<ol style="padding:5px;"><li> Instalación del equipo <ol style="padding:5px;"><li> Selecciona un área en el laboratorio donde la temperatura ambiente es igual o superior a 20 ° C. </li><li> Evite los lugares cerca de los respiraderos de aire acondicionado y / o un fuerte ruido. </li><li> Evite la exposición del animal a los olores que pueden causar irritación y / o el estrés, tales como perfumes, colonias, productos químicos, y productos de limpieza. </li><li> Encienda el controlador de CODA. </li><li> A su vez en la plataforma de calentamiento y se puso a nivel 3. Si la temperatura ambiente es superior a 25 º C, empieza en los niveles 1 o 2. </li><li> Prueba de diagnóstico de controlador de <ol style="padding:5px;"><li> Abra el software de CODA. </li><li> Seleccione el dispositivo de CODA haciendo clic en él. </li><li> Haga clic en &quot;dispositivo de prueba seleccionado&quot;. </li><li> Seleccione los puños y los canales para poner a prueba y haga clic en &quot;Test&quot;. </li><li> Después de la prueba se haya completado, cierre la ventana de prueba de dispositivos. </li></ol></li></ol></li><li> Software de Configuración <ol style="padding:5px;"><li> Gestión de personal y los animales <ol style="padding:5px;"><li> Gestión de personal <ol style="padding:5px;"><li> Seleccione Herramientas&gt; Administrar personal. </li><li> Haga clic en &quot;Investigador 1&quot; para cambiar el nombre. </li><li> Haga clic en las filas posteriores de añadir nuevos investigadores. </li><li> Haga clic en la pestaña &quot;técnicos&quot; para modificar o agregar los técnicos. </li><li> Haga clic en la pestaña &quot;muestras&quot; para el nombre del animal para facilitar su identificación y para seleccionar el tipo de animal, la rata o el ratón. </li><li> Haga clic en la pestaña &quot;animales&quot; para modificar el tipo de animal y la sensibilidad. </li></ol></li></ol></li><li> Nuevo experimento <ol style="padding:5px;"><li> Seleccione Archivo&gt; Nuevo Experimento&gt; </li><li> Comenzar un nuevo experimento <ol style="padding:5px;"><li> Escriba el nombre del experimento. </li><li> Seleccione el investigador clave. </li><li> Seleccione una fecha de inicio y haga clic en &quot;Siguiente&quot;. </li></ol></li><li> Información básica de sesión <ol style="padding:5px;"><li> Escriba el nombre de sesión </li><li> Establecer &quot;ciclos de aclimatación&quot; a 5. Si prefiere no incluir los ciclos de aclimatación, a cero. Para ayudar a aclimatar al animal a la titular y en los puños, añadir ciclos adicionales a su experimento y ejecutar el experimento durante 5 días consecutivos. </li><li> Ajuste &quot;Número de Sets&quot; a 1. </li><li> Conjunto &quot;El tiempo entre grupos&quot; a 30 segundos para los sistemas múltiples </li><li> Establecer &quot;ciclos por Set&quot; a 20 ciclos. </li><li> Conjunto &quot;El tiempo entre los ciclos&quot; a 5 segundos y haga clic en &quot;Siguiente&quot;. </li></ol></li></ol></li><li> SELECCIÓN DE LA MUESTRA <ol style="padding:5px;"><li style="list-style-type:none;"><ol style="padding:5px;"><li> Seleccionar la muestra de la piscina de la muestra y la asigna a la primer canal disponible y haga clic en &quot;Siguiente&quot;. </li></ol></li><li> Parámetros de la sesión <ol style="padding:5px;"><li> Establecer &quot;la presión de oclusión máxima&quot; a 250mmHg. </li><li> Set &quot;Tiempo deflación&quot; a 20 segundos para los ratones, 15 segundos para las ratas. </li><li> Set &quot;Volumen mínimo&quot; a 15ul. </li><li> Set &quot;Estilo de visualización&quot; para un canal por el gráfico. </li><li> Añadir investigadores y técnicos adicionales y haga clic en &quot;Siguiente&quot;. </li></ol></li><li> Sesión de secuencias de comandos <ol style="padding:5px;"><li> Revisar la secuencia de comandos de sesión y haga clic en &quot;Siguiente&quot;. </li><li> No seleccione Finalizar hasta que se complete la sección Preparación del animal. </li></ol></li></ol></li></ol></li><li> Preparación de los animales <ol style="padding:5px;"><li> Animal Set-up <ol style="padding:5px;"><li> Animal titular <ol style="padding:5px;"><li> Para entrenar y mejorar la aclimatación de los animales, añadir ciclos adicionales a su experimento y ejecutar el experimento durante cinco días consecutivos. También puede ayudar con la aclimatación se coloca el animal en el soporte por 15 minutos durante 3 días consecutivos antes del estudio actual. </li><li> Lugar de cada animal en un soporte al recoger el animal por la cola y con cuidado colocar al animal en la parte trasera del titular que se enfrenta el extremo abierto de la ojiva. Nota: el tamaño del animal en relación con el tamaño del titular seleccionado. El animal debe ser capaz de entrar libremente en el titular. </li><li> Con cuidado, seguro la puerta trasera al titular girando el tapón de rosca roja en la puerta trasera. Se debe tener cuidado de evitar pellizcar la cola o en cualquier otra parte del cuerpo garantizando al mismo tiempo la puerta trasera. </li><li> Deslice el cono de la nariz hacia la puerta trasera, lo que limita el movimiento de los animales. El cono de la nariz debe estar en condiciones de limitar el animal darse vuelta, mientras que en el interior del soporte. </li><li> Coloque el soporte a la plataforma de calentamiento en la posición designada. Si la prueba el mismo grupo de animales durante un período de tiempo, asegúrese de rotar la posición de titular en la plataforma de calentamiento para mejorar la consistencia y precisión de sus mediciones de la PA. Permitir que el animal por lo menos 5 minutos para aclimatarse a la titular. </li><li> No tocar o manipular el animal, mientras que en el interior del soporte. El mayor contacto podría irritar al animal. </li><li> Nunca deje a los animales en los titulares de vigilancia. </li></ol></li><li> Colocación del manguito <ol style = &quot;padding: 5px;&quot;&gt; <li> Tema de la cola a través del &quot;manguito de oclusión&quot;. Coloque esta banda tan cerca de la base de la cola como sea posible sin la fuerza. </li><li> Asegurar el &quot;Tubo de oclusión del manguito&quot; en la muesca en la parte posterior superior del soporte. </li><li> Tema de la cola a través de los &quot;Gemelos VPR&quot;, colocándolo dentro de 2 mm de la &quot;manguito de oclusión&quot;. </li><li> Asegurar el &quot;Tubo de VPR espontáneas&quot; en la muesca en la parte posterior superior del soporte. </li><li> Coloque los puños al controlador de CODA. </li></ol></li><li> Animal de temperatura <ol style="padding:5px;"><li> Permitir que el animal por lo menos 5 minutos para regular la temperatura. </li><li> Registrar la temperatura del animal con frecuencia con un termómetro infrarrojo. La temperatura de los animales debe ser de 32 a 35 ° C. </li></ol></li></ol></li></ol></li><li> Experimento <ol style="padding:5px;"><li> Comience Experimento <ol style="padding:5px;"><li> Haga clic en &quot;Finalizar&quot; y el experimento se iniciará de inmediato. </li></ol></li><li> Final Experiment <ol style="padding:5px;"><li> Animal <ol style="padding:5px;"><li> Quitarse de inmediato al animal de los puños y el soporte. </li><li> Devolver el animal a su jaula después de que el &quot;Informe simple resumen de la sesión&quot; en la pantalla. </li></ol></li><li> Recopilación de datos <ol style="padding:5px;"><li> Después de completar el último ciclo los ciclos aceptados se mostrarán automáticamente en formato de hoja de cálculo dentro de la aplicación CODA, haga clic en &quot;Exportar a Excel&quot; botón. </li><li> Seleccione la carpeta para colocar el archivo, el nombre del archivo y haga clic en &quot;Guardar&quot;. </li><li> Abra la aplicación Excel y haga clic en &quot;Abrir&quot;. </li><li> Ver todos los tipos de archivo para mostrar la &quot;. Csv salvo&quot;, haga clic en el archivo. </li></ol></li><li> Procesamiento de Datos <ol style="padding:5px;"><li> Hay varias maneras de procesar los datos en el archivo de Excel. Una práctica común es para obtener el promedio y desviación estándar. </li><li> Eliminar las medidas si la desviación estándar es mayor que 30. </li><li> Además, los ciclos rechazado puede ser visto y la base de datos puede ser fácilmente exportado a Excel. Consulte el manual del usuario para obtener información adicional. </li></ol></li></ol></li></ol></li></ol>

The authors have nothing to disclose.

measuring blood pressure in mice using volume pressure recording, a tail-cuff method

El CODA de 8 canales de alto rendimiento sistema no invasivo de presión arterial mide la presión arterial hasta en 8 ratones o ratas de forma simultánea. El CODA cola del manguito sistema utiliza registro del volumen de presión (VPR) para medir la presión arterial mediante la determinación del volumen de sangre cola. Un diferencial especialmente diseñado transductor de presión y una oclusión cola del manguito medir el volumen total de sangre en la cola, sin la necesidad de obtener la señal de pulso individual. Especial atención se brinda a la longitud del manguito de oclusión a fin de obtener las lecturas de presión sanguínea más precisa. VPR pueden obtener lecturas de piel oscura los roedores, como ratones C57BL6 y es compatible con MRI. El sistema CODA proporciona mediciones de seis (6) diferentes parámetros de la presión arterial, presión arterial sistólica y diastólica, frecuencia cardíaca, presión arterial media, el flujo de sangre cola, la cola y el volumen sanguíneo. Las mediciones se pueden realizar en cualquiera de los ratones despiertos o anestesiados o ratas. El sistema incluye un controlador de CODA, un ordenador portátil, el software, los puños, los titulares de los animales, las almohadillas de infrarrojos calentamiento, y un termómetro de infrarrojos. Hay siete tamaños diferentes titular de los ratones tan pequeños como de 8 gramos de ratas tan grandes como 900 gramos.

Watch this Scientific Journal Video about Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method at JoVE.com

Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method

El CODA de 8 canales de alto rendimiento sistema no invasivo de presión arterial mide la presión arterial hasta en 8 ratones o ratas de forma simultánea. Este sistema utiliza la cola del manguito de registro del volumen de presión (VPR) para medir la presión arterial mediante la determinación del volumen de sangre cola.

Medición de la presión arterial en ratones con registro del volumen de presión, un método de la cola del manguito

The CODA 8-Channel High Throughput Non-Invasive Blood Pressure system measures the blood pressure in up to 8 mice or rats simultaneously. The CODA ...

measuring-blood-pressure-mice-using-volume-pressure-recording-tail

Research

JoVE Journal

Biology

51.3K vistas.  University of Kentucky. El CODA de 8 canales de alto rendimiento sistema no invasivo de presión arterial mide la presión arterial hasta en 8 ratones o ratas de forma simultánea. Este sistema utiliza la cola del manguito de registro del volumen de presión (VPR) para medir la presión arterial mediante la determinación del volumen de sangre cola.

Video: Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method

Pressure-polishing Pipettes for Improved Patch-clamp Recording

Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording. We first developed this method for fabricating pipettes suitable for recording from small (&lt;3 m) neuronal cell bodies. The basic principal is similar to glass-blowing and combines air pressure and heat to modify the shape of patch pipettes prepared by a conventional micropipette puller. It can be applied to so-called soft (soda lime) and hard (borosilicate) glasses. Generally speaking, pressure polishing can reduce pipette resistance by 25% without decreasing the diameter of the tip opening (Goodman and Lockery, 2000). It can be applied to virtually any type of glass and requires only the addition of a high-pressure valve and fitting to a microforge. This technique is essential for recording from ultrasmall cells (&lt;5 m) and can also improve single-channel recording by minimizing pipette resistance. The blunt shape is also useful for perforated-patch clamp recording since this tip shape results in a larger membrane bleb available for perforation.

This is a guide to modifying the shape of glass micropipettes. Specifically, by using heat and air pressure the taper is widened without increasing the tip opening, leading to lower pipette resistance. This is critical to obtain low noise recordings of small cells but is useful in many applications.

Pulido a presión para la Mejora de pipetas de patch-clamp grabación

Pressure-polishing is a method for shaping glass pipettes for patch-clamp recording.  We first developed this method for fabricating pipettes suitable for ...

Investigación

Biología

Making Gynogenetic Diploid Zebrafish by Early Pressure

Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. Methods that produce viable homozygous diploid offspring directly from heterozygous females allow F1 mutagenized females to be screened directly for deleterious mutations in an accelerated forward genetic screen. Streisinger et al.1,2 described methods for making gynogenetic (homozygous) diploid zebrafish by activating zebrafish eggs with ultraviolet light-inactivated sperm and preventing either the second meiotic or the first zygotic cell division using physical treatments (heat or pressure) that deploymerize microtubules. The "early pressure" (EP) method blocks the meiosis II, which occurs shortly after fertilization. The EP method produces a high percentage of viable embryos that can develop to fertile adults of either sex. The method generates embryos that are homozygous at all loci except those that were separated from their centromere by recombination during meiosis I. Homozygous mutations are detected in EP clutches at between 50% for centromeric loci and less than 1% for telomeric loci. This method is reproduced verbatim from the Zebrafish Book3.

This is a method for generating gynogenetic diploid zebrafish embryos (embryos whose only genetic contribution comes from the mother) by blocking the second meiotic division immediately after fertilization with ultraviolet light-inactivated sperm. EP embryos are not fully homozygous due to recombination during the first meiotic division, however they are homozygous at all loci that have not been separated from their centromere by recombination.

Hacer pez cebra Gynogenetic diploides por presión inicial

Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. Methods that produce viable homozygous diploid offspring directly from ...

Ambulatory ECG Recording in Mice

Telemetric ECG recording in mice is essential to understanding the mechanisms behind arrhythmias, conduction disorders, and sudden cardiac death. Although the surface ECG is utilized for short-term measurements of waveform intervals, it is not practical for long-term studies of heart rate variability or the capture of rare episodes of arrhythmias. Implantable ECG telemeters offer the advantages of simple surgical implantation, long-term recording of electrograms in ambulatory mice, and scalability with simultaneous recordings of multiple animals. Here, we present a step-by-step guide to the implantation of telemeters for ambulatory ECG recording in mice. Careful adherence to aseptic technique is required for favorable survival results with the possibility of implantation and recording from weeks to months. Thus, implantable ECG telemetry is a valuable tool for detection of critical information on cardiac electrophysiology in ambulatory animal models such as the mouse. 

Telemetric ECG has emerged as an essential tool in evaluating animal models for cardiac arrhythmias and sudden cardiac death. Here, we present a stepwise guide to telemetric ECG recordings for application in long-term ambulatory ECG monitoring in mice.

Ambulatorio de grabación ECG en ratones

Telemetric ECG recording in mice is essential to understanding the mechanisms behind arrhythmias, conduction disorders, and sudden cardiac death. Although ...

Isolation and Culture of Pulmonary Endothelial Cells from Neonatal Mice

Endothelial cells provide a useful research model in many areas of vascular biology. Since its first isolation 1, human umbilical vein endothelial cells (HUVECs) have shown to be convenient, easy to obtain and culture, and thus are the most widely studied endothelial cells. However, for research focused on processes like angiogenesis, permeability or many others, microvascular endothelial cells (ECs) are a much more physiologically relevant model to study 2. Furthermore, ECs isolated from knockout mice provide a useful tool for analysis of protein function ex vivo. Several approaches to isolate and culture microvascular ECs of different origin have been reported to date 3-7, but consistent isolation and culture of pure ECs is still a major technical problem in many laboratories. Here, we provide a step-by-step protocol on a reliable and relatively simple method of isolating and culturing mouse lung endothelial cells (MLECs). In this approach, lung tissue obtained from 6- to 8-day old pups is first cut into pieces, digested with collagenase/dispase (C/D) solution and dispersed mechanically into single-cell suspension. MLECS are purified from cell suspension using positive selection with anti-PECAM-1 antibody conjugated to Dynabeads using a Magnetic Particle Concentrator (MPC). Such purified cells are cultured on gelatin-coated tissue culture (TC) dishes until they become confluent. At that point, cells are further purified using Dynabeads coupled to anti-ICAM-2 antibody. MLECs obtained with this protocol exhibit a cobblestone phenotype, as visualized by phase-contrast light microscopy, and their endothelial phenotype has been confirmed using FACS analysis with anti-VE-cadherin 8 and anti-VEGFR2 9 antibodies and immunofluorescent staining of VE-cadherin. In our hands, this two-step isolation procedure consistently and reliably yields a pure population of MLECs, which can be further cultured. This method will enable researchers to take advantage of the growing number of knockout and transgenic mice to directly correlate in vivo studies with results of in vitro experiments performed on isolated MLECs and thus help to reveal molecular mechanisms of vascular phenotypes observed in vivo.

Here, we describe a protocol for isolation and culture of murine pulmonary endothelial cells. This method comprises mechanic and enzymatic lung tissue dissociation as well as a 2-step purification process using anti-PECAM-1 and anti-ICAM-2 antibodies conjugated to magnetic beads, which produces a pure endothelial cell population of mostly microvascular origin.

Aislamiento y cultivo de células endoteliales pulmonares de ratones recién nacidos

Endothelial cells provide a useful research model in many areas of vascular biology. Since its first isolation 1, human umbilical vein endothelial cells ...

Assessing Murine Resistance Artery Function Using Pressure Myography

Pressure myograph systems are exquisitely useful in the functional assessment of small arteries, pressurized to a suitable transmural pressure. The near physiological condition achieved in pressure myography permits in-depth characterization of intrinsic responses to pharmacological and physiological stimuli, which can be extrapolated to the in vivo behavior of the vascular bed. Pressure myograph has several advantages over conventional wire myographs. For example, smaller resistance vessels can be studied at tightly controlled and physiologically relevant intraluminal pressures. Here, we study the ability of 3rd order mesenteric arteries (3-4 mm long), preconstricted with phenylephrine, to vaso-relax in response to acetylcholine. Mesenteric arteries are mounted on two cannulas connected to a pressurized and sealed system that is maintained at constant pressure of 60 mmHg. The lumen and outer diameter of the vessel are continuously recorded using a video camera, allowing real time quantification of the vasoconstriction and vasorelaxation in response to phenylephrine and acetylcholine, respectively.
	To demonstrate the applicability of pressure myography to study the etiology of cardiovascular disease, we assessed endothelium-dependent vascular function in a murine model of systemic hypertension. Mice deficient in the &alpha;1 subunit of soluble guanylate cyclase (sGC&alpha;1-/-) are hypertensive when on a 129S6 (S6) background (sGC&alpha;1-/-S6) but not when on a C57BL/6 (B6) background (sGC&alpha;1-/-B6). Using pressure myography, we demonstrate that sGC&alpha;1-deficiency results in impaired endothelium-dependent vasorelaxation. The vascular dysfunction is more pronounced in sGC&alpha;1-/-S6 than in sGC&alpha;1-/-B6 mice, likely contributing to the higher blood pressure in sGC&alpha;1-/-S6 than in sGC&alpha;1-/-B6 mice. 
	Pressure myography is a relatively simple, but sensitive and mechanistically useful technique that can be used to assess the effect of various stimuli on vascular contraction and relaxation, thereby augmenting our insight into the mechanisms underlying cardiovascular disease.

In pressure myography, an intact small segment of a vessel is mounted onto two small cannulas and pressurized to a suitable luminal pressure. Here, we describe the method to measure vasorelaxation response of the mouse 3rd order mesenteric arteries in c57 and sGC&alpha;1-/- mice using pressure myography.

La evaluación de la resistencia murino Función Arterial Utilizando miografía Presión

Pressure myograph systems are  exquisitely useful in the functional assessment of small arteries, pressurized  to a suitable transmural pressure. The  ...

One-channel Cell-attached Patch-clamp Recording

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate depends on properties intrinsic to each channel protein as well as the mechanism by which it is generated and controlled and represents an important area of current research. Information about the operational dynamics of ion channel proteins can be obtained by observing long stretches of current produced by a single molecule. Described here is a protocol for obtaining one-channel cell-attached patch-clamp current recordings for a ligand gated ion channel, the NMDA receptor, expressed heterologously in HEK293 cells or natively in cortical neurons. Also provided are instructions on how to adapt the method to other ion channels of interest by presenting the example of the mechano-sensitive channel PIEZO1. This method can provide data regarding the channel&#8217;s conductance properties and the temporal sequence of open-closed conformations that make up the channel&#8217;s activation mechanism, thus helping to understand their functions in health and disease.

Described here is a procedure for obtaining long stretches of current recording from one ion channel with the cell-attached patch-clamp technique. This method allows for observing, in real time, the pattern of open-close channel conformations that underlie the biological signal. These data inform about channel properties in undisturbed biological membranes.

Un canal celular conectado a Patch-clamp grabación

Ion channel proteins are universal devices for fast communication across biological membranes. The temporal signature of the ionic flux they generate ...

Measuring Respiratory Function in Mice Using Unrestrained Whole-body Plethysmography

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative assessment of lung function in rodent models is an important tool in the development of future therapies. Commonly used techniques for assessing respiratory function including invasive plethysmography and forced oscillation. While these techniques provide valuable information, data collection can be fraught with artefacts and experimental variability due to the need for anesthesia and/or invasive instrumentation of the animal. In contrast, unrestrained whole-body plethysmography (UWBP) offers a precise, non-invasive, quantitative way by which to analyze respiratory parameters. This technique avoids the use of anesthesia and restraints, which is common to traditional plethysmography techniques. This video will demonstrate the UWBP procedure including the equipment set up, calibration and lung function recording. It will explain how to analyze the collected data, as well as identify experimental outliers and artefacts that results from animal movement. The respiratory parameters obtained using this technique include tidal volume, minute volume, inspiratory duty cycle, inspiratory flow rate and the ratio of inspiration time to expiration time. UWBP does not rely on specialized skills and is inexpensive to perform. A key feature of UWBP, and most appealing to potential users, is the ability to perform repeated measures of lung function on the same animal.

The assessment of respiratory physiology has traditionally relied upon techniques, which require restraint or sedation of the animal. Unrestrained whole-body plethysmography, however, provides precise, non-invasive, quantitative analysis of respiratory physiology in animal models. In addition, the technique allows repeated respiratory assessment of mice allowing for longitudinal studies.

Medición de la función respiratoria en ratones usando irrestricta pletismografía corporal total

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative ...

A Method for Screening and Validation of Resistant Mutations Against Kinase Inhibitors

The discovery of BCR/ABL as a driver oncogene in chronic myeloid leukemia (CML) resulted in the development of Imatinib, which, in fact, demonstrated the potential of targeting the kinase in cancers by effectively treating the CML patients. This observation revolutionized drug development to target the oncogenic kinases implicated in various other malignancies, such as, EGFR, B-RAF, KIT and PDGFRs. However, one major drawback of anti-kinase therapies is the emergence of drug resistance mutations rendering the target to have reduced or lost affinity for the drug. Understanding the mechanisms employed by resistant variants not only helps in developing the next generation inhibitors but also gives impetus to clinical management using personalized medicine. We reported a retroviral vector based screening strategy to identify the spectrum of resistance conferring mutations in BCR/ABL, which has helped in developing the next generation BCR/ABL inhibitors. Using Ruxolitinib and JAK2 as a drug target pair, here we describe in vitro screening methods that utilizes the mouse BAF3 cells expressing the random mutation library of JAK2 kinase.

Emergence of genetic resistance against kinase inhibitor therapy poses significant challenge for effective cancer therapy. Identification and characterization of resistant mutations against a newly developed drug helps in better clinical management and next generation drug design. Here, we describe our protocol for in vitro screening and validation of resistant mutations.

Un método para la detección y validación de mutaciones de resistencia contra la quinasa Inhibidores

The discovery of BCR/ABL as a driver oncogene in chronic myeloid leukemia (CML) resulted in the development of Imatinib, which, in fact, demonstrated the ...

Cardiac Pressure-Volume Loop Analysis Using Conductance Catheters in Mice

Cardiac pressure-volume loop analysis is the &#8220;gold-standard&#8221; in the assessment of load-dependent and load-independent measures of ventricular systolic and diastolic function. Measures of ventricular contractility and compliance are obtained through examination of cardiac response to changes in afterload and preload. These techniques were originally developed nearly three decades ago to measure cardiac function in large mammals and humans. The application of these analyses to small mammals, such as mice, has been accomplished through the optimization of microsurgical techniques and creation of conductance catheters. Conductance catheters allow for estimation of the blood pool by exploiting the relationship between electrical conductance and volume. When properly performed, these techniques allow for testing of cardiac function in genetic mutant mouse models or in drug treatment studies. The accuracy and precision of these studies are dependent on careful attention to the calibration of instruments, systematic conduct of hemodynamic measurements and data analyses. We will review the methods of conducting pressure-volume loop experiments using a conductance catheter in mice.

Cardiac pressure-volume loop analysis is the most comprehensive way to measure cardiac function in the intact heart. We describe a technique to perform and analyze cardiac pressure volume loops, using conductance catheters.

Cardiaca Presión-Volumen Análisis Loop Utilizando conductancia catéteres en ratones

Cardiac pressure-volume loop analysis is the &#8220;gold-standard&#8221; in the assessment of load-dependent and load-independent measures of ventricular ...

Measuring Pressure Volume Loops in the Mouse

Understanding the causes and progression of heart disease presents a significant challenge to the biomedical community. The genetic flexibility of the mouse provides great potential to explore cardiac function at the molecular level. The mouse&#39;s small size does present some challenges in regards to performing detailed cardiac phenotyping. Miniaturization and other advancements in technology have made many methods of cardiac assessment possible in the mouse. Of these, the simultaneous collection of pressure and volume data provides a detailed picture of cardiac function that is not available through any other modality. Here a detailed procedure for the collection of pressure-volume loop data is described. Included is a discussion of the principles underlying the measurements and the potential sources of error. Anesthetic management and surgical approaches are discussed in great detail as they are both critical to obtaining high quality hemodynamic measurements. The principles of hemodynamic protocol development and relevant aspects of data analysis are also addressed.

This manuscript describes a detailed protocol for the collection of pressure-volume data from the mouse.

La medición de volumen de bucles de presión en el ratón

Understanding the causes and progression of heart disease presents a significant challenge to the biomedical community. The genetic flexibility of the ...