Die Autoren danken Christiane Pahrmann für technische Unterstützung. SS wurde mit Mitteln der Deutschen Forschungsgemeinschaft (DFG) (SCHR992/3-2 und SCHR992/4-2).

Alle Tier Arbeiten sollten nach den einschlägigen Tierpflege-Richtlinien durchgeführt werden. Erhalten institutionelle Zulassung für Tier Arbeit vor Beginn Protokoll. 1. Myointimale Hyperplasie Modell (A) Für die Intimahyperplasie Modell kaufen C57BL/6J (Artikelnummer 000664, C57BL/6J)-Mäuse im Alter von 8 Wochen mit einem Gewicht von ca. 25 g. Hausmäuse für diese Versuche unter herkömmlichen Bedingungen; Mäuse füttern Standardmäusefutter und Wasser ad libitum zur Verfügung stellen. <ol><li> Maus Zubereitung: Führen Sie die Maus, um ein Narkosezustand mit Isofluran (2%) unter Verwendung eines Induktionskammer. <ol><li> Entfernen Sie die Bauch Haare mit einem Haarschneider, legen Sie die Maus auf den Rücken und decken ihre Nase und Mund mit der Gesichtsmaske, um die Narkose zu gewährleisten. </li><li> Mit Provo-Jod, desinfizieren Sie die Bauch-Bereich allgemein, gefolgt von 80% Ethanol, in zwei Zyklen. </li><li> Sichern Sie die Abwesenheitvon Reflexen durch Kneifen der Hintergliedmaße, die ausreichende Narkosetiefe zu überwachen. </li><li> Trennen Sie die Haut und Muskeln entlang der Linea alba in zwei Schritten auf die Bauchorgane aus. </li><li> Legen Sie den Darm in einer Kochsalzlösung befeuchtet Handschuh. Wickeln Sie den Handschuh um den Darm, um sie feucht zu halten. </li><li> Aufmerksam wegzuräumen das Ausrollen des Bauchaorta Fettgewebe. Expose der infrarenalen Aorta bis auf die Bifurkation, kümmert sich nicht um alle Zweiggefäße verletzen. </li><li> Zuerst ein mikro Klemme an der proximalen, infrarenalen Aorta zum Herunterfahren des Blutflusses. Nun tragen Sie eine zweite distale Klemm neben der Bifurkation. </li><li> Wenn der Blutstrom unterbrochen wird, einen kleinen Einschnitt in der Aorta in der Nähe des proximalen Klemme. </li><li> Legen Sie die Ballonkatheters, der mit 0,9% Salzlösung befeuchtet wurde, und vorab in Richtung des distalen Klemme der Aorta. </li><li> Für endotheliale Verletzung, entblößen die Aorta durch Aufblasen des balloon vorsichtig und langsam Ziehen des Katheters in Richtung des proximalen Klemme. </li><li> Wiederholen Sie zweimal die Entblößung Manöver. </li><li> Entfernen Sie den Katheter und schließen Sie die Aorten-Schnitt mit 10-0 Prolene laufenden Nähten. </li><li> Öffnen Sie vorsichtig das distale Klemme. Im Falle von Blutungen an der Aortotomie Website, schließen Sie die Klemme wieder, suchen die Blutung und setzen zusätzliche unterbrochen Stiche, wie nötig. </li><li> Wenn keine Blutungen beobachtet werden kann, öffnen Sie langsam das proximale Klemme. </li><li> Eine Aortapuls sollte distal von dem Einschnitt sichtbar. Jetzt kann der Darm wieder in situ angeordnet werden. </li><li> Spülen Sie die Bauchhöhle Verwendung vorgewärmten sterilen Kochsalzlösung. </li><li> Schließen Sie die Bauchdecke beginnend mit der Muskelschicht mit 6-0 Prolene laufenden Nähten. </li><li> Weiter die Haut anpassen unter der Verwendung von 5-0 Prolene Durchführung laufenden Nähten. </li><li> Während der Zeit, die Maus ist immer noch betäubt, gelten 4-5 mg / kg Carprofen über eine subkutane injektion. </li><li> In Metamizol dem Trinkwasser (50 mg Metamizol pro 100 ml) zur Schmerzkontrolle und fahren für 3 Tage nach der Operation. Überwachung der gesamten Tier Erholung. Hinweis: Der Beobachtungszeitraum für dieses Modell beträgt 28 Tage. </li></ol></li></ol> 2. Atherosklerose-Modell (B) Für die Atherosklerose-Modell, kaufen Apolipoprotein E-defizienten (ApoE - / -)-Mäusen (Artikelnummer 002052, B6.129P2-Apoetm1Unc / J) im Alter von 4 Wochen, und dann für 4-6 Monate zugeführt. Hausmäuse für diese Versuche unter herkömmlichen Bedingungen; Mäuse füttern hohen Lipid westlichen Ernährung bei der Ankunft (Harlan Laboratories TD.88137) und bieten Wasser ad libitum für eine Dauer von 4 bis 6 Monaten. <ol><li> Maus Zubereitung: <ol><li> Führen Sie das ApoE - / - Mäusen, die mit westlichen Ernährung bei Alter von 4 Wochen ab. Diätetische Manipulation sollte direkt nach der Ankunft beginnen. </li><li> Pflegen westlichen Ernährung gesamten Dauer des Experiments. &lt;/ Li&gt; </ol></li></ol> 3. Analyse <ol><li> Myointimale Hyperplasie Modell (A) Die Region von Interesse in diesem Modell ist der &quot;Läsion Bereich&quot;. Optionen für die Analyse sind luminalen Auslöschung, I / M-Verhältnis (Intima / Media-Verhältnis), die maximale Dicke myointima, myointimale Bereich Trichromfärbung für Fibrose und H &amp; E-Färbung für mononuklearen Zellen (wie in den 1A-1C dargestellt). Nach der wissenschaftlichen Fragestellung zahlreiche andere Färbungen wie Picrosirius rot für Kollagen, Immunhistochemie und konfokale Methoden verwendet werden. </li><li> Atherosklerose-Modell (B) Die Bereiche von Interesse in diesem Modell sind die Aortenklappe Aortenbogen und die absteigende Aorta. Optionen für die Analyse: Sudan-Rot-Färbung für Lipide und Trichromfärbung der Aortenklappe, Aortenbogen und absteigender Aorta (wie in 1D dargestellt). Dieses Modell liefert auch eine Vielzahl von anderen Optionen, wie Färbungbeispielsweise Oil Red O für neutrale Lipide, Picrosirius rot, Immunhistochemie und konfokale Methoden. <ol><li> Sudan-Rot-Färbung: Führen Sie die Sudan-Rot-Färbung auf frisch geernteten Aortengewebe. <ol><li> Entfernen Sie die Diät-6h vor der Ernte. </li><li> Führen Sie die Maus, um ein Narkosezustand mit Isofluran (2%) unter Verwendung eines Induktionskammer. Assure das Fehlen von Reflexen durch Kneifen der Hintergliedmaße, die ausreichende Narkosetiefe zu überwachen. </li><li> Öffnen Sie die Brust; bricht die Spitze des Herzens und Perfusion über den linken Ventrikel zuerst mit Kochsalzlösung (3 ml), gefolgt von 4% PFA (3 ml) durch die Aortawurzel an der Aorta zu befestigen. </li><li> Befreie die Aorta von der Wurzel zu ihrer Bifurkation. Versuchen, Fettgewebe zu entfernen, so viel wie möglich. </li><li> Schneiden Sie das Herz und die Aortenwurzel, und speichern sie in 4% PFA für die Histologie. </li><li> Für die Histologie ist die Aortenwurzel sorgfältig in dem Paraffinblock in eine aufrechte Position, um die Schneiden der AOR ermöglichen gelegt werdentic Wurzel Erzeugungs Folien, die alle drei Klappenflugblätter anzuzeigen. </li><li> Schneiden Sie ein Ende des Aorten-Baum und befestigen es auf der Silikonschicht mit feinen Stiften. Öffnen Sie die Aorta in Längsrichtung und fixieren Sie die Aorten-Gewebe. </li><li> Spülen Sie das Gewebe mit 50% Ethanol. Tauchen Sie das Gewebe mit Sudan III-Färbung Lösung für ca. 15 min. </li><li> Leicht Spülung mit 50% Ethanol, um die überschüssige Sudan III zu entfernen. (Anmerkung: 50% Ethanol kann der Farbstoff weg zu waschen.) Spülen mit dH 2 O, und Fotos für die weitere Analyse. </li></ol></li></ol></li></ol>

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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. 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Die Autoren haben nichts zu offenbaren.

Obwohl beide Erkrankungen führen zu ähnlichen Symptomen des Patienten, sind die zugrunde liegenden Mechanismen der Plaqueentwicklung und damit die Behandlungsansätze sehr unterschiedlich 2. In verschiedenen Formen der arteriellen Stenose die klinischen Befunde bei den Patienten als auch der Zeitrahmen von Plaqueentwicklung hängt von der zugrunde liegenden Mechanismus. Je nach den klinischen Befunden im Patienten müssen verschiedene Methoden der Probenanalyse durchgeführt wer...

In den Industrieländern bleibt die ischämische Herzkrankheit eine der Hauptursachen der Sterblichkeit ein. Die Ursachen für Koronarstenosen sind vielfältig und umfassen myointimale Hyperplasie sowie Arteriosklerose, mit Rest die häufigste Behandlungsstrategie 2 Revaskularisation. Forschungsmodelle, die klar zwischen den Reaktionswege der Intimahyperplasie und Atherosklerose sind unerlässlich, um die pathobiologischen und pathophysiologische Prozesse im arteriellen Plaqueentwicklung zu untersuchen. In diesem Video zwei verschiedene Mausmodelle verwendet werden, um entweder die Entwicklung von myointimale Hyperplasie oder Atherosklerose studieren werden eingeführt. Myointimale Hyperplasie wird postuliert, dass über die &quot;response-to-injury&quot; Paradigma ursprünglich für Atherosklerose 3 beschrieben zu bilden. Die mechanische Störung der endothelialen Schicht führt zu einem intensiven Umbau durch parakrine Effekte verbessert. Es gibt mehrere different Tiermodellen häufig verwendet, um myointimale Hyperplasie zu studieren. Einige Gruppen verwenden Metall Geräte in der aufsteigenden Aorta von Ratten 4. Die Aorten Denudation Modell mit einem Ballonkatheter durchgeführt wird häufig auch in Laborratten 5,6 verwendet. Die Intimahyperplasie Modell in Labormäusen durchgeführt, 7 implementiert Ligation der A. carotis und induziert myointimale Läsionen 8. In unserem Labor, die Bauch-Aorten-Entblößung Modell - sowie einen humanisierten Stent-Restenose-Modell 9 werden häufig verwendet, - die Entwicklung von myointimale Hyperplasie zu studieren. Dieses Video wird betont, dass die Wahl der richtigen experimentellen Tiermodell ist entscheidend für die mechanistische oder pathophysiologische Studien der arteriellen Stenose. Myointimale Hyperplasie Modelle müssen von Atherosklerose-Modellen unterscheiden. Für letztere Apolipoprotein E-defizienten (ApoE - / -)-Mäuse sind in Kombination mit westlichen Ernährung häufig verwendet, um atheroscl induzierenErotik Läsionen 10,11,12,13. Beispiele für die Induktion von beiden dieser Arten von myointimale Krankheit in Mäusen werden hier gezeigt zusammen mit verschiedenen Modelloptionen angepasst, um Gefäßverengung 14 zu analysieren.

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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>
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			<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>
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		<tr height="17">
			<td height="17" style="height:17px;">Western Diet</td>
			<td>Harlan Laboratories</td>
			<td>TD.88137</td>
			<td></td>
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		<tr height="17">
			<td height="17" style="height:17px;">hair clipper</td>
			<td>WAHL</td>
			<td>8786-451A ARCO SE</td>
			<td></td>
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		<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>
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			<td height="17" style="height:17px;">catheter</td>
			<td></td>
			<td></td>
			<td></td>
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			<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>
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		<tr height="17">
			<td height="17" style="height:17px;">Metacam 1.5mg/ml</td>
			<td>Boehringer Ingelheim</td>
			<td></td>
			<td>Metamizol</td>
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inducing myointimal hyperplasia versus atherosclerosis in mice: an introduction of two valid models

Verschiedene in vivo-Labor-Nagetiermodellen für die Induktion der Stenose festgelegt worden, um Krankheiten, die arterielle Plaque-Bildung und Stenose umfassen, wie z. B. bei ischämischen Herzkrankheiten beobachtet imitieren. Zwei hoch reproduzierbare Mausmodelle - sowohl was Stenose, aber jeder einen anderen Weg der Entwicklung zu Grunde liegenden - hier werden eingeführt. Die Modelle stellen die beiden häufigsten Ursachen der Stenose; nämlich ein Maus-Modell für jeden myointimale Hyperplasie und Atherosklerose gezeigt. Um myointimale Hyperplasie wird ein Ballonkatheterverletzung der abdominalen Aorta. Für die Entwicklung von atherosklerotischen Plaques, die ApoE - / - Maus-Modell wird in Kombination mit westlichen fettreiche Ernährung verwendet werden. Verschiedene Modelloptionen angepasst für die Messung und Auswertung der Ergebnisse benannt und in diesem Manuskript beschrieben. Die Einführung und Vergleich dieser beiden Modelle bietet InformatIonen für Wissenschaftler, um die entsprechende Stenose Modell in Übereinstimmung mit der wissenschaftlichen Frage gestellt zu wählen.

Abbildung 1 zeigt verschiedene Analysemöglichkeiten und die induzierte Krankheitszustand für beide Modelle. Für die Intima-Hyperplasie Modell wird die Auswertung von repräsentativen Querschnitte in Trichromfärbung gezeigt. Aus diesen Querschnitten, können die Ergebnisse wie I / M-Verhältnisse, die maximale Plaquedicke, maximale Dicke der Intima in das Lumen, luminalen Auslöschung in Prozent, sowie Plaque-Bereich gemessen und berechnet werden. Um das Ergebnis der Athe...

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

Dieses Video zeigt zwei Modelle der Intima Plaqueentwicklung in murinen Arterien und betont die Unterschiede in myointimale Hyperplasie und Atherosklerose.

Induktion Myointimale Hyperplasie Versus Atherosklerose in Mäuse: Eine Einführung von zwei gültigen Modelle

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

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12.0K Aufrufe.  University Hospital Hamburg.  Dieses Video zeigt zwei Modelle der Intima Plaqueentwicklung in murinen Arterien und betont die Unterschiede in myointimale Hyperplasie und Atherosklerose. 

Serie: 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 einer Karotis Cuff für Triggering Shear-Stress-Induced Arteriosklerose bei Mäusen

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

Forschung

Medizin

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.

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.

Akute Brain Trauma in Mäusen Gefolgt durch Längs Zwei-Photonen-Imaging

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-invasive chirurgische Verfahren zur Induktion Myokardinfarkt 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 ...

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.

Chirurgische Modelle aus gastroösophagealen Reflux mit Mäusen

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.

Ein verbessertes Verfahren zur schnellen Intubation der Luftröhre in Mice

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.

Glaucoma auslösende Verfahren in ein<em&gt; In Vivo</em&gt; Rat Modell und Whole-mount Retina Vorbereitung

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.

Induktion von beschleunigter Atherosklerose bei Mäusen: Das "Wire-Injury"-Modell

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.

Ballon-basierte Verletzungen induzieren Myointimal Hyperplasie in der Bauchschlagader Maus

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.

Induzieren akute Lungenschädigung bei Mäusen durch direkte intratracheale Lipopolysaccharid-Instillation

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