We are supported by research grants from the National Natural Science Foundation of China (NO. 81400590), National Institutes of Health (U54 CA156735) and Takeda Pharmaceutical Company Ltd. (MA-NC-D-156).

Tutti gli esperimenti sugli animali sono stati approvati dalla cura degli animali e Usa Comitato Istituzionale. 1. Preparazione del mouse <ol><li> Utilizzare i mouse sopra ~ 20 g di peso corporeo o superiori a ~ 6 settimane di età per la chirurgia. </li><li> Prima dell&#39;intervento, dare topi un laboratorio chow e acqua ad libitum e mantenere su un ciclo luce-buio 00:12 h. </li><li> Radere l&#39;area chirurgica con un tagliatore di capelli. Anestetizzare topi mediante iniezione intraperitoneale di 80 mg / kg di ketamina e 12 mg / kg xilazina. Questa dose mette topi nel sonno in pochi minuti e fornisce anestesia sufficienti per le seguenti procedure chirurgiche. </li><li> Confermare anestesia corretta basata sull&#39;assenza di riflesso corneale e l&#39;incolumità fisica retrazione quando la zampa viene pizzicato. Applicare lubrificante pomata oftalmica sugli occhi per prevenire la secchezza. </li><li> Sterilizzare pelle con la soluzione betadine. </li></ol> 2. reflusso gastrico Model (Figura 1B) <ol><li> Fare un superioreincisione mediana di ~ 2 cm a partire dal xifoide indicando l&#39;ano. </li><li> Aprire la cavità addominale attraverso la linea mediana. Rimuovere la xifoideo per migliorare l&#39;esposizione con un paio di forbici. </li><li> Separare e tagliare il tessuto connettivo tra il fegato e lo stomaco. Legare e tagliare il fascio nave tra la milza e il fondo, al fine di liberare completamente il fondo. </li><li> Ruotare il fondo leggermente a sinistra ed esporre il lato sinistro della giunzione gastroesofageo nel campo. Effettuare una incisione longitudinale 5 mm sul muscolo lungo l&#39;esofago distale usando un paio di forbici affilate per esporre l&#39;epitelio. </li><li> Tagliare aprire l&#39;epitelio lungo la stessa direzione (incisione 1, Figura 1B). Fare tre incisioni sulla il prestomaco (incisione 2, 3 e 4, Figura 1B) per tagliare più del prestomaco con le forbici operativi taglienti. Se lo stomaco non è vuoto, rimuovere con attenzione contenuto gastrico. </li><li> Inserire un Sutu 8-0 prolenere (con un punto conico ago) per il punto A (in esofago) e un punto &#39;(sulla il prestomaco) con una precisione di mucosa mucosa opposizione. Allo stesso modo luogo suture per il punto B e il punto B &#39;, e altre coppie di punti, rispettivamente. Uniformemente spazio queste suture 3-4 mm l&#39;una dall&#39;altra. </li><li> Lavare la cavità addominale con soluzione fisiologica per pulire sangue e contenuto gastrico. Chiudere la parete addominale con suture in seta e la pelle con clip metalliche. </li></ol> 3. Misto reflusso Modello (Figura 1C) <ol><li> Seguire i passaggi da 2.1 a 2.4 per esporre la giunzione gastroesofagea. </li><li> Separare delicatamente il lato dorsale dell&#39;esofago dai vasi sanguigni dietro l&#39;esofago. Passare una piccola punta di cotone tra i vasi sanguigni e dell&#39;esofago. Nota: Il cotone può essere parzialmente rimosso dalla punta per ridurne le dimensioni. Questa punta di cotone due scopi, sollevando l&#39;esofago e proteggere i vasi sanguigni. </li><li> Fai inc due 5 millimetri longitudinaleisions ciascuno sulla giunzione gastroesofageo e l&#39;estremità prossimale del duodeno adiacente al piloro con le forbici operativi taglienti. Per l&#39;incisione sul duodeno, evitare i vasi sanguigni e immettere sul confine anti-mesenterica. </li><li> Anastomose le incisioni con mucosa preciso per mucosa opposizione con interrotti 8-0 punti di sutura Prolene. Solitamente posizionare 3-4 sutura sul lato dorsale e 2-3 suture sul lato anteriore. </li><li> Rimuovere la punta di cotone. Lavare la cavità addominale con soluzione salina normale e chiudere la parete addominale e la pelle. </li></ol> 4. duodenale reflusso Modello (Figura 1D) <ol><li> Seguire il passaggio da 3.1 a 3.4 per generare reflusso misto. </li><li> Sollevare delicatamente lo stomaco per esporre il suo lato posteriore. Un lobo del fegato può essere catturato da tessuto connettivo tra il fegato e la parte posteriore dello stomaco. Asportare accuratamente i tessuti connettivi, adeguatamente proteggere il fegato, ed esporre i vasi sanguigni sul lato dorsale dell &#39;esofago. </li><li> Legare etagliare i vasi sanguigni. Legare e tagliare l&#39;esofago all&#39;incrocio gastroesofageo. </li><li> Legare duodeno al piloro. Legare e tagliare il mesentere. Rimuovere l&#39;intero stomaco. </li><li> Lavare la cavità addominale con soluzione fisiologica. Chiudere la parete addominale e la pelle. </li></ol> Il trattamento post-chirurgico 5. <ol><li> Dopo l&#39;intervento chirurgico mantenere la temperatura corporea con una piastra elettrica. Solo dopo aver ripreso decubito sternale, posizionare il mouse posteriore con altri animali. </li><li> Dare antibiotici e analgesici per prevenire l&#39;infezione e alleviare il dolore. Iniettare Baytril (10 mg / kg, ip, qd, per 3 giorni) per prevenire l&#39;infezione, e buprenorfina cloridrato (0,05 mg / kg, sc, offerta. Per due giorni) come antidolorifico. Il digiuno non è necessario dopo l&#39;intervento chirurgico. Una dieta liquida o morbido può essere dato. </li><li> Valutare lo stato di salute generale basis.If giornaliere topi mostrano i seguenti segni (drammatica perdita di peso&gt; 15%, la mancanza di appetite, vocalizzazione, scaricare dalla bocca, il naso o gli occhi, gesto curvo, l&#39;inattività, l&#39;attività anormale grooming), l&#39;eutanasia i topi. </li></ol>

<ol>
	<li>Kandulski, A., Malfertheiner, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastroesophageal+reflux+disease--from+reflux+episodes+to+mucosal+inflammation.">Gastroesophageal reflux disease--from reflux episodes to mucosal inflammation.</a> Nat Rev Gastroenterol Hepatol. 9 (1), 15-22 (2012).</li><li>Orlando, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+integrity+of+the+esophageal+mucosa.+Balance+between+offensive+and+defensive+mechanisms.">The integrity of the esophageal mucosa. Balance between offensive and defensive mechanisms.</a> Best Pract Res Clin Gastroenterol. 24 (6), 873-882 (2010).</li><li>Stairs, D. B., Kong, J., Lynch, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cdx+genes,+inflammation,+and+the+pathogenesis+of+intestinal+metaplasia.">Cdx genes, inflammation, and the pathogenesis of intestinal metaplasia.</a> Prog Mol Biol Transl Sci. 96, 231-270 (2010).</li><li>Spechler, S. J., Fitzgerald, R. C., Prasad, G. A., History Wang, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=molecular+mechanisms,+and+endoscopic+treatment+of+Barrett's+esophagus.">molecular mechanisms, and endoscopic treatment of Barrett's esophagus.</a> Gastroenterology. 138 (3), 854-869 (2010).</li><li>Fang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+origins+and+molecular+mechanisms+of+Barrett's+esophagus+and+esophageal+adenocarcinoma.">Cellular origins and molecular mechanisms of Barrett's esophagus and esophageal adenocarcinoma.</a> Ann N Y Acad Sci. 1300, 187-199 (2013).</li><li>Garman, K. S., Orlando, R. C., Chen, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Review:+Experimental+models+for+Barrett's+esophagus+and+esophageal+adenocarcinoma.">Review: Experimental models for Barrett's esophagus and esophageal adenocarcinoma.</a> Am J Physiol Gastrointest Liver Physiol. 302 (11), G1231-G1243 (2012).</li><li>Baruah, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Translational+research+on+Barrett's+esophagus.">Translational research on Barrett's esophagus.</a> Ann N Y Acad Sci. 1325, 170-186 (2014).</li><li>Lechpammer, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Flavopiridol+reduces+malignant+transformation+of+the+esophageal+mucosa+in+p27+knockout+mice.">Flavopiridol reduces malignant transformation of the esophageal mucosa in p27 knockout mice.</a> Oncogene. 24 (10), 1683-1688 (2005).</li><li>Hao, J., Liu, B., Yang, C. S., Chen, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastroesophageal+reflux+leads+to+esophageal+cancer+in+a+surgical+model+with+mice.">Gastroesophageal reflux leads to esophageal cancer in a surgical model with mice.</a> BMC Gastroenterol. 9, 59 (2009).</li><li>Mari, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+pSMAD/CDX2+complex+is+essential+for+the+intestinalization+of+epithelial+metaplasia.">A pSMAD/CDX2 complex is essential for the intestinalization of epithelial metaplasia.</a> Cell Rep. 7 (4), 1197-1210 (2014).</li><li>Fang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastroesophageal+reflux+activates+the+NF-kappaB+pathway+and+impairs+esophageal+barrier+function+in+mice.">Gastroesophageal reflux activates the NF-kappaB pathway and impairs esophageal barrier function in mice.</a> Am J Physiol Gastrointest Liver Physiol. 305 (1), G58-G65 (2013).</li><li>Chen, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nrf2+deficiency+impairs+the+barrier+function+of+mouse+oesophageal+epithelium.">Nrf2 deficiency impairs the barrier function of mouse oesophageal epithelium.</a> Gut. 63 (5), 711-719 (2014).</li></ol>

One of the authors (XC) received funding that was provided by Takeda Pharmaceutical Company Ltd which supports basic research associated with gastroesophageal reflux disease. None of the reagents or instruments used in this article is produced by this company.

Vari modelli chirurgici sono stati stabiliti per imitare gastrica, duodenale e reflusso misto nei roditori. Queste tre procedure descritte sono adatte per esperimenti a lungo termine con tassi ragionevoli di sopravvivenza postoperatoria. Un ricercatore con esperienza chirurgica dovrebbe essere in grado di cogliere la tecnica, dopo un breve periodo di pratica. Il sanguinamento può derivare da iniezione intraperitoneale di anestetici prima dell&#39;intervento chirurgico, la lacerazione del fe...

Gastroesophageal reflux disease (GERD) is a chronic disorder caused by the prolonged exposure of distal esophagus to gastric or gastroduodenal contents1. Prolonged exposure to these noxious refluxates impairs the intrinsic defenses within the esophageal epithelium and thus results in esophagitis2. Barrett&#8217;s esophagus arises in the setting of chronic reflux, and is a premalignant lesion with increased risk of esophageal adenocarcinoma3,4. Despite the clinical importance, the mechanisms of GERD, Barrett&#8217;s esophagus and adenocarcinoma have not been well understood. 
Animal models are essential for research on etiology, pathology, molecular mechanisms, prevention and treatment of human diseases. Up to date, various animal models of GERD, Barrett&#8217;s esophagus and adenocarcinoma have been developed using model animals5,6. Mouse esophagus is lined with stratified squamous epithelium which is histologically similar to that in human esophagus. Although a mouse esophagus is different from human esophagus in terms of keratinization and the absence of submucosal glands, the mouse is still an appealing model animal because of its relatively low cost of maintenance and its potential of sophisticated genetic modifications. Two approaches are commonly used to model GERD, Barrett&#8217;s esophagus and adenocarcinoma in mice: reflux surgery and genetic modification. Reflux surgery is the best way to induce reflux and genetic modifications mimics molecular alterations5,7. Reflux surgery can be combined with genetic modifications to further understand disease mechanisms8. 
Many surgical procedures have been reported by us and others6,9: (1) gastric reflux: pyloric ligation, pyloric constriction with forestomach ligation, Wendel cardioplasty, and esophagogastric anastomosis; (2) mixed reflux: esophagogastroduodenal anastomosis, esophagoduodenostomy (or esophagojejunostomy); (3) duodenal reflux: esophagogastroduodenal anastomosis plus gastrectomy; (4) reflux of chemical components: bilious reflux, pancreatic reflux, esophageal perfusion; and (5) esophageal transplantation5. Recently a microsurgical mouse model was reported to produce jejunal reflux via an esophagojejunostomy with magnets10. These surgical models have advantages over in vitro cell culture or organotypic culture models. In vitro, esophageal cells cannot tolerate a medium with high acidity or high concentrations of bile acids. Unconjugated bile acids which are commonly used to produce changes in esophageal epithelial cells in vitro are usually not present in the duodenal refluxate in vivo. Thus conclusions drawn from such in vitro studies should be taken with caution.
Surgery on the mouse esophagus remains a technical challenge because of its small size. A low rate of postoperative survival does not allow experiments which require certain sample size to reach statistically sound conclusions. In the past we have successfully developed and characterized surgical models of gastric reflux, mixed reflux, duodenal reflux with mice in long-term experiments9,11,12. We have also provided consultation to several other groups in their mouse surgery. Herein, we describe three surgical procedures in mice in order to help the community to establish these models in their labs.

<table><tbody><tr><td>Dumont #1 Forceps Dumostar Tip&amp;nbsp;</td><td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td><td></td><td></td></tr><tr><td>Micro Clip Applying Forceps 5.5&quot;</td><td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td><td></td><td></td></tr><tr><td>Bonn Scissors 3.5&quot; Straight 15 mm Sharp/Sharp Tungsten Carbide Blades</td><td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td><td></td><td></td></tr><tr><td>Operating Scissors 5.5&quot; Straight Sharp/Sharp SureCut</td><td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td><td></td><td></td></tr><tr><td>4-0 Silk Black Braid 100 Yard Spool</td><td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td><td></td><td></td></tr><tr><td>Surgeon&#039;s Needle 1/2 Circle Cutting Edge Size 12 (25 mm Chord Length) Pack 12</td><td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td><td></td><td></td></tr><tr><td>Halsey Needle Holder 5&quot; Smooth</td><td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td><td></td><td></td></tr><tr><td>Micro Needle Holder 5.125&quot; Curved Lock .6 mm</td><td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td><td></td><td></td></tr><tr><td>Reflex 9 mm Wound Clip Applier</td><td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td><td></td><td></td></tr><tr><td>Reflex 9 mm Wound Clips Box Of 100</td><td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td><td></td><td></td></tr><tr><td>PRONOVA Poly (hexafluoropropylene-VDF) Suture 8-0</td><td>Ethicon US, LLC</td><td></td><td></td></tr><tr><td>Ringer&#039;s solution</td><td>Henry Schein, Inc.</td><td></td><td></td></tr><tr><td>ketamine</td><td>Henry Schein, Inc.</td><td></td><td></td></tr><tr><td>xylazine</td><td>Henry Schein, Inc.</td><td></td><td></td></tr></tbody></table>

surgical models of gastroesophageal reflux with mice

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to induce reflux of gastric contents, duodenal contents or mixed contents. Surgical procedures and general principles have been described in detail. A researcher with surgical experience should be able to grasp the technique after a short period of practice. After surgery, most mice can survive and develop reflux esophagitis similar to that in humans. However, it should be noted that histological differences between mouse and human esophagus are the inherent limitations of these surgical models. If used for research on Barrett&#8217;s esophagus and adenocarcinoma, these procedures may need to be combined with genetic modifications.

La maggior parte dei topi (&gt; 95%) possono sopravvivere chirurgia. Durante il periodo perioperatorio, le principali cause di morte includono sovradosaggio di anestetici, sanguinamento e motivi sconosciuti. Quattro settimane dopo l&#39;intervento,&gt; 90% topi con reflusso gastrico o reflusso misto e&gt; 80% topi con reflusso duodeno possono sopravvivere. Durante questo periodo, i topi muoiono principalmente da stenosi esofagea e l&#39;incapacità di mangiare. Questi topi mostrano segni di ...

Watch this Scientific Journal Video about Surgical Models of Gastroesophageal Reflux with Mice at JoVE.com

Surgical Models of Gastroesophageal Reflux with Mice

This article demonstrates surgical procedures of gastroesophageal reflux with mice. These models are useful tools for research on mechanisms and treatment of gastroesophageal reflux disease and potentially Barrett&#8217;s esophagus and esophageal adenocarcinoma.

I modelli chirurgici di reflusso gastroesofageo con Mice

Multiple surgical procedures have been reported to induce gastroesophageal reflux in animals. Herein, we report three surgical models with mice aiming to ...

surgical-models-of-gastroesophageal-reflux-with-mice

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

10.1K Views.  Ningxia Medical University General Hospital. This article demonstrates surgical procedures of gastroesophageal reflux with mice. These models are useful tools for research on mechanisms and treatment of gastroesophageal reflux disease and potentially Barrett’s esophagus and esophageal adenocarcinoma.

Video: Surgical Models of Gastroesophageal Reflux with Mice

Non-invasive Assessment of the Efficacy of New Therapeutics for Intestinal Pathologies Using Serial Endoscopic Imaging of Live Mice

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic mechanisms that contribute to the onset and progression of intestinal diseases. The identification of new molecules that promote these pathologies has led to a flurry of activity focused on the development of potential new therapies to inhibit their function. As a result, various pre-clinical mouse models with an intact immune system and stromal microenvironment are now heavily used. Here we describe three experimental protocols to test the efficacy of new therapeutics in pre-clinical models of (1) acute mucosal damage, (2) chronic colitis and/or colitis-associated colon cancer, and (3) sporadic colorectal cancer. We also outline procedures for serial endoscopic examination that can be used to document the therapeutic response of an individual tumor and to monitor the health of individual mice. These protocols provide complementary experimental platforms to test the effectiveness of therapeutic compounds shown to be well tolerated by mice.

We describe methods for longitudinal monitoring of the efficacy of therapeutics for the treatment of colonic pathologies in mice using a rigid endoscope. This protocol can be readily used for the characterization of the therapeutic response of an individual tumor in live mice and also for monitoring potential disease relapse.

Valutazione non invasiva della efficacia di nuove terapie per patologie intestinali Utilizzando Serial Imaging endoscopica Live Mice

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic ...

Ricerca

Medicina

Mouse Models Of Helicobacter Infection And Gastric Pathologies

Helicobacter pylori is a gastric pathogen that is present in half of the global population and is a significant cause of morbidity and mortality in humans. Several mouse models of gastric Helicobacter infection have been developed to study the molecular and cellular mechanisms whereby H. pylori bacteria colonize the stomach of human hosts and cause disease. Herein, we describe protocols to: 1) prepare bacterial suspensions for the in vivo infection of mice via intragastric gavage; 2) determine bacterial colonization levels in mouse gastric tissues, by polymerase chain reaction (PCR) and viable counting; and 3) assess pathological changes, by histology. To establish Helicobacter infection in mice, specific pathogen-free (SPF) animals are first inoculated with suspensions (containing &#8805;105 colony-forming units, CFUs) of mouse-colonizing strains of either Helicobacter pylori or other gastric Helicobacter spp. from animals, such as Helicobacter felis. At the appropriate time-points post-infection, stomachs are excised and dissected sagittally into two equal tissue fragments, each comprising the antrum and body regions. One of these fragments is then used for either viable counting or DNA extraction, while the other is subjected to histological processing. Bacterial colonization and histopathological changes in the stomach may be assessed routinely in gastric tissue sections stained with Warthin-Starry, Giemsa or Haematoxylin and Eosin (H&#38;E) stains, as appropriate. Additional immunological analyses may also be undertaken by immunohistochemistry or immunofluorescence on mouse gastric tissue sections. The protocols described below are specifically designed to enable the assessment in mice of gastric pathologies resembling those in human-related H. pylori diseases, including inflammation, gland atrophy and lymphoid follicle formation. The inoculum preparation and intragastric gavage protocols may also be adapted to study the pathogenesis of other enteric human pathogens that colonize mice, such as Salmonella Typhimurium or Citrobacter rodentium.

Mouse Models Of Helicobacter Infection And Gastric Pathologies

Mice represent an invaluable in vivo model to study infection and diseases caused by gastrointestinal microorganisms. Here, we describe the methods used to study bacterial colonization and histopathological changes in mouse models of Helicobacter pylori-related disease.

Modelli murini di patologie gastriche e infezione da Helicobacter

Helicobacter pylori is a gastric pathogen that is present in half of the global population and is a significant cause of morbidity and mortality in ...

Immunologia e infezioni

Techniques of Sleeve Gastrectomy and Modified Roux-en-Y Gastric Bypass in Mice

Obesity is a major public health issue, with a prevalence of 4 to 28% for men and 6.2 to 36.5% for women in Europe (from 2003 to 2008). Morbid obesity is frequently associated with metabolic complications, such as type 2 diabetes, hypertension, and dyslipidemia, reducing life expectancy and quality. In the absence of any effective noninvasive treatments, bariatric surgery is a valuable therapeutic option for patients with morbid obesity (body mass index (BMI) &#62;40 kg/m2), leading to long-term, sustained weight loss and improvements in metabolic complications. However, the underlying cellular and molecular mechanisms sustaining the beneficial effects of bariatric surgery are not yet fully understood. Due to the numerous genetically-modified strains available, the mouse model is the most convenient animal model to explore the molecular mechanisms behind the pleiotropic beneficial effects of bariatric surgeries. Here, we detailed the optimized healthcare methods and surgical protocols in mice for the two most widely-used bariatric surgeries: the sleeve gastrectomy and the modified Roux-en-Y gastric bypass. Deciphering the molecular mechanisms underlying the therapeutic effects of bariatric surgeries offers the promise of identifying new therapeutics targets.

Bariatric surgery is the most efficient way to reduce body weight and the deadly metabolic complications (diabetes, obesity, and dyslipidemia) frequently associated with morbid obesity. Mouse models of bariatric surgery represent a unique asset for deciphering molecular mechanisms behind the beneficial effects of these surgeries on diabetes, hypertension, and dyslipidemia.

Tecniche di sleeve gastrectomy e modificata Roux-en-Y bypass gastrico nei topi

Obesity is a major public health issue, with a prevalence of 4 to 28% for men and 6.2 to 36.5% for women in Europe (from 2003 to 2008). Morbid obesity is ...

Creation of Colonic Anastomosis in Mice

Intestinal anastomoses are commonly performed in both elective and emergent operations. Even so, anastomotic leaks are a highly feared complications of colonic surgeries and can occur in up to 26% of surgical anastomoses, with mortality being up to 39% for patients with such a leak. Currently, there remains a paucity of data detailing the cellular mechanisms of anastomotic healing. Devising preventative strategies and treatment modalities for anastomotic leak could be greatly potentiated by a better understanding of appropriate anastomotic healing.
A murine model is ideal as previous studies have shown that the murine anastomosis is the most clinically similar to the human case as compared with other animal models. We offer an easily reproducible murine model of colonic anastomosis in mice that will allow for further illustration of anastomotic healing.

Anastomotic leak or breakdown after surgery is a major cause of postoperative morbidity and mortality. Our surgery for creating a colonic anastomosis is a reliable and reproducible method for studying the healing mechanisms of anastomoses.

Creazione dell'anastomosi colica in topi

Intestinal anastomoses are commonly performed in both elective and emergent operations. Even so, anastomotic leaks are a highly feared complications of ...

Sleeve Gastrectomy in Mice using Surgical Clips

The number of people who are overweight and obese is continually increasing both in the adult and adolescent populations. This coincides with the increased universal phenomenon of type 2 diabetes (T2D) and other metabolic problems. Bariatric surgery, such as SG, is currently one of the most effective and commonly used long-term treatment for obesity and T2D, but the association between them is not completely explored yet. The mechanisms underlying the outcomes seen after bariatric surgery in humans can be investigated based on preclinical animal studies. The SG reduces body weight, glucose levels and many metabolic parameters, and is easy to perform with a low incidence of complications. The goal of this work is to provide a simple method and an uncomplicated preclinical model of bariatric surgery in animals for researchers.

The prevalence of diabetes and obesity is continuously increasing worldwide. The mechanisms between diabetes, obesity and their associated mortality and co-morbidities need to be further investigated. Here, we present a protocol for sleeve gastrectomy (SG) in animals as an uncomplicated preclinical model of bariatric surgery.

Gastrectomia a maniche nei topi con clip chirurgiche

The number of people who are overweight and obese is continually increasing both in the adult and adolescent populations. This coincides with the ...

Single-Anastomosis Duodeno-Ileal Bypass with Sleeve Gastrectomy Model in Mice

Obesity is a major health issue worldwide. As a response, bariatric surgeries have emerged to treat obesity and its related comorbidities (e.g., diabetes mellitus, dyslipidemia, non-alcoholic steatohepatitis, cardiovascular events, and cancers) through restrictive and malabsorptive mechanisms. Understanding the mechanisms by which these procedures allow such improvements often require their transposition into animals, especially in mice, because of the ease of generating genetically modified animals. Recently, the single-anastomosis duodeno-ileal bypass with sleeve gastrectomy (SADI-S) has emerged as a procedure that uses both restrictive and malabsorptive effects, which is being used as an alternative to gastric bypass in case of major obesity. Thus far, this procedure has been associated with strong metabolic improvements, which has led to a marked increase in its use in daily clinical practice. However, the mechanisms underlying these metabolic effects have been poorly studied as a result of a lack of animal models. In this article, we present a reliable and reproducible model of SADI-S in mice, with a special focus on perioperative management. The description and use of this new rodent model will be helpful for the scientific community to better understand the molecular, metabolic, and structural changes induced by the SADI-S and to better define the surgical indications for clinical practice.

Single-anastomosis duodeno-ileal bypass (SADI-S) is an emerging bariatric procedure with important metabolic effects. In this article, we present a reliable and reproducible model of SADI-S in mice.

Bypass duodeno-ileale a singola anastomosi con modello di gastrectomia a manica nei topi

Obesity is a major health issue worldwide. As a response, bariatric surgeries have emerged to treat obesity and its related comorbidities (e.g., diabetes ...

A Murine Model of Vertical Sleeve Gastrectomy

Bariatric surgery, such as vertical sleeve gastrectomy (VSG), is a surgery of the gastrointestinal tract that is performed for the purpose of weight loss. Bariatric surgery is currently the most effective long-term treatment for obesity. In addition to weight loss, bariatric surgery produces additional health benefits such as remission of type 2 diabetes, remission of hypertension, and decreased risk of developing certain types of cancer. The mechanisms beyond weight loss for these benefits remain incompletely defined. Therefore, animal models of bariatric surgery are being developed and validated to identify the mechanisms leading to these benefits, with the goal of improving understanding of gastrointestinal physiology and identifying new therapeutic targets. VSG has become the most commonly performed bariatric procedure in the clinic in the United States because it is highly effective at producing weight loss and metabolic improvement, and is simpler to perform than other bariatric procedures. Therefore, we have developed and validated a murine model of VSG. This murine VSG model recapitulates many of the effects of VSG seen in humans, including improved glucose and blood pressure regulation. The method is based on isolation of the stomach, ligation of gastric vessels, and removal of 70% of the stomach by transecting along the greater curvature of the stomach. We have successfully applied this surgical protocol to various genetically modified mouse lines to define the mechanistic contributors to the benefits of VSG. Furthermore, this murine VSG model has been combined with other surgical techniques, to achieve deeper mechanistic insight. Therefore, this is a simple and versatile model for studying gastrointestinal physiology and the health benefits of bariatric surgery.

The following describes the performance of vertical sleeve gastrectomy in mice. This is a type of weight-loss surgery that involves removal of approximately 70% of the stomach.

Un modello murino di gastrectomia verticale

Bariatric surgery, such as vertical sleeve gastrectomy (VSG), is a surgery of the gastrointestinal tract that is performed for the purpose of weight loss. ...

Biologia

A Mouse Model of Intestinal Partial Obstruction

Intestinal obstructions, that impede or block peristaltic movement, can be caused by abdominal adhesions and most gastrointestinal (GI) diseases including tumorous growths. However, the cellular remodeling mechanisms involved in, and caused by, intestinal obstructions are poorly understood. Several animal models of intestinal obstructions have been developed, but the mouse model is the most cost/time effective. The mouse model uses the surgical implantation of an intestinal partial obstruction (PO) that has a high mortality rate if it is not performed correctly. In addition, mice receiving PO surgery fail to develop hypertrophy if an appropriate blockade is not used or not properly placed. Here, we describe a detailed protocol for PO surgery which produces reliable and reproducible intestinal obstructions with a very low mortality rate. This protocol utilizes a surgically placed silicone ring that surrounds the ileum which partially blocks digestive movement in the small intestine. The partial blockage makes the intestine become dilated due to the halt of digestive movement. The dilation of the intestine induces smooth muscle hypertrophy on the oral side of the ring that progressively develops over 2 weeks until it causes death. The surgical PO mouse model offers an in vivo model of hypertrophic intestinal tissue useful for studying pathological changes of intestinal cells including smooth muscle cells (SMC), interstitial cells of Cajal (ICC), PDGFR&#945;+, and neuronal cells during the development of intestinal obstruction.

Intestinal obstructions are a partial or complete blockage of the intestine that can cause severe abdominal pain, nausea, vomiting, and preventing the passage of stool. This procedure for creating intestinal partial obsructions in mice is reliable in studying the mechanisms underlying pathological cell growth and death in the intestine.

Un modello murino di ostruzione intestinale parziale

Intestinal obstructions, that impede or block peristaltic movement, can be caused by abdominal adhesions and most gastrointestinal (GI) diseases including ...

Murine Vertical Sleeve Gastrectomy: A Surgical Procedure to Downsize Stomach in Experimental Mouse Models

Source: Garibay, D., Cummings, B. P. <a href="https://jove.unicartagenaproxy.elogim.com/video/56534" target="_blank">A Murine Model of Vertical Sleeve Gastrectomy.</a> J. Vis. Exp. (2017)
In this video, a vertical sleeve gastrectomy procedure is performed on an experimental mouse model. This procedure helps remove a significant portion of the stomach along its greater curvature.

Gastrectomia a manica verticale murina: una procedura chirurgica per ridimensionare lo stomaco in modelli murini sperimentali

Source: Garibay, D., Cummings, B. P. A Murine Model of Vertical Sleeve Gastrectomy. J. Vis. Exp. (2017)
In this video, a vertical sleeve gastrectomy ...

JoVE Encyclopedia of Experiments

Ricerca sul cancro

Gastrointestinal Cancer

Animal model studies

Roux-en-Y Gastric Bypass Procedure: A Bariatric Surgical Procedure for Gastric Resectioning in Murine Model

For a Roux-en-Y gastric bypass, make a midline skin incision from the sternum to the middle of the abdomen, protecting the skin with a sterile compress soaked with 37 degrees Celsius saline solution.
Externalize the intestine and measure 8 centimeters from the pyloris. Using 5-0 non-absorbable sutures, place two ligatures on the intestine on either side of the 8-centimeter mark, and cut the tissue between the sutures.
Then, place the proximal limb of the tissue in the upper left quadrant of the abdomen, and the distal limb facing the alimentary limb, 6 centimeters below the proximal limb. Using micro scissors, cut both the proximal limb and the intestinal loop, and make two anti-mesenteric incisions of the same length. Next, use two 8-0 non-absorbable sutures to make a side-to-side anastomosis; placing the dorsal side anastomosis first, followed by the ventral side anastomosis.
Gently roll two moistened cotton swabs toward each side of the anastomosis, to confirm that the suture is leakproof, and carefully externalizing the stomach as just demonstrated. Place a re-absorbable hemostatic collagen compress behind the organ, and used curved micro forceps to pass a 5-0 non-absorbable suture through the omentum to ligate the pylorus.
Cut the ventral side of the stomach 1.5 centimeters from the pylorus and the distal limb, creating two incisions of the same length, followed by a dorsal to ventral side to side anastomosis with two 8-0 non-absorbable sutures. Confirm that the suture does not leak.
Then, close the muscle layer of the abdominal wall and the skin, with 5-0 non-absorbable sutures, and administer 25 milliliters per kilogram of warm saline solution, via subcutaneous injection.
When the mouse has fully recovered from the anesthesia, place the animal alone in a cage in 30 degrees Celsius incubator for 5 days, with daily iron and vitamin supplementation and analgesia, and free access to gel diet food for the first two days. On the third day after surgery, reintroduce solid food to the animal&#39;s diet.