Los autores no tienen reconocimientos.

Todos los animales procedimientos fueron realizados con la aprobación del Comité de Cuidado y Uso de Animales Institucional de la Universidad de Cincinnati y de conformidad con la Guía para el Cuidado y Uso de Animales de Laboratorio de los Institutos Nacionales de Salud (NIH Publication No. 85-23, Revisado 1996). 1. Preparación del ratón <ol><li> Practicar la eutanasia mediante la exposición del animal a una dosis de anestésico supraterapéutica, isoflurano inhalado al efecto. Verifique la eutanasia primario a través pizca de pie, como un estímulo nocivo. Método secundario de la eutanasia, de corte diafragma, tendrá lugar en una próxima etapa. Los métodos alternativos pueden ser utilizados se obtiene la eutanasia sentado. </li><li> Sanitizie la superficie externa del ratón por pulverización de la piel a lo largo de la región abdominal, donde se harán los cortes iniciales, con etanol al 70% hasta el punto de humedad por lo que la piel está húmeda con etanol y los pelos sueltos / seco no entran la región. </li><li> Coloque el ratón sobre un surtablero de lógica en la posición supina con apéndices superiores e inferiores se extendió hacia el exterior. </li><li> Apéndices seguros a la junta con cinta quirúrgica. </li></ol> 2. Aislamiento del corazón y la aorta <ol><li> Después de la colocación apropiada del ratón, el uso de fórceps para localizar y aislar la piel abdominal justo por debajo de la apófisis xifoides del esternón. </li><li> Crear tensión levantando la piel hacia arriba y utilizar las tijeras para quitar la piel superficial, dejando al descubierto la parte superior del peritoneo y la parte inferior de la cavidad torácica. </li><li> Introduzca la cavidad peritoneal levantando el apéndice xifoides y la realización de incisiones laterales apenas inferiores al proceso a lo largo de los márgenes subcostal. </li><li> Diseccionar hasta la cavidad torácica, pasando a través del diafragma, asegurándose de no lacerar el corazón o cualquier principales vasos sanguíneos. </li><li> Extender las incisiones laterales hechas en el paso 2.3 cranealmente para eliminar la porción anterior de la caja torácica. Sinecesario, liberar el corazón de la pared anterior del tórax mediante disección roma. </li><li> Limpie la cavidad torácica de la sangre ajena y líquido utilizando una gasa estéril para absorber el material. Una vez que el área es más visible, retire los pulmones para exponer mejor el corazón y la aorta. </li></ol> 3. La perfusión de corazón y la aorta NOTA: Para obtener la sangre mediante punción cardíaca, hacerlo justo antes de este paso. <ol><li> Llenar una jeringa de 10 cc con 10 ml de hielo frío estéril 1x solución de tampón fosfato (PBS), y adjuntar una aguja de calibre 25 a la jeringa. </li><li> Suavemente inserte la aguja en el ventrículo izquierdo del corazón. </li><li> Cortar la aurícula derecha para aliviar la acumulación de la presión de perfusión. Perfundir el contenido de la jeringa en el ratón más de 2 - 3 minutos. </li><li> Utilice una gasa estéril en la abertura en la aurícula derecha para absorber líquido de perfusión. </li></ol> 4. El aislamiento y la escisión de la aorta <ol><li> Al término de PErfusion, utilizar una gasa estéril para absorber cualquier líquido restante enturbiamiento del campo de visión dentro de la cavidad torácica. </li><li> Exponer los contenidos gastrointestinales cortando caudalmente a través de la pared abdominal, la incisión se extiende a la zona suprapúbica. Extienda la incisión más hacia los miembros inferiores bilateralmente para crear colgajos de piel que pueden ser puestas abajo o extirpados. </li><li> Retire los lóbulos del hígado, el páncreas, el estómago, el bazo y los intestinos para visualizar mejor la aorta. Tenga cuidado cuando la disección de cerca de la región perirrenal, como la aorta es superficial en las ramas de las arterias renales. NOTA: Realice este paso en un preciso y matricularse manera como el tracto gastrointestinal es rico en bacterias que aumenta la propensión a la contaminación. </li><li> Enjuague el área con 1x PBS y retirar todo el fluido por absorción con una gasa estéril. </li><li> El uso de micro tijeras estériles y microforceps, separar la aorta dorsal de la columna vertebral y el esófago ventralmente.Lo mejor es usar una disección roma y la utilización de un microscopio de disección para esto. NOTA: En el aislamiento de la aorta, iniciar caudalmente en la bifurcación ilíaca y mover cranealmente limpieza y separación de la aorta de la superficie dorsal de la cavidad o iniciar cranealmente en las carótidas y arterias braquiocefálica de disección caudalmente. Cualquier método es apropiado y es hasta la comodidad investigadores. </li><li> Retire el tejido adiposo perivascular utilizando finas micro tijeras asegurándose de no eliminar una parte de la pared aórtica. Esto es esencial para minimizar el potencial de contaminación de fibroblastos cuando el cultivo de células del músculo liso aórtico. </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>

Los autores no tienen nada que revelar.

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...

La aorta juega un papel fundamental en la función cardiovascular suministrar al cuerpo con sangre oxigenada y las porciones similares a los demás del cuerpo, es susceptible a la enfermedad. Enfermedades de la aorta comunes son la aterosclerosis y el aneurisma, que son el resultado de factores genéticos y ambientales como la dieta, el tabaquismo y el sedentarismo 1. La aterosclerosis es la deposición de calcio y una placa base de lípidos a la pared aórtica típicamente encontrado en pacientes con hiperlipidemia crónica 2. Los aneurismas se caracterizan por la degradación de los componentes estructurales de la aorta y el adelgazamiento de la pared del vaso, seguido por un aumento del diámetro lo que finalmente puede conducir a la ruptura 3. Los modelos animales son herramientas importantes utilizadas para estudiar el mecanismo de la enfermedad y la eficacia de los tratamientos potenciales. Modelos animales más comunes utilizados en la investigación cardiovascular, investigar específicamente la investigación de los trastornos vasculares y metabólicasincluir ratones modificados genéticamente para alterar los niveles de lípidos, tales como golpe de gracia gen de la apolipoproteína E y lipoproteínas de densidad baja ratones knockout del gen del receptor 4. La mayoría de los métodos para evaluar los mecanismos de la enfermedad y la eficacia de la terapia incluiría el aislamiento y la escisión de la aorta. Una vez que la aorta está localizado y aislado, el análisis morfométrico se puede determinar, como la presencia de un aneurisma, típicamente definido como un incremento mayor que 50% en el diámetro 5. Después necesario el análisis in situ es completa, la aorta se puede escindir para su posterior análisis. Una aorta extirpada puede ser congelado instantáneamente para llevar a cabo estudios moleculares, tales como proteínas y / o ensayos de expresión de genes o fijo utilizando 4% de paraformaldehído y posteriormente incrustado, se seccionaron y se tiñeron para el análisis histológico. El análisis histológico de la aorta puede mostrar las características comunes de la enfermedad aórtica, como la degradación estructural, la formación de placa, y la infiltración de leucocitos &lt;sup&gt; 6,7. Además, una aorta extirpada se puede utilizar para aislar líneas celulares primarias que incluyen células endoteliales y de músculo liso que posteriormente se pueden utilizar para una variedad de estudios in vitro 7. Actualmente, no existen otros métodos para proporcionar esta caracterización en profundidad de la enfermedad de la aorta, así como brindar a los investigadores herramientas para estudiar más a fondo la enfermedad cardiovascular. Técnicas de imagen como la resonancia magnética, tomografía computarizada y la ecografía son los métodos más cercanos para evaluar morfológicamente la aorta, sin embargo esto es difícil en pequeños animales y la obtención de un dispositivo con tecnología adecuada es caro 8. Las líneas celulares inmortalizadas se pueden comprar para investigar los mecanismos potenciales de la enfermedad y la eficacia de las terapias, sin embargo, la naturaleza artificial de estas células son limitantes en el estudio de los efectos sobre el ciclo de vida celular y la apoptosis 9. El objetivo general deeste manuscrito es demostrar el aislamiento estéril y la escisión de la aorta murino en la investigación de la enfermedad cardiovascular.

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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 finalizar el procedimiento, habrá una aorta intacta procedente del corazón, descendiendo en las cavidades torácica y abdominal con las arterias renales todavía conectados (Figura 1A). Desde aquí, la aorta se pueden obtener imágenes in situ para cuantificar los cambios morfométricos que son de diagnóstico en el estudio de los aneurismas aórticos abdominales (Figura 1B y 1C). Posteriormente, la aorta se puede quitar, fijadas y teñidas para observar l...

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.

Aislamiento y Escisión de murino Aorta; Una técnica versátil en el Estudio de la Enfermedad Cardiovascular

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 vistas.  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 modelo murino de la implantación de un stent en la arteria carótida para el Estudio de la reestenosis

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

Investigación

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.

Fotografía del fondo como una herramienta conveniente para estudiar las respuestas microvasculares a factores de riesgo cardiovascular en los estudios epidemiológicos

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.

Localización, identificación y Escisión de depósitos adiposos murino

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.

Adaptación videofluoroscópicos Swallow Métodos de Estudio Humanos para detectar y caracterizar disfagia en modelos murinos de enfermedades

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.

Un movimiento versátil modelo murino de la sustancia blanca subcortical para el Estudio de la degeneración axonal y la sustancia blanca Neurobiología

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.

Murino de Ecocardiografía del atrio izquierdo, la aorta y arteria pulmonar

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.

Lesión basada en globo para inducir la hiperplasia Myointimal de la Aorta Abdominal de ratón

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.

Imágenes por ultrasonido de la Aorta torácica y Abdominal en ratones para determinar dimensiones de aneurisma

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 modelo de criolesiones para estudiar el infarto de miocardio en el ratón

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.

Trasplante intramiocardio de hidrogeles inyectables msc-loading después de infarto de miocardio en un modelo 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 ...