Questo lavoro è stato sostenuto in parte da un finanziamento della American Society otologico di CAM e da una sovvenzione dalla sentenza University Hospitals Medical Center a KNA e CAM. Vorremmo ringraziare Jeff Churney a Focus Medical Group, Inc. (Hinckley, OH) per averci fornito una fotocamera microscopio. Servizi di produzione video per gentile concessione di marketing creativo Fusion, Inc. (Valdosta, GA) Ringraziamo Jonathan Frankel e Alex Levitt a CWRU per i loro commenti durante la preparazione del video.

Parte 1: Anestesia e preparazione per la chirurgia <ol><li> Un adulto, di sesso femminile Duncan-Hartley cavia da Charles River International Laboratories, Inc. (Wilmington, MA) è anestetizzato con ketamina 60 mg / kg e xilazina 12 mg / kg per via intramuscolare. </li><li> Lubrificante petrolio viene applicato agli occhi per proteggere l&#39;animale da lesione corneale a causa di essiccazione. </li><li> L&#39;animale è rasata dalla cima della testa alle spalle e tra le orecchie. </li><li> L&#39;area rasata viene preparato in maniera sterile prima con 7,5% di iodio-povidone seguita del 70% di alcool etilico. </li><li> L&#39;animale viene posto su un tavolo operatorio con coperta riscaldamento e su una piattaforma che eleva la testa e si flette il collo. La testa dell&#39;animale è diretta dal chirurgo. Flessione del collo adeguata è necessaria per esporre adeguatamente la cresta occipitale. </li><li> L&#39;animale è avvolta in un modo sterile. </li></ol> 2 parti: Incisione ed esposizione dell&#39;occipite ossea <ol><li> Bupivacaina epinefrina più (Marcaine) 0,25% è applicato lungo il sito di incisione prospettico (linea mediana incisione cutanea dorsale lungo la cresta occipitale). Questo provoca vasocostrizione locale, limitando sanguinamento, e fornisce duraturo analgesia post-operatoria dopo che l&#39;animale si sveglia. </li><li> Incisione cutanea mediana dorsale è fatta sul cuoio capelluto lungo all&#39;occipite. <img src="/files/ftp_upload/1728/1728fig1.jpg" alt="Figura 1" /> Figura 1: incisione cutanea è fatta lungo la linea mediana dorsale, come mostrato sopra. </li><li> Per ottenere un&#39;adeguata esposizione, la pelle è retratto lateralmente mettendo un hemostat piccola curva su entrambi i lati dell&#39;incisione. </li><li> Una incisione orizzontale è fatto lungo il muscolo del collo per esporre all&#39;occipite ossea. In questa fase l&#39;animale può contrazione, nonostante l&#39;anestesia adeguata a causa di resezione del nervo grande occipitale. Transezione midollo spinale è un pericolo con incisione orizzontale nella regione suboccipitale. Questa complicanza può essere evitata facendo in modo che l&#39;osso sottostante è sempre sentito sotto il bisturi. </li><li> Con la muscolatura suboccipitale incise fino all&#39;osso, un hemostat terzo è usata per ritrarre il bordo superiore del muscolo. Questo hemostat è tenuto in posizione da l&#39;assistente chirurgico. </li><li> Un ascensore Lempert periostale viene utilizzato per esporre l&#39;articolazione temporo-occipitale sutura. </li></ol> Parte 3: Foratura temporo-occipitale sutura <ol><li> Al momento dell&#39;esposizione della sutura temporo-occipitale (Figura 2), un trapano otologica con 1,5 millimetri diamante radica è usato per esercitarsi inferio-lateralmente alla sutura. </li><li> Sotto la tabella esterna del cranio, la consistenza spugnosa del diploë (L&#39;osso spugnoso intervenendo tra i tavoli del cranio) diventerà visibile. Una volta che il diploë è stato perforato via, la tabella interna del cranio appare, insieme a una tonalità tenue blu che è il seno sigmoideo. </li><li> I margini del seno sigmoideo sono definiti senza penetrare la nave. </li><li> Una volta che l&#39;osso sovrastante il seno sigmoideo è stato sufficientemente assottigliato, più alto ingrandimento viene utilizzata e un pick rettilineo viene utilizzato per rimuovere con attenzione l&#39;osso sovrastante il seno sigmoideo. </li></ol><img src="/files/ftp_upload/1728/1728fig2.jpg" alt="Figura 2" /> Figura 2 L&#39;area da forare in parte si sovrappone la sutura temporo-occipitale. Parte 4: ostruzione del dotto <ol><li> Il seno sigmoideo scheletrato è retratto medialmente a rivelare l&#39;opercolo. </li><li> Un mm 0,5 diamante radica viene utilizzato per perforare l&#39;opercolo mediale e nel sacco endolinfatico e condotto. </li><li> Il dotto endolinfatico è poi ricco di cera ossea con un pick diritto. </li><li> Il difetto del cranio viene poi rinforzata con Gelfoam (Pfizer, New York, NY). </li><li> 4-0 Polysorb (United States Surgical, Norwalk, CT) sutura è utilizzato per portare il muscolo insieme. Vi è poi una transizione senza soluzione di continuità ad una tecnica di sutura sottocutanea per chiudere l&#39;incisione cutanea. Con questa chiusura non è necessario rimuovere punti di sutura dopo e non vi è alcun rischio che l&#39;animale artiglio sul sito e tirare fuori una sutura. </li></ol> Parte 5: assistenza post-operatoria La cavia viene messa in un incubatore per mantenere la temperatura corporea e controllati ogni 30 minuti fino a quando sveglio e in grado di mantenere una posizione sternale. L&#39;animale è ispezionato per determinare il tasso di respirazione e il colore del corpo. Questo centro usa enrofloxacina (0,2 ml a 22,7 mg / ml, per via sottocutanea) e la buprenorfina (0,05 ml a 0.3mg/mL, per via sottocutanea) per la profilassi post-operatoria l&#39;infezione e il controllo del dolore, rispettivamente. Analgesico è somministrato una volta al giorno dell&#39;intervento e due volte in giornata post-operatoria uno e due. Sebbene gli antibiotici non sono necessari per su asettichergery, cinque giorni, due volte al giorno di corso la profilassi antibiotica può essere givenonce post-operatoria di tre giorni. Complicanze e gestione <ol><li> Depressione respiratoria Il chirurgo deve sempre essere consapevoli della respirazione degli animali s. Se la respirazione rallenta o si ferma, interrompere l&#39;intervento chirurgico e ispezionare visivamente il movimento del torace. Se l&#39;animale è in difficoltà respiratoria o respiro affannoso, controllare che la bocca e le vie aeree non sono occluse. </li><li> Sigma penetrazione Gelfoam Usa e la compressione del seno con la punta di aspirazione fino a quando il sanguinamento è fermato. </li><li> Penetrazione durale coprire il difetto con Gelfoam. </li><li> Danno vestibolare Non praticare più di 1 mm lateralmente al seno sigmoideo. In questo modo rischi di ferire il canale semicircolare posteriore, tali da inficiare tutti i dati per la cavia. </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>

Gli esperimenti sugli animali sono stati eseguiti secondo le linee guida e regolamenti stabiliti dalla cura degli animali e del Comitato Istituzionale Usa presso la Case Western Reserve University.

Nelle settimane successive ostruzione chirurgica del dotto endolinfatico, il soggetto animale esperienze fluttuante perdita uditiva neurosensoriale misurata come risposta evocati uditivi del tronco encefalico. Un campione di risposta uditivi del tronco encefalico (ABR), la registrazione è mostrata in Figura 3. L&#39;orecchio sinistro serve come controllo non operati. <a href="https://www.jove.com/files/ftp_upload/1728/1728fig4.jpg"><img border="0" src="/files/ftp_upload/1728/1728fig3tmb.jpg" alt="Figura 3" /></a...

surgical induction of endolymphatic hydrops by obliteration of the endolymphatic duct

Induzione chirurgica del idrope endolinfatico (ELH) nella cavia da obliterazione e ostruzione del dotto endolinfatico è ben accettato modello animale della malattia e un importante correlazione per le malattie umane di Meniere. Nel 1965, Robert Kimura e Harold Schuknecht descrisse per primo un approccio intradurale per ostruzione del dotto endolinfatico (Kimura, 1965). Anche se efficace, questa tecnica, che richiede la penetrazione del rivestimento di protezione del cervello, sostenute a livello indesiderabili di morbilità e mortalità nei soggetti animali. Di conseguenza, Andrews e Bohmer sviluppato un approccio extradurale, che produce prevedibile un minor numero di complicanze associate del sistema nervoso centrale (SNC) penetrazione. (Andrews e Bohmer 1989) L&#39;approccio extradurale qui descritto richiede prima una incisione sulla linea mediana nella regione dell&#39;occipite per esporre lo strato muscolare sottostante. Operiamo solo sul lato destro. Dopo la ritrattazione appropriato del tessuto sovrastante, una incisione orizzontale è fatto nella muscolatura del occipite diritto di esporre il diritto temporo-occipitale linea di sutura. L&#39;osso immediatamente inferio-laterali la linea di sutura (Fig. 1) viene poi forato con un trapano otologica fino a quando il seno sigmoideo diventa visibile. Retrazione mediale del seno sigmoideo rivela l&#39;opercolo del condotto endolinfatico, che ospita il sacco endolinfatico. Foratura mediale per l&#39;opercolo nell&#39;area del sacco endolinfatico rivela il dotto endolinfatico, che viene poi confezionato con cera osseo di produrre l&#39;ostruzione e, in definitiva ELH. Nelle settimane successive, l&#39;animale sarà dimostrare la progressiva perdita dell&#39;udito e fluttuante evidenza istologica di 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

Questo video mostra come per ostacolare chirurgicamente condotto endolinfatico la cavia di produrre idrope endolinfatico.

Induzione chirurgica di Idrope endolinfatico da Obliterazione del dotto endolinfatico

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 Views.  Case Western Reserve University. Questo video mostra come per ostacolare chirurgicamente condotto endolinfatico la cavia di produrre idrope endolinfatico.

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.

Imaging Effector cellule T di memoria nell&#39;orecchio dopo l&#39;induzione di DTH adottivi

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

Ricerca

Biologia

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.

Test di induzione neurale nell&#39;embrione Chick

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.

Microchirurgico Obliteration clip del Medio aneurisma cerebrale mediante valutazione del flusso intraoperatoria

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.

Isolamento delle cellule basali e sottomucosa cellule del dotto di Gland 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. ...

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.

Coltura primaria Rat midollare interna Raccolta cellule del dotto

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.

Induzione e Analisi dei epiteliale a Mesenchymal Transizione

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.

L&#39;isolamento di epatociti umani da un Two-step Collagenase perfusione procedura

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 inseguimento 4-dimensionale di emopoietiche staminali e progenitrici celle di topo adulto cranica di Midollo Osseo

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.

Fecale Analisi glucocorticoidi: non invasivo di monitoraggio surrenale in equidi

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.

Legatura del dotto biliare parziale nel topo: un modello radiocomandato di colestasi ostruttiva localizzata

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