cuore e dell&#39;aorta. </li></ol> 3. perfusione di Cuore e Aorta NOTA: Per ottenere il sangue tramite puntura cardiaca, farlo appena prima di questo passaggio. <ol><li> Riempire una siringa da 10 cc con 10 ml di ghiaccio sterile 1x freddo tampone fosfato (PBS), e inserire un ago calibro 25 alla siringa. </li><li> Inserire delicatamente l&#39;ago nel ventricolo sinistro del cuore. </li><li> Tagliare l&#39;atrio destro per alleviare l&#39;accumulo di pressione da perfusione. Profumato il contenuto della siringa nel mouse sopra 2 - 3 min. </li><li> Utilizzare garza sterile all&#39;apertura nell&#39;atrio destro di assorbire liquido di perfusione. </li></ol> 4. L&#39;isolamento e l&#39;escissione del Aorta <ol><li> Al termine del perfusion, utilizzare garza sterile per assorbire eventuali residui di liquido opacità il campo di vista nella cavità toracica. </li><li> Esporre contenuto gastrointestinale tagliando caudalmente attraverso la parete addominale, estendendo l&#39;incisione alla zona sovrapubica. Estendere l&#39;incisione più verso gli arti inferiori bilateralmente per creare lembi di pelle che possono essere risolti o escisse. </li><li> Rimuovere i lobi del fegato, pancreas, stomaco, milza e intestino per visualizzare meglio l&#39;aorta. Fare attenzione quando sezionare vicino alla regione perirenale, l&#39;aorta è superficiale ai rami delle arterie renali. NOTA: Eseguire questo passaggio in una precisa e immatricolarsi modo il tratto gastrointestinale è ricco di batteri che aumenta la propensione per la contaminazione. </li><li> Risciacquare la zona con PBS 1x e rimuovere tutto il fluido per assorbimento con una garza sterile. </li><li> Utilizzando microscissors sterili e microforceps, separare l&#39;aorta dalla dorsale spina dorsale e l&#39;esofago ventrale.E &#39;meglio usare scollamento e utilizzando un microscopio dissezione per questo. NOTA: In isolare l&#39;aorta, avviare caudalmente alla biforcazione iliaca e spostare cranialmente pulizia e separazione l&#39;aorta dalla superficie dorsale della cavità o iniziare craniale le carotidi e le arterie brachiocefalici dissezione caudale. Entrambi i metodi è appropriato ed è fino al comfort ricercatori. </li><li> Rimuovere il tessuto adiposo perivascolare utilizzando sottili microscissors assicurandosi di non rimuovere una porzione della parete aortica. Questo è essenziale per ridurre al minimo la possibilità di contaminazione dei fibroblasti quando la coltura di cellule muscolari lisce aortiche. </li></ol>

<ol>
	<li>Go, A. 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=Heart+disease+and+stroke+statistics--2014+update:+a+report+from+the+American+Heart+Association.">Heart disease and stroke statistics--2014 update: a report from the American Heart Association.</a> Circulation. 129 (3), 28-292 (2014).</li><li>Ross, R., Harker, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperlipidemia+and+atherosclerosis.">Hyperlipidemia and atherosclerosis.</a> Science. 193 (4258), 1094-1100 (1976).</li><li>Clouse, W. D., 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=Acute+aortic+dissection:+population-based+incidence+compared+with+degenerative+aortic+aneurysm+rupture.">Acute aortic dissection: population-based incidence compared with degenerative aortic aneurysm rupture.</a> Mayo Clin Proc. 79 (2), 176-180 (2004).</li><li>Daugherty, A., Cassis, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+abdominal+aortic+aneurysms.">Mouse models of abdominal aortic aneurysms.</a> Arterioscler Thromb Vasc Biol. 24 (3), 429-434 (2004).</li><li>Pleumeekers, H. J., 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=Aneurysms+of+the+abdominal+aorta+in+older+adults.+The+Rotterdam+Study.">Aneurysms of the abdominal aorta in older adults. The Rotterdam Study.</a> Am J Epidemiol. 142 (12), 1291-1299 (1995).</li><li>Brophy, C. M., Reilly, J. M., Smit, G. J., Tilson, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+inflammation+in+nonspecific+abdominal+aortic+aneurysm+disease.">The role of inflammation in nonspecific abdominal aortic aneurysm disease.</a> Ann Vasc Surg. 5 (3), 229-233 (1991).</li><li>Ray, J. L., Leach, R., Herbert, J. M., Benson, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+vascular+smooth+muscle+cells+from+a+single+murine+aorta.">Isolation of vascular smooth muscle cells from a single murine aorta.</a> Methods Cell Sci. 23 (4), 185-188 (2001).</li><li>Weiss, R. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Imaging+the+murine+cardiovascular+system+with+magnetic+resonance.">Imaging the murine cardiovascular system with magnetic resonance.</a> Circ Res. 88 (6), 550-551 (2001).</li><li>Graham, E. L., Balla, C., Franchino, H., Melman, Y., del Monte, F., Das, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+culture,+and+functional+characterization+of+adult+mouse+cardiomyoctyes.">Isolation culture, and functional characterization of adult mouse cardiomyoctyes.</a> J Vis Exp. (79), 50289 (2013).</li><li>Wassef, M., Upchurch, G. R., Kuivaniemi, H., Thompson, R. W., Tilson, M. D. 3. r. d. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Challenges+and+opportunities+in+abdominal+aortic+aneurysm+research.">Challenges and opportunities in abdominal aortic aneurysm research.</a> J Vasc Surg. 45 (1), 192-198 (2007).</li></ol>

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Aortic disease can lead to significant systemic pathology and possibly death in severe cases. Treatment options are currently limited therefore continued research in the field is necessary10. Murine models are ideal to further study disease progression and potentially therapeutic options4. Therefore, the successful completion of the method described in this manuscript will provide investigators with the tools to measure disease progression and drug efficacy. 
This techniq...

L&#39;aorta gioca un ruolo centrale in funzione cardiovascolare fornendo il corpo con sangue ossigenato e simili ad altri parti del corpo, è suscettibile di malattia. Malattie dell&#39;aorta comuni includono l&#39;aterosclerosi e aneurisma, che sono il risultato di fattori genetici e ambientali, tra cui la dieta, il fumo e la sedentarietà 1. L&#39;aterosclerosi è la deposizione di un calcio- e la placca base lipidica alla parete aortica tipicamente riscontrati in pazienti con iperlipidemia cronica 2. Gli aneurismi sono caratterizzati dalla degradazione dei componenti strutturali della aorta e assottigliamento della parete del vaso, seguite da un maggiore diametro che potrebbe infine portare alla rottura 3. Modelli animali sono strumenti importanti utilizzati per studiare il meccanismo della malattia e l&#39;efficacia di potenziali trattamenti. Modelli animali comuni utilizzati nella ricerca cardiovascolare, in particolare di ricerca sui disturbi vascolari e metabolicheincludere i topi geneticamente modificati per alterare i livelli di lipidi, come knockout gene apolipoproteina E e lipoproteine ​​a bassa densità gene del recettore knockout mice 4. La maggior parte dei metodi per valutare meccanismi della malattia e l&#39;efficacia della terapia includerebbe l&#39;isolamento e l&#39;escissione dell&#39;aorta. Una volta che l&#39;aorta è localizzata e isolata, analisi morfometrica può essere determinata, come la presenza di un aneurisma, tipicamente definita come un aumento superiore al 50% del diametro 5. Dopo necessarie analisi in situ è completa, l&#39;aorta può essere asportato per ulteriori analisi. Un aorta asportato può essere congelato lampeggerà condurre studi molecolari come proteine ​​e / o saggi di espressione genica o fisso utilizzando 4% paraformaldeide e successivamente incorporato, sezionati e colorati per l&#39;analisi istologica. L&#39;analisi istologica dell&#39;aorta può dimostrare le caratteristiche comuni della malattia aortica come il degrado strutturale, la formazione di placca, e l&#39;infiltrazione dei leucociti &lt;sup&gt; 6,7. Inoltre, l&#39;aorta asportato può essere utilizzata per isolare linee cellulari primarie comprese cellule muscolari lisce endoteliali e che possono successivamente essere utilizzati per una varietà di studi in vitro 7. Attualmente, non ci sono altri metodi per fornire questa caratterizzazione approfondita della patologia aortica nonché fornire ricercatori con strumenti che studiare ulteriormente malattie cardiovascolari. Modalità di imaging come la risonanza magnetica, tomografia computerizzata e l&#39;ecografia sono i metodi più vicini per valutare morfologicamente l&#39;aorta, ma questo è difficile nei piccoli animali e ottenere un dispositivo con tecnologia adeguata è costosa 8. Linee cellulari immortali possono essere acquistati per studiare potenziali meccanismi della malattia e dell&#39;efficacia delle terapie, ma la natura artificiale di queste cellule limitano a studiare gli effetti sul ciclo di vita delle cellule e l&#39;apoptosi 9. L&#39;obiettivo generale diquesto manoscritto è quello di dimostrare l&#39;isolamento sterile e escissione dell&#39;aorta murine nella ricerca delle malattie cardiovascolari.

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isolation and excision of murine aorta; a versatile technique in the study of cardiovascular disease

Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated blood is the aorta. The aorta may become affected in diseases such as atherosclerosis and aneurysm. Researchers investigating these diseases would benefit from direct observation of the aorta to characterize disease progression as well as to evaluate efficacy of potential therapeutics. The goal of this protocol is to describe proper isolation and excision of the aorta to aid investigators researching cardiovascular disease. Isolation and excision of the aorta allows investigators to look at gross morphometric changes as wells as allowing them to preserve and stain the tissue to look at histologic changes if desired. The aorta may be used for molecular studies to evaluate protein and gene expression to discover targets of interest and mechanisms of action. This technique is superior to imaging modalities as they have inherent limitations in technology and cost. Additionally, primary isolated cells from a freshly isolated and excised aorta can allowing researchers to perform further in situ and in vitro assays. The isolation and excision of the aorta has the limitation of having to sacrifice the animal however, in this case the benefits outweigh the harm as it is the most versatile technique in the study of aortic disease.

Al termine della procedura, ci sarà un aorta intatta proveniente dal cuore, scendendo nella cavità toracica e addominale, con le arterie renali ancora attaccate (Figura 1A). Da qui, l&#39;aorta può essere ripreso in situ di quantificare cambiamenti morfometrici che sono diagnostica nello studio degli aneurismi dell&#39;aorta addominale (Figura 1B e 1C). Successivamente, l&#39;aorta può essere rimosso, fisso e tinto a guardare i cambiamenti istologici. Una m...

Watch this Scientific Journal Video about Isolation and Excision of Murine Aorta; A Versatile Technique in the Study of Cardiovascular Disease at JoVE.com

Isolation and Excision of Murine Aorta; A Versatile Technique in the Study of Cardiovascular Disease

Pathology of the aorta can lead to severe morbidity and mortality, therefore research of disease progression and potential therapies is warranted. Here, we present a protocol to isolate and excise the murine aorta to aid researchers in their investigation of cardiovascular disease.

Isolamento e Asportazione di Murine Aorta; Una tecnica versatile nello studio delle malattie cardiovascolari

Cardiovascular disease is a broad term describing disease of the heart and/or blood vessels. The main blood vessel supplying the body with oxygenated ...

isolation-excision-murine-aorta-versatile-technique-study

Research

JoVE Journal

Medicine

32.0K Views.  University of Cincinnati College of Medicine. Pathology of the aorta can lead to severe morbidity and mortality, therefore research of disease progression and potential therapies is warranted. Here, we present a protocol to isolate and excise the murine aorta to aid researchers in their investigation of cardiovascular disease. 

Video: Isolation and Excision of Murine Aorta; A Versatile Technique in the Study of Cardiovascular Disease

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

Un modello murino di impianto di stent a livello dell&#39;arteria carotidea per lo Studio della restenosi

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Ricerca

Medicina

Fundus Photography as a Convenient Tool to Study Microvascular Responses to Cardiovascular Disease Risk Factors in Epidemiological Studies

The microcirculation consists of blood vessels with diameters less than 150 &#181;m. It makes up a large part of the circulatory system and plays an important role in maintaining cardiovascular health. The retina is a tissue that lines the interior of the eye and it is the only tissue that allows for a non-invasive analysis of the microvasculature. Nowadays, high-quality fundus images can be acquired using digital cameras. Retinal images can be collected in 5 min or less, even without dilatation of the pupils. This unobtrusive and fast procedure for visualizing the microcirculation is attractive to apply in epidemiological studies and to monitor cardiovascular health from early age up to old age.
Systemic diseases that affect the circulation can result in progressive morphological changes in the retinal vasculature. For example, changes in the vessel calibers of retinal arteries and veins have been associated with hypertension, atherosclerosis, and increased risk of stroke and myocardial infarction. The vessel widths are derived using image analysis software and the width of the six largest arteries and veins are summarized in the Central Retinal Arteriolar Equivalent (CRAE) and the Central Retinal Venular Equivalent (CRVE). The latter features have been shown useful to study the impact of modifiable lifestyle and environmental cardiovascular disease risk factors.
The procedures to acquire fundus images and the analysis steps to obtain CRAE and CRVE are described. Coefficients of variation of repeated measures of CRAE and CRVE are less than 2% and within-rater reliability is very high. Using a panel study, the rapid response of the retinal vessel calibers to short-term changes in particulate air pollution, a known risk factor for cardiovascular mortality and morbidity, is reported. In conclusion, retinal imaging is proposed as a convenient and instrumental tool for epidemiological studies to study microvascular responses to cardiovascular disease risk factors.

Retinal image analysis is an unobtrusive procedure for visualizing the microcirculation. The impact of cardiovascular disease risk factors can result in changes of retinal vessel calibers. The procedures to acquire fundus images and the steps for calculating the vessel calibers are described.

Fundus Fotografia come un comodo strumento per studiare le risposte microvascolari a malattie cardiovascolari Fattori di rischio in studi epidemiologici

The microcirculation consists of blood vessels with diameters less than 150 &#181;m. It makes up a large part of the circulatory system and plays an ...

Localization, Identification, and Excision of Murine Adipose Depots

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 reached a staggering sum of $190.2 billion in direct medical costs alone. Obesity is a major risk factor for a wide host of diseases. Historically, little was known regarding adipose and its major and essential functions in the body. Brown and white adipose are the two main types of adipose but current literature has identified a new type of fat called brite or beige adipose. Research has shown that adipose depots have specific metabolic profiles and certain depots allow for a propensity for obesity and other related disorders. The goal of this protocol is to provide researchers the capacity to identify and excise adipose depots that will allow for the analysis of different factorial effects on adipose; as well as the beneficial or detrimental role adipose plays in disease and overall health. Isolation and excision of adipose depots allows investigators to look at gross morphological changes as well as histological changes. The adipose isolated can also be used for molecular studies to evaluate transcriptional and translational change or for in vitro experimentation to discover targets of interest and mechanisms of action. This technique is superior to other published techniques due to the design allowing for isolation of multiple depots with simplicity and minimal contamination.

Due to the drastic and negative connection between obesity and other comorbidities, research on the role adipose plays in disease and overall health is warranted. We present a protocol for the isolation and excision of adipose depots allowing for the study of adipose using in situ and in vitro methods.

Localizzazione, identificazione, e l&#39;escissione dei Depositi Murine adipose

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 ...

Adapting Human Videofluoroscopic Swallow Study Methods to Detect and Characterize Dysphagia in Murine Disease Models

This study adapted human videofluoroscopic swallowing study (VFSS) methods for use with murine disease models for the purpose of facilitating translational dysphagia research. Successful outcomes are dependent upon three critical components: test chambers that permit self-feeding while standing unrestrained in a confined space, recipes that mask the aversive taste/odor of commercially-available oral contrast agents, and a step-by-step test protocol that permits quantification of swallow physiology. Elimination of one or more of these components will have a detrimental impact on the study results. Moreover, the energy level capability of the fluoroscopy system will determine which swallow parameters can be investigated. Most research centers have high energy fluoroscopes designed for use with people and larger animals, which results in exceptionally poor image quality when testing mice and other small rodents. Despite this limitation, we have identified seven VFSS parameters that are consistently quantifiable in mice when using a high energy fluoroscope in combination with the new murine VFSS protocol. We recently obtained a low energy fluoroscopy system with exceptionally high imaging resolution and magnification capabilities that was designed for use with mice and other small rodents. Preliminary work using this new system, in combination with the new murine VFSS protocol, has identified 13 swallow parameters that are consistently quantifiable in mice, which is nearly double the number obtained using conventional (i.e., high energy) fluoroscopes. Identification of additional swallow parameters is expected as we optimize the capabilities of this new system. Results thus far demonstrate the utility of using a low energy fluoroscopy system to detect and quantify subtle changes in swallow physiology that may otherwise be overlooked when using high energy fluoroscopes to investigate murine disease models.

This study successfully adapted human videofluoroscopic swallowing study (VFSS) methods for use with murine disease models for the purpose of facilitating translational dysphagia research.

Adattamento umani videofluoroscopia Rondine metodi di studio per individuare e caratterizzare disfagia in modelli di malattia Murine

This study adapted human videofluoroscopic swallowing study (VFSS) methods for use with murine disease models for the purpose of facilitating ...

A Versatile Murine Model of Subcortical White Matter Stroke for the Study of Axonal Degeneration and White Matter Neurobiology

Stroke affecting white matter accounts for up to 25% of clinical stroke presentations, occurs silently at rates that may be 5-10 fold greater, and contributes significantly to the development of vascular dementia. Few models of focal white matter stroke exist and this lack of appropriate models has hampered understanding of the neurobiologic mechanisms involved in injury response and repair after this type of stroke. The main limitation of other subcortical stroke models is that they do not focally restrict the infarct to the white matter or have primarily been validated in non-murine species. This limits the ability to apply the wide variety of murine research tools to study the neurobiology of white matter stroke. Here we present a methodology for the reliable production of a focal stroke in murine white matter using a local injection of an irreversible eNOS inhibitor. We also present several variations on the general protocol including two unique stereotactic variations, retrograde neuronal tracing, as well as fresh tissue labeling and dissection that greatly expand the potential applications of this technique. These variations allow for multiple approaches to analyze the neurobiologic effects of this common and understudied form of stroke.

Here we present methodology for the production of a focal stroke in murine white matter by local injection of an irreversible endothelial nitric oxide synthase (eNOS) inhibitor (L-Nio). Presented are two stereotactic variations, retrograde neuronal tracing, and fresh tissue labeling and dissection that expand the potential applications of this technique.

A Corsa Versatile Modello murino di Subcortical bianco Materia per lo Studio della assonale degenerazione e bianco Materia Neurobiologia

Stroke affecting white matter accounts for up to 25% of clinical stroke presentations, occurs silently at rates that may be 5-10 fold greater, and ...

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 ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

Basato su palloncino ferita per indurre iperplasia Myointimal nell'Aorta addominale di Mouse

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...

Ultrasound Imaging of the Thoracic and Abdominal Aorta in Mice to Determine Aneurysm Dimensions

Contemporary high-resolution ultrasound instruments have sufficient resolution to facilitate the measurement of mouse aortas. These instruments have been widely used to measure aortic dimensions in mouse models of aortic aneurysms. Aortic aneurysms are defined as permanent dilations of the aorta, which occur most frequently in the ascending and abdominal regions. Sequential measurements of aortic dimensions by ultrasound are the principal approach for assessing the development and progression of aortic aneurysms in vivo. Although many reported studies used ultrasound imaging to measure aortic diameters as a primary endpoint, there are confounding factors, such as probe position and cardiac cycle, that may impact the accuracy of data acquisition, analysis, and interpretation. The purpose of this protocol is to provide a practical guide on the use of ultrasound to measure the aortic diameter in a reliable and reproducible manner. This protocol introduces the preparation of mice and instruments, the acquisition of appropriate ultrasound images, and data analysis.

Ultrasound imaging has become a common modality to determine the luminal dimensions of thoracic and abdominal aortic aneurysms in mice. This protocol describes the procedure to acquire reliable and reproducible two-dimensional ultrasound images of the ascending and abdominal aorta in mice.

Formazione immagine di ultrasuono dell'Aorta toracica ed addominale in topi per determinare le dimensioni di Aneurysm

Contemporary high-resolution ultrasound instruments have sufficient resolution to facilitate the measurement of mouse aortas. These instruments have been ...

A Cryoinjury Model to Study Myocardial Infarction in the Mouse

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we demonstrate a murine cryoinjury infarct model that generates precise infarct sizes with high reproducibility and replicability. In brief, after intubation and sternotomy of the animal, the heart is lifted from the thorax. The probe of a handheld liquid nitrogen delivery system is applied onto the myocardial wall to induce cryoinjury. Impaired ventricular function and electrical conduction can be monitored with echocardiography or optical mapping. Transmural myocardial remodeling of the infarcted area is characterized by collagen deposition and loss of cardiomyocytes. Compared to other models (e.g., LAD-ligation), this model utilizes a handheld liquid nitrogen delivery system to generate more uniform infarct sizes.

This article demonstrates a model to study cardiac remodeling after myocardial cryoinjury in mice.

Un modello di criolesionismo per studiare l'infarto miocardico nel mouse

The use of animal models is essential for developing new therapeutic strategies for acute coronary syndrome and its complications. In this article, we ...

Intramyocardial Transplantation of MSC-Loading Injectable Hydrogels after Myocardial Infarction in a Murine Model

One of the major issues facing current cardiac stem cell therapies for preventing postinfarct heart failure is the low retention and survival rates of transplanted cells within the injured myocardium, limiting their therapeutic efficacy. Recently, the use of scaffolding biomaterials has gained attention for improving and maximizing stem cell therapy. The objective of this protocol is to introduce a simple and straightforward technique to transplant bone marrow-derived mesenchymal stem cells (MSCs) using injectable hydroxyphenyl propionic acid (GH) hydrogels; the hydrogels are favorable as a cell delivery platform for cardiac tissue engineering applications due to their ability to be cross-linked in situ and high biocompatibility. We present a simple method to fabricate MSC-loading GH hydrogels (MSC/hydrogels) and evaluate their survival and proliferation in three-dimensional (3D) in vitro culture. In addition, we demonstrate a technique for intramyocardial transplantation of MSC/hydrogels in mice, describing a surgical procedure to induce myocardial infarction (MI) via left anterior descending (LAD) coronary artery ligation and subsequent MSC/hydrogels transplantation.

Stem cell-based therapy has emerged as an efficient strategy to repair injured cardiac tissues after myocardial infarction. We provide an optimal in vivo application for stem cell transplantation using gelatin hydrogels that are able to be enzymatically cross-linked.

Trapianto intramiocardico di idrogel iniettabili a carico MSC dopo infarto del miocardio in un modello murino

One of the major issues facing current cardiac stem cell therapies for preventing postinfarct heart failure is the low retention and survival rates of ...