The present study was supported by grants from Kidney Research UK (ST4/2011), the Cunningham Trust (CT11/14), the Mrs AE Hogg Charitable Trust for Kidney Research and the Renal Endowment Fund of the Royal Infirmary of Edinburgh.

Directives générales: expériences sur les animaux ont été effectuées en conformité avec les directives et règlements imposés par les animaux (Scientific Procedures) de Loi 1986. Ce protocole et le protocole de vidéo qui l&#39;accompagne sont à la fois un UUO standard et un R-UUO, qui peut être effectuée sur plusieurs souches de souris. Dans la vidéo d&#39;accompagnement, les deux procédures sont effectuées sur des souris C57BL / 6 mâles âgées de 8 semaines les souris. Les données présentées dans la section des résultats représentatifs ont été obtenus auprès de sexe masculin souris FVB / n. NOTE: Ce protocole et accompagner détails de la vidéo comment effectuer une UUO standard et R-UUO utilisant l&#39;uretère gauche, mais les mêmes techniques peuvent facilement être appliqués à l&#39;uretère droit. 1. Préparation des animaux et la laparotomie <ol><li> Effectuer toutes les procédures avec les instruments et consommables stériles (autoclave). </li><li> Injecter chlorhydrate de kétamine (70 mg / kg) et du chlorhydrate de médétomidine (1 mg / kg) par voie intraperitoneale à anesthésierla souris. Remarque: La durée de l&#39;anesthésie plan résultant est de 4 heures et pas d&#39;anesthésie supplémentaire ne est nécessaire. </li><li> Confirmer la profondeur de l&#39;anesthésie par la perte des réflexes, par exemple pincement de l&#39;orteil. </li><li> Retirez tous les cheveux entourant la zone d&#39;incision et de préparer la peau pour la chirurgie abdominale via l&#39;application d&#39;une solution diluée de chlorhexidine. </li><li> Placez la souris sur un bloc chirurgical chauffé dans une position couchée et fixer les membres à l&#39;aide de ruban adhésif pad à faible adhérence. </li><li> Durant la procédure, surveiller la souris les signes de brûlures thermiques en raison du tampon chirurgical chauffé. Si possible, utilisez une source de chaleur non-électrique. </li><li> Administrer un analgésique par injection sous-cutanée de chlorhydrate de buprénorphine (0,06 mg / kg) et appliquer du lubrifiant oculaire pour éviter un dessèchement de la cornée. </li><li> Faire une laparotomie médiane et une incision de la linea alba avasculaire l&#39;aide de ciseaux de tissu de séparation pour accéder à la cavité péritonéale. </li><li> Drapé de la souris et insérer un colibrirétracteur dans l&#39;incision. </li></ol> 2. unilatérale obstruction urétérale <ol><li> Utilisation de bourgeons de coton stérilisés exposer l&#39;uretère gauche en déplaçant les intestins vers le côté droit de la cavité abdominale et les couvrir avec des rideaux humides. </li><li> En utilisant des pinces angulaires isoler et soulever l&#39;uretère gauche. </li><li> Pour créer une obstruction urétérale, ligaturer l&#39;uretère gauche deux fois avec 6 / O noir tressé de soie suture ne importe où entre la vessie et du bassinet du rein. Pour les expériences à long terme, utiliser suture absorbable pour toutes les chirurgies abdominales. En variante, une pince de ligature peut être appliquée à l&#39;uretère. </li><li> Pour isoler la vessie de l&#39;uretère, diviser l&#39;uretère entre les deux points de suture. </li><li> Replacez soigneusement les intestins dans la cavité péritonéale. </li><li> Suivez les étapes énumérées à la section 4 - récupération post-opératoire et des Soins de fermer l&#39;incision et d&#39;inverser l&#39;anesthésie. </li></ol> 3. Préparation pour Reversible unilatérale obstruction urétérale <ol><li> Préparer une souris pour la chirurgie et d&#39;isoler l&#39;uretère gauche comme indiqué dans les étapes 1.1 à 2.2 ci-dessus. </li><li> Pour créer une obstruction urétérale qui peut être inversée, ligaturer l&#39;uretère gauche deux fois avec 6 / O noir tressé de soie suture près de la vessie. Laissez une extrémité de la suture supérieure tant que ce sera utilisé pour ancrer le tube en plastique à paroi souple en place autour de l&#39;uretère. </li><li> Faire une fente longitudinale sur une longueur de 5 mm de tube en plastique à paroi souple de manière à pouvoir être évasée ouverte pour lui permettre d&#39;être appliqué à l&#39;uretère. De préférence un tube en plastique à paroi souple en silicone, avec un diamètre interne de 1 mm et un diamètre externe de 2 mm, peut être utilisé. </li><li> Placez le tube en plastique à paroi souple doucement autour de l&#39;uretère. Se assurer que le fil de suture longue émerge du centre de la fente dans le tube une fois qu&#39;il est fermé autour de l&#39;uretère. </li><li> Placez une longueur de 6 / O noire tressée suture de soie autour du tube à paroi souple enfermant l&#39;uretère et cravateune fois. Maintenant, placer l&#39;extrémité longue du fil de suture, émergeant du centre de la fente dans le tube, longitudinalement à travers le tube de telle sorte qu&#39;il se trouve au-dessus de la suture attachée précédemment. </li><li> Attacher le fil de suture autour du tube situé à deux reprises pour ancrer le tube et l&#39;extrémité de fil de suture longue en place autour de l&#39;uretère. Remarque: Pour éviter la formation d&#39;adhérences sur l&#39;uretère et le tube en plastique à paroi souple, d&#39;une solution de réduction de l&#39;adhérence peut être appliqué à la zone autour du tube. </li><li> Replacez soigneusement les intestins dans la cavité péritonéale. </li></ol> 4. récupération post-opératoire et des Soins <ol><li> Fermez le péritoine avec point de feston utilisant 5 / O noire tressée suture de soie et de se rapprocher de la peau à l&#39;aide des clips métalliques de la peau. </li><li> Pour minimiser le risque d&#39;infection post-opératoire, appliquez un antiseptique comme solution d&#39;iode / alcool pour la peau abdominale. </li><li> Partiellement anesthésie inverse avec le chlorhydrate d&#39;atipamézole (2 mg / kg) sous-cutanée. </li&gt; <li> Administrer des fluides par une injection sous-cutanée de 1 ml de solution saline chauffée. </li><li> Surveiller la souris jusqu&#39;à ce qu&#39;il ait repris connaissance. </li><li> Laisser la souris pour récupérer dans une boîte chauffée maintenue à 29 ° C pendant 24 heures. Nourriture humide peut également être fournie pour encourager l&#39;apport hydrique et de la nutrition. </li><li> Laissez la souris pour récupérer pour induire un niveau souhaité de l&#39;obstruction, généralement sept jours. </li><li> Pour les expériences de reconstitution à long terme, fournir des analgésiques en cours. Si la souris est à récupérer pendant plus de sept jours, retirez les clips de la peau 7 jours après la chirurgie. </li><li> Une fois le niveau de l&#39;obstruction désiré a été induite, soit inverser la UUO, comme décrit ci-dessous, ou euthanasier la souris par dislocation cervicale et de recueillir les reins pour l&#39;analyse de l&#39;histopathologie. </li></ol> 5. réversible unilatérale obstruction urétérale <ol><li> Pour effectuer une R-UUO préparer une souris, qui a subi une préparation pour le R-UUO, pour une chirurgie tel que décrit dans les étapes 1De 0,1 à 1,7. </li><li> Se il est présent, retirez les clips de la peau et de diviser ou de retirer les points de suture dans le péritoine pour accéder à la cavité abdominale. Remarque: Si l&#39;uretère a été obstrué pendant long terme clips de la peau de l&#39;expérience aurait dû être retiré sept jours suivant l&#39;application. </li><li> Préparer la souris et isoler l&#39;uretère gauche de la manière décrite dans les étapes 1,8 à 2,2. </li><li> En utilisant des pinces angulaires, libérer le tube en plastique à paroi souple de tout tissu de granulation qui peuvent avoir formé. </li><li> Couper le fil de suture en tenant le tuyau plastique autour de l&#39;uretère obstrué avec un scalpel et retirer le tube. </li><li> Confirmez succès UUO en évaluant la présence d&#39;hydronéphrose dans le rein gauche, le rein devrait également apparaître pâle. </li><li> Diviser l&#39;uretère entre les sutures. </li><li> Passer la longueur restante de l&#39;uretère, le rein attaché à, sur un petit morceau de gaze stérile. Il sera utilisé pour recueillir le sédiment urinaire et les cellules mortes qui se écouler de l&#39;uretère unee bassinet une fois la suture sont enlevés. </li><li> Diviser l&#39;uretère ci-dessus, mais à proximité, la suture et permettre le rein se écouler sur la gaze. Laissez la suture inférieure, plus proche de la vessie, intacte. Il se agit de se assurer qu&#39;aucune fuite urine provenant de la vessie et dans la cavité peritoneale. </li><li> Une fois l&#39;uretère et bassinet du rein ont été drainés, appliquez une longue 6 / O noire tressée suture de soie à la fin de la durée restante de l&#39;uretère. Il sera utilisé pour aider l&#39;uretère anastomose de la vessie réalisée plus tard. </li><li> Suivez les étapes ci-dessous pour anastomoser la longueur restante de l&#39;uretère dans la vessie: <ol><li> Mettre l&#39;uretère tel qu&#39;il se trouve en avant de sa position d&#39;origine et se trouve sur le rein. </li><li> Placer un unique 9 / O polyamide monofilament pointage suture 2 mm de l&#39;extrémité de l&#39;uretère de telle sorte qu&#39;il émerge dans la direction de la vessie. Veillez à ce que la suture reste dans la couche musculaire et ne entre pas dans la lumière de l&#39;uretère. Le wil pointage de suturel être utilisé pour ancrer l&#39;uretère dans la vessie. </li><li> Pour créer un canal à travers la vessie, passer une aiguille de 21G en diagonale à travers la vessie de telle sorte que sa sortie vers l&#39;avant (ventrale) paroi de la vessie. </li><li> Reste une aiguille à chas dans le biseau de l&#39;aiguille 21G. Utilisez l&#39;aiguille de 21G pour guider l&#39;aiguille yeux à travers la vessie. L&#39;aiguille à chas sera utilisé pour prendre l&#39;uretère à travers la vessie. </li><li> Faire passer le fil de suture 9 adhésivité / O, appliqué à l&#39;uretère à l&#39;étape 5.11.2, à travers la première incision dans la vessie et à travers la paroi de la vessie adjacente au point d&#39;entrée. Une fois l&#39;uretère est passé à travers la vessie ce sera liée à ancrer l&#39;uretère à la vessie. </li><li> Placer la longue 6 / O noire tressée suture de soie appliqué à la fin de l&#39;uretère à l&#39;étape 5.10 borgne à travers l&#39;aiguille. </li><li> Retirer délicatement l&#39;aiguille yeux hors de la vessie tout en garantissant que l&#39;uretère est également tiré à travers la vessie. </li><li> Une fois l&#39;uretère se dégage de the vessie retirer l&#39;aiguille yeux et appliquer une pince à suture long à la fin de l&#39;uretère pour empêcher la rétraction en arrière dans la vessie. </li><li> Pour ancrer au sein de l&#39;uretère dans la vessie, attacher le 9 / O polyamide monofilament amure suture, appliquée à l&#39;étape 5.11.5. </li><li> Appliquer sutures monofilament de liaison supplémentaires simples 9 / O de polyamide comme décrit dans les étapes 5.11.2 et 5.11.5 à deux endroits à travers le point d&#39;entrée pour ancrer fermement l&#39;uretère dans la vessie. </li><li> Avec le dos de ciseaux ouverts, pousser la paroi de la vessie légèrement en arrière pour exposer davantage de l&#39;uretère émergente. </li><li> Diviser l&#39;uretère-dessus de la longue suture située à l&#39;extrémité de l&#39;uretère. L&#39;uretère doit se rétracter dans le corps de la vessie. Il est habituel d&#39;observer l&#39;urine se écoulant hors de l&#39;ouverture dans la vessie, ce qui confirme la lumière de l&#39;uretère ouverts. </li><li> Fermez la sortie des plaies dans la vessie avec un polyamide suture monofilament 9 / O. </li></ol></li><li> Replacez soigneusement les intestins enla cavité péritonéale. </li><li> Fournir des soins post-opératoire comme indiqué dans les étapes 4.1 à 4.8. </li><li> Laisser la souris pour récupérer jusqu&#39;à ce que le rein est décompressé, généralement sept jours. </li><li> Au point de fin expérimentale, euthanasier la souris par dislocation cervicale et de recueillir les reins pour l&#39;analyse histopathologique. </li></ol>

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S., 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=Chronic+kidney+disease+induced+in+mice+by+reversible+unilateral+ureteral+obstruction+is+dependent+on+genetic+background.">Chronic kidney disease induced in mice by reversible unilateral ureteral obstruction is dependent on genetic background.</a> Am J Physiol Renal Physiol. 298, F1024-F1032 (2010).</li><li>Heikkila, J., Holmberg, C., Kyllonen, L., Rintala, R., Taskinen, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+risk+of+end+stage+renal+disease+in+patients+with+posterior+urethral+valves.">Long-term risk of end stage renal disease in patients with posterior urethral valves.</a> J Urol. 186, 2392-2396 (2011).</li><li>Ravanan, R., Tomson, C. 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C., 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=Postobstructive+regeneration+of+kidney+is+derailed+when+surge+in+renal+stem+cells+during+course+of+unilateral+ureteral+obstruction+is+halted.">Postobstructive regeneration of kidney is derailed when surge in renal stem cells during course of unilateral ureteral obstruction is halted.</a> Am J Physiol Renal Physiol. 298, F357-F364 (2010).</li><li>Manson, S. R., Niederhoff, R. A., Hruska, K. A., Austin, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endogenous+BMP-7+is+a+critical+molecular+determinant+of+the+reversibility+of+obstruction-induced+renal+injuries.">Endogenous BMP-7 is a critical molecular determinant of the reversibility of obstruction-induced renal injuries.</a> Am J Physiol Renal Physiol. 301, F1293-F1302 (2011).</li><li>Manson, S. R., Niederhoff, R. A., Hruska, K. A., Austin, P. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+BMP-7-Smad1/5/8+pathway+promotes+kidney+repair+after+obstruction+induced+renal+injury.">The BMP-7-Smad1/5/8 pathway promotes kidney repair after obstruction induced renal injury.</a> J Urol. 185, 2523-2530 (2011).</li><li>Chaves, L. D., 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=Contrasting+effects+of+systemic+monocyte/macrophage+and+CD4++T+cell+depletion+in+a+reversible+ureteral+obstruction+mouse+model+of+chronic+kidney+disease.">Contrasting effects of systemic monocyte/macrophage and CD4+ T cell depletion in a reversible ureteral obstruction mouse model of chronic kidney disease.</a> Clin Dev Immunol. 2013, 836-989 (2013).</li><li>Haque, M. E., 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=Longitudinal+changes+in+MRI+markers+in+a+reversible+unilateral+ureteral+obstruction+mouse+model:+preliminary+experience.">Longitudinal changes in MRI markers in a reversible unilateral ureteral obstruction mouse model: preliminary experience.</a> J Magn Reson Imaging. 39, 835-841 (2014).</li><li>Bai, Z. 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=Arterially+transplanted+mesenchymal+stem+cells+in+a+mouse+reversible+unilateral+ureteral+obstruction+model:+in+vivo+bioluminescence+imaging+and+effects+on+renal+fibrosis.">Arterially transplanted mesenchymal stem cells in a mouse reversible unilateral ureteral obstruction model: in vivo bioluminescence imaging and effects on renal fibrosis.</a> Chinese Med J. 126, 1890-1894 (2013).</li><li>Chaabane, W., 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=Renal+functional+decline+and+glomerulotubular+injury+are+arrested+but+not+restored+by+release+of+unilateral+ureteral+obstruction+(UUO).">Renal functional decline and glomerulotubular injury are arrested but not restored by release of unilateral ureteral obstruction (UUO).</a> Am J Physiol Renal Physiol. 304, F432-F439 (2013).</li><li>Thornhill, B. A., Burt, L. E., Chen, C., Forbes, M. S., Chevalier, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variable+chronic+partial+ureteral+obstruction+in+the+neonatal+rat:+a+new+model+of+ureteropelvic+junction+obstruction.">Variable chronic partial ureteral obstruction in the neonatal rat: a new model of ureteropelvic junction obstruction.</a> Kidney Int. 67, 42-52 (2005).</li><li>Thornhill, B. A., Chevalier, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variable+partial+unilateral+ureteral+obstruction+and+its+release+in+the+neonatal+and+adult+mouse.">Variable partial unilateral ureteral obstruction and its release in the neonatal and adult mouse.</a> Methods Mol Biol. 886, 381-392 (2012).</li></ol>

Les auteurs ne ont pas des intérêts concurrents ou de divulguer contradictoires.

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. The R-UUO model is of relevance to human disease as patients often present with obstruction of the renal tract secondary to prostatic hypertrophy, posterior urethral valves etc. that requires decompression with patients commonly left with significant residual kidney impairment that has no current treatment options and may eventually lead to end stage kidney disease.8,9 The models detailed here enable ...

The overall aim of the experimental model described here is to induce obstructive nephropathy by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). A simple irreversible model of UUO is presented in which the left ureter is permanently obstructed by ligation with a suture or by the application of a ligating clip. This results in marked dilatation of the ureter together with reduced renal blood flow and glomerular filtration. Renal histology demonstrates tubular dilatation and increasingly severe interstitial renal inflammation and fibrosis. Irreversible UUO is a useful model and has been adopted by many researchers in the study of both renal inflammation and fibrosis.1-4 Although the irreversible model of UUO requires some surgical expertise it is relatively straightforward and is often used to seek insights into the pathogenesis of interstitial renal injury and the ensuing fibrosis. Also presented is a less frequently used R-UUO model using a modification of the method originally described by Tapmeier et al.5 The R-UUO model has much future potential for the study of inflammatory and immune processes, cellular and tissue regeneration as well as the subsequent tissue remodeling following the removal of an injurious stimulus. 
The more challenging R-UUO model has been used by a limited number of investigators with some groups employing a significantly different surgical technique to that described here6,7 though with interesting results. In the R-UUO model presented, the ureter is ligated to induce complete ureteric obstruction for a period of time sufficient to induce the level of injury and fibrosis desired: 7 days of UUO was chosen in the method described here. The ureteric obstruction is reversed and the kidney is allowed to decompress for a period of time determined by the investigator before the mice are culled and the kidney removed for analysis: 7 days of decompression was chosen in the method described here though a longer period would be chosen if the resolution of inflammation and fibrosis was being studied. Although the method described here requires significant surgical expertise, it offers several advantages over other R-UUO models. The application of soft walled plastic tubing to the obstructed ureter prevents excessive ureteric dilation and this facilitates the subsequent manipulation and anastomosis of the ureter. Furthermore, in the described R-UUO model the ureter is divided thereby allowing the removal of any residual urinary sediment and debris. This confirms that the remaining ureter lumen is de-obstructed and patent prior to anastomosis to the bladder. 
Experiments incorporating both the irreversible and reversible UUO models can provide researchers with a powerful insight into the molecular and cellular mechanisms of both injury and subsequent resolution and regeneration. Thus, the model of R-UUO described here would be highly relevant to researchers interested in post-inflammatory tissue remodeling and how this can be modulated.

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Blunt Dissecting Scissors</td>
			<td>Fine Science Tools</td>
			<td>14072-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Spring Scissors &#8211; straight</td>
			<td>Fine Science Tools</td>
			<td>15000-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Toothed forceps</td>
			<td>Fine Science Tools</td>
			<td>11021-12</td>
			<td></td>
		</tr>
		<tr>
			<td>Angled forceps x 2</td>
			<td>Fine Science Tools</td>
			<td>00649-11</td>
			<td></td>
		</tr>
		<tr>
			<td>Straight forceps</td>
			<td>Fine Science Tools</td>
			<td>00632-11</td>
			<td></td>
		</tr>
		<tr>
			<td>Colibri 3cm wire retractor</td>
			<td>Fine Science Tools</td>
			<td>17000-03</td>
			<td></td>
		</tr>
		<tr>
			<td>Castroviejo needle holder with lock</td>
			<td>Fine Science Tools</td>
			<td>12565-14</td>
			<td></td>
		</tr>
		<tr>
			<td>Wound clip applicator</td>
			<td>Fine Science Tools</td>
			<td>12031-07</td>
			<td></td>
		</tr>
		<tr>
			<td>7mm wound clips</td>
			<td>Fine Science Tools</td>
			<td>12032-07</td>
			<td></td>
		</tr>
		<tr>
			<td>Castroviejo micro needle holder with lock</td>
			<td>Fine Science Tools</td>
			<td>12060-01</td>
			<td></td>
		</tr>
		<tr>
			<td>OPMI pico microscope</td>
			<td>Carl Zeiss</td>
			<td>S100</td>
			<td></td>
		</tr>
		<tr>
			<td>Heat electronic pad</td>
			<td>Cozee Comfort</td>
			<td>n/a</td>
			<td></td>
		</tr>
		<tr>
			<td>6/O silk braided suture</td>
			<td>Harvard Apparatus</td>
			<td>72-3287</td>
			<td></td>
		</tr>
		<tr>
			<td>9/O Dafilon (polyamide) suture</td>
			<td>B-Braun</td>
			<td>G1111434</td>
			<td></td>
		</tr>
		<tr>
			<td>5/O braided silk suture</td>
			<td>Harvard Apparatus</td>
			<td>51-7680</td>
			<td></td>
		</tr>
		<tr>
			<td>Regular bevel needle, 1 inch, 21G</td>
			<td>Becton, Dickinson and Company</td>
			<td>305175</td>
			<td></td>
		</tr>
		<tr>
			<td>1 ml syringe slip tip</td>
			<td>Becton, Dickinson and Company</td>
			<td>300184</td>
			<td></td>
		</tr>
		<tr>
			<td>Wypall paper swabs</td>
			<td>Kimberley-Clark</td>
			<td>L40</td>
			<td>Sterilised (Autoclave)</td>
		</tr>
		<tr>
			<td>Cotton wool buds</td>
			<td>Johnson and Johnson</td>
			<td>n/a</td>
			<td>Sterilised (Autoclave)</td>
		</tr>
		<tr>
			<td>Plain drapes</td>
			<td>Guardian</td>
			<td>CB03</td>
			<td>Sterilised (Autoclave)</td>
		</tr>
		<tr>
			<td>Soft wall silicone rubber tubing</td>
			<td></td>
			<td></td>
			<td>Silicone tubing - internal diameter 1.0 mm; external diameter 2.0 mm; wall thickness 0.6 mm</td>
		</tr>
		<tr>
			<td>(Lacri-Lube) White soft paraffin 57.3%, mineral oil 42.5% and lanolin alcohols 0.2%</td>
			<td>Allergan Ltd</td>
			<td>21956GB10X</td>
			<td></td>
		</tr>
		<tr>
			<td>(Videne) Povidone-iodine 10%</td>
			<td>Ecolab Ltd</td>
			<td>PL 04509/0041</td>
			<td></td>
		</tr>
		<tr>
			<td>(Vetalar V) Ketamine hydrochloride</td>
			<td>Pfizer Animal Health</td>
			<td>Vm 42058/4165</td>
			<td>100mg/ml solution</td>
		</tr>
		<tr>
			<td>(Domitor) Medetomidine hydrochloride</td>
			<td>Orion Pharma</td>
			<td>Vm 06043/4003</td>
			<td>1mg/ml</td>
		</tr>
		<tr>
			<td>(Vetergesic) Bupernorphine hydrochloride</td>
			<td>Alsto Animal Health</td>
			<td>Vm 00063/4002</td>
			<td>0.3mg/ml</td>
		</tr>
		<tr>
			<td>(Antisedan) Atipamezole hydrochoride</td>
			<td>Orion Pharma</td>
			<td>Vm 06043/4004</td>
			<td>5mg/ml</td>
		</tr>
		<tr>
			<td>(Adept) 4% Icodextrin</td>
			<td>Baxter</td>
			<td></td>
			<td>Adhesion reduction solution</td>
		</tr>
		<tr>
			<td>NaCl 0.9%</td>
			<td>Baxter</td>
			<td>FKE1323</td>
			<td></td>
		</tr>
	</tbody>
</table>

a murine model of irreversible and reversible unilateral ureteric obstruction

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive nephropathy induced by unilateral ureteric obstruction (UUO), which can either be irreversible (UUO) or reversible (R-UUO). In the irreversible UUO model, the ureter may be obstructed for variable periods of time in order to induce increasingly severe renal inflammation and interstitial fibrotic scarring. In the reversible R-UUO model the ureter is obstructed to induce hydronephrosis, tubular dilation and inflammation. After a suitable period of time the ureteric obstruction is then surgically reversed by anastomosis of the severed previously obstructed ureter to the bladder in order to allow complete decompression of the kidney and restoration of urinary flow to the bladder. The irreversible UUO model has been used to investigate various aspects of renal inflammation and scarring including the pathogenesis of disease and the testing of potential anti-inflammatory or anti-fibrotic therapies. The more challenging model of R-UUO has been used by some investigators and does offer significant research potential as it allows the study of inflammatory and immune processes and tissue remodeling in an injured and scarred kidney following the removal of the injurious stimulus. As a result, the R-UUO model offers investigators the opportunity to explore the resolution of kidney inflammation together with key aspects of tissue repair. These experimental models are of relevance to human disease as patients often present with obstruction of the renal tract that requires decompression and are commonly left with significant residual kidney impairment that has no current treatment options and may lead to eventual end stage kidney failure.

L&#39;apparition des modifications rénales nettement suivante obstruction urétérale et il devient plus pâle et tendu à la palpation avec le temps (Figure 1). Il est de plus en plus la dilatation de l&#39;urètre proximal à l&#39;obstruction et le bassinet du rein. Le rein devient de plus en plus atrophique que la durée de néphropathie obstructive augmente avec obstruction prolongée entraîne un amincissement notable de la moelle et le cortex rénal. Après le renversement de l&#39;obstruction d...

Watch this Scientific Journal Video about A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction at JoVE.com

A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

Un modèle murin d&#39;irréversible et réversible unilatérale obstruction urétérale

Obstruction of the kidney may affect native or transplanted kidneys and results in kidney injury and scarring. Presented here is a model of obstructive ...

a-murine-model-irreversible-reversible-unilateral-ureteric

Research

JoVE Journal

Medicine

30.0K vues.  University of Edinburgh. The murine model of irreversible unilateral ureteric obstruction (UUO) is presented together with the model of reversible UUO in which the ureteric obstruction is relieved by anastomosis of the severed ureter into the bladder. These models enable the study of renal inflammation and scarring as well as tissue remodeling.

Vidéo: A Murine Model of Irreversible and Reversible Unilateral Ureteric Obstruction

Creation of Reversible Cholestatic Rat Model

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Cholestasis can cause various physiological changes and affect the nutritional status and surgical outcomes. Study of the pathophysiological changes occurring in the liver and other organs is of importance. Various studies have been done in cholestatic rat models.
We used a reversible cholestatic rat model in our recent study looking at the role of methylprednisolone in the ischemia reperfusion injury. Various techniques for creation of a reversible cholestatic model have been described. Creation of a reversible cholestatic rat model can be challenging in view of the smaller size and unique hepatopancreatobiliary anatomy in rats. This video article demonstrates the creation of a reversible cholestatic model.
This model can be used in various studies, such as looking at the changes in nutritional, physiological, pathological, histological and immunological changes in the gastrointestinal tract. This model can also be used to see the effects of cholestasis and various therapeutic interventions on major hepatic surgeries.

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Creation of a reversible cholestatic rat model can be challenging in view of the smaller size and unique hepatopancreatobiliary anatomy in rats. This video article demonstrates the creation of a reversible cholestatic model.

Création du modèle de rat réversible cholestatique

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Cholestasis can cause various physiological changes and ...

Recherche

Médecine

A Contusive Model of Unilateral Cervical Spinal Cord Injury Using the Infinite Horizon Impactor

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of spinal cord injury (SCI) in which the thoracic spinal cord is injured. Additionally, because most human cord injuries occur as the result of blunt, non-penetrating trauma (e.g. motor vehicle accident, sporting injury) where the spinal cord is violently struck by displaced bone or soft tissues, the majority of SCI researchers are of the opinion that the most clinically relevant injury models are those in which the spinal cord is rapidly contused.1 Therefore, an important step in the preclinical evaluation of novel treatments on their way to human translation is an assessment of their efficacy in a model of contusion SCI within the cervical spinal cord. Here, we describe the technical aspects and resultant anatomical and behavioral outcomes of an unilateral contusive model of cervical SCI that employs the Infinite Horizon spinal cord injury impactor.
Sprague Dawley rats underwent a left-sided unilateral laminectomy at C5. To optimize the reproducibility of the biomechanical, functional, and histological outcomes of the injury model, we contused the spinal cords using an impact force of 150 kdyn, an impact trajectory of 22.5&deg; (animals rotated at 22.5&deg;), and an impact location off of midline of 1.4 mm. Functional recovery was assessed using the cylinder rearing test, horizontal ladder test, grooming test and modified Montoya's staircase test for up to 6 weeks, after which the spinal cords were evaluated histologically for white and grey matter sparing.
The injury model presented here imparts consistent and reproducible biomechanical forces to the spinal cord, an important feature of any experimental SCI model. This results in discrete histological damage to the lateral half of the spinal cord which is largely contained to the ipsilateral side of injury. The injury is well tolerated by the animals, but does result in functional deficits of the forelimb that are significant and sustained in the weeks following injury. The cervical unilateral injury model presented here may be a resource to researchers who wish to evaluate potentially promising therapies prior to human translation.

A reliable and repeatable way to produce a cervical unilateral spinal cord injury using the Infinite Horizon impactor is described. The method takes advantage of a custom designed frame and clamp to stabilize the spine. The standardized procedure and biomechanical injury parameters result in sufficient and sustained injuries.

Un modèle de contusion unilatérale des blessures moelle épinière cervicale aide de l&#39;impacteur Infinite Horizon

While the majority of human spinal cord injuries occur in the cervical spinal cord, the vast majority of laboratory research employs animal models of ...

A Murine Closed-chest Model of Myocardial Ischemia and Reperfusion

Surgical trauma by thoracotomy in open-chest models of coronary ligation induces an immune response which modifies different mechanisms involved in ischemia and reperfusion. Immune response includes cytokine expression and release or secretion of endogenous ligands of innate immune receptors. Activation of innate immunity can potentially modulate infarct size. We have modified an existing murine closed-chest model using hanging weights which could be useful for studying myocardial pre- and postconditioning and the role of innate immunity in myocardial ischemia and reperfusion. This model allows animals to recover from surgical trauma before onset of myocardial ischemia.
Volatile anesthetics have been intensely studied and their preconditioning effect for the ischemic heart is well known. However, this protective effect precludes its use in open chest models of coronary artery ligation. Thus, another advantage could be the use of the well controllable volatile anesthetics for instrumentation in a chronic closed-chest model, since their preconditioning effect lasts up to 72 hours. Chronic heart diseases with intermittent ischemia and multiple hit models are other possible applications of this model.
For the chronic closed-chest model, intubated and ventilated mice undergo a lateral blunt thoracotomy via the 4th intercostal space. Following identification of the left anterior descending a ligature is passed underneath the vessel and both suture ends are threaded through an occluder. Then, both suture ends are passed through the chest wall, knotted to form a loop and left in the subcutaneous tissue. After chest closure and recovery for 5 days, mice are anesthetized again, chest skin is reopened and hanging weights are hooked up to the loop under ECG control.
At the end of the ischemia/reperfusion protocol, hearts can be stained with TTC for infarct size assessment or undergo perfusion fixation to allow morphometric studies in addition to histology and immunohistochemistry.

Surgical trauma induces an inflammatory response. Cytokines and endogenous ligands are known to modulate myocardial infarct size following ischemia and reperfusion. We present a modified closed-chest model of murine ischemia and reperfusion using hanging weights to minimize effects of thoracotomy.

Un murin à thorax fermé Modèle d&#39;ischémie myocardique et la reperfusion

Surgical trauma by thoracotomy in open-chest models of coronary ligation induces an immune response which modifies different mechanisms involved in ...

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

Modèle murin de la cicatrisation

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

A Murine Model of Subarachnoid Hemorrhage

In this video publication a standardized mouse model of subarachnoid hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of Willis perforation (CWp) and proven by intracranial pressure (ICP) monitoring. Thereby a homogenous blood distribution in subarachnoid spaces surrounding the arterial circulation and cerebellar fissures is achieved. Animal physiology is maintained by intubation, mechanical ventilation, and continuous on-line monitoring of various physiological and cardiovascular parameters: body temperature, systemic blood pressure, heart rate, and hemoglobin saturation. Thereby the cerebral perfusion pressure can be tightly monitored resulting in a less variable volume of extravasated blood. This allows a better standardization of endovascular filament perforation in mice and makes the whole model highly reproducible. Thus it is readily available for pharmacological and pathophysiological studies in wild type and genetically altered mice.

A standardized mouse model of subarachnoid hemorrhage by intraluminal Circle of Willis perforation is described. Vessel perforation and subarachnoid bleeding are monitored by intracranial pressure monitoring. In addition various vital parameters are recorded and controlled to maintain physiologic conditions.

Un modèle murin d&#39;hémorragie sous-arachnoïdienne

In this video publication a standardized mouse model of subarachnoid  hemorrhage (SAH) is presented. Bleeding is induced by endovascular Circle of  Willis ...

A Preclinical Murine Model of Hepatic Metastases

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, murine pancreatic tumor model of hepatic metastases via a hemispleen injection of syngeneic murine pancreatic tumor cells is described. This model mimics many of the clinical conditions in patients with metastatic disease to the liver. Mice consistently develop metastases in the liver allowing for investigation of the metastatic process, experimental therapy testing, and tumor immunology research.

A preclinical, murine model of hepatic metastases performed via a hemispleen injection technique.

Un préclinique modèle murin de métastases hépatiques

Numerous murine models have been developed to study human cancers and advance the understanding of cancer treatment and development. Here, a preclinical, ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information about this disease. The human orthotopic prostate cancer xenograft mouse model provides a useful alternative approach for understanding the specific interactions between genetically and molecularly altered tumor cells, their organ microenvironment, and for evaluation of efficacy of therapeutic regimens. This is a well characterized model designed to study the molecular events of primary tumor development and it recapitulates the early events in the metastatic cascade prior to embolism and entry of tumor cells into the circulation. Thus it allows elucidation of molecular mechanisms underlying the initial phase of metastatic disease. In addition, this model can annotate drug targets of clinical relevance and is a valuable tool to study prostate cancer progression. In this manuscript we describe a detailed procedure to establish a human orthotopic prostate cancer xenograft mouse model.

Prostate cancer is the second most common cause of cancer-related deaths in the United States. An orthotopic cancer model provides a useful approach to understand the biology of prostate cancer and to evaluate the efficacy of therapeutic regimens. This protocol describes detailed steps necessary to establish an orthotopic prostate cancer mouse model.

Pré-clinique Orthotopique modèle murin de cancer de la prostate humaine

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

Generating a Murine Orthotopic Metastatic Breast Cancer Model and Performing Murine Radical Mastectomy

In vivo mouse models to assess breast cancer progression are essential for cancer research, including preclinical drug developments. However, the majority of the practical and technical details are commonly omitted in published manuscripts which, therefore, makes it challenging to reproduce the models, particularly when it involves surgical techniques. Bioluminescence technology allows for the evaluation of small amounts of cancer cells even when a tumor is not palpable. Utilizing luciferase-expressing cancer cells, we establish a breast cancer orthotopic inoculation technique with a high tumorigenesis rate. Lung metastasis is assessed utilizing an ex vivo technique. We, then, establish a mastectomy model with a low local recurrence rate to assess the metastatic tumor burden. Herein, we describe, in detail, the surgical techniques of orthotopic implantation and mastectomy for breast cancer with a high tumorigenesis rate and low local recurrence rates, respectively, to improve breast cancer model efficiency.

We introduce a murine orthotopic breast cancer model and radical mastectomy model with bioluminescence technology to quantify the tumor burden to mimic human breast cancer progression.

Génération d’un modèle de Cancer du sein métastatique orthotopique murin et effectuant une mastectomie radicale Murine

In vivo mouse models to assess breast cancer progression are essential for cancer research, including preclinical drug developments. However, the majority ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Mesure de la contractilité isométrique de l’artère mésentérique de souris à l’aide de fil myographie

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

A Reversible Silicon Oil-Induced Ocular Hypertension Model in Mice

Elevated intraocular pressure (IOP) is a well-documented risk factor for glaucoma. Here we describe a novel, effective method for consistently inducing stable IOP elevation in mice that mimics the post-operative complication of using silicone oil (SO) as a tamponade agent in human vitreoretinal surgery. In this protocol, SO is injected into the anterior chamber of the mouse eye to block the pupil and prevent inflow of aqueous humor. The posterior chamber accumulates aqueous humor and this in turn increases the IOP of the posterior segment. A single SO injection produces reliable, sufficient, and stable IOP elevation, which induces significant glaucomatous neurodegeneration. This model is a true replicate of secondary glaucoma in the eye clinic. To further mimic the clinical setting, SO can be removed from the anterior chamber to reopen the drainage pathway and allow inflow of aqueous humor, which is drained through the trabecular meshwork (TM) at the angle of the anterior chamber. Because IOP quickly returns to normal, the model can be used to test the effect of lowering IOP on glaucomatous retinal ganglion cells. This method is straightforward, does not require special equipment or repeat procedures, closely simulates clinical situations, and may be applicable to diverse animal species. However, minor modifications may be required.

Here, we present a protocol to induce ocular hypertension and glaucomatous neurodegeneration in mouse eyes by intracameral injection of silicone oil and the procedure for silicone oil removal from the anterior chamber to return elevated intraocular pressure to normal.

Un modèle d'hypertension oculaire induite par l'huile de silicium réversible chez les souris

Elevated intraocular pressure (IOP) is a well-documented risk factor for glaucoma. Here we describe a novel, effective method for consistently inducing ...