Queremos agradecer a Francesca Polverino MD, investigador en el Hospital Brigham y de la Mujer, por su contribución a este artículo, y también Mónica Yao, BS, y Kate Rydell, BS por su ayuda con la cría de murino y la exposición de los ratones al humo del cigarrillo. Este trabajo fue apoyado por el Servicio Público de Salud, Instituto Nacional del Corazón, los Pulmones y la Sangre Instituto Subvenciones HL111835, HL105339, HL114501, asistentes de vuelo Instituto de Investigación Médica Grant # CIA123046, el Brigham and Institute Hospital Lovelace respiratoria Consorcio de Investigación de la Mujer, y el Cambridge NIHR Biomédica Centro de Investigación.

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We have no conflicts of interest to disclose.

Most researcher use mice to model the main chronic lung pathologies and abnormal lung physiology in COPD (airspace enlargement, SAR, and increases in lung compliance) present in the human disease. A comprehensive approach to assess the effect of molecules of interest on both emphysema development and SAR is needed in mice in order to comprehensively assess the activities of molecules of interest in these chronic disease processes. 
There are several critical steps in this protocol. First, dur...

El uso de modelos animales para estudiar la EPOC es un reto porque ningún modelo puede replicar perfectamente todas las características de la enfermedad humana (2). La mayoría de los investigadores utilizan ratones para modelar la EPOC debido a las similitudes entre los ratones y los seres humanos en sus pulmonares fisiología, patología, genética y metabolitos. Además, los ratones son relativamente baratos para estudiar, y tanto el enfisema y la remodelación de la vía aérea pequeña desarrollan dentro de los 6 meses de exposición CS (5,7-9). Cigarrillos EPOC inducida por el humo: Varios métodos pueden inducir la EPOC en ratones. La mayoría de los investigadores exponen ratones para CS, que es el principal factor etiológico para la EPOC humano. La exposición CS durante 6 meses causa el desarrollo de enfisema y la remodelación de la vía aérea pequeña (SAR) en ratones, pero la gravedad de la enfermedad que es inducida varía dependiendo de la cepa murina estudiado. Por ejemplo, los ratones NZWLacZ son resistentes al desarrollo de enfisema inducido-CS mientras que AKR / J ratones son extremely sensible (10). La mayoría de los investigadores estudian C57BL / 6 ratones de la cepa en el modelo de exposición CS como muchos ratones de genes dirigidos están disponibles en esta cepa. Después de 6 meses de exposición CS, el enfisema y la fibrosis de la pequeña vía aérea a desarrollar en el tipo salvaje (WT) C57BL / 6 ratones, y ambas lesiones son relativamente leves en gravedad (5,10). Los investigadores utilizan dos tipos de exposición CS: sólo de la nariz y de todo el cuerpo exposiciones. Las principales desventajas de la única nariz técnica de exposición son que: 1) es un método más mano de obra; y 2) los ratones tienen que ser contenido en pequeñas cámaras que pueden inducir una respuesta de estrés y la hipertermia en los animales (11). La principal desventaja de exposición de cuerpo entero (que se describe en la presente memoria) es que los animales pueden ingerir (así como inhalar) nicotina y alquitrán productos cuando limpian su piel. Los ratones expuestos a todo el cuerpo CS también tienen niveles de carboxihemoglobina inferiores y pérdida de peso corporal reducido en comparación con los animales expuestos a la nariz de sólo CS (12). prueba de función pulmonar (PFT): Las medidas de la distensibilidad pulmonar y elastancia suelen ser similares en ratones C57BL / 6 de tipo salvaje (WT) ratones expuestos al aire o CS durante 6 meses debido al enfisema relativamente leve que se desarrolla cuando este cepa se expone a CS (10). Sin embargo, cuando la destrucción enfisematosa es más grave, el aumento de la distensibilidad pulmonar y desplazamientos a la izquierda en la presión-volumen (PV) fluyen bucles pueden ser detectados. Este último se puede observar, por ejemplo, en las cepas murinas que son más susceptibles a los efectos de CS (10), en / 6 ratones de cepas de genes dirigidos expuestas-CS C57BL que tienen un tipo enfisema más severo que C57BL / 6 ratones WT (13), o en ratones expuestos-CS sometidos a cambios ambientales que las hacen más susceptibles a los efectos de CS (14). Este protocolo utiliza un pequeño ventilador para animales para medir la reducción en el retroceso elástico del pulmón (aumento de la distensibilidad pulmonar cuasiestático [Cst] y las reducciones en el tejidoelastancia [H]), los bucles de flujo de PV, y los cambios en la resistencia de las vías respiratorias y el tejido en ratones anestesiados (15,16). Medidas de enfisema pulmonar: Análisis del enfisema en expuestos al CS C56BL / 6 ratones cepa es un reto, ya que su distribución es espacialmente heterogénea. Varios métodos diferentes cuantificar ampliación del espacio aéreo en los ratones. El primer método utilizado fue la media de intercepción lineal (L m) (17). Sin embargo, el método L m es un proceso lento, manual que puede no captar la heterogeneidad de la enfermedad (a menos que todas las secciones del pulmón se seleccionan aleatoriamente muestras) y su uso por lo tanto, puede introducir un sesgo de observador en el análisis. El índice de destrucción [DI, (18)] también cuantifica ampliación del espacio aéreo utilizando una lámina transparente con 50 puntos igualmente distribuidos colocados sobre una imagen digitalizada impresa de un hematoxilina y eosina sección de pulmón manchados. El método PI puntajes de la zona que rodea a cada punto de acacuerdo con el grado en que se destruyen los conductos alveolares y paredes alveolares dentro de esta área. La principal desventaja del método de DI es que es mucho tiempo y no más exacto que otros métodos (19,20). Este protocolo mide significan longitud de la cuerda alveolar y zona alveolar de pulmón secciones incluidas en parafina teñidos con tinción de Gill. Software Morfometría convierte las imágenes de secciones de pulmón a imágenes binarias (en la que el tejido es de color blanco y el espacio aéreo es negro), y luego se superpone una rejilla uniforme de líneas horizontales y verticales (acordes) y el software entonces cuantifica la duración de cada acorde dentro de las áreas identificadas por software como espacio aéreo. Usando este método, es posible medir el tamaño de los alvéolos en todas las partes del pulmón de una manera estandarizada y relativamente automatizado (21). La remodelación de la vía aérea pequeña (SAR): El aumento de la deposición de proteínas ECM (especialmente intersticiosl colágenos) alrededor de las pequeñas vías aéreas se produce en los animales expuestos al CS y contribuye a la obstrucción al flujo aéreo. Los investigadores no estudian SAR en modelos animales de EPOC con tanta frecuencia como el desarrollo de enfisema (22). Para cuantificar SAR en ratones expuestos-CS, este protocolo utiliza el software de análisis de imagen para medir el grosor de la capa de proteínas ECM que se deposita alrededor de las pequeñas vías respiratorias (vías respiratorias que tiene un diámetro medio entre 300 y 899 m) en secciones de pulmón incluidos en parafina teñidas con tricrómico de Masson.

<table border="0" cellpadding="0" cellspacing="0" width="579">
	<tbody>
		<tr height="34">
			<td height="34" style="height:34px;width:223px;">Whole-body smoke exposure device</td>
			<td style="width:103px;">Teague Enterprise</td>
			<td style="width:101px;">TE-10z</td>
			<td rowspan="2" style="width:153px;">Chronic Smoke exposures to induce chronic lung disease in mice</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:223px;">Research Cigarette</td>
			<td style="width:103px;">University of Kentucky</td>
			<td style="width:101px;">3R4F reference cigarettes</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:223px;">Pallflex&#174; Air Monitoring Filters, Emfab Filters TX40HI20WW, 25 mm</td>
			<td style="width:103px;">Pall Corporation</td>
			<td style="width:101px;">7219</td>
			<td rowspan="4" style="width:153px;">For measurement of TPMs</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:223px;">25 mm filter holder</td>
			<td style="width:103px;">Pall Corporation</td>
			<td style="width:101px;"></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:223px;">Filter sampler</td>
			<td style="width:103px;">Intermatic</td>
			<td style="width:101px;">Metal T100</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:223px;">Gas meter</td>
			<td style="width:103px;">AEM</td>
			<td style="width:101px;">Gas meters G1.6; G2.5; G4</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:223px;">Tracheal Cannula for mouse 18 gauge</td>
			<td style="width:103px;">Labinvention</td>
			<td style="width:101px;"></td>
			<td rowspan="2" style="width:153px;">Analysis of pulmonary function</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:223px;">Mechanical ventilator</td>
			<td style="width:103px;">Scireq</td>
			<td style="width:101px;">FlexiVent</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:223px;">Gill&#39;s hematoxylin solution&#160;</td>
			<td style="width:103px;">Sigma-Aldrich</td>
			<td style="width:101px;">GSH316</td>
			<td rowspan="3" style="width:153px;">For Gill staining, work under a fume hood</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:223px;">Hematoxylin solution, Harris modified</td>
			<td style="width:103px;">Sigma-Aldrich</td>
			<td style="width:101px;">HHS16</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:223px;">Cytoseal-60</td>
			<td style="width:103px;">Thermo Scientific</td>
			<td>8310-16</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:223px;">Micro-Slide-Field-Finder</td>
			<td style="width:103px;">Andwin Scientific INC</td>
			<td>50-949-582</td>
			<td rowspan="2" style="width:153px;">For analysis of emphysema</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:223px;">Scion Image Program</td>
			<td style="width:103px;">Scion Corporation</td>
			<td style="width:101px;"></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;width:223px;">Mason&#39;s trichrome stain</td>
			<td style="width:103px;">Sigma-Aldrich</td>
			<td style="width:101px;">HT15</td>
			<td rowspan="2" style="width:153px;">For analysis of small airway fibrosis</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:223px;">MetaMorp Offline version 7.0</td>
			<td style="width:103px;">Molecullar Devices LLC</td>
			<td style="width:101px;">31032</td>
		</tr>
	</tbody>
</table>

automated measurement of pulmonary emphysema and small airway remodeling in cigarette smoke-exposed mice

COPD is projected to be the third most common cause of mortality world-wide by 2020(1). Animal models of COPD are used to identify molecules that contribute to the disease process and to test the efficacy of novel therapies for COPD. Researchers use a number of models of COPD employing different species including rodents, guinea-pigs, rabbits, and dogs(2). However, the most widely-used model is that in which mice are exposed to cigarette smoke. Mice are an especially useful species in which to model COPD because their genome can readily be manipulated to generate animals that are either deficient in, or over-express individual proteins. Studies of gene-targeted mice that have been exposed to cigarette smoke have provided valuable information about the contributions of individual molecules to different lung pathologies in COPD(3-5). Most studies have focused on pathways involved in emphysema development which contributes to the airflow obstruction that is characteristic of COPD. However, small airway fibrosis also contributes significantly to airflow obstruction in human COPD patients(6), but much less is known about the pathogenesis of this lesion in smoke-exposed animals. To address this knowledge gap, this protocol quantifies both emphysema development and small airway fibrosis in smoke-exposed mice. This protocol exposes mice to CS using a whole-body exposure technique, then measures respiratory mechanics in the mice, inflates the lungs of mice to a standard pressure, and fixes the lungs in formalin. The researcher then stains the lung sections with either Gill&#8217;s stain to measure the mean alveolar chord length (as a readout of emphysema severity) or Masson&#8217;s trichrome stain to measure deposition of extracellular matrix (ECM) proteins around small airways (as a readout of small airway fibrosis). Studies of the effects of molecular pathways on both of these lung pathologies will lead to a better understanding of the pathogenesis of COPD.

Este protocolo comienza con la exposición de todo el cuerpo de los ratones para CS. Supervisión y mantenimiento del dispositivo y seguimiento de TPM adecuada recuentos garantizar exposiciones de humo consistentes (Figura 1). Es importante que el investigador practica la técnica de la inflación pulmonar usando el dispositivo de inflado Este protocolo comienza con la exposición de todo el cuerpo de los ratones para CS. Supervisión y mantenimiento del dispositivo y seguim...

Watch this Scientific Journal Video about Automated Measurement of Pulmonary Emphysema and Small Airway Remodeling in Cigarette Smoke-exposed Mice at JoVE.com

Automated Measurement of Pulmonary Emphysema and Small Airway Remodeling in Cigarette Smoke-exposed Mice

The goal of this protocol is to provide automated methods to quantify chronic lung pathologies in a murine model of COPD. The protocol includes exposing mice to cigarette smoke (CS), measuring pulmonary function, inflating the lungs, and using morphometry methods to measure emphysema and small airway remodeling in mice.

Medición automática de enfisema pulmonar y la pequeña vía aérea Remodelación de cigarrillos sin humo Ratones expuestos

COPD is projected to be the third most common cause of mortality world-wide by 2020(1).  Animal models of COPD are used to identify molecules that ...

automated-measurement-pulmonary-emphysema-small-airway-remodeling

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JoVE Journal

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13.1K vistas.  Brigham and Women's Hospital - Harvard Medical School. The goal of this protocol is to provide automated methods to quantify chronic lung pathologies in a murine model of COPD. The protocol includes exposing mice to cigarette smoke (CS), measuring pulmonary function, inflating the lungs, and using morphometry methods to measure emphysema and small airway remodeling in mice. 

Video: Automated Measurement of Pulmonary Emphysema and Small Airway Remodeling in Cigarette Smoke-exposed Mice

Isolation of Mouse Respiratory Epithelial Cells and Exposure to Experimental Cigarette Smoke at Air Liquid Interface

Pulmonary epithelial cells can be isolated from the respiratory tract of mice and cultured at air-liquid interface (ALI) as a model of differentiated respiratory epithelium. A protocol is described for isolating and exposing these cells to mainstream cigarette smoke (CS), in order to study epithelial cell responses to CS exposure. The protocol consists of three parts: the isolation of airway epithelial cells from mouse trachea, the culturing of these cells at air-liquid interface (ALI) as fully differentiated epithelial cells, and the delivery of calibrated mainstream CS to these cells in culture. The ALI culture system allows the culture of respiratory epithelia under conditions that more closely resemble their physiological setting than ordinary liquid culture systems. The study of molecular and lung cellular responses to CS exposure is a critical component of understanding the impact of environmental air pollution on human health. Research findings in this area may ultimately contribute towards understanding the etiology of chronic obstructive pulmonary disease (COPD), and other tobacco-related diseases, which represent major global health problems.

Pulmonary epithelial cells can be isolated from the respiratory tract of mice and cultured at air-liquid interface as a model of differentiated respiratory epithelium. A protocol is described for isolating, culturing and exposing these cells to mainstream cigarette smoke, in order to study molecular responses to this environmental toxin.

Aislamiento de ratón las células epiteliales respiratorias y la exposición al humo del cigarrillo Experimental de la interfaz de aire líquido

Pulmonary epithelial cells can be isolated from the respiratory tract of mice and cultured at air-liquid interface (ALI) as a model of differentiated ...

Investigación

Medicina

Protocol for Long Duration Whole Body Hyperthermia in Mice

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body temperature that is observed during fever. The use of hyperthermia as an adjuvant has emerged as a promising procedure for tumor regression in the field of cancer biology. For this purpose, the most important requirement is to have reliable and uniform heating protocols. We have developed a protocol for hyperthermia (whole body) in mice. In this protocol, animals are exposed to cycles of hyperthermia for 90 min followed by a rest period of 15 min. During this period mice have easy access to food and water. High body temperature spikes in the mice during first few hyperthermia exposure cycles are prevented by immobilizing the animal. Additionally, normal saline is administered in first few cycles to minimize the effects of dehydration. This protocol can simulate fever like conditions in mice up to 12-24 hr. We have used 8-12 weeks old BALB/Cj female mice to demonstrate the protocol.

This paper describes a protocol for whole body hyperthermia in mice that can stimulate fever like conditions up to 12-24 hr.

Protocolo para la hipertermia corporal Larga Duración Total en ratones

Hyperthermia is a general term used to define the increase in core body temperature above normal. It is often used to describe the increased core body ...

In vivo Measurement of the Mouse Pulmonary Endothelial Surface Layer

The endothelial glycocalyx is a layer of proteoglycans and associated glycosaminoglycans lining the vascular lumen. In vivo, the glycocalyx is highly hydrated, forming a substantial endothelial surface layer (ESL) that contributes to the maintenance of endothelial function. As the endothelial glycocalyx is often aberrant in vitro and is lost during standard tissue fixation techniques, study of the ESL requires use of intravital microscopy. To best approximate the complex physiology of the alveolar microvasculature, pulmonary intravital imaging is ideally performed on a freely-moving lung. These preparations, however, typically suffer from extensive motion artifact. We demonstrate how closed-chest intravital microscopy of a freely-moving mouse lung can be used to measure glycocalyx integrity via ESL exclusion of fluorescently-labeled high molecular weight dextrans from the endothelial surface. This non-recovery surgical technique, which requires simultaneous brightfield and fluorescent imaging of the mouse lung, allows for longitudinal observation of the subpleural microvasculature without evidence of inducing confounding lung injury.

In vivo Measurement of the Mouse Pulmonary Endothelial Surface Layer

The endothelial glycocalyx/endothelial surface layer is ideally studied using intravital microscopy. Intravital microscopy is technically challenging in a moving organ such as the lung. We demonstrate how simultaneous brightfield and fluorescent microscopy may be used to estimate endothelial surface layer thickness in a freely-moving in vivo mouse lung.

Medición in vivo de la capa de ratón pulmonar superficie endotelial

The endothelial glycocalyx is a layer of proteoglycans and associated glycosaminoglycans lining the vascular lumen. In vivo, the glycocalyx is highly ...

Echocardiographic Assessment of the Right Heart in Mice

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential therapies. Given the expense and time involved in creating animal models of disease, it is critical that researchers have tools to accurately assess phenotypic expression of disease. Right ventricular dysfunction is the major manifestation of pulmonary hypertension. Echocardiography is the mainstay of the noninvasive assessment of right ventricular function in rodent models and has the advantage of clear translation to humans in whom the same tool is used. Published echocardiography protocols in murine models of PAH are lacking.
In this article, we describe a protocol for assessing RV and pulmonary vascular function in a mouse model of PAH with a dominant negative BMPRII mutation; however, this protocol is applicable to any diseases affecting the pulmonary vasculature or right heart. We provide a detailed description of animal preparation, image acquisition and hemodynamic calculation of stroke volume, cardiac output and an estimate of pulmonary artery pressure.

This article provides a protocol for the echocardiographic assessment of right ventricular size and pulmonary hypertension in mice. Applications include phenotype determination and serial assessment in transgenic and toxin-induced mouse models of cardiomyopathy and pulmonary vascular disease.

Evaluación ecocardiográfica de la parte derecha del corazón en ratones

Transgenic and toxic models of pulmonary arterial hypertension (PAH) are widely used to study the pathophysiology of PAH and to investigate potential ...

Automated Measurement of Microcirculatory Blood Flow Velocity in Pulmonary Metastases of Rats

Because the lung is a major target organ of metastatic disease, animal models to study the physiology of pulmonary metastases are of great importance. However, very few methods exist to date to investigate lung metastases in a dynamic fashion at the microcirculatory level, due to the difficulty to access the lung with a microscope. Here, an intravital microscopy method is presented to functionally image and quantify the microcirculation of superficial pulmonary metastases in rats, using a closed-chest pulmonary window and automated analysis of blood flow velocity and direction. The utility of this method is demonstrated to measure increases in blood flow velocity in response to pharmacological intervention, and to image the well-known tortuous vasculature of solid tumors. This is the first demonstration of intravital microscopy on pulmonary metastases in a closed-chest model. Because of its minimized invasiveness, as well as due to its relative ease and practicality, this technology has the potential to experience widespread use in laboratories that specialize on pulmonary tumor research.

A method is presented to measure microcirculatory blood flow velocity in pulmonary cancer metastases of the pleural surface in rats in an automated fashion, using closed-chest pulmonary intravital microscopy. This model has potential to be used as a widespread tool to perform physiologic research on pulmonary metastases in rodents.

Medición Automatizada de la microcirculación de la sangre Velocidad del caudal en metástasis pulmonar de ratas

Because the lung is a major target organ of metastatic disease, animal models to study the physiology of pulmonary metastases are of great importance. ...

An Improved Method for Rapid Intubation of the Trachea in Mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

Un método mejorado para la rápida intubación de la tráquea en ratones

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and fluid retention. Changes in myocardial structure, electrical conduction, and energy metabolism develop with heart failure, leading to contractile dysfunction, increased risk of arrhythmias, and sudden death. Hypertensive heart disease is one of the key contributing factors of cardiac remodeling associated with heart failure. The most commonly-used animal model mimicking hypertensive heart disease is created via surgical interventions, such as by narrowing the aorta. Abdominal aortic constriction is a useful experimental technique to induce a pressure overload, which leads to heart failure. The surgery can be easily performed, without the need for chest opening or mechanical ventilation. Abdominal aortic constriction-induced cardiac pathology progresses gradually, making this model relevant to clinical hypertensive heart failure. Cardiac injury and remodeling can be observed 10 weeks after the surgery. The method described here provides a simple and effective approach to produce a hypertensive heart disease animal model that is suitable for studying disease mechanisms and for testing novel therapeutics.

A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

Un modelo de remodelamiento cardiaco A través de la constricción de la aorta abdominal en ratas

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and ...

Comparing the Effects of Electronic Cigarette Vapor and Cigarette Smoke in a Novel In Vivo Exposure System

Electronic cigarettes (E-cigarettes) are being widely used, and growing in popularity. It is estimated that more than 9 million adults use them regularly. The potential adverse health effects of electronic cigarette vapor (E-vapor) exposure are poorly defined. While several animal models of E-vapor exposure have been developed, few models expose rodents to clinically relevant quantities of nicotine and make direct comparisons to cigarette smoke within the same exposure system. Here, we present a method for constructing and operating an E-vapor chamber and cigarette smoke chamber. The chambers are constructed by outfitting anesthesia chambers with a computer controlled pumping system that delivers consistent amounts of E-vapor or cigarette smoke to rodents. Nicotine exposure is measured indirectly by quantifying pre and post-exposure serum cotinine levels. This exposure system can be modified to accommodate various types of E-cigarettes&#160;and tobacco cigarettes, and can be used to compare the effects of E-vapor&#160;and cigarette smoke in vivo.

Comparing the Effects of Electronic Cigarette Vapor and Cigarette Smoke in a Novel In Vivo Exposure System

This protocol describes a method for exposing rodents to electronic cigarette vapor (E-vapor) and cigarette smoke. Exposure chambers are constructed by modifying anesthesia chambers with an automated pumping system that delivers E-vapor or cigarette smoke to rodents. This system can easily be modified to accommodate many experimental endpoints.

Comparación de los efectos del vapor de cigarrillo electrónico y el humo del cigarrillo en una novela<em&gt; In Vivo</em&gt; Sistema de exposición

Electronic cigarettes (E-cigarettes) are being widely used, and growing in popularity. It is estimated that more than 9 million adults use them regularly. ...

Lung Fixation under Constant Pressure for Evaluation of Emphysema in Mice

Emphysema is a significant feature of chronic obstructive pulmonary disease (COPD). Studies involving an emphysematous mouse model require optimal lung fixation to produce reliable histological specimens of the lung. Due to the nature of the lung&rsquo;s structural composition, which consists largely of air and tissue, there is a risk that it collapses or deflates during the fixation process. Various lung fixation methods exist, each of which has its own advantages and disadvantages. The lung fixation method presented here utilizes constant pressure to enable optimal tissue evaluation for studies using an emphysematous mouse lung model. The main advantage is that it can fix many lungs with the same condition at one time. Lung specimens are obtained from chronic cigarette smoke-exposed mice. Lung fixation is performed using specialized equipment that enables the production of constant pressure. This constant pressure maintains the lung in a reasonably inflated state. Thus, this method generates a histological specimen of the lung that is suitable to evaluate cigarette smoke-induced mild emphysema.

Presented here is a useful protocol for lung fixation that creates a stable condition for histological evaluate of lung specimens from a mouse model of emphysema. The main advantage of this model is that it can fix many lungs with the same constant pressure without lung collapse or deflation.

Fijación pulmonar bajo presión constante para la evaluación del enfisema en ratones

Emphysema is a significant feature of chronic obstructive pulmonary disease (COPD). Studies involving an emphysematous mouse model require optimal lung ...

Transdermal Measurement of Glomerular Filtration Rate in Mice

Transdermal analysis of glomerular filtration rate (GFR) is an established technique that is used to assess renal function in mouse and rat models of acute kidney injury and chronic kidney disease. The measurement system consists of a miniaturized fluorescence detector that is directly attached to the skin on the back of conscious, freely moving animals, and measures the excretion kinetics of the exogenous GFR tracer, fluorescein-isothiocyanate (FITC) conjugated sinistrin (an inulin analog). This system has been described in detail in rats. However, because of their smaller size, measurement of transcutaneous GFR in mice presents additional technical challenges. In this paper we therefore provide the first detailed practical guide to the use of transdermal GFR monitors in mice based on the combined experience of three different investigators who have been performing this assay in mice over a number of years.

Here we describe a protocol to measure glomerular filtration rate (GFR) in conscious, freely moving mice using a transdermal GFR monitor.

Transdérmica de medición de la tasa de filtración Glomerular en los ratones

Transdermal analysis of glomerular filtration rate (GFR) is an established technique that is used to assess renal function in mouse and rat models of ...