We are grateful to Dr. Clowes for first developing and describing this method. We are also thankful to Dr. Tulis for his detailed protocol which has been fundamentally helpful to our previous, current and future work. This work was supported by grants R01HL097111 and R01HL123364 from the NIH to M.T., and by American Heart Association grant 14GRNT18880008 to M.T.
We would like to thank Rachel Newton for her expert technical support and for her valuable help during the filming process.

NOTA: El uso de animales para los siguientes experimentos ha sido revisado y aprobado por el Cuidado y Uso de Animales Comité Institucional (IACUC). 1. Procedimientos preoperatorios <ol style="margin-left: 40px;"><li> Esterilizar instrumentos quirúrgicos antes de su uso. <ol><li> Autoclave todos los instrumentos quirúrgicos de 24 horas o menos antes de la cirugía. Si varias cirugías se realizan en el mismo día, esterilizar los instrumentos por un esterilizador de cuentas en seco entre cirugías. </li></ol></li><li> Filter-esterilizar la solución salina antes de su uso. </li><li> Pesar la rata y calcular la dosis de los fármacos anestésicos ketamina (80 mg / kg y xilazina 7 mg / kg). </li><li> Administrar los fármacos anestésicos por vía intraperitoneal (ip). <ol><li> Verificar la adecuación de la sedación por pizca dedo del pie en 5-10 minutos. Administrar una pequeña dosis adicional de drogas (ketamina 7 mg / kg y xilazina 0,6 mg / kg) si la sedación no es completa. </li><li> Asegúrese de que la aguja es lo suficientemente profundo como para suministrar fármacos intra-peritoneally ya falta de entrega de la solución entera de drogas en la cavidad peritoneal causará sedación inadecuada. NOTA: Las úlceras y pérdida de cabello en la piel en el lugar de la inyección serán visibles varios días después de la cirugía, debido a inadvertida subcutánea (sc) de inyección de solución de fármaco. </li></ol></li><li> Inyectar 3 ml de solución salina estéril por vía subcutánea (sc). Con un hisopo de algodón estéril, aplique una pequeña cantidad de pomada oftálmica de ambos ojos para evitar que las córneas se sequen. </li><li> Preparar el equipo de calefacción. Almohadillas de calor Pre-caliente por microondas o al baño maría. </li><li> Ponga el animal supina en la plataforma quirúrgica. <ol><li> Retire el cabello en la región del cuello ventral. Aplicar el removedor de pelo con un hisopo de algodón, esperar 30 segundos y limpie completamente con una gasa. </li><li> Limpie la nuca con matorrales de povidona yodada y etanol al 70%. </li></ol></li><li> Use el equipo de protección personal, incluyendo el vestido, cubierta de pelo, máscara quirúrgica y gafas. Póngase sterile guantes quirúrgicos al final antes de tocar los instrumentos y suministros quirúrgicos estériles. </li><li> Coloque la rata con una hoja quirúrgica estéril con sólo la región del cuello expuesta. </li></ol> 2. Procedimientos Quirúrgicos <ol style="margin-left: 40px;"><li> Durante el procedimiento quirúrgico, verificar la profundidad de la sedación de los subproductos animales pizca dedo del pie en cada 15 minutos. Si animal responde a la pizca dedo del pie, añadir pequeñas dosis adicional (10% de la dosis inicial) de ketamina y xilazina. </li><li> Diseccionar la arteria carótida común izquierda (CCA) <ol><li> Utilice un bisturí para realizar una incisión longitudinal recta en el medio de cuello. La longitud aproximada de incisión de la piel es de 1.5-2 cm, con el propósito de aislamiento de una porción de 1,5-2 cm de la arteria. La longitud puede variar depende de la finalidad del estudio. </li><li> Sin rodeos diseccionar el tejido conectivo de la piel. Mantenga las puntas de las pinzas y asegúrese de no perforar la piel o los tejidos subyacentes. </li><li> Diseccionar capas musculares longitudinal along el lado izquierdo de la tráquea. </li><li> Al abrir la capa muscular, visualizar la CCA izquierda con el nervio vago estrechamente unida. Sin rodeos diseccionar al lado de la arteria carótida izquierda con extrema precaución para separar el nervio vago con el estiramiento mínimo. </li><li> Diseccionar CCA distalmente hasta la bifurcación. Diseccionar cuidadosamente la bifurcación y dos ramas - la arteria carótida interna (ACI) y la arteria carótida externa (ACE). </li><li> Mantenga la disección alrededor de la CCA, hasta que aproximadamente una parte de 1,5-2 cm de la arteria está aislado de los tejidos circundantes. </li></ol></li><li> Lesión por balón <ol><li> Permanentemente hacer una ligadura en el Tribunal de Cuentas en aproximadamente 5 mm de distancia de bifurcación. Permanentemente ligar la rama occipital de la CEPA, que está cerca de la bifurcación de la CEPA y el ICA. También ligar permanentemente otras ramas, en su caso, la localización de la bifurcación entre la ECA y ligadura - por ejemplo, la rama tiroidea superior. Sutura que utiliza para todas las ligaduras es 4-0 negroseda. Clip en el extremo proximal de la CCA y el extremo distal de la ACI. NOTA: Ahora, el flujo de sangre se ha detenido de forma permanente a través de la ligadura (en ECA) o temporalmente a través de recorte (en la CCA y el ICA). Contenido luminal en el área de bifurcación se han aislado de la circulación sistémica. </li><li> Hacer una incisión arteriotomía en CEPA por pequeña micro-tijeras. Asegúrese de que la incisión está cerca del nudo de sutura distal. La sangre limpia con solución salina y con palitos de algodón. </li><li> Inserte el catéter de balón 2F desinflado en el lumen de la CEPA. Avance el catéter de globo proximal a la luz del CCA. Mantenga avanzar el catéter proximal hasta que la punta llegue a donde el clip permanece. </li><li> Conectar el dispositivo de inflado del balón a un bloqueo luer hembra en una llave de paso de 3 vías y conectar la cerradura Luer macho de la llave de paso de 3 vías para el catéter de balón. </li><li> Lentamente inflar el globo con aproximadamente 1,5 atm de presión, con el fin de dilatar la arteria carótida a 1,5 veces el diámetro. </li&gt; <li> Tire suavemente el globo en rotación hacia atrás hasta la bifurcación. </li><li> Desinflar el globo y hacer avanzar de nuevo al extremo proximal. Infle de nuevo y repita el procedimiento tirando hacia atrás dos veces más. </li><li> Retirar el catéter de balón desde la luz de la arteria. </li></ol></li><li> La administración intraluminal de reactivos (por ejemplo, siRNA, drogas) NOTA: Aquí, el reactivo utilizado fue una solución que contiene partículas lentivirales que codifican shRNA orientación del estroma de la molécula Interacción 1 (STIM1) o no la orientación de control shRNA. STIM1 es un endoplasmático proteína transmembrana única retículo (ER) que es un 2 + sensor ER Ca controlar la activación de la membrana plasmática de Ca 2 + canales y es regulada hasta durante la desdiferenciación del músculo liso vascular en un fenotipo migratorio proliferativa 12,30-33. <ol><li> Adjunte una intravascular catéter sobre la aguja de una jeringa (24 G, 1,6 cm). Aspirar solución 30 l de la reag pruebasentos. Inserte el catéter en la misma incisión en la CEPA. </li><li> Avanzar en la punta del catéter en la CCA y atar un trozo de sutura con un solo nudo en la CEPA para fijar el catéter y cerrar la incisión temporal. </li><li> Se inyecta la solución reactivo de prueba en el lumen del CCA. Conservar la solución en la luz del vaso durante 30 minutos. Mantenga el tejido expuesto húmeda con solución salina y se cubre con una gasa húmeda. </li><li> Después de la incubación, aspirar la solución restante. Afloje que solo nudo y retirar el catéter. </li><li> Ate la ECA con un pedazo de sutura proximal a la incisión arteriotomía. Hacer el nudo lo más cerca posible a la bifurcación. </li></ol></li><li> Cierre de la herida. <ol><li> Desde la lesión con balón y la introducción del catéter puede causar una fuga o la punción del vaso sanguíneo, retire el clip de la ICA y comprobar si hay alguna fuga. Si se observa sangrado, aplique un trozo de gasa y aplicar una suave presión para detener el sangrado. </li><li> QuitarPor otro clip en CCA. </li><li> Asegúrese de que no hay signos de sangrado y retire todas las abrazaderas y otros instrumentos quirúrgicos. Corte el exceso de suturas. </li><li> Cerrar suturas de la piel de la herida utilizando (4-0 seda negro). Swab en todos los lados de la herida cerrada con el Povidon-yodo u otro antiséptico / bactericida / agente virucida. Inyectar 3 ml de solución salina estéril sc </li></ol></li></ol> 3. Procedimientos postoperatorios <ol style="margin-left: 40px;"><li> Mantenga rata en el cojín de calor después de la cirugía. Administrar una dosis de 0,05 mg / kg de buprenorfina a través de inyección intramuscular (im) a la rata. Durante el procedimiento de recuperación, mantenga los ojos y la boca de los animales húmedo. Monitorear el animal hasta que esté despierto y ambulante. </li><li> Colocar el animal en una jaula limpia sin la ropa de cama hasta que esté totalmente recuperado. Después de la recuperación, el animal será devuelto a la sala de los animales y alojado individualmente. </li></ol>

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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rat+carotid+artery+balloon+injury+model.">Rat carotid artery balloon injury model.</a> Methods In Molecular Medicine. 139, 1-30 (2007).</li><li>Zhang, W., Trebak, M., Szallasi, A., B&#237;r&#243;, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Balloon+Injury+in+Rats+as+a+Model+for+Studying+TRP+Channel+Contribution+to+Vascular+Smooth+Muscle+Remodeling.">Balloon Injury in Rats as a Model for Studying TRP Channel Contribution to Vascular Smooth Muscle Remodeling.</a> T TRP Channels in Drug DiscoveryMethods in Pharmacology and Toxicology. , 101-111 (2012).</li><li>Tulis, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histological+and+morphometric+analyses+for+rat+carotid+balloon+injury+model).">Histological and morphometric analyses for rat carotid balloon injury model).</a> Methods In Molecular Medicine. 139, 31-66 (2007).</li></ol>

The authors declare that they have no competing financial interests.

La lesión de balón carótida de rata ha sido bien descrita por Tulis en 2007 34. Se ha discutido exhaustivamente todos los detalles de este procedimiento por el Dr. Tulis. Los lectores que estén interesados ​​en realizar este procedimiento son muy recomendables para leer protocolo Tulis &#39;. Sin embargo, hay una cosa que no estamos de acuerdo con el Dr. Tulis: En vez de inflar el globo con solución salina o cualquier tipo de líquido, lo sugerimos para inflar con aire. De acuerdo con nuestra experiencia personal, inflan...

Los catéteres de balón son dispositivos médicos utilizados en el procedimiento de la angioplastia, con el fin de ampliar sitio obstruido (s) de ateroma o trombo en un vaso sanguíneo. La luz del vaso estrechado se ve obligado a abrir por el globo inflado y suministro de sangre serían restaurados de forma secuencial para aliviar los síntomas de isquemia posteriores, como la angina de pecho, infarto de miocardio y dolor en las piernas. Sin embargo, el gran éxito de la angioplastia ha disminuido debido a las complicaciones postoperatorias, como resulta de la fuerza que causa barotrauma vascular (lesión globo), a saber, remodelación de la pared del vaso y en muchos casos re-estrechamiento de la luz del vaso (restenosis) 1. Un número de modelos animales se han desarrollado imitando el procedimiento de angioplastia para ayudar a los investigadores a comprender los mecanismos subyacentes de la pared remodelado vascular relacionada con el globo de la lesión 2. Entre todas las especies de animales utilizados para el modelado, la rata es el más utilizado. Compared a los conejos, los perros y los cerdos, las ventajas de las ratas son su bajo coste, su relativa facilidad de uso y el conocimiento actual de la fisiología de la rata. Aunque los ratones tienen una ventaja añadida en una amplia gama de cepas manipuladas genéticamente, el recipiente de ratones es demasiado pequeña para insertar un catéter de globo. Durante las últimas tres décadas, las ratas experimentales han permitido a los investigadores obtener una mejor comprensión de los mecanismos moleculares y celulares que sustentan la formación de neoíntima vascular y remodelación de 3-6. Más allá de la lesión por balón, el remodelado vascular también están involucrados en la mayoría de las principales enfermedades vasculares, tales como aterosclerosis 7,8, hipertensión 9, 10 y aneurisma. Por lo tanto, el conocimiento obtenido a través del modelo de lesión de globo es en general beneficiosa para los estudios generales de la enfermedad de la pared vascular. El objetivo general del modelo de lesión globo rata es no sólo para entender mejor las enfermedades vasculares, sino también para poner a prueba la potencia de los nuevos agentes paracontrol de la enfermedad 11,12. Tratamiento farmacológico clínico actual a la reestenosis es aplicada por los stents liberadores de fármacos realizados a través de la luz del vaso justo después de la angioplastia. En modelos animales, de una manera eficiente aún más económico para nuevas pruebas agente es un método de perfusión intraluminal local bien desarrollada. Los agentes candidatos que han sido probados a través de este método incluyen fármacos de molécula pequeña 13,14, citoquinas o factores de crecimiento 15,16, agentes de genes (la manipulación de clones de ADNc, siRNA, etc.) 17-20, y nuevas formulaciones farmacéuticas 21,22. Hasta ahora, el modelo de lesión globo rata sigue siendo uno de los modelos más útiles para el estudio de enfermedades vasculares / trastornos. Es el paso fundamental del banco a la cama, por lo general como la primera etapa de movimiento de in vitro a in vivo, pero no debe ser el último. El resultado de los experimentos con ratas necesita ser deliberado y caracterizado además antes de traducción al humanouso clínico, debido a la diferencia en los lechos vasculares y anatomía de los vasos, así como las especies intrínsecas diferencias entre humano y de rata 23-26. Sin embargo, sigue siendo una herramienta esencial en la investigación médica traslacional. Si bien este tipo de investigación solía ser limitada por la falta de ratas genéticamente modificados, ya no ha sido un problema ya que los enfoques genómicos novedosos como zinc-finger nucleasas 27, 28 y Talens CRISPR-Cas 29 han hecho ratas knockout de fácil acceso.

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			<td height="21" style="height:21px;">Nair Baby Oil Hair Removal Lotion-9 oz</td>
			<td>Amazon/Walmart/CVS</td>
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			<td>Demax Medical</td>
			<td>DID30</td>
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			<td height="21" style="height:21px;">D300 3-way Stopcock</td>
			<td>B.Braun Medical Inc.</td>
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			<td height="21" style="height:21px;">Artificial Tears Ointment&#160;</td>
			<td>Rugby Laboratories, Duluth, GA</td>
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vascular balloon injury and intraluminal administration in rat carotid artery

El modelo de lesión por balón de la arteria carótida en ratas ha sido bien establecida desde hace más de dos décadas. Sigue siendo un método importante para estudiar los mecanismos moleculares y celulares implicados en la desdiferenciación del músculo liso vascular, formación de la neoíntima y la remodelación vascular. Hombre ratas Sprague-Dawley son los animales más frecuentemente empleado para este modelo. Las ratas hembra no se prefieren como hormonas femeninas son protectores contra las enfermedades vasculares y por lo tanto introducir una variación en este procedimiento. La carótida izquierda normalmente se lesiona con la carótida derecha sirve como un control negativo. Lesión de carótida izquierda es causada por el globo inflado que despoja el endotelio y distender s la pared del vaso. Después de una lesión, las posibles estrategias terapéuticas tales como el uso de compuestos farmacológicos y cualquiera de los genes o transferencia shRNA pueden ser evaluados. Normalmente para el gen o transferencia shRNA, la sección de heridos de la luz del vaso se transduce localmente durante 30 minutos con vpartículas víricas que codifican una proteína o shRNA para la entrega y la expresión en la pared del vaso lesionado. Engrosamiento neointimal que representa las células musculares lisas vasculares proliferativos generalmente picos a las 2 semanas después de la lesión. Los buques se cosechan sobre todo en este punto de tiempo para el análisis celular y molecular de las vías de señalización celular, así como de genes y la expresión de la proteína. Los buques también pueden ser cosechadas en puntos de tiempo anteriores para determinar el inicio de la expresión y / o activación de una proteína o vía específica, dependiendo de los objetivos experimentales destinados. Los buques pueden ser caracterizados y evaluados utilizando la tinción histológica, inmunohistoquímica, ensayos de proteína / ARNm, y ensayos de actividad. La arteria carótida derecha intacta del mismo animal es un control interno ideal. Cambios inducidos por la lesión en los parámetros moleculares y celulares pueden ser evaluados mediante la comparación de la arteria lesionada a la arteria control interno derecho. Del mismo modo, las modalidades terapéuticas se pueden evaluar mediante la comparación de la injuredy arteria tratada con el control herido única arteria.

Dos semanas después de la lesión, las arterias carótidas se cosechan, se seccionaron y se sometieron a análisis morfológico. Las arterias son en sección transversal y teñidas con H &amp; E (Figuras 1, 2 B, C y 3). Pared de la arteria carótida de rata contiene cuatro capas de lámina elástica, que aparecen como líneas de color rosa. El área entre la lámina más externa, la lámina elástica externa (EEL) y la lámina más interior, lámina elástica interna (IEL) es la capa de músculo liso m...

Watch this Scientific Journal Video about Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery at JoVE.com

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

Este protocolo utiliza un catéter de balón para causar una lesión intraluminal de la arteria carótida de rata y en adelante provocar hiperplasia neointimal. Este es un modelo bien establecido para el estudio de los mecanismos de remodelación vascular en respuesta a una lesión. También es ampliamente utilizado para determinar la validez de los enfoques terapéuticos potenciales.

Lesión Vascular Globo y Administración intraluminal en rata arteria carótida

The carotid artery balloon injury model in rats has been well established for over two decades. It remains an important method to study the molecular and ...

vascular-balloon-injury-intraluminal-administration-rat-carotid

Research

JoVE Journal

Medicine

16.3K vistas.  State University of New York College of Nanoscale Science and Engineering (SUNY CNSE).  Este protocolo utiliza un catéter de balón para causar una lesión intraluminal de la arteria carótida de rata y en adelante provocar hiperplasia neointimal. Este es un modelo bien establecido para el estudio de los mecanismos de remodelación vascular en respuesta a una lesión. También es ampliamente utilizado para determinar la validez de los enfoques terapéuticos potenciales. 

Video: Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery

Training a Sophisticated Microsurgical Technique: Interposition of External Jugular Vein Graft in the Common Carotid Artery in Rats

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in peripheral arterial bypass surgery as well as in arteriovenous fistulas.1-5 The experimental procedure of vein graft interposition in the common carotid artery by using the cuff-technique has been applied in several research projects to examine the aetiology of neointimal hyperplasia and therapeutic options to address it. 6-8 The cuff prevents vessel anastomotic remodeling and induces turbulence within the graft and thereby the development of neointimal hyperplasia.
Using the superior caval vein graft is an established small-animal model for venous arterialization experiment.9-11 This current protocol refers to an established jugular vein graft interposition technique first described by Zou et al., 9 as well as others.12-14 Nevertheless, these cited small animal protocols are complicated.
To simplify the procedure and to minimize the number of experimental animals needed, a detailed operation protocol by video training is presented. This video should help the novice surgeon to learn both the cuff-technique and the vein graft interposition. Hereby, the right external jugular vein was grafted in cuff-technique in the common carotid artery of 21 female Sprague Dawley rats categorized in three equal groups that were sacrificed on day 21, 42 and 84, respectively. Notably, no donor animals were needed, because auto-transplantations were performed. The survival rate was 100 % at the time point of sacrifice. In addition, the graft patency rate was 60 % for the first 10 operated animals and 82 % for the remaining 11 animals. The blood flow at the time of sacrifice was 8&plusmn;3 ml/min. In conclusion, this surgical protocol considerably simplifies, optimizes and standardizes this complicated procedure. It gives novice surgeons easy, step-by-step instruction, explaining possible pitfalls, thereby helping them to gain expertise fast and avoid useless sacrifice of experimental animals.

Neointimal hyperplasia is the primary cause of stenosis in arterialized veins. We propose a new protocol whereby the right external jugular vein is grafted using the cuff technique in the common carotid artery of Sprague Dawley rats. The survival rate was 100 % at the time point of sacrifice.

La formación de una sofisticada técnica de microcirugía: La interposición de injerto de la vena yugular externa en la arteria carótida común en ratas

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in ...

Investigación

Medicina

Optimized System for Cerebral Perfusion Monitoring in the Rat Stroke Model of Intraluminal Middle Cerebral Artery Occlusion

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal models of acute ischemic stroke allows to confirm successful arterial occlusion and exclude subarachnoid hemorrhage. Cerebral perfusion monitoring can also be used to study intracranial collateral circulation, which is emerging as a powerful determinant of stroke outcome and a possible therapeutic target. Despite a recognized role of Laser Doppler perfusion monitoring as part of the current guidelines for experimental cerebral ischemia, a number of technical difficulties exist that limit its widespread use. One of the major issues is obtaining a secure and prolonged attachment of a deep-penetration Laser Doppler probe to the animal skull. In this video, we show our optimized system for cerebral perfusion monitoring during transient middle cerebral artery occlusion by intraluminal filament in the rat. We developed in-house a simple method to obtain a custom made holder for twin-fibre (deep-penetration) Laser Doppler probes, which allow multi-site monitoring if needed. A continuous and prolonged monitoring of cerebral perfusion could easily be obtained over the intact skull.

Cerebral perfusion monitoring has been demonstrated to improve accuracy in ischemic stroke models. Technical difficulties often limit the use of this essential tool for cerebrovascular research. In this video, an optimized system is shown to obtain a single or multi-site hemodynamic monitoring during intraluminal middle cerebral artery occlusion in rats.

Sistema optimizado para la monitorización de la perfusión cerebral en el modelo de rata de Carrera intraluminal oclusión arteria cerebral media

The translational potential of pre-clinical stroke research depends on the accuracy of experimental modeling. Cerebral perfusion monitoring in animal ...

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. Intra-tracheal inoculation of a known amount of bacteria has been successfully used to reproduce the pathogenesis of pneumonia which then develops into sepsis. Monitoring hemodynamic parameters and providing standard clinical treatment including infusion of antibiotics, fluids and drugs to maintain blood pressure is critical to simulate routine supportive care in this model but to do so requires both arterial and venous vascular access. The video details the surgical technique for implanting carotid artery and common jugular vein catheters in an anesthetized rat. Following a 72 hr recovery period, the animals will be re-anesthetized and connected to a tether and swivel setup attached to the rodent housing which connects the implanted catheters to the hemodynamic monitoring system. This setup allows free movement of the rat during the study while continuously monitoring pressures, infusing fluids and drugs (antibiotics, vasopressors) and performing blood sampling.

Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

La cateterización de la arteria carótida y la vena yugular para realizar mediciones hemodinámicas, Infusiones y muestras de sangre en un modelo de rata Consciente

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

Infusiones de la arteria carótida para farmacocinéticas y farmacodinámicas Análisis de taxanos en ratones

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Trombosis modelo de ratón de la arteria carótida y Mesenterio Embarcaciones inducida por cloruro férrico-

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is also a valuable model system to test, and to evaluate therapeutics and drugs that negate maladaptive remodeling in the vessel. The injury, or barotrauma, in the vessel lumen caused by an inflated balloon via an inserted catheter induces subsequent neointimal growth, often leading to hyperplasia or thickening of the vessel wall that narrows, or obstructs the lumen. The method described here is sufficiently sensitive, and the results can be obtained in relatively short time (2 weeks after the surgery). The efficacy of the drug or therapeutic against the induced-remodeling can be evaluated either by the post-mortem pathological and histomorphological analysis, or by ultrasound sonography in live animals. In addition, this model system has also been used to determine the therapeutic window or the time course of the administered drug. These studies can leadto the development of a better administrative strategy and a better therapeutic outcome. The procedure described here provides a tool for translational studies that bring drug and therapeutic candidates from bench research to clinical applications.

The rat carotid balloon injury model described below allows researchers to evaluate drugs or therapeutics that negate injury-induced arterial hyperplasia. Detailed pre-surgical preparation, surgical procedure, and post-surgical cares of the animal are described.

Una rata carótida lesión con el balón modelo para probar Terapéutica Remodelación Anti-vasculares

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is ...

Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The simple murine AVF model was combined with intraluminal administration of drug solution to the venous endothelium at the same time as fistula creation. Technical aspects of this model are discussed. Under general anesthesia, an abdominal incision is made and the aorta and inferior vena cava (IVC) are exposed. The infra-renal aorta and IVC are dissected for clamping. After proximal and distal clamping, the puncture site is exposed and a 25 G needle is used to puncture both walls of the aorta and into the IVC. Immediately after the puncture, a reporter gene-expressing viral vector was infused in the IVC via the same needle, followed by 15 min of incubation. The intraluminal administration method enabled more robust viral gene delivery to the venous endothelium compared to administration by the external route. This novel method of delivery will facilitate studies that explore the role of the endothelium in AVF maturation and enable intraluminal drug delivery at the time of surgical operation.

After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

Intraluminal entrega de la droga a la fístula arteriovenosa ratón endotelio

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

The Rabbit Model of Accelerated Atherosclerosis: A Methodological Perspective of the Iliac Artery Balloon Injury

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the industrialized world. New Zealand White (NZW) rabbits are widely used as an animal model for the study of atherosclerosis. They develop spontaneous lesions when fed with atherogenic diet; however, this requires long time of 4 - 8 months. To further enhance and accelerate atherogenesis, a combination of atherogenic diet and mechanical endothelial injury is often employed. The presented procedure for inducing atherosclerotic plaques in rabbits uses a balloon catheter to disrupt the endothelium in the left iliac artery of NZW rabbits fed with atherogenic diet. Such mechanical damage caused by the balloon catheter induces a chain of inflammatory reactions initiating neointimal lipid accumulation in a time dependent fashion. Atherosclerotic plaque following balloon injury show neointimal thickening with extensive lipid infiltration, high smooth muscle cell content and presence of macrophage derived foam cells. This technique is simple, reproducible and produces plaque of controlled length within the iliac artery. The whole procedure is completed within 20 - 30 min. The procedure is safe with low mortality and also offers high success in obtaining substantial intimal lesions. The procedure of balloon catheter induced arterial injury results in atherosclerosis within two weeks. This model can be used for investigating the disease pathology, diagnostic imaging and to evaluate new therapeutic strategies.

Animal models of atherosclerosis are essential to understand the mechanism and to investigate newer approaches to prevent plaque development or rupture, a leading cause of death in the industrialized world. This protocol uses a combination of balloon injury and cholesterol rich diet to induce atherosclerotic plaques in rabbit iliac artery.

El modelo de conejo de ateroesclerosis acelerada: una perspectiva metodológica de la lesión de globo de la arteria ilíaca

Acute coronary syndrome resulting from coronary occlusion following atherosclerotic plaque development and rupture is the leading cause of death in the ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

Fase contraste resonancia magnética en la arteria carótida común de rata

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, we have translated a vascular injury model from a small animal into a large animal (canine), with slight modifications that can be used for pre-clinical screening of prophylactic and thrombolytic agents. In addition to the surgical methods, the modified protocol describes the step-by-step methods to assess carotid artery canalization by angiography, detailed instructions to process both the brain and carotid artery for histological analysis to verify carotid canalization and cerebral hemorrhage, and specific parameters to complete an assessment of downstream thromboembolic events by utilizing magnetic resonance imaging (MRI). In addition, specific procedural changes from the previously well-established small animal model necessary to translate into a large animal (canine) vascular injury are discussed.

Here, we present the modifications necessary to a well characterized and commonly used small animal ferric chloride-induced (FeCl3) carotid artery injury model for use in a large animal vascular injury model. The resulting model can be utilized for pre-clinical trial assessment of both prophylactic and thrombolytic pharmacological and mechanical interventions.

Trombosis de la arteria carótida canina cloruro férrico-inducida: Un modelo Animal grande de lesión Vascular

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, ...