Este trabajo fue apoyado en parte por una beca de la Sociedad Americana otológico a CAM y por una beca de la Universidad de casos del Centro Médico de los Hospitales de KNA y CAM. Nos gustaría agradecer a Jeff Churney en Focus Medical Group, Inc. (Hinckley, OH) para que nos proporciona una cámara de microscopio. Servicios de producción de video cortesía de Fusión Creativa Marketing, Inc. (Valdosta, GA) Damos las gracias a Jonathan Frankel y Alex Levitt en CWRU por sus comentarios durante la preparación del vídeo.

Parte 1: Anestesia y preparación para la cirugía <ol><li> Un adulto, mujer Duncan-Hartley conejillo de Indias de Charles River Internacional Laboratories, Inc. (Wilmington, MA) es anestesiados con ketamina 60 mg / kg y xilacina 12 mg / kg por vía intramuscular. </li><li> Lubricantes derivados del petróleo se aplica a los ojos para proteger al animal de lesión en la córnea debido a la desecación. </li><li> El animal se afeita desde la parte superior de la cabeza a los hombros y entre las orejas. </li><li> El área afeitada es preparado de una manera aséptica primero con el 7,5% de povidona yodada seguido de 70% de alcohol etílico. </li><li> El animal se coloca sobre una mesa de operaciones con manta térmica y en una plataforma que eleva la cabeza y flexiona el cuello. La cabeza del animal se dirige fuera de su cirujano. Flexión adecuada del cuello se requiere a fin de exponer adecuadamente la cresta occipital. </li><li> El animal se cubre de una manera aséptica. </li></ol> 2 Parte: La incisión y la exposición del occipucio ósea <ol><li> Epinefrina más bupivacaína (bupivacaína) 0,25% se aplica a lo largo del sitio de la incisión posibles (incisión media piel dorsal a lo largo de la protuberancia occipital). Esto provoca vasoconstricción local, lo que limita el sangrado, y proporciona duradera analgesia postoperatoria después de que el animal se despierta. </li><li> Incisión de la línea media dorsal está hecha de piel en el cuero cabelludo a lo largo del occipucio. <img src="/files/ftp_upload/1728/1728fig1.jpg" alt="Figura 1" /> Figura 1: La piel está formada incisión a lo largo de la línea media dorsal, como se muestra arriba. </li><li> Para obtener una adecuada exposición, la piel se retrae lateralmente mediante la colocación de una pinza hemostática curva pequeñas a cada lado de la incisión. </li><li> Una incisión horizontal a lo largo del músculo del cuello para exponer el occipucio ósea. En esta etapa, el animal puede contracción a pesar de la anestesia adecuada debido a la transección del nervio occipital mayor. Sección transversal de la médula espinal es un peligro con la incisión horizontal en la región suboccipital. Esta complicación se puede evitar asegurándose de que el hueso subyacente siempre se siente debajo de la hoja de bisturí. </li><li> Con la musculatura suboccipital incisión hasta el hueso, una pinza hemostática tercero se utiliza para retraer el borde del músculo superior. Esta pinza de hemostasia se mantiene en su lugar por el ayudante de cirugía. </li><li> Un elevador de periostio Lempert se utiliza para exponer la sutura temporo-occipital. </li></ol> Parte 3: Perforación de la sutura temporo-occipital <ol><li> Tras la exposición de la sutura temporo-occipital (Figura 2), un taladro otológico con 1,5 mm fresa de diamante se utiliza para perforar inferio-lateralmente a la sutura. </li><li> Debajo de la tabla externa del cráneo, la textura esponjosa del diploe (El hueso esponjoso entre las tablas que intervienen en el cráneo) se hará visible. Una vez que el diploe se ha perforado de distancia, la tabla interna del cráneo aparece, junto con un color azul pálido que es el seno sigmoideo. </li><li> Los márgenes del seno sigmoideo se definen sin penetrar en el buque. </li><li> Una vez que el hueso que cubre el seno sigmoideo ha sido lo suficientemente diluida, un aumento mayor y se utiliza una selección directa se utiliza para quitar con cuidado el hueso que cubre el seno sigmoideo. </li></ol><img src="/files/ftp_upload/1728/1728fig2.jpg" alt="Figura 2" /> Figura 2 El área a ser perforados en parte se superpone a la sutura temporo-occipital. Parte 4: La obstrucción del conducto <ol><li> El seno sigmoideo esqueleto se retrae medialmente para revelar el opérculo. </li><li> Un diamante de 0,5 mm de fresa se utiliza para perforar por dentro de la opérculo y en el saco endolinfático y el conducto. </li><li> El conducto endolinfático es después cubierta con cera de hueso con un pico recto. </li><li> El defecto del cráneo se refuerza entonces con Gelfoam (Pfizer, Nueva York, NY). </li><li> 4-0 Polysorb (United States Surgical, Norwalk, CT) de sutura se utiliza para poner los músculos en conjunto. Hay entonces una transición sin problemas a una técnica de sutura subcutánea para el cierre de la incisión de la piel. Con este cierre no hay necesidad de retirar las suturas después y no hay riesgo de que el animal garra en el lugar y sacar una sutura. </li></ol> Parte 5: cuidado post-operatorio El cuy es colocado en una incubadora para mantener la temperatura corporal y se revisa a intervalos de 30 minutos hasta que despierta y capaz de mantener una posición esternal. El animal es inspeccionado por la respiración y el color de la carrocería. Este centro utiliza la enrofloxacina (0,2 ml a 22,7 mg / ml, vía subcutánea) y la buprenorfina (0,05 ml a 0.3mg/mL, por vía subcutánea) para la profilaxis de la infección post-operatoria y el control del dolor, respectivamente. Analgésico se administra una vez en el día de la cirugía y dos veces en el post-operatorio día uno y dos. Aunque los antibióticos no son necesarios para Su asépticargery, uno de cinco días, dos veces al día un curso de antibióticos profilácticos pueden ser givenonce en el post-operatorio de tres días. Complicaciones y Gestión <ol><li> La depresión respiratoria, el cirujano debe estar siempre al tanto de respiración del animal s. Si la respiración se hace más lento o se detiene, suspender la cirugía y una inspección visual para el movimiento torácico. Si el animal está experimentando dificultad respiratoria o dificultad para respirar, compruebe que la boca y las vías respiratorias no están ocluidas. </li><li> La penetración del seno sigmoideo Gelfoam uso y compresión a lo largo de la punta de succión hasta que el sangrado se detiene. </li><li> Penetración dural cubrir el defecto con Gelfoam. </li><li> Daño vestibular No perforar más de 1 mm lateral al seno sigmoideo. Si lo hace, los riesgos de dañar el conducto semicircular posterior, lo que invalida todos los datos para el conejillo de indias. </li></ol>

<ol>
	<li>Andrews, J. C., Bohmer, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+surgical+approach+to+the+endolymphatic+sac+and+the+cochlear+aqueduct+in+the+guinea+pig.">The surgical approach to the endolymphatic sac and the cochlear aqueduct in the guinea pig.</a> Am J Otolaryngol. 10, 61-66 (1989).</li><li>Kimura, R., Schuknecht, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Membranous+hydrops+in+the+inner+ear+of+the+guinea+pig+after+obliteration+of+the+endolymphatic+sac.">Membranous hydrops in the inner ear of the guinea pig after obliteration of the endolymphatic sac.</a> Pract Otolaryngol. , (1965).</li></ol>

Los experimentos con animales se realizaron de acuerdo con las directrices y normas establecidas por el Cuidado de Animales institucional y el empleo de la Case Western Reserve University.

En las semanas siguientes a la obstrucción quirúrgica del conducto endolinfático, el tema animal experimenta pérdida de audición neurosensorial fluctuante, medida por la respuesta de tronco cerebral evocados auditivos. Una muestra de la respuesta auditiva del tronco cerebral (ABR) la grabación se muestra en la Figura 3. La oreja izquierda sirve como un control no operado. <a href="https://www.jove.com/files/ftp_upload/1728/1728fig4.jpg"><img border="0" src="/files/ftp_upload/1728/1728fig3tmb.jpg" alt="Figur...

surgical induction of endolymphatic hydrops by obliteration of the endolymphatic duct

Inducción quirúrgica del edema endolinfático (ELH) en el conejillo de indias por la obliteración y la obstrucción del conducto endolinfático es un modelo animal ampliamente aceptado de la enfermedad y una importante correlación entre la enfermedad humana de Meniere. En 1965, Robert Kimura y Schuknecht Harold describió por primera vez un enfoque intradural para la obstrucción del conducto endolinfático (Kimura 1965). Aunque eficaz, esta técnica, que requiere la penetración de la cubierta protectora del cerebro, incurrió en un nivel no deseado de la morbilidad y la mortalidad en las materias de origen animal. En consecuencia, Andrews y Bohmer desarrollado un enfoque extradural, que previsiblemente produce menos de las complicaciones asociadas con el sistema nervioso central (SNC) la penetración. (Andrews y Bohmer 1989) El enfoque extradural se describe aquí primero requiere una incisión en la región del occipucio para exponer la capa muscular subyacente. Nosotros operamos sólo en el lado derecho. Después de la retracción adecuada del tejido que recubre, una incisión horizontal en la musculatura del occipital derecho a exponer el derecho temporo-occipital línea de sutura. El hueso inmediatamente inferio-lateral de la línea de sutura (Fig. 1) es perforado con un taladro otológico hasta el seno sigmoideo se hace visible. Retracción medial del seno sigmoideo revela el opérculo del conducto endolinfático, que alberga el saco endolinfático. Perforación por dentro de la opérculo en el área del saco endolinfático revela el conducto endolinfático, que luego se llena con cera de hueso para producir la obstrucción y ELH en última instancia. En las semanas siguientes, el animal demostrará la pérdida progresiva, auditiva fluctuante y evidencia histológica de la ELH.

Watch this Scientific Journal Video about Surgical Induction of Endolymphatic Hydrops by Obliteration of the Endolymphatic Duct at JoVE.com

Surgical Induction of Endolymphatic Hydrops by Obliteration of the Endolymphatic Duct

Este video muestra la forma de obstruir el conducto quirúrgicamente el conejillo de indias de endolinfático para producir edema endolinfático.

Inducción quirúrgica de edema endolinfático por la obliteración del conducto endolinfático

Surgical induction of endolymphatic hydrops (ELH) in the guinea pig by obliteration and obstruction of the endolymphatic duct is a well-accepted animal ...

surgical-induction-endolymphatic-hydrops-obliteration-endolymphatic

Research

JoVE Journal

Biology

11.3K vistas.  Case Western Reserve University. Este video muestra la forma de obstruir el conducto quirúrgicamente el conejillo de indias de endolinfático para producir edema endolinfático.

Video: Surgical Induction of Endolymphatic Hydrops by Obliteration of the Endolymphatic Duct

Imaging Effector Memory T cells in the Ear After Induction of Adoptive DTH

Delayed type hypersensitivity (DTH) is an immune reaction in which the main players are CCR7- effector / memory T lymphocytes. Here, we demonstrate a method for inducing and recording the progress of a DTH reaction in the rat ear. This is followed by a demonstration of the preparation of rat ear tissue for two-photon imaging of the CCR7- effector / memory T cell response.
An adoptive DTH is induced by the intraperitoneal injection of GFP-labeled Ova-specific CCR7- effector / memory T cell line (Beeton, C J. Visualized Experiments, Issue 8). Cells are then allowed to equilibrate in the rat for 48 hours before challenge by injecting one ear with saline (control ear) and the other with a 1:1 mix of Ova and Ova conjugated to Texas-Red (Ova-TR) to allow visualization of resident antigen-presenting cells.
We describe a method of tissue preparation useful for imaging the motility of cells within the deep dermal layer during an immune response, in conjunction with visualization of collagen fibers by second harmonic generation. Ear tissue is cut into 5 x 5 mm squares (slightly larger is better) and mounted onto plastic cover slips using Vetbond&trade;, which are then secured using silicone grease in an imaging chamber and superfused by oxygen-bubbled tissue culture medium at 37&deg;C.

Here we demonstrate a method for inducing and recording the progress of a delayed type-hypersensitivity (DTH) reaction in the rat ear. This is followed by a demonstration of the preparation of rat ear tissue for two-photon imaging of the effector / memory T cell response.

Imágenes de las células efectoras T de memoria en el oído después de la inducción de DTH adoptivos

Delayed type hypersensitivity (DTH) is an immune reaction in which the main players are CCR7- effector / memory T lymphocytes. Here, we demonstrate a ...

Investigación

Biología

Assay for Neural Induction in the Chick Embryo

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ideal tool for studying the formation and initial patterning of the nervous system. This video demonstrates how to graft organizer tissue into a host, a method by which Hensen s node (the organizer in the chick embryo) is grafted to a host competent ectoderm. The organizer graft instructs overlying na ve tissue to adopt a neural fate via neural inducing signals. This mechanism is referred to as neural induction, and constitutes the initial step in the formation of brain and spinal cord in amniotes. This method is essentially used for the characterization of putative neural inducing molecules in chick. This video demonstrates the different steps in the assay for neural induction; First, the donnor embryo is explanted and pinned on a dish. Then, the host embryo is prepared for New culture. The graft is excised and transplanted to the host area pellucida margin. The host is cultured for 18-22 hrs. The assembly is fixed and processed for further applications (e.g. in situ hybridization). This method was originally devised by Waddington 1,2 and Gallera 3,4.

Neural induction is the first step in the formation of the brain. It is a mechanism by which Hensen's node (organizer), instructs adjacent tissue to adopt a neural fate, i.e. to give rise to the nervous system. This video demonstrates an assay for neural induction in chick embryo.

Ensayo de inducción neural en el embrión de pollo

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ...

Microsurgical Clip Obliteration of Middle Cerebral Aneurysm Using Intraoperative Flow Assessment

Cerebral aneurysms are abnormal widening or ballooning of a localized segment of an intracranial blood vessel.  Surgical clipping is an important treatment for aneurysms which attempts to exclude blood from flowing into the aneurysmal segment of the vessel while preserving blood flow in a normal fashion.  Improper clip placement may result in residual aneurysm with the potential for subsequent aneurysm rupture or partial or full occlusion of distal arteries resulting in cerebral infarction.  Here we describe the use of an ultrasonic flow probe to provide quantitative evaluation of arterial flow before and after microsurgical clip placement at the base of a middle cerebral artery aneurysm.  This information helps ensure adequate aneurysm reconstruction with preservation of normal distal blood flow.

Description of the surgical obliteration of a cerebral aneurysm utilizing an ultrasonic flow probe to assess arterial flow prior to and after aneurysm clip placement.

La obliteración de microcirugía Clip de Aneurisma Cerebral Media uso de la evaluación intraoperatoria de flujo

Cerebral aneurysms are abnormal widening or ballooning of a localized segment of an intracranial blood vessel.  Surgical clipping is an important ...

Isolation of Basal Cells and Submucosal Gland Duct Cells from Mouse Trachea

The large airways are directly in contact with the environment and therefore susceptible to injury from toxins and infectious agents that we breath in 1. The large airways therefore require an efficient repair mechanism to protect our bodies. This repair process occurs from stem cells in the airways and isolating these stem cells from the airways is important for understanding the mechanisms of repair and regeneration. It is also important for understanding abnormal repair that can lead to airway diseases 2. The goal of this method is to isolate a novel stem cell population from the mouse tracheal submucosal gland ducts and to place these cells in in vitro and in vivo model systems to identify the mechanisms of repair and regeneration of the submucosal glands 3. This production shows methods that can be used to isolate and assay the duct and basal stem cells from the large airways 3.This will allow us to study diseases of the airway, such as cystic fibrosis, asthma and chronic obstructive pulmonary disease. Currently, there are no methods for isolation of submucosal gland duct cells and there are no in vivo models to study the regeneration of submucosal glands.

Here we demonstrate our protocol for isolation of basal and submucosal gland duct cells from mouse tracheas. We also demonstrate the method of injecting stem cells into the dorsal mouse fat pad to create an in vivo model of submucosal gland regeneration.

Aislamiento de células basales y de células submucosas conducto de la glándula de ratón tráquea

The large airways are directly in contact with the environment and therefore susceptible to injury from toxins and infectious agents that we breath in 1. ...

Culturing Primary Rat Inner Medullary Collecting Duct Cells

Arginine-vasopressin (AVP) facilitates water reabsorption by renal collecting duct principal cells and thereby fine-tunes body water homeostasis. AVP binds to vasopressin V2 receptors (V2R) on the surface of the cells and thereby induces synthesis of cAMP. This stimulates cellular signaling processes leading to changes in the phosphorylation of the water channel aquaporin-2 (AQP2). Protein kinase A phoshorylates AQP2 and thereby triggers the translocation of AQP2 from intracellular vesicles into the plasma membrane facilitating water reabsorption from primary urine. Aberrations of AVP release from the pituitary or AVP-activated signaling in principal cells can cause central or nephrogenic diabetes insipidus, respectively; an elevated blood plasma AVP level is associated with cardiovascular diseases such as chronic heart failure and the syndrome of inappropriate antidiuretic hormone secretion.
Here, we present a protocol for cultivation of primary rat inner medullary collecting duct (IMCD) cells, which express V2R and AQP2 endogenously. The cells are suitable for elucidating molecular mechanisms underlying the control of AQP2 and thus to discover novel drug targets for the treatment of diseases associated with dysregulation of AVP-mediated water reabsorption. IMCD cells are obtained from rat renal inner medullae and are used for experiments six to eight days after seeding. IMCD cells can be cultured in regular cell culture dishes, flasks and micro-titer plates of different formats, the procedure only requires a few hours, and is appropriate for standard cell culture laboratories.

Arginine-vasopressin (AVP) controls fine-tuning of body water homeostasis through facilitating water reabsorption by renal principal cells. Here, we present a protocol for the cultivation of primary rat inner medullary collecting duct cells suitable for the elucidation of molecular mechanisms underlying AVP-mediated water reabsorption.

El cultivo primarios de rata medular interna recolección de células del conducto

Arginine-vasopressin (AVP) facilitates water reabsorption by renal collecting duct principal cells and thereby fine-tunes body water homeostasis. AVP ...

Induction and Analysis of Epithelial to Mesenchymal Transition

Epithelial to mesenchymal transition (EMT) is essential for proper morphogenesis during development. Misregulation of this process has been implicated as a key event in fibrosis and the progression of carcinomas to a metastatic state. Understanding the processes that underlie EMT is imperative for the early diagnosis and clinical control of these disease states. Reliable induction of EMT in vitro is a useful tool for drug discovery as well as to identify common gene expression signatures for diagnostic purposes. Here we demonstrate a straightforward method for the induction of EMT in a variety of cell types. Methods for the analysis of cells pre- and post-EMT induction by immunocytochemistry are also included. Additionally, we demonstrate the effectiveness of this method through antibody-based array analysis and migration/invasion assays.

A straightforward method is described for the induction of epithelial to mesenchymal transition (EMT) in a variety of cell types. Methods for the analysis of cells in EMT states by immunocytochemistry are included.

Inducción y Análisis de epitelial a mesenquimal de transición

Epithelial  to mesenchymal transition (EMT) is essential for proper morphogenesis during  development. Misregulation of this  process has been implicated ...

Isolation of Human Hepatocytes by a Two-step Collagenase Perfusion Procedure

The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. As such, hepatocytes are an important model for a large variety of research questions. In particular, the use of human hepatocytes is especially important in the fields of pharmacokinetics, toxicology, liver regeneration and translational research. Thus, this method presents a modified version of a two-step collagenase perfusion procedure to isolate hepatocytes as described by Seglen 1.
Previously, hepatocytes have been isolated by mechanical methods. However, enzymatic methods have been shown to be superior as hepatocytes retain their structural integrity and function after isolation. This method presented here adapts the method designed previously for rat livers to human liver pieces and results in a large yield of hepatocytes with a viability of 77&plusmn;10%. The main difference in this procedure is the process of cannulization of the blood vessels. Further, the method described here can also be applied to livers from other species with comparable liver or blood vessel sizes.

A modified two-step collagenase perfusion procedure for isolation of human hepatocytes is described. This method can also be applied to other mammalian livers. The isolated hepatocytes are available in high yield and viability, making them a suitable model for scientific research in areas such as liver regeneration, pharmacokinetics and toxicology.

El aislamiento de hepatocitos humanos por una colagenasa perfusión Procedimiento de dos pasos

The liver, an organ with an exceptional regeneration capacity, carries out a wide range of functions, such as detoxification, metabolism and homeostasis. ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) maintain the turnover of all mature blood cell lineages. The coordination of the complex signals leading to specific HSC fates relies upon the interaction between HSCs and the intricate bone marrow microenvironment, which is still poorly understood[1-2].
We describe how by combining a newly developed specimen holder for stable animal positioning with multi-step confocal and two-photon in vivo imaging techniques, it is possible to obtain high-resolution 3D stacks containing HSPCs and their surrounding niches and to monitor them over time through multi-point time-lapse imaging. High definition imaging allows detecting ex vivo labeled hematopoietic stem and progenitor cells (HSPCs) residing within the bone marrow. Moreover, multi-point time-lapse 3D imaging, obtained with faster acquisition settings, provides accurate information about HSPC movement and the reciprocal interactions between HSPCs and stroma cells.
Tracking of HSPCs in relation to GFP positive osteoblastic cells is shown as an exemplary application of this method. This technique can be utilized to track any appropriately labeled hematopoietic or stromal cell of interest within the mouse calvarium bone marrow space.

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

The nature of the interactions between hematopoietic stem and progenitor cells (HSPCs) and bone marrow niches is poorly understood. Custom hardware modifications and a multi-step acquisition protocol allow the use of two-photon and confocal microscopy to image ex vivo labeled HSPCs homed within bone marrow areas, tracking interactions and movement.

En Vivo Tracking 4-Dimensional de madre hematopoyéticas y células progenitoras en ratones adultos de calota de Médula Ósea

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Fecal Glucocorticoid Analysis: Non-invasive Adrenal Monitoring in Equids

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, corticotrophin releasing hormone (CRH) is released from the hypothalamus in the brain. This stimulates the release of adrenocorticotrophic hormone (ACTH) from the pituitary gland, which in turn stimulates release of glucocorticoids from the adrenal gland. In horses the glucocorticoid corticosterone is responsible for several adaptations needed to support equine flight behaviour and subsequent removal from the aversive situation. Corticosterone metabolites can be detected in the feces of horses and assessment offers a non-invasive option to evaluate long term patterns of adrenal activity. Fecal assessment offers advantages over other techniques that monitor adrenal activity including blood plasma and saliva analysis. The non-invasive nature of the method avoids sampling stress which can confound results. It also allows the opportunity for repeated sampling over time and is ideal for studies in free ranging horses. This protocol describes the enzyme linked immunoassay (EIA) used to assess feces for corticosterone, in addition to the associated biochemical validation.

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. The method offers a non-invasive option to assess long term patterns in both domestic and free ranging horses. This protocol describes the enzyme linked immunoassay involved and the associated biochemical validation.

Análisis de glucocorticoides fecales: Monitoreo suprarrenal no invasiva en los équidos

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, ...

Partial Bile Duct Ligation in the Mouse: A Controlled Model of Localized Obstructive Cholestasis

In rodents, complete bile duct ligation (cBDL) of the common bile duct is an established surgical technique for studying obstructive cholestasis and bile duct proliferation. However, long-term experiments can lead to increased morbidity and mortality. In select mouse strains with underlying liver disease, meaningful comparisons can be made even with ligation of a single lobe of the liver, which can reduce animal losses and expenses. Here, we describe partial bile duct ligation (pBDL) in the mouse, in which only the left hepatic bile duct is ligated, causing biliary obstruction in the left lobe but not the remaining lobes. With careful microsurgical technique, pBDL experiments can be cost-effective, since the unligated lobe serves as an internal control to the ligated lobes, when subjected to the same conditions in the same animal. Unlike cBDL, a separate sham-operated control group is not necessary. pBDL is highly useful to directly compare localized versus systemic effects of cholestasis and other retained bile components. pBDL can also be repurposed as a novel method to investigate mechanisms related to medications and cell migration.

Here, we present partial bile duct ligation as a surgical model of liver injury and regeneration in rodents.

Ligadura del conducto biliar parcial en el ratón: un modelo controlado de la colestasis obstructiva localizada

In rodents, complete bile duct ligation (cBDL) of the common bile duct is an established surgical technique for studying obstructive cholestasis and bile ...