Les auteurs remercient Christiane Pahrmann pour l&#39;assistance technique. SS a reçu des fonds de la Deutsche Forschungsgemeinschaft (DFG) (SCHR992/3-2 et SCHR992/4-2).

Tous les travaux des animaux doit être effectué selon les directives de protection des animaux concernés. Obtenir l&#39;approbation institutionnelle pour le travail des animaux avant protocole de début. 1. Myointimale hyperplasie modèle (A) Pour le modèle de l&#39;hyperplasie intimale, acheter C57BL/6J (Stock nombre 000664, C57BL/6J) souris à l&#39;âge de 8 semaines pesant environ 25 g. Souris Maison à ces expériences dans des conditions classiques; nourrir les souris bouffe de souris standard et de fournir l&#39;eau ad libitum. <ol><li> Préparation de la souris: Présentez la souris à un état ​​d&#39;anesthésie avec de l&#39;isoflurane (2%) en utilisant une chambre d&#39;induction. <ol><li> Enlever les poils abdominale en utilisant une tondeuse à cheveux, poser la souris sur le dos et couvrir le nez et la bouche avec le masque facial pour assurer l&#39;anesthésie. </li><li> Avec Provo-iode, désinfecter la zone abdominale universellement, suivie par 80% d&#39;éthanol, en deux cycles. </li><li> Assurer l&#39;absencedes réflexes de pincement de la patte arrière pour surveiller la profondeur suffisante de l&#39;anesthésie. </li><li> Séparer la peau et les muscles le long de la ligne blanche en deux étapes pour exposer les organes abdominaux. </li><li> Disposez les intestins dans un gant de solution saline hydratée. Enroulez le gant autour des intestins pour les garder humides. </li><li> Attention déblayer le tissu adipeux bâches l&#39;aorte abdominale. Exposer la aorte jusqu&#39;à sa bifurcation, en prenant soin de ne pas blesser des navires de la branche. </li><li> Tout d&#39;abord, placer une pince de microchirurgie sur la proximale, aorte pour arrêter le flux de sang. Maintenant, appliquer une deuxième pince distale à côté de la bifurcation aortique. </li><li> Lorsque le flux de sang est interrompu, une petite incision dans l&#39;aorte à proximité de la pince proximale. </li><li> Insérer le cathéter à ballonnet, qui a été hydratée avec une solution saline à 0,9%, et de faire avancer la pince en direction distale de l&#39;aorte. </li><li> Pour lésion endothéliale, dénuder l&#39;aorte en gonflant le balloon prudemment et lentement en tirant le cathéter dans la direction proximale de la pince. </li><li> Répéter deux fois la manœuvre de la dénudation. </li><li> Retirer le cathéter et fermer l&#39;incision aortique par 10-0 prolène sutures de fonctionnement. </li><li> Ouvrez avec précaution la pince distale. En cas de saignement au site de aortotomie, fermer la pince à nouveau, localiser le saignement et placer des points interrompus supplémentaires, si nécessaire. </li><li> Si aucun saignement peut être observé, ouvrir lentement la pince proximale. </li><li> Une impulsion aortique doit être visible de manière distale à partir de l&#39;incision. Maintenant, les intestins peuvent être organisées retour in situ. </li><li> Rincer la cavité abdominale en utilisant une solution saline stérile préchauffée. </li><li> Fermer la paroi abdominale en commençant par la couche de muscles en utilisant 6-0 sutures prolène de fonctionnement. </li><li> Adapter ensuite la peau sous l&#39;utilisation de 5-0 prolène effectuer des sutures de fonctionnement. </li><li> Pendant le temps de la souris est toujours anesthésié, appliquer 4-5 mg / kg Carprofen via un Inje sous-cutanéection. </li><li> Ajouter Metamizol à l&#39;eau potable (50 mg par 100 ml Metamizol) pour le contrôle de la douleur et continuer pendant 3 jours après la chirurgie. Surveillance des animaux tout au long de la récupération. Note: La période d&#39;observation pour ce modèle est de 28 jours. </li></ol></li></ol> 2. L&#39;athérosclérose modèle (B) Pour le modèle de l&#39;athérosclérose, l&#39;achat d&#39;apolipoprotéine E déficient (ApoE - / -) chez la souris (Stock nombre 002052, B6.129P2-Apoetm1Unc / J) à l&#39;âge de 4 semaines, puis nourris pendant 4-6 mois. Souris Maison à ces expériences dans des conditions classiques; nourrir des souris haut régime occidental lipides à l&#39;arrivée (Harlan Laboratories TD.88137) et fournir de l&#39;eau ad libitum pour une durée de 4 à 6 mois. <ol><li> Préparation de la souris: <ol><li> Nourrir la ApoE - / - souris avec un régime alimentaire occidental à partir de 4 semaines d&#39;âge. Modification du régime alimentaire devrait commencer directement à l&#39;arrivée. </li><li> Maintenir une alimentation occidentale tout au long de la durée de l&#39;expérience. &lt;/ Li&gt; </ol></li></ol> 3. Analyse <ol><li> Modèle de l&#39;hyperplasie myo-intimale (A) La région d&#39;intérêt de ce modèle est la &quot;zone de lésion&quot;. Options pour analyse comprennent oblitération luminale, rapport I / M (rapport intima / média), épaisseur maximale de myointima, zone myointimale, coloration au trichrome de la fibrose, et coloration H &amp; E pour les cellules mononucléaires (comme illustré dans les figures 1A-1C). Selon la question scientifique comme de nombreux autres colorations rouge de picrosirius collagène, l&#39;immunohistochimie, et des procédés peuvent être appliqués à foyer commun. </li><li> modèle de l&#39;athérosclérose (B) Les régions d&#39;intérêt dans ce modèle sont la valve aortique, l&#39;arc aortique ainsi que l&#39;aorte descendante. Options pour analyse: Soudan coloration rouge pour les lipides et coloration au trichrome de la valve aortique, l&#39;arc aortique et de l&#39;aorte descendante (comme illustré à la figure 1D). Ce modèle donne également une variété d&#39;autres options de coloration commepar exemple Oil Red O pour les lipides neutres, picrosirius rouge, immunohistochimie, et les méthodes confocale. <ol><li> Soudan coloration rouge: Effectuer la coloration rouge Soudan, le tissu aortique fraîchement récoltées. <ol><li> Retirez le 6h de régime avant la récolte. </li><li> Introduire la souris à un état d&#39;anesthésie avec de l&#39;isoflurane (2%) en utilisant une chambre d&#39;induction. Assurer l&#39;absence de réflexes en pinçant la patte arrière pour surveiller la profondeur suffisante de l&#39;anesthésie. </li><li> Ouvrez le coffre; couper l&#39;apex du cœur et perfuser le ventricule gauche par l&#39;intermédiaire, d&#39;abord avec une solution saline (3 ml), suivie de 4% de PFA (3 ml) à travers la racine de l&#39;aorte à fixer l&#39;aorte. </li><li> Libérez l&#39;aorte de la racine à la bifurcation. Essayez d&#39;enlever les tissus gras autant que possible. </li><li> Coupez le coeur et la racine de l&#39;aorte, et les stocker dans 4% PFA pour l&#39;histologie. </li><li> Pour l&#39;histologie de la racine de l&#39;aorte doit être soigneusement placé dans le bloc de paraffine dans une position verticale pour permettre à la pointe de l&#39;aortic diapositives génératrices de racine qui affichent tous les trois dépliants valvulaires. </li><li> Couper une extrémité de l&#39;arbre de l&#39;aorte et le fixer sur la couche de silicone avec des épingles fines. Ouvrez l&#39;aorte longitudinalement et fixer le tissu aortique. </li><li> Rincer le tissu avec 50% d&#39;éthanol. Immerger le tissu avec une solution de coloration au Soudan III pendant environ 15 min. </li><li> Rincez légèrement avec 50% d&#39;éthanol pour éliminer l&#39;excès Soudan III. (Remarque: 50% d&#39;éthanol peut laver le colorant loin.) Rincer avec dH 2 O, et prendre des photos pour une analyse ultérieure. </li></ol></li></ol></li></ol>

<ol>
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B., Ma, X., Simard, T., Pourdjabbar, A., Hibbert, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathogenesis+of+neointima+formation+following+vascular+injury.">Pathogenesis of neointima formation following vascular injury.</a> Cardiovasc Hematol Disord Drug Targets. 11, 30-39 (2011).</li><li>Ross, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+pathogenesis+of+atherosclerosis:+a+perspective+for+the+1990s.">The pathogenesis of atherosclerosis: a perspective for the 1990s.</a> Nature. 362, 801-809 (1993).</li><li>Meng, X., Zhang, F., Valji, K., 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=Ultrasound+guidance+for+the+creation+of+a+small+animal+model+of+aortic+injury.">Ultrasound guidance for the creation of a small animal model of aortic injury.</a> J Vasc Interv Radiol. 22, 1193-1197 (2011).</li><li>Deuse, T., Koyanagi, T., Erben, R. G., 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=Sustained+inhibition+of+epsilon+protein+kinase+C+inhibits+vascular+restenosis+after+balloon+injury+and+stenting.">Sustained inhibition of epsilon protein kinase C inhibits vascular restenosis after balloon injury and stenting.</a> Circulation. 122, 170-178 (2010).</li><li>Schrepfer, S., Deuse, T., Sultan, K. R., 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=Inhibition+of+restenosis+development+after+mechanical+injury:+a+new+field+of+application+for+malononitrilamides.">Inhibition of restenosis development after mechanical injury: a new field of application for malononitrilamides.</a> Cardiology. 108, 128-137 (2007).</li><li>Tao, M., Mauro, C. R., Yu, P., 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+simplified+murine+intimal+hyperplasia+model+founded+on+a+focal+carotid+stenosis.">A simplified murine intimal hyperplasia model founded on a focal carotid stenosis.</a> Am J Pathol. 182, 277-287 (2013).</li><li>Kumar, A., Lindner, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Remodeling+with+neointima+formation+in+the+mouse+carotid+artery+after+cessation+of+blood+flow.">Remodeling with neointima formation in the mouse carotid artery after cessation of blood flow.</a> Arterioscler Thromb Vasc Biol. 17, 2238-2244 (1997).</li><li>Hua, X., Deuse, T., Michelakis, E. 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=Human+internal+mammary+artery+(IMA)+transplantation+and+stenting:+a+human+model+to+study+the+development+of+in-stent+restenosis.">Human internal mammary artery (IMA) transplantation and stenting: a human model to study the development of in-stent restenosis.</a> J Vis Exp. , 3663 (2012).</li><li>Daugherty, A., Whitman, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+atherosclerosis+in+mice.">Quantification of atherosclerosis in mice.</a> Methods Mol Biol. , (2003).</li><li>Nakashima, Y., Plump, A. S., Raines, E. W., Breslow, J. L., Ross, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=ApoE-deficient+mice+develop+lesions+of+all+phases+of+atherosclerosis+throughout+the+arterial+tree.">ApoE-deficient mice develop lesions of all phases of atherosclerosis throughout the arterial tree.</a> Arterioscler Thromb. 14, 133-1340 (1994).</li><li>Zhang, Y., Li, L., You, J., Cao, J., Fu, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+7-difluoromethyl-5,+4'-dimethoxygenistein+on+aorta+atherosclerosis+in+hyperlipidemia+ApoE(-/-)+mice+induced+by+a+cholesterol-rich+diet.">Effect of 7-difluoromethyl-5, 4'-dimethoxygenistein on aorta atherosclerosis in hyperlipidemia ApoE(-/-) mice induced by a cholesterol-rich diet.</a> Drug Des Devel Ther. 7, 233-242 (2013).</li><li>Rudolph, T. K., Rudolph, V., Edreira, M. 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=Nitro-fatty+acids+reduce+atherosclerosis+in+apolipoprotein+E-deficient+mice.">Nitro-fatty acids reduce atherosclerosis in apolipoprotein E-deficient mice.</a> Arterioscler Thromb Vasc Biol. 30, 938-9345 (2010).</li><li>Rondelet, B., Kerbaul, F., Motte, 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=Bosentan+for+the+prevention+of+overcirculation-induced+experimental+pulmonary+arterial+hypertension.">Bosentan for the prevention of overcirculation-induced experimental pulmonary arterial hypertension.</a> Circulation. 107, 1329-1335 (2003).</li><li>Zeibig, S., Li, Z., Wagner, 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=Effect+of+the+oxLDL+binding+protein+Fc-CD68+on+plaque+extension+and+vulnerability+in+atherosclerosis.">Effect of the oxLDL binding protein Fc-CD68 on plaque extension and vulnerability in atherosclerosis.</a> Circ Res. 108, 695-703 (2011).</li><li>Painter, T. 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Les auteurs n&#39;ont rien à révéler.

Bien que ces deux maladies entraînent des symptômes similaires chez le patient, les mécanismes sous-jacents de développement de la plaque et par conséquent, les approches de traitement sont très différents 2. Dans différentes formes de sténose artérielle les résultats cliniques des patients ainsi que la période de développement de la plaque dépendent du mécanisme sous-jacent. Selon les résultats cliniques chez le patient, différentes méthodes d&#39;analyse de l&#...

Dans les pays industrialisés cardiopathie ischémique reste une des principales causes de la mortalité 1. Les causes de sténose de l&#39;artère coronaire sont multiples et comprennent l&#39;hyperplasie myo-intimale ainsi que l&#39;athérosclérose, une revascularisation restant stratégie de traitement la plus courante 2. Les modèles de recherche qui distinguent clairement les voies mécanistiques de l&#39;hyperplasie intimale et l&#39;athérosclérose sont essentiels pour enquêter sur la pathobiologique et processus physiopathologiques en développement de la plaque artérielle. Dans cette vidéo, deux modèles différents de souris utilisées pour étudier soit le développement de l&#39;hyperplasie myo-intimale ou l&#39;athérosclérose sont introduits. Hyperplasie myo-intimale est postulé pour former via le paradigme &quot;réponse à des blessures&quot; décrit à l&#39;origine de l&#39;athérosclérose 3. La rupture mécanique de la couche endothéliale conduit à un remodelage intense renforcée par des effets paracrines. Il existe plusieurs dimodèles animaux fférents couramment utilisées pour étudier l&#39;hyperplasie myo-intimale. Certains groupes utilisent des dispositifs de métaux dans l&#39;aorte ascendante de 4 rats. Le modèle de dénudation aortique réalisée avec un cathéter à ballonnet est également communément utilisé dans des rats de laboratoire 5,6. Le modèle de l&#39;hyperplasie intimale réalisée chez des souris de laboratoire 7 met en oeuvre la ligature de l&#39;artère carotide et induit des lésions myo-intimales 8. Dans notre laboratoire, le modèle de la dénudation de l&#39;aorte abdominale - pour étudier le développement de l&#39;hyperplasie myo-intimale - ainsi que d&#39;un modèle humanisé stent resténose 9 sont couramment utilisés. Cette vidéo souligne que le choix du modèle animal expérimental adéquat est essentiel pour des études mécanistiques ou physiopathologiques de la sténose artérielle. Modèles hyperplasie myo-intimale doivent être distinguées des modèles d&#39;athérosclérose. Pour ces derniers, l&#39;apolipoprotéine E-deficient (ApoE - / -) chez la souris en combinaison avec un régime alimentaire occidental sont couramment utilisés pour induire atheroscllésions érotiques 10,11,12,13. Des exemples pour l&#39;induction de ces deux types de la maladie chez les souris myointimal sont présentés ici, avec différentes options de modèle adapté pour analyser vaisseau sténose 14.

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		<col />
		<col />
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			<td height="17" style="height:17px;width:156px;">Name</td>
			<td style="width:183px;">Company</td>
			<td style="width:139px;">Catalog number</td>
			<td style="width:140px;">Comments</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Thyoglycollate Broth 3%</td>
			<td>Fluka</td>
			<td align="right">70157</td>
			<td>powder</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">PFA 4%</td>
			<td>Electron Microscopy Sciences</td>
			<td>#157135S</td>
			<td align="right">20%</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Sudan III staining solution</td>
			<td>Sigma Aldrich</td>
			<td>S4131</td>
			<td>powder</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">mouse C57BL/6J</td>
			<td>Jackson Laboratories</td>
			<td>&#160;Stock # 0006664</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">mouse ApoE-/-</td>
			<td>Jackson Laboratories</td>
			<td>&#160;Stock #002052</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Western Diet</td>
			<td>Harlan Laboratories</td>
			<td>TD.88137</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">hair clipper</td>
			<td>WAHL</td>
			<td>8786-451A ARCO SE</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Forene</td>
			<td>Abbott</td>
			<td></td>
			<td>Isoflurane</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">microsurgical clamp</td>
			<td>Fine Science Tools</td>
			<td>18055-04</td>
			<td>Micro-Serrefine - 4mm</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">clamp applicator</td>
			<td>Fine Science Tools</td>
			<td>18056-14</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">catheter</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">10-0 prolene suture</td>
			<td>Ethicon</td>
			<td>788G</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">6-0 prolene suture</td>
			<td>Ethicon</td>
			<td>8709H</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">5-0 prolene suture</td>
			<td>Ethicon</td>
			<td>EH7229H</td>
			<td></td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Rimadyl</td>
			<td>Pfizer&#160;</td>
			<td></td>
			<td>Carprofen</td>
		</tr>
		<tr height="17">
			<td height="17" style="height:17px;">Metacam 1.5mg/ml</td>
			<td>Boehringer Ingelheim</td>
			<td></td>
			<td>Metamizol</td>
		</tr>
	</tbody>
</table>

inducing myointimal hyperplasia versus atherosclerosis in mice: an introduction of two valid models

Différents modèles in vivo laboratoire de rongeurs pour l&#39;induction de la sténose de l&#39;artère ont été créés pour imiter les maladies qui comprennent la formation de la plaque artérielle et de la sténose, comme on l&#39;observe par exemple dans la cardiopathie ischémique. Deux modèles de souris hautement reproductibles - à la fois entraînant sténose de l&#39;artère sous-jacents, mais chacun un chemin différent de développement - sont introduits ici. Les modèles représentent les deux causes les plus fréquentes de sténose de l&#39;artère; à savoir un modèle de souris pour chaque hyperplasie myointimale et l&#39;athérosclérose sont présentés. Provoquer l&#39;hyperplasie myo-intimale, une lésion par cathéter à ballonnet de l&#39;aorte abdominale est effectuée. Pour le développement de la plaque athéroscléreuse, l&#39;ApoE - / - modèle de souris en combinaison avec un régime alimentaire occidental gras est utilisé. Options de modèle adapté différentes pour la mesure et l&#39;évaluation des résultats sont nommées et décrites dans ce manuscrit. L&#39;introduction et la comparaison de ces deux modèles fournit information pour les scientifiques de choisir le modèle de sténose de l&#39;artère appropriée, conformément à la question scientifique posée.

La figure 1 montre les différentes options d&#39;analyse et l&#39;état de la maladie induite pour les deux modèles. Pour le modèle de l&#39;hyperplasie intimale, l&#39;analyse des sections représentatives de coloration au trichrome est affiché. A partir de ces sections transversales, de tels résultats ratios E / S, l&#39;épaisseur maximale de la plaque, l&#39;épaisseur maximale de l&#39;intima dans le lumen, l&#39;oblitération luminale en pour cent, ainsi que la zone de la plaque peuvent être...

Watch this Scientific Journal Video about Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models at JoVE.com

Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

Cette vidéo montre deux modèles de l&#39;intima développement de la plaque dans les artères de souris et met l&#39;accent sur les différences dans l&#39;hyperplasie myo-intimale et l&#39;athérosclérose.

Induisant myointimale hyperplasie Versus athérosclérose chez la souris: An Introduction de deux modèles valides

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque ...

inducing-myointimal-hyperplasia-versus-atherosclerosis-mice-an

Research

JoVE Journal

Medicine

12.0K vues.  University Hospital Hamburg.  Cette vidéo montre deux modèles de l'intima développement de la plaque dans les artères de souris et met l'accent sur les différences dans l'hyperplasie myo-intimale et l'athérosclérose. 

Vidéo: Inducing Myointimal Hyperplasia Versus Atherosclerosis in Mice: An Introduction of Two Valid Models

Implantation of a Carotid Cuff for Triggering Shear-stress Induced Atherosclerosis in Mice

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce atherosclerosis (reviewed in 1 and 2). The role of shear stress has been extensively studied in vitro investigating the influence of flow dynamics on cultured endothelial cells 1,3,4 and in vivo in larger animals and humans 1,5,6,7,8. However, highly reproducible small animal models allowing systematic investigation of the influence of shear stress on plaque development are rare. Recently, Nam et al. 9 introduced a mouse model in which the ligation of branches of the carotid artery creates a region of low and oscillatory flow. Although this model causes endothelial dysfunction and rapid formation of atherosclerotic lesions in hyperlipidemic mice, it cannot be excluded that the observed inflammatory response is, at least in part, a consequence of endothelial and/or vessel damage due to ligation.
In order to avoid such limitations, a shear stress modifying cuff has been developed based upon calculated fluid dynamics, whose cone shaped inner lumen was selected to create defined regions of low, high and oscillatory shear stress within the common carotid artery 10. By applying this model in Apolipoprotein E (ApoE) knockout mice fed a high cholesterol western type diet, vascular lesions develop upstream and downstream from the cuff. Their phenotype is correlated with the regional flow dynamics 11 as confirmed by in vivo Magnetic Resonance Imaging (MRI) 12: Low and laminar shear stress upstream of the cuff causes the formation of extensive plaques of a more vulnerable phenotype, whereas oscillatory shear stress downstream of the cuff induces stable atherosclerotic lesions 11. In those regions of high shear stress and high laminar flow within the cuff, typically no atherosclerotic plaques are observed.
 
In conclusion, the shear stress-modifying cuff procedure is a reliable surgical approach to produce phenotypically different atherosclerotic lesions in ApoE-deficient mice.

The constricting cuff presented in this article is designed to induce atherosclerosis in the murine common carotid artery. Due to the conical shape of its inner lumen the implanted cuff generates well-defined regions of low, high and oscillatory shear stress triggering the development of atherosclerotic lesions of different inflammatory phenotypes.

Implantation d&#39;une manchette carotidienne pour le déclenchement de cisaillement-stress athérosclérose induite chez la souris

It is widely accepted that alterations in vascular shear stress trigger the expression of inflammatory genes in endothelial cells and thereby induce ...

Recherche

Médecine

Quantitative Analysis and Characterization of Atherosclerotic Lesions in the Murine Aortic Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have been developed to facilitate the study of the molecular and cellular pathophysiology of this disease. In this manuscript we describe specific techniques for the quantification and characterization of atherosclerotic lesions in the murine aortic sinus and ascending aorta. The advantage of this procedure is that it provides an accurate measurement of the cross-sectional area and total volume of the lesion, which can be used to compare atherosclerotic progression across different treatment groups. This is possible through the use of the valve leaflets as an anatomical landmark, together with careful adjustment of the sectioning angle. We also describe basic staining methods that can be used to begin to characterize atherosclerotic progression. These can be further modified to investigate antigens of specific interest to the researcher. The described techniques are generally applicable to a wide variety of existing and newly created dietary and genetically-induced models of atherogenesis.

We describe procedures to quantify and characterize atherosclerotic lesions in mouse models using precision sectioning of the aortic sinus and ascending aorta combined with histochemical and immunohistochemical analysis.

Analyse quantitative et caractérisation des lésions d&#39;athérosclérose dans le murin aortique Sinus

Atherosclerosis is a disease of the large arteries and a major underlying cause of myocardial infarction and stroke. Several different mouse models have ...

Acute Brain Trauma in Mice Followed By Longitudinal Two-photon Imaging

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no effective treatment for it yet. Limitations of currently used animal models impede understanding of the pathology mechanism. Multiphoton microscopy allows studying cells and tissues within intact animal brains longitudinally under physiological and pathological conditions. Here, we describe two models of acute brain injury studied by means of two-photon imaging of brain cell behavior under posttraumatic conditions. A selected brain region is injured with a sharp needle to produce a trauma of a controlled width and depth in the brain parenchyma. Our method uses stereotaxic prick with a syringe needle, which can be combined with simultaneous drug application. We propose that this method can be used as an advanced tool to study cellular mechanisms of pathophysiological consequences of acute trauma in mammalian brain in vivo. In this video, we combine acute brain injury with two preparations: cranial window and skull thinning. We also discuss advantages and limitations of both preparations for multisession imaging of brain regeneration after trauma.

Acute brain trauma is a severe injury that has no adequate treatment to date. Multiphoton microscopy allows studying longitudinally the process of acute brain trauma development and probing therapeutical strategies in rodents. Two models of acute brain trauma studied with in vivo two-photon imaging of brain are demonstrated in this protocol.

Aiguë traumatisme cérébral chez la souris Suivi par longitudinale à deux photons imagerie

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no ...

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 intervention chirurgicale invasive d&#39;induire un infarctus du myocarde chez des souris

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

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.

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.

Les modèles de chirurgie de reflux gastro-œsophagien avec des souris

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

An Improved Method for Rapid Intubation of the Trachea in Mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

Une méthode améliorée pour Rapid intubation de la trachée chez la souris

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the brain for visual perception. Damage to RGCs and their axons leads to vision loss and/or blindness. Although the specific cause of glaucoma is unknown, the primary risk factor for the disease is an elevated intraocular pressure. Glaucoma-inducing procedures in animal models are a valuable tool to researchers studying the mechanism of RGC death. Such information can lead to the development of effective neuroprotective treatments that could aid in the prevention of vision loss. The protocol in this paper describes a method of inducing glaucoma - like conditions in an in vivo rat model where 50 &#181;l of 2 M hypertonic saline is injected into the episcleral venous plexus. Blanching of the vessels indicates successful injection. This procedure causes loss of RGCs to simulate glaucoma. One month following injection, animals are sacrificed and eyes are removed. Next, the cornea, lens, and vitreous are removed to make an eyecup. The retina is then peeled from the back of the eye and pinned onto sylgard dishes using cactus needles. At this point, neurons in the retina can be stained for analysis. Results from this lab show that approximately 25% of RGCs are lost within one month of the procedure when compared to internal controls. This procedure allows for quantitative analysis of retinal ganglion cell death in an in vivo rat glaucoma model.

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is characterized by damage to retinal ganglion cells. Inducing glaucoma in animal models can provide insight into the study of this disease. Here, we outline a procedure that induces loss of RGCs in an in vivo rat model and demonstrates the preparation of whole-mount retinas for analysis.

Glaucomes induisant une procédure dans un<em&gt; In Vivo</em&gt; Modèle de rat et Whole-mount Retina Préparation

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

Induction de l’athérosclérose accélérée chez les souris : le modèle « Wire-Injury »

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

Blessure axée sur le ballon pour provoquer l’hyperplasie Myointimal dans l’aorte abdominale de souris

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...

Inducing Acute Lung Injury in Mice by Direct Intratracheal Lipopolysaccharide Instillation

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. Various approaches have been described, such as the inhalation of aerosolized LPS as well as nasal or intratracheal instillation. The presented protocol describes a detailed step-by-step procedure to induce ALI in mice by direct intratracheal LPS instillation and perform FACS analysis of blood samples, bronchoalveolar lavage (BAL) fluid, and lung tissue. After intraperitoneal sedation, the trachea is exposed and LPS is administered via a 22 G venous catheter. A robust and reproducible inflammatory reaction with leukocyte invasion, upregulation of proinflammatory cytokines, and disruption of the alveolo-capillary barrier is induced within hours to days, depending on the LPS dosage used. Collection of blood samples, BAL fluid, and lung harvesting, as well as the processing for FACS analysis, are described in detail in the protocol. Although the use of the sterile LPS is not suitable to study pharmacologic interventions in infectious diseases, the described approach offers minimal invasiveness, simple handling, and good reproducibility to answer mechanistic immunological questions. Furthermore, dose titration as well as the use of alternative LPS preparations or mouse strains allow modulation of the clinical effects, which can exhibit different degrees of ALI severity or early vs. late onset of disease symptoms.

Presented here is a step-by-step procedure to induce acute lung injury in mice by direct intratracheal lipopolysaccharide instillation and to perform FACS analysis of blood samples, bronchoalveolar lavage fluid, and lung tissue. Minimal invasiveness, simple handling, good reproducibility, and titration of disease severity are advantages of this approach.

Induire une lésion pulmonaire aigue chez la souris par instillation directe du lipopolysaccharide intratrachéal

Airway administration of lipopolysaccharide (LPS) is a common way to study pulmonary inflammation and acute lung injury (ALI) in small animal models. ...