Ce travail a été soutenu en partie par une subvention de l&#39;American Society otologique au CAM et par une subvention du Centre de l&#39;University Hospitals Case Medical KNA et CAM. Nous tenons à remercier Jeff Churney à Focus Medical Group, Inc (Hinckley, OH) pour nous avoir fourni une caméra microscope. Services de production vidéo est une gracieuseté de fusion créative Marketing, Inc (Valdosta) Nous remercions Jonathan Frankel et Alex Levitt au CWRU pour leurs commentaires lors de la préparation de la vidéo.

Partie 1: Anesthésie et préparation à la chirurgie <ol><li> Un adulte, femelle Duncan-Hartley de porc Guinée auprès de Charles River Laboratories international, Inc (Wilmington, MA) est anesthésié à la kétamine 60 mg / kg et de xylazine à 12 mg / kg par voie intramusculaire. </li><li> Lubrifiant Petroleum est appliqué sur les yeux afin de protéger l&#39;animal de lésion cornéenne due à la dessiccation. </li><li> L&#39;animal est rasé par le haut de la tête sur les épaules et entre les oreilles. </li><li> La zone rasée est préparée de façon stérile, d&#39;abord avec 7,5% de povidone-iode suivie par 70% d&#39;alcool éthylique. </li><li> L&#39;animal est placé sur une table d&#39;opération avec couverture chauffante et sur une plate-forme qui élève la tête et du cou fléchit. La tête de l&#39;animal est dirigé loin de l&#39;opérateur. La flexion de la nuque adéquate est requise afin de répondre adéquatement exposer la crête occipitale. </li><li> L&#39;animal est drapé d&#39;une façon stérile. </li></ol> Partie 2: L&#39;incision et l&#39;exposition de l&#39;occiput osseuse <ol><li> La bupivacaïne, plus d&#39;adrénaline (marcaïne) 0,25% est appliquée le long du site d&#39;incision prospective (ligne médiane dorsale incision de la peau le long de la protubérance occipitale). Cela provoque une vasoconstriction locale, limitant saignements, et fournit une durée analgésie postopératoire après que l&#39;animal se réveille. </li><li> Incision médiane peau dorsale est faite sur le cuir chevelu le long de la protubérance occipitale. <img src="/files/ftp_upload/1728/1728fig1.jpg" alt="Figure 1" /> Figure 1: L&#39;incision cutanée est faite le long de la ligne médiane dorsale, comme indiqué ci-dessus. </li><li> Pour obtenir une exposition adéquate, la peau est écarté en dehors en plaçant une petite pince hémostatique courbe sur chaque côté de l&#39;incision. </li><li> Une incision horizontale est pratiquée le long du muscle du cou pour exposer l&#39;occiput osseux. A ce stade, l&#39;animal peut contraction, malgré une anesthésie adéquate en raison de section transversale du nerf grand occipital. Section de la moelle épinière est un danger avec incision horizontale dans la région sous-occipitale. Cette complication peut être évitée en veillant à ce que l&#39;os sous-jacent est toujours senti sous le bistouri. </li><li> Avec la musculature sous-occipitale incisées à l&#39;os, un hémostatique troisième est utilisé pour rétracter le bord du muscle de qualité supérieure. Cette hémostatique est maintenu en place par l&#39;assistant en chirurgie. </li><li> Un Lempert rugine est utilisée pour exposer la suture temporo-occipitale. </li></ol> Partie 3: Forage de la suture temporo-occipitale <ol><li> Lors de l&#39;exposition de la suture temporo-occipital (figure 2), une perceuse otologiques avec 1,5 mm de bavures diamant est utilisée pour forer inferio-latéralement à la suture. </li><li> Ci-dessous la table externe du crâne, la texture spongieuse du diploé (L&#39;os spongieux intervenant entre les tables du crâne) va devenir visible. Une fois le diploé a été foré loin, la table interne du crâne apparaît, avec une teinte bleu pâle qui est le sinus sigmoïde. </li><li> Les marges du sinus sigmoïde sont définies sans pénétrer dans le navire. </li><li> Une fois l&#39;os recouvrant le sinus sigmoïde a été suffisamment éclaircis, plus fort grossissement est utilisée et un choix de droite est utilisé pour enlever soigneusement l&#39;os recouvrant le sinus sigmoïde. </li></ol><img src="/files/ftp_upload/1728/1728fig2.jpg" alt="Figure 2" /> Figure 2 La zone à être forés partie recouvre la suture temporo-occipitale. Partie 4: l&#39;obstruction du canal <ol><li> Le sinus sigmoïde squelette est rentré dedans pour révéler l&#39;opercule. </li><li> Un diamant de 0,5 mm bavure est utilisée pour forer dedans de l&#39;opercule et dans le sac endolymphatique et le canal. </li><li> Le canal endolymphatique est ensuite emballé avec de la cire d&#39;os à l&#39;aide un pick droite. </li><li> Le défaut crâne est ensuite renforcée avec Gelfoam (Pfizer, New York, NY). </li><li> 4-0 POLYSORB (United States Surgical, Norwalk, CT) de suture est utilisée pour amener le muscle ensemble. Il ya donc une transition en douceur à une technique de suture sous-cutanée pour la fermeture de l&#39;incision cutanée. Avec cette fermeture il n&#39;est pas nécessaire d&#39;enlever les sutures après et il n&#39;ya aucun risque que l&#39;animal griffe sur le site et tirez un fil de suture. </li></ol> Partie 5: Soins postopératoires Le cochon de Guinée est mis dans un incubateur pour maintenir la température du corps et vérifié à intervalles de 30 minutes jusqu&#39;à ce éveillé et capable de maintenir une position sternale. L&#39;animal est inspecté pour le taux de respiration et de la couleur du corps. Ce centre utilise l&#39;enrofloxacine (0,2 mL à 22,7 mg / ml, sous-cutanée) et la buprénorphine (0,05 ml à 0.3mg/mL, sous-cutanée) pour la prophylaxie des infections post-opératoires et contrôle de la douleur, respectivement. Analgésique est administré une fois le jour de la chirurgie et deux fois le jour post-opératoire un et deux. Bien que les antibiotiques ne sont pas nécessaires pour su aseptiquesrgery, une durée de cinq jours, deux fois par jour bien sûr des antibiotiques à titre prophylactique peut être givenonce sur le post-opératoire de trois jours. Complications et de gestion <ol><li> La dépression respiratoire Le chirurgien doit toujours être conscient de la respiration de l&#39;animal s. Si la respiration ralentit ou s&#39;arrête, arrêter la chirurgie et d&#39;inspecter visuellement le mouvement thoracique. Si l&#39;animal éprouve de la difficulté respiratoire ou une respiration laborieuse, vérifier que la bouche et des voies respiratoires ne sont pas obstruées. </li><li> Utilisez des sinus sigmoïde Gelfoam pénétration et de compression ainsi que la pointe d&#39;aspiration jusqu&#39;à ce saignement est arrêté. </li><li> Pénétration de Dural couvrir le défaut d&#39;Gelfoam. </li><li> Lésions vestibulaires Ne pas percer plus de 1 mm latéral du sinus sigmoïde. Cela risque en blessant le canal semi-circulaire postérieur, ce qui invaliderait toutes les données pour le cochon de Guinée. </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>

Expériences sur les animaux ont été effectuées en conformité avec les directives et les règlements énoncés par le soin des animaux et du Comité institutionnel Utilisez à la Case Western Reserve University.

Dans les semaines qui suivent l&#39;obstruction chirurgicale du canal endolymphatique, le sujet des expériences animales fluctuant perte auditive neurosensorielle telle que mesurée par la réponse évoquée auditive évoquée. Un échantillon de auditifs du tronc cérébral (ABR) l&#39;enregistrement est montré dans la figure 3. L&#39;oreille gauche sert de contrôle non opérés. <a href="https://www.jove.com/files/ftp_upload/1728/1728fig4.jpg"><img border="0" src="/files/ftp_upload/1728/1728fig3tmb.jpg" alt=...

surgical induction of endolymphatic hydrops by obliteration of the endolymphatic duct

L&#39;induction chirurgicale d&#39;hydrops endolymphatique (ELH) dans le cochon Guinée par l&#39;oblitération et l&#39;obstruction du canal endolymphatique est un modèle animal bien accepté de l&#39;état et une importante corrélation entre la maladie humaine de Ménière. En 1965, Robert Kimura et Harold Schuknecht décrite pour la première une approche intradurale d&#39;obstruction du canal endolymphatique (Kimura 1965). Bien qu&#39;efficace, cette technique, qui nécessite la pénétration du revêtement protecteur du cerveau, engagés un niveau indésirable de la morbidité et de mortalité chez les sujets animaux. Par conséquent, Andrews et Bohmer développé une approche extradural, qui produit moins de façon prévisible des complications associées au système nerveux central (SNC) la pénétration. (Andrews et Bohmer 1989) L&#39;approche décrite ici extradural exige d&#39;abord une incision médiane dans la région de l&#39;occiput pour exposer la couche musculaire sous-jacent. Nous intervenons uniquement sur le côté droit. Après rétraction du tissu appropriés sus-jacente, une incision horizontale est pratiquée dans la musculature de l&#39;occiput droit d&#39;exposer la ligne de suture temporo-occipital droit. Les os immédiatement inferio-latérale de la ligne de suture (Fig. 1) est ensuite percé avec une perceuse otologiques jusqu&#39;à ce que le sinus sigmoïde devient visible. Rétraction médiale du sinus sigmoïde révèle l&#39;opercule du conduit endolymphatique, qui abrite le sac endolymphatique. Forage en dedans de l&#39;opercule dans la région du sac endolymphatique révèle le canal endolymphatique, qui est ensuite emballée avec de la cire osseuse à produire une obstruction et finalement ELH. Dans les semaines suivantes, l&#39;animal montrent le développement progressif, la perte d&#39;audition fluctuante et la preuve histologique d&#39;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

Cette vidéo montre comment chirurgicalement obstruction conduit endolymphatique le cochon de Guinée pour produire hydrops endolymphatique.

Induction chirurgicale d&#39;hydrops endolymphatique par oblitération du canal endolymphatique

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

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11.3K vues.  Case Western Reserve University. Cette vidéo montre comment chirurgicalement obstruction conduit endolymphatique le cochon de Guinée pour produire hydrops endolymphatique.

Vidéo: 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.

Imagerie Les cellules T effectrices de mémoire dans l&#39;oreille après induction de SRD adoptifs

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

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Biologie

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.

Dosage de l&#39;induction neurale dans l&#39;embryon de poulet

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.

Oblitération clip microchirurgie de l&#39;anévrisme cérébral Moyen Utilisation de l&#39;évaluation de débit peropératoire

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.

Isolement de cellules basales et les cellules sous-muqueux du canal de la glande de la souris Trachée

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.

Culture primaire Rat collecteur médullaire interne les cellules des canaux

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.

Induction et analyse des épithéliale à mésenchymateuses transition

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.

Isolement des hépatocytes humains par un deux-étape collagénase procédure de perfusion

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.

In Vivo suivi 4-Dimensional de hématopoïétiques cellules souches et progénitrices adultes dans la voûte du crâne de souris de moelle osseuse

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.

Fecal glucocorticoïdes Analyse: Surveillance surrénale non invasive en Equidés

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.

La ligature partielle biliaires chez la souris : un modèle contrôlé de la cholestase Obstructive localisée

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