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 border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Instruments</td>
			<td>Source</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont #1 Forceps Dumostar Tip&#160;</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Clip Applying Forceps 5.5&#34;</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Bonn Scissors 3.5&#34; Straight 15mm 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&#34; 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&#39;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&#34; Smooth</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Needle Holder 5.125&#34; Curved Lock .6mm</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Reflex 9mm Wound Clip Applier</td>
			<td>Roboz Surgical Instrument Co. (Gaithersburg, MD)</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Reflex 9mm 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></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Reagents</td>
			<td>Source</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ringer&#39;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

Research

JoVE Journal

Medicine

10.0K 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

Surgical Management of Meatal Stenosis with Meatoplasty

Meatal stenosis is a common urologic complication after circumcision. Children present to their primary care physicians with complaints of deviated urinary stream, difficult-to-aim, painful urination, and urinary frequency. Clinical exam reveals a pinpoint meatus and if the child is asked to urinate, he will usually have an upward, thin, occasionally forceful urinary stream with incomplete bladder emptying. The mainstay of management is meatoplasty (reconstruction of the distal urethra /meatus). This educational video will demonstrate how this is performed.

Meatoplasty, surgical management of meatal stenosis.

Trattamento chirurgico della stenosi Meatal con Meatoplasty

Meatal stenosis is a common urologic complication after circumcision.  Children present to their primary care physicians with complaints of deviated ...

Ricerca

Medicina

Implantation of Radiotelemetry Transmitters Yielding Data on ECG, Heart Rate, Core Body Temperature and Activity in Free-moving Laboratory Mice

The laboratory mouse is the animal species of choice for most biomedical research, in both the academic sphere and the pharmaceutical industry. Mice are a manageable size and relatively easy to house. These factors, together with the availability of a wealth of spontaneous and experimentally induced mutants, make laboratory mice ideally suited to a wide variety of research areas.
In cardiovascular, pharmacological and toxicological research, accurate measurement of parameters relating to the circulatory system of laboratory animals is often required. Determination of heart rate, heart rate variability, and duration of PQ and QT intervals are based on electrocardiogram (ECG) recordings. However, obtaining reliable ECG curves as well as physiological data such as core body temperature in mice can be difficult using conventional measurement techniques, which require connecting sensors and lead wires to a restrained, tethered, or even anaesthetized animal. Data obtained in this fashion must be interpreted with caution, as it is well known that restraining and anesthesia can have a major artifactual influence on physiological parameters1, 2.
Radiotelemetry enables data to be collected from conscious and untethered animals. Measurements can be conducted even in freely moving animals, and without requiring the investigator to be in the proximity of the animal. Thus, known sources of artifacts are avoided, and accurate and reliable measurements are assured. This methodology also reduces interanimal variability, thus reducing the number of animals used, rendering this technology the most humane method of monitoring physiological parameters in laboratory animals3, 4. Constant advancements in data acquisition technology and implant miniaturization mean that it is now possible to record physiological parameters and locomotor activity continuously and in realtime over longer periods such as hours, days or even weeks3, 5.
Here, we describe a surgical technique for implantation of a commercially available telemetry transmitter used for continuous measurements of core body temperature, locomotor activity and biopotential (i.e. onelead ECG), from which heart rate, heart rate variability, and PQ and QT intervals can be established in freeroaming, untethered mice. We also present pre-operative procedures and protocols for post-operative intensive care and pain treatment that improve recovery, well-being and survival rates in implanted mice5, 6.

A surgical technique for implantation of commercially available telemetry transmitters used for continuous measurement of biopotential (one-lead ECG), heart rate, core body temperature and locomotor activity in freely moving mice is shown. Recommendations and protocols for post-operative care and pain relief, improving recovery, well being and survival rate are also presented.

L&#39;impianto di trasmettitori Radiotelemetry Cedendo dati su ECG, frequenza cardiaca, temperatura corporea e attività di movimento libero topi di laboratorio

The laboratory mouse is the animal species of choice for most biomedical research, in both the academic sphere and the pharmaceutical industry. Mice are a ...

Minimal Invasive Surgical Procedure of Inducing Myocardial Infarction in Mice

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last decades. For clarification of the underlying mechanisms and the development of new therapeutic strategies, the availability of valid animal models are mandatory. Since we need new insights into pathomechanisms of cardiovascular diseases under in vivo conditions to combat myocardial infarction, the validity of the animal model is a crucial aspect. However, protection of animals are highly relevant in this context. Therefore, we establish a minimally invasive and simple model of myocardial infarction in mice, which assures a high reproducibility and survival rate of animals. Thus, this models fulfils the requirements of the 3R principle (Replacement, Refinement and Reduction) for animal experiments and assure the scientific information needed for further developing of therapeutical strategies for cardiovascular diseases.

A highly reproducible model for myocardial infarction in mice with minimal invasive manipulations is described. The model can be easily performed, resulting in a high reproducibility and survival rate. Thus, the described model will reduce the number of required animals as requested by the 3R principle (Replacement, Refinement and Reduction).

Minimal procedura chirurgica invasiva di indurre infarto miocardico nei topi

Myocardial infarction still remains the main cause of death in western countries, despite considerable progress in the stent development area in the last ...

Using Saccadometry with Deep Brain Stimulation to Study Normal and Pathological Brain Function

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including Parkinson's and Huntington's can disrupt it. People with Parkinson's disease, for example, tend to have increased saccadic latencies. Consequently, the quantitative measurement of saccadic eye movements has received considerable attention as a potential biomarker for neurodegenerative conditions. A lot more can be learned about the brain in both health and disease by observing what happens to eye movements when the function of specific brain areas is perturbed. Deep brain stimulation is a surgical intervention used for the management of a range of neurological conditions including Parkinson's disease, in which stimulating electrodes are placed in specific brain areas including several sites in the basal ganglia. Eye movement measurements can then be made with the stimulator systems both off and on and the results compared. With suitable experimental design, this approach can be used to study the pathophysiology of the disease being treated, the mechanism by which DBS exerts it beneficial effects, and even aspects of normal neurophysiology.

This paper describes the use of quantitative measurement of eye movements in conjunction with stimulation of focal areas of the deep brain in order to study physiology, pathophysiology, and the mechanisms of deep brain stimulation.

Utilizzando Saccadometry con stimolazione cerebrale profonda per studiare normali e patologiche funzione cerebrale

The oculomotor system involves a large number of brain areas including parts of the basal ganglia, and various neurodegenerative diseases including ...

Cutaneous Surgical Denervation:  A Method for Testing the Requirement for Nerves in Mouse Models of Skin Disease

Cutaneous somatosensory nerves function to detect diverse stimuli that act upon the skin. In addition to their established sensory roles, recent studies have suggested that nerves may also modulate skin disorders including atopic dermatitis, psoriasis and cancer. Here, we describe protocols for testing the requirement for nerves in maintaining a cutaneous mechanosensory organ, the touch dome (TD). Specifically, we discuss methods for genetically labeling, harvesting and visualizing TDs by whole-mount staining, and for performing unilateral surgical denervation on mouse dorsal back skin. Together, these approaches can be used to directly compare TD morphology and gene expression in denervated as well as sham-operated skin from the same animal. These methods can also be readily adapted to examine the requirement for nerves in mouse models of skin pathology. Finally, the ability to repeatedly sample the skin provides an opportunity to monitor disease progression at different stages and times after initiation.

This article includes detailed protocols for genetic labeling of mouse skin, surgical denervation, skin biopsy and visualizing labeled epithelia by whole-mount &#946;-galactosidase staining. These methods can be used to test the requirement for nerves in mouse models of normal and pathological skin.

Cutanea denervazione chirurgica: Un metodo per il test il requisito per i nervi in ​​modelli murini di malattia della pelle

Cutaneous somatosensory nerves function to detect diverse stimuli that act upon the skin. In addition to their established sensory roles, recent studies ...

Intravital Microscopy of Tumor-associated Vasculature Using Advanced Dorsal Skinfold Window Chambers on Transgenic Fluorescent Mice

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static information and is often not sufficient for a correct interpretation. Intravital evaluation, in which a process is followed in time, provides extra and often unexpected information. With the creation of transgenic animals expressing cell-specific markers and live cell tracers, improvements to imaging equipment, and the development of several imaging chambers, intravital microscopy has become an important tool to better understand biological processes. This paper describes an experimental design for the investigation of tumor vessel development and of therapeutic effects in a spatial and temporal manner. Using this setup, the stage of vessel development, tip cell and lumen formation, blood flow, extravasation, an established vascular bed, and vascular destruction can be visualized and followed. Furthermore, therapeutic effects, intratumoral fate, and the localization of chemotherapeutic compounds can also be followed.

This protocol describes the intravital imaging of transgenic mice expressing cell-specific fluorescent markers. Intravital imaging provides a non-invasive method for high-resolution observations in living animals using implantable windows through which the microscopic visualization of processes is possible. This method is especially useful to study longitudinal processes.

Videomicroscopia microscopia del sistema vascolare del tumore-associati utilizzando avanzate dorsale dello Skinfold finestra Chambers su topi transgenici fluorescenti

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static ...

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 ...

Vacuum-Sealed Hot Water Bath Immersion for the Preparation of Anatomical and Surgical Cadaveric Bone Models

Bone models serve many purposes, including improving anatomical understanding, preoperative surgical planning, and intraoperative referencing. Several techniques for the maceration of soft tissues have been described, mainly for forensic analysis. For clinical research and medical use, these methods have been superseded by three-dimensional (3D) printed models, which require substantial equipment and expertise, and are costly. Here, cadaveric sheep vertebral bone was cleaned by vacuum sealing the specimen with commercial dishwashing detergent, immersing in a hot water bath, and subsequently manually removing the soft tissue. This eliminated the disadvantages of the previously existing maceration methods, such as the existence of foul odors, usage of hazardous chemicals, substantial equipment, and high costs. The described technique produced clean, dry samples while maintaining anatomical detail and structure to accurately model the osseous structures that can be useful for preoperative planning and intraoperative referencing. The method is simple, low-cost, and effective for bone model preparation for education and surgical planning in veterinary and human medicine.

The present protocol describes the maceration and cleaning of cadaveric bone with a vacuum-sealed, hot water bath immersion technique. This is a low-cost, safe, and effective method to produce anatomical specimens for surgical planning and medical education as an alternative to three-dimensional (3D) printed models.

Immersione con bagno di acqua calda sottovuoto per la preparazione di modelli anatomici e chirurgici in osso cadaverico

Bone models serve many purposes, including improving anatomical understanding, preoperative surgical planning, and intraoperative referencing. Several ...

Surgical Procedure for Intrapulmonary Tracheal Transplantation in Mice

To begin, record the weight of each mouse, then position the euthanized mouse in a supine orientation and secure its limbs with tape. Use 70%isopropyl alcohol to sterilize the surgical area. Next, create a midline incision on the skin starting from the mid-abdomen to the anterior cervical region.To access the trachea, carefully retract the fat pads, laterally move the strap muscles and separate the trachea from the surrounding connective tissue. Using forceps, create a space between the trachea and the esophagus. Then lift the xiphoid and cut the diaphragm.Insert a hemostat to raise the sternum for an unobstructed path from the sternum to the neck region. Clamp the rib cage on both sides and cut through the sternum extending through the neck muscles. Now remove the thymus and any fat or muscle obstructing the trachea to expose the tracheal bifurcation.Cut the main bronchi and carefully separate the airway from the esophagus. Afterward, cut the larynx and remove it. Spray the dissected trachea with a sterile saline solution or a preservation solution.Use sterile gauze soaked in sterile saline or preservation solution and place it on ice to maintain viability. After lightly anesthetizing the mouse and an induction chamber, intraperitoneally inject a cocktail consisting of xylazine and ketamine. Return the mouse to the induction chamber with 2 to 3%isoflurane maintained.Then shave the fur at the surgical site. Confirm the absence of reflex response to a toe pinch before orotracheal intubation. Using a 20 gauge intravenous catheter, intubate the mouse orotracheally and connect it to a ventilator with 50%oxygen and 2%isoflurane.Now, activate a heating pad and position the mouse in a right lateral position on the pad with the head away from the surgeon and the tail towards the surgeon. Then secure the limbs with tape. Apply veterinary ointment to the eyes to prevent dryness while under anesthesia.Scrub the surgical area first with 7.5 povidone iodine, then 70%isopropyl alcohol, and finally, with 10%povidone iodine. During this time, load the donor trachea into a 16 gauge intravenous catheter. After making an incision in the recipient's skin, cauterize the muscle and connective tissue.Using two retractors, open the fifth or sixth intercostal space and hold the rib cage open. Then use cotton swabs and scissors to dissect the inferior pulmonary ligament. Simulate the creation of the pathway for the donor trachea.To secure the ventilator outflow tube, partially occlude it with a three-way stopcock to facilitate inflation of the left lung. Puncture the left lung with a 20 gauge needle to create a pathway. Then insert the 16 gauge intravenous catheter into the left lung and extrude the donor trachea into it.After insertion of the tracheal allograft, release the three-way stopcock to allow unobstructed expiratory flow through the outflow tube. To close the pleural injection site, position the clip precisely onto the puncture site with its edge aligned to the contour of the lung's edge. Then fill the thoracic cavity with saline solution and absorb the saline with gauze.Reinflate the left lung and close the ribs using a running suture. Close the muscle and skin with interrupted sutures. Now, turn off isoflurane and administer meloxicam analgesic subcutaneously at the end of the surgery.Observe the recipient mouse until it is awake. Then remove the tracheal tube and put the recipient mouse in a cage. A tracheal allograft exhibited complete obstruction with fibroblastic tissue, and the epithelial cells were visibly destroyed.Conversely, a tracheal isograft remained patent, and the epithelial cells were preserved. Lymphoid aggregates were observed in the lung with the transplanted tracheal allograft.

Procedura chirurgica per il trapianto tracheale intrapolmonare nei topi

<meta charset="utf8"></head><body>Chirurgia in un'unica fase per indurre il coma ischemico cerebrale nei ratti

Cerebral Ischemic Coma Model Induced by Modified Four-Vessel Occlusion

Coma caused by cerebral ischemia is the most serious complication of cerebral ischemia. Four-vessel occlusion can establish a cerebral ischemic coma model for disease research and drug development. However, the commonly used four-vessel occlusion method mainly involves inserting an electrocoagulation pen into the bilateral pterygoid foramen of the first cervical vertebra behind the neck to electrocoagulate the vertebral arteries. This process carries the risk of incomplete electrocoagulation, bleeding, and damage to the brainstem and spinal cord. Twenty-four hours after surgery, re-anesthetized rats undergo carotid artery ligation in front of the neck. Two surgeries expose the rats to a higher risk of infection and increase the experimental period. In this study, during a single surgical procedure, an anterior cervical incision was used to locate the key site where the vertebral artery penetrates the first cervical vertebra. The bilateral vertebral arteries were electrocauterized under visual conditions, while the bilateral common carotid arteries were separated to place loose knots. When the limbs of the rats began twitching, the bilateral common carotid arteries were quickly ligated to induce ischemic coma. This method can avoid the risk of infection caused by two surgical operations and is easy to perform with a high success rate, providing a useful reference for relevant practitioners.

<meta charset="utf8"></head><body>Autore in primo piano: Migliorare la ricerca sull'ischemia cerebrale con un modello di ratto semplificato

This protocol describes the process for inducing a cerebral ischemic coma model using a modified four-vessel occlusion method.

Modello di coma ischemico cerebrale indotto da occlusione a quattro vasi modificata

Coma caused by cerebral ischemia is the most serious complication of cerebral ischemia. Four-vessel occlusion can establish a cerebral ischemic coma model ...