Das Projekt wird teilweise durch die NIH T32 GM008440 zu Espen Walker, R01 NS27713 William L.Young, P01 NS44155 William L.Young und Hua Su, R21 NS070153 Hua SU und von der American Heart Association AHA 10GRNT3130004 Hua Su unterstützt. Dr. Ana Rodríguez-Hernández wird durch einen Zuschuss von &quot;Obra Social La Caixa&quot; unterstützt

1. Vorbereitung der Maus <ol><li> Induzieren Vollnarkose in der Maus mit Isofluran Gas. Injizieren 0,15 ml intraperitoneale brupenorphine für die Schmerztherapie. Überprüfen Sie zunächst, ob die Narkoseniveau ist durch Einstechen Pfoten die Maus zufriedenstellend. </li><li> Sie mit der Maus in Rückenlage mit den vier von Klebeband fixiert Gliedmaßen. Entfernen Sie die Haare des Halses und der oberen Brustbereich mit einer Schere. Subkutane Injektion verabreichen 0,2-0,4 ml 0,9% Kochsalzlösung, um die Maus während des chirurgischen Eingriffs mit Feuchtigkeit versorgt. </li><li> Bereiten Sie das Operationsfeld nach einem strengen sterilen Verfahren. Das Gebiet der Hautschnitt sollte mit 90% Alkohol gereinigt werden. </li></ol> 2. Präparieren der Arteria carotis communis und die äußere Jugularvene <ol><li> Machen Sie eine horizontale Mittellinie Hals-Schnitt über den unteren Halsbereich der Maus. Nachdem die Vertiefung der Wunde, erhöhen die Speicheldrüsen und Halsweichteile unter Verwendung vonraction Fadens (Abbildung 1). Setzen Sie die rechte äußere Jugularvene (EJV) lateral des M. sternocleidomastoideus (SCM). Dieser Schritt sollte unter dem Mikroskop durchgeführt werden, da zu hohe Traktion über die Vene zu Beschädigungen und induzieren ihre Thrombose. </li><li> Das Recht EJV entlang seinem Kurs sorgfältig sezieren vom Schlüsselbein zur Schädelbasis. Normalerweise elektrischen bipolaren Zange, gerinnen und teilen alle Branchen, um eine ausreichende Länge für die spätere temporäre Klammer Platzierung und Anastomose vorzubereiten. </li><li> Lateral der Luftröhre und medial des SCM, erkunden Sie die A. carotis communis (CCA). Es sollte vorsichtig aus dem Schlüsselbein gerade jenseits ihrer Gabelung in die externen und internen Karotisarterien ausgesetzt werden. Während dieser Stufe sollte die Aufmerksamkeit wieder gegeben übermäßigen Zug am EJVs die später beeinträchtigen könnte die Durchgängigkeit der Anastomose zu vermeiden. </li></ol> 3. Vorbereitung des Anastomose <ol><li> Ligieren die CCA mit 10-0 Nylon proximal ihrer Bifurkation. Anschließend eine temporäre Clip über das proximale CCA so nahe dem Schlüsselbein wie möglich. </li><li> Sobald die Strömung unterbrochen wurde, durchschneiden die Arterie knapp Bifurkation Ligatur und bewässern sie mit Kochsalzlösung in das Innere des Lumens auszuwaschen verbleibende Blut. Vermeiden bipolare Koagulation in diesem Schritt, da die thermische Schädigung der Arterienwand könnte die Zukunft Anastomose in Gefahr bringen. </li><li> Um die Sichtbarkeit der Kanten des EJV verbessern, wird die mediale Wand mit einem blauen Markierungsstift entlang des Verlaufs der geplanten Venotomie markiert. Sobald die EJVs markiert ist, mit einem 10-0 Naht um das distale Ende als Schwanz möglichst ligieren und Anwendung einer temporären Gefäßclip an dem proximalen Ende als Schädel wie möglich. </li><li> Mit einer feinen Nadel 30 G und 0,5 ml Spritze, vereinbaren Sie einen Anfangsöffnung über den markierten Bereich der EJV und sofortige Spülung mit Kochsalzlösung in das Lumen zur Thrombusbildung zu vermeiden. Als nächstes erweitern die Venenschnitt mit den Mikroscherebis die Länge ist etwa 2-3 mal der Durchmesser des CCA. Vermeiden Sie gewaltsame Dehnung und achten Sie auf eine scharfe und saubere Schnittkante zu halten. </li><li> Näherung für das Ende des CCA in die EJV. Einen Taillierung Einschnitt im Spenderende des CCA um den Durchmesser zu Länge zum Venotomie Größe (2) einzustellen. </li></ol> 4. End-zu-Seit-Anastomose <ol><li> Verwenden Sie einen 11-0 Monofilament Nylonfaden für das CCA-to-EJV Ende-zu-Seit-Anastomose. Naht der medialen Wand der Anastomose in einem craneo-kaudal ist der erste Schritt. Jeder Stich sollte von außen in die Venenwand ersten (Abbildung 3) von innen nach außen der Arterienwand (Abbildung 4) neben platziert und. Dadurch wird der Knoten auf der Außenfläche der anastomotischen Gefäße zu allen Zeiten (Abbildung 5) zu halten. Entweder unterbrochen oder kontinuierliche Fäden verwendet werden können, aber alle kontinuierliche Nähte sollten als Endstufe angezogen werden. </li><li> Once der medialen Wand wurde genäht hat, wiederholen Sie den Vorgang von kaudal mit der Seitenwand kranial. Nun sollte jeder Stich von außen in die Arterienwand erste und von innen nach außen der Venenwand neben platziert werden. Saline Bewässerung wird dazu beitragen, das Lumen der Anastomose jederzeit sichtbar während des Verfahrens. </li><li> Nach Abschluss aller Schritte der Anastomose erster und aus der Arterie entfernen zunächst die temporäre Clip aus der Vene. Das arterielle Blut wird in den EJV mit keiner oder wenig Nässen aus der Anastomose fließen (siehe Videos VH und VH-Modell Modell 2). Die minimale Blutung sollte ohne Baumwolle Kompression über die Anastomose zu stoppen. Diese müssen, um eine Thrombose des empfindlichen Vene verhindert vermeiden. </li><li> Einmal durch die pulsierende Anastomose fließen wird bestätigt und keine sichtbare Blutungen beobachtet wird, bewässern das Operationsfeld mit Kochsalzlösung und schließen Sie den Hals-Schnitt mit 6-0 Nylonnaht. Schließlich verwalten 0,15 ml mehr intraperitoneal brupenorphine zur postoperativen Schmerztherapie und 0,2-0,4 ml subkutane 0,9% Salzlösung, jede Blutverlust während der Operation wieder aufzufüllen. </li></ol>

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	<li>Bederson, J. B., Wiestler, O. D., Br&#252;stle, O., Roth, P., Frick, R., Ya&#351;argil, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intracranial+venous+hypertension+and+the+effects+of+venous+outflow+obstruction+in+a+rat+model+of+arteriovenous+fistula.">Intracranial venous hypertension and the effects of venous outflow obstruction in a rat model of arteriovenous fistula.</a> Neurosurgery. 29, 341-350 (1991).</li><li>Gao, P., Zhu, Y., Ling, F., Shen, F., Lee, B., Gabriel, R. A., Hao, Q., Yang, G. Y., Su, H., Young, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nonischemic+cerebral+venous+hypertension+promotes+a+pro-angiogenic+stage+through+HIF-1+downstream+genes+and+leukocyte-derived+MMP-9.">Nonischemic cerebral venous hypertension promotes a pro-angiogenic stage through HIF-1 downstream genes and leukocyte-derived MMP-9.</a> J. Cereb. Blood Flow Metab. 29, 1482-1490 (2009).</li><li>Kim, H., Su, H., Weinsheimer, S., Pawlikowska, L., Brain Young, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=arteriovenous+malformation+pathogenesis:+a+response-to-injury+paradigm.">arteriovenous malformation pathogenesis: a response-to-injury paradigm.</a> Acta Neurochir. Suppl. 111, 83-92 (2011).</li><li>Lawton, M. T., Arnold, C. M., Kim, Y. J., Bogarin, E. A., Stewart, C. L., Wulfstat, A. A., Derugin, N., Deen, D., Young, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiation+arteriopathy+in+the+transgenic+arteriovenous+fistula+model.">Radiation arteriopathy in the transgenic arteriovenous fistula model.</a> Neurosurgery. 62, 1129-1138 (2008).</li><li>Lawton, M. T., Stewart, C. L., Wulfstat, A. A., Derugin, N., Hashimoto, T., Young, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+transgenic+arteriovenous+fistula+in+the+rat:+an+experimental+model+of+gene+therapy+for+brain+arteriovenous+malformations.">The transgenic arteriovenous fistula in the rat: an experimental model of gene therapy for brain arteriovenous malformations.</a> Neurosurgery. 54, 1463-1471 (2004).</li><li>Schaller, B., Graf, R., Sanada, Y., Tolnay, M., Rosner, G., Wienhard, K., Heiss, W., D, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemodynamic+changes+after+occlusion+of+the+posterior+superior+sagittal+sinus:+an+experimental+PET+study+in+cats.">Hemodynamic changes after occlusion of the posterior superior sagittal sinus: an experimental PET study in cats.</a> AJNR Am J Neuroradiol. 24, 1876-1880 (2003).</li><li>Zhu, Y., Lawton, M. T., Du, R., Shwe, Y., Chen, Y., Shen, F., Young, W. L., Yang, G. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+hypoxia-inducible+factor-1+and+vascular+endothelial+growth+factor+in+response+to+venous+hypertension.">Expression of hypoxia-inducible factor-1 and vascular endothelial growth factor in response to venous hypertension.</a> Neurosurgery. 59, 687-696 (2006).</li><li>Herman, J. M., Spetzler, R. F., Bederson, J. B., Kurbat, J. M., Zabramski, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genesis+of+a+dural+arteriovenous+malformation+in+a+rat+model.">Genesis of a dural arteriovenous malformation in a rat model.</a> J. Neurosurg. 83, 539-545 (1995).</li><li>Terada, T., Higashida, R. T., Halbach, V. V., Dowd, C. F., Tsuura, M., Komai, N., Wilson, C. B., Hieshima, G. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+acquired+arteriovenous+fistulas+in+rats+due+to+venous+hypertension.">Development of acquired arteriovenous fistulas in rats due to venous hypertension.</a> J. Neurosurg. 80, 884-889 (1994).</li><li>Yassari, R., Sayama, T., Jahromi, B. S., Aihara, Y., Stoodley, M., Macdonald, 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=Angiographic,+hemodynamic+and+histological+characterization+of+an+arteriovenous+fistula+in+rats.">Angiographic, hemodynamic and histological characterization of an arteriovenous fistula in rats.</a> Acta Neurochir (Wien). 146, 495-504 (2004).</li><li>Solheim, O., Skeidsvoll, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transient+global+amnesia+may+be+caused+by+cerebral+vein+thrombosis.">Transient global amnesia may be caused by cerebral vein thrombosis.</a> Med. Hypotheses. 65, 1142-1149 (2005).</li><li>Yang, B., Shergill, U., Fu, A. A., Knudsen, B., Misra, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mouse+arteriovenous+fistula+model.">The mouse arteriovenous fistula model.</a> J Vasc Interv Radiol. 20, 946-950 (2009).</li><li>Choi, E. J., Choi, E. J., Walker, E. J., Shen, F., Oh, S. P., Arthur, H. M., Young, W. L., Su, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Minimal+Homozygous+Endothelial+Deletion+of+Eng+with+VEGF+Stimulation+Is+Sufficient+to+Cause+Cerebrovascular+Dysplasia+in+the+Adult+Mouse.">Minimal Homozygous Endothelial Deletion of Eng with VEGF Stimulation Is Sufficient to Cause Cerebrovascular Dysplasia in the Adult Mouse.</a> Cerebrovascular diseases. 33, 540-547 (2012).</li><li>Hao, Q., Su, H., Marchuk, D. A., Rola, R., Wang, Y., Liu, W., Young, W. L., Yang, G. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+tissue+perfusion+promotes+capillary+dysplasia+in+the+ALK1-deficient+mouse+brain+following+VEGF+stimulation.">Increased tissue perfusion promotes capillary dysplasia in the ALK1-deficient mouse brain following VEGF stimulation.</a> Am. J. Physiol. Heart Circ. Physiol. 295, H2250-H2256 (2008).</li><li>Su, H., Hao, Q., Shen, F., Zhu, Y., Lee, C. Z., Young, W. L., Yang, G. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+cerebral+microvascular+dysplasia+model+in+rodents.">Development of a cerebral microvascular dysplasia model in rodents.</a> Acta Neurochir. Suppl. 105, 185-189 (2008).</li><li>Walker, E. J., Su, H., Shen, F., Degos, V., Jun, K., Young, W. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bevacizumab+Attenuates+VEGF-Induced+Angiogenesis+and+Vascular+Malformations+in+the+Adult+Mouse+Brain.">Bevacizumab Attenuates VEGF-Induced Angiogenesis and Vascular Malformations in the Adult Mouse Brain.</a> Stroke; a journal of cerebral circulation. , (2012).</li></ol>

Die experimentellen Verfahren mit Labortieren wurden von der Institutional Animal Care und Verwenden Committee der University of California, San Francisco (UCSF) zugelassen. Die Autoren haben keine potenziellen Interessenkonflikte auf die Medikamente und Materialien in diesem Verfahren verwendet verwandt.

Anhaltende Hirnvenenbluthochdruck hat eng mit schweren klinischen Manifestationen und schlechter Prognose bei Patienten mit Dura AVFs und Gehirn AVMs 3 zusammen. Diese Wirkungen von CVH wurden weitgehend in Rattenmodellen 1,2,8 sucht. Eine entsprechende Modell in der Maus würde die Verwendung von gentechnisch veränderten Tieren, die letztlich erlauben würde die Analyse der molekularen Signalwege auf die Pathogenese der venösen Bluthochdruck und seine Beziehung mit der Dura AVF und Gehirn AVM be...

Tiermodelle der Hirnvenenbluthochdruck haben sich als ein wichtiges Instrument für das Verständnis der Pathophysiologie des Gehirns arteriovenöse Malformationen und arteriovenöse Fisteln 1-7 sein. Das am häufigsten verwendete ist die Ratten-Modell durch eine künstliche Fistel zwischen der Arteria carotis communis (CCA) und die äußere Jugularvene (EJVs), die eine konsistente cerebralvenösen Hypertonie (CVH) bei der Ratte 1,8-10 provoziert erstellt. Äquivalent Mäusemodellen, indem die Möglichkeit der Verwendung verschiedener transgener Mäuse Stämme wären weitere Studie über nicht nur Pathophysiologie sondern auch möglichen genetischen Therapien für diese zerebrovaskuläre Erkrankungen ermöglichen. Ferner sind die Versuchs Konstrukt könnte auch auf die Untersuchung von anderen zerebrovaskulären Krankheiten mit venöser Hypertonie, wie Migräne, transienten globalen Amnesie, transiente monokulare Blindheit usw. 11 Frühere Versuche im Zusammenhang geeignet ist, diese Mäusemodellen h konstruierenave demonstriert die Schwierigkeiten mit der Durchgängigkeit der Fistel aufgrund der winzigen anatomischen 5,12. Hier beschreiben wir die Schritt-für-Schritt-Protokoll für eine erfolgreiche Anastomose des murinen CCA und EJVs die in einen langfristigen Patent Fistel und einem haltbaren venöse Hypertonie in der Maus übersetzt.

<table border="1" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>10-0 Sterile Microsuture</td>
			<td>Arosurgical Ic.</td>
			<td>VT5A010Q10</td>
		</tr>
		<tr>
			<td>11-0 Sterile Microsuture</td>
			<td>Arosurgical Ic</td>
			<td>VT4A00N07</td>
		</tr>
		<tr>
			<td>DUROTIP Scissors</td>
			<td>Aesculap</td>
			<td>BC210R</td>
		</tr>
		<tr>
			<td>Micro-Adson Tissue Forceps</td>
			<td>Aesculap</td>
			<td>BD510R</td>
		</tr>
		<tr>
			<td>Microscissors</td>
			<td>Aesculap</td>
			<td>OC496R</td>
		</tr>
		<tr>
			<td>Micro Forceps #5 Jewelers</td>
			<td>Aesculap</td>
			<td>BD331R</td>
		</tr>
		<tr>
			<td>Angled Jewelers Forceps</td>
			<td>Aesculap</td>
			<td>BD329R</td>
		</tr>
		<tr>
			<td>Micro Suture Forceps</td>
			<td>Aesculap</td>
			<td>BD338R</td>
		</tr>
		<tr>
			<td>DUROGRIP Needle Holder</td>
			<td>Aesculap</td>
			<td>BM009R</td>
		</tr>
	</tbody>
</table>

adult mouse venous hypertension model: common carotid artery to external jugular vein anastomosis.

Das Verständnis der Pathophysiologie von Gehirn arteriovenösen Fehlbildungen und arteriovenösen Fisteln hat dank Tiermodellen verbessert. Ein Rattenmodell die Schaffung eines künstlichen Fistel zwischen der A. carotis communis (CCA) und der externen Halsschlagader (EJV) ist vielfältig beschrieben und bewiesen technisch möglich ist. Dieses Konstrukt provoziert eine konsistente cerebralvenösen Hypertonie (CVH), und damit dazu beigetragen hat die Untersuchung der Beteiligung von venöser Hypertonie, um die Bildung, die klinischen Symptome und der Prognose von Gehirn AVMs und Dura AVFs. Entspricht Mäusen Modelle wurden nur knapp beschrieben und haben Probleme mit Stenose der Fistel gezeigt. Ein etabliertes Mausmodell würde das Studium nicht nur Pathophysiologie, sondern auch potenzielle genetische Therapien für diese zerebrovaskuläre Erkrankungen ermöglichen. Wir präsentieren ein Modell der arteriovenöse Fistel, die eine dauerhafte Hirnvenenbluthochdruck in der Maus erzeugt. Mikrochirurgische Anastomose of murinen CCA und EJV kann schwierig sein, aufgrund von winzigen Anatomie und in einem nicht-Patent Fistel häufig führen. In diesem Schritt-für-Schritt-Protokoll sprechen wir alle wichtigen Herausforderungen bei diesem Vorgang auftreten. Vermeidung übermäßiger Zurückziehen der Vene während der Belichtung mit 11-0 Nähte statt 10-0, und machen eine sorgfältig geplante End-zu-Seit-Anastomose sind einige der wichtigsten Schritte. Obwohl diese Methode erfordert erweiterte mikro Fähigkeiten und eine längere Lernkurve, die den Gegenwert in der Ratte, es konsequent weiterentwickelt werden kann. Dieses neuartige Modell wurde entworfen, um Transgene Maus Techniken mit einer zuvor etablierten experimentellen Systems, das auf die Gehirn AVMs und Dura AVFs untersuchen erwiesen integrieren. Durch Öffnen der Möglichkeit der Verwendung von transgenen Mäusen kann ein breiteres Spektrum von gültigen Modellen erzielt werden und genetische Anwendungen können ebenfalls getestet werden. Die experimentelle Konstrukt könnte auch auf die Untersuchung von ot angepasst werdenihr zerebrovaskuläre Erkrankungen mit venösen Bluthochdruck wie Migräne, transiente globale Amnesie, transiente monokulare Blindheit usw. verwandt

Ein erfolgreiches Ergebnis des Modells ist ein Patent arteriovenöse Fistel, die venöse Hypertonie im murinen Gehirn induziert. Um das Modell zu validieren wir zunächst den intrakraniellen Venendruck in der sagittalen Sinus der Mäuse gemessen bei 2, 3 und 4 Wochen nach der Operation. 6 verschiedene Mäuse wurden für jede Zeitgruppe zugewiesen. Der Sinus Druck betrug 8,8 ± 1,2 mmHg in der Gruppe, gemessen zwei Wochen nach der Operation. In den 6-Mäusen gemessen 3 Wochen nach der Operation, die Sinus Druck betrug 4,...

Watch this Scientific Journal Video about Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis. at JoVE.com

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

Wir beschreiben ein Verfahren zum Erzeugen einer zuverlässigen Modell der zerebralen venöse Hypertonie in der adulten Maus. Dieses Modell ist vielfältig beschrieben und geprüft bei der Ratte ist. Diese neue Gegenstück in den Mäusen eröffnet die Möglichkeit der Verwendung von gentechnisch veränderten Tieren und verbreitert, wodurch die Anwendungen des Modells.

Erwachsene Maus venöse Bluthochdruckmodell: A. carotis communis, um äußere Jugularvene Anastomose.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

adult-mouse-venous-hypertension-model-common-carotid-artery-to

Research

JoVE Journal

Medicine

14.0K Aufrufe.  University of California, San Francisco.  Wir beschreiben ein Verfahren zum Erzeugen einer zuverlässigen Modell der zerebralen venöse Hypertonie in der adulten Maus. Dieses Modell ist vielfältig beschrieben und geprüft bei der Ratte ist. Diese neue Gegenstück in den Mäusen eröffnet die Möglichkeit der Verwendung von gentechnisch veränderten Tieren und verbreitert, wodurch die Anwendungen des Modells. 

Serie: Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

Training a Sophisticated Microsurgical Technique: Interposition of External Jugular Vein Graft in the Common Carotid Artery in Rats

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in peripheral arterial bypass surgery as well as in arteriovenous fistulas.1-5 The experimental procedure of vein graft interposition in the common carotid artery by using the cuff-technique has been applied in several research projects to examine the aetiology of neointimal hyperplasia and therapeutic options to address it. 6-8 The cuff prevents vessel anastomotic remodeling and induces turbulence within the graft and thereby the development of neointimal hyperplasia.
Using the superior caval vein graft is an established small-animal model for venous arterialization experiment.9-11 This current protocol refers to an established jugular vein graft interposition technique first described by Zou et al., 9 as well as others.12-14 Nevertheless, these cited small animal protocols are complicated.
To simplify the procedure and to minimize the number of experimental animals needed, a detailed operation protocol by video training is presented. This video should help the novice surgeon to learn both the cuff-technique and the vein graft interposition. Hereby, the right external jugular vein was grafted in cuff-technique in the common carotid artery of 21 female Sprague Dawley rats categorized in three equal groups that were sacrificed on day 21, 42 and 84, respectively. Notably, no donor animals were needed, because auto-transplantations were performed. The survival rate was 100 % at the time point of sacrifice. In addition, the graft patency rate was 60 % for the first 10 operated animals and 82 % for the remaining 11 animals. The blood flow at the time of sacrifice was 8&plusmn;3 ml/min. In conclusion, this surgical protocol considerably simplifies, optimizes and standardizes this complicated procedure. It gives novice surgeons easy, step-by-step instruction, explaining possible pitfalls, thereby helping them to gain expertise fast and avoid useless sacrifice of experimental animals.

Neointimal hyperplasia is the primary cause of stenosis in arterialized veins. We propose a new protocol whereby the right external jugular vein is grafted using the cuff technique in the common carotid artery of Sprague Dawley rats. The survival rate was 100 % at the time point of sacrifice.

Trainieren eines Sophisticated mikrochirurgische Technik: Interposition der Vena jugularis externa Graft in der Arteria carotis communis bei Ratten

Neointimal hyperplasia is one the primary causes of stenosis in arterialized veins that are of great importance in arterial coronary bypass surgery, in ...

Forschung

Medizin

Bilateral Common Carotid Artery Occlusion as an Adequate Preconditioning Stimulus to Induce Early Ischemic Tolerance to Focal Cerebral Ischemia

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We have established bilateral common carotid artery occlusion as a preconditioning stimulus to induce early ischemic tolerance to transient focal cerebral ischemia in C57Bl6/J mice. In this video, we will demonstrate the methodology used for this study.

There is accumulating evidence, that ischemic preconditioning (PC) &#x2013; a non-damaging ischemic challenge to the brain - confers a transient protection to a subsequent damaging ischemic insult. We established bilateral common carotid artery occlusion (BCCAO) as a preconditioning stimulus to induce early ischemic tolerance (IT) to transient focal cerebral ischemia (induced by middle cerebral artery occlusion, MCAO) in C57Bl6/J mice.

Bilaterale Arteria carotis communis Occlusion als adäquate Vorbehandlung Stimulus zu frühe ischämische Toleranz zu Focal zerebraler Ischämie induzieren

There is accumulating evidence, that ischemic preconditioning - a non-damaging ischemic challenge to the brain - confers a transient protection to a ...

Catheterization of the Carotid Artery and Jugular Vein to Perform Hemodynamic Measures, Infusions and Blood Sampling in a Conscious Rat Model

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. Intra-tracheal inoculation of a known amount of bacteria has been successfully used to reproduce the pathogenesis of pneumonia which then develops into sepsis. Monitoring hemodynamic parameters and providing standard clinical treatment including infusion of antibiotics, fluids and drugs to maintain blood pressure is critical to simulate routine supportive care in this model but to do so requires both arterial and venous vascular access. The video details the surgical technique for implanting carotid artery and common jugular vein catheters in an anesthetized rat. Following a 72 hr recovery period, the animals will be re-anesthetized and connected to a tether and swivel setup attached to the rodent housing which connects the implanted catheters to the hemodynamic monitoring system. This setup allows free movement of the rat during the study while continuously monitoring pressures, infusing fluids and drugs (antibiotics, vasopressors) and performing blood sampling.

Vascular accesses to measure hemodynamics, provide fluids and perform blood sampling are important to any small animal model study. We present a technique for implanting catheters into the carotid artery and the common jugular vein in an anesthetized rat for connecting to a system to perform monitoring, infusions and sampling.

Katheterisierung der Halsschlagader und Vena jugularis zur hämodynamischen Maßnahmen im Conscious Rattenmodell durchführen, Infusionen und Blutentnahme

The success of a small animal model to study critical illness is, in part, dependent on the ability of the model to simulate the human condition. ...

Ferric Chloride-induced Thrombosis Mouse Model on Carotid Artery and Mesentery Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric chloride injury is now a well-established technique to rapidly and accurately induce the formation of thrombi in exposed veins or artery of small and large diameter. This model has played a key role in the study of the pathophysiology of thrombosis, in the discovery and validation of novel antithrombotic drugs and in the understanding of the mechanism of action of these new agents. Here, the implementation of this technique on a mesenteric vessel and carotid artery in mice is presented. The method describes how to label circulating leukocytes and platelets with a fluorescent dye and to observe, by intravital microscopy on the exposed mesentery, their accumulation at the injured vessel wall which leads to the formation of a thrombus. On the carotid artery, the occlusion caused by the clot formation is measured by monitoring the blood flow with a Doppler probe.

The FeCl3 induced thrombosis model in mice is described herein. A method to monitor thrombus growth by intravital microscopy observation on a mesenteric vessel and by blood flow measurement in the carotid artery is presented.

Eisenchlorid-induzierte Thrombose Mausmodell für Halsschlagader und Mesentery Vessel

Severe thrombosis and its ischemic consequences such as myocardial infarction, pulmonary embolism and stroke are major worldwide health issues. The ferric ...

Performing Permanent Distal Middle Cerebral with Common Carotid Artery Occlusion in Aged Rats to Study Cortical Ischemia with Sustained Disability

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, obesity and diabetes. Accordingly, when evaluating therapies for stroke in animals, it is important to select a model with excellent face validity. Ischemic stroke accounts for 80% of all strokes, and the majority of these occur in the territory of the middle cerebral artery (MCA), often inducing infarcts that affect the sensorimotor cortex, causing persistent plegia or paresis on the contralateral side of the body. We demonstrate in this video a method for producing ischemic stroke in elderly rats, which causes sustained sensorimotor disability and substantial cortical infarcts. Specifically, we induce permanent distal middle cerebral artery occlusion (MCAO) in elderly female rats by using diathermy forceps to occlude a short segment of this artery. The carotid artery on the ipsilateral side to the lesion was then permanently occluded and the contralateral carotid artery was transiently occluded for 60 min. We measure the infarct size using structural T2-weighted magnetic resonance imaging (MRI) at 24 hr and 8 weeks after stroke. In this study, the mean infarct volume was 4.5% &#177; 2.0% (standard deviation) of the ipsilateral hemisphere at 24 hr (corrected for brain swelling using Gerriet&#8217;s equation, n = 5). This model is feasible and clinically relevant as it permits the induction of sustained sensorimotor deficits, which is important for the elucidation of pathophysiological mechanisms and novel treatments.

Here we present a protocol to produce permanent distal middle cerebral artery occlusion in elderly female rats with simultaneous occlusion of the carotid arteries to generate large cortical infarcts and sustained deficits. We show confirmation of the lesion size using structural MRI at 24 hr and 8 weeks after stroke.

Darstellende Permanent Distal cerebri mit Arteria carotis communis Occlusion bei alten Ratten zu studieren kortikale Ischämie mit anhaltenden Behinderungs

Stroke typically occurs in elderly people with a range of comorbidities including carotid (or other arterial) atherosclerosis, high blood pressure, ...

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves focal endothelial-selective transgenesis, thereby allowing an investigator to determine the biological roles of endothelial-expressed transgenes and to quantify the in vivo transcriptional activity of DNA sequences in large artery endothelial cells. The method uses surgical isolation of rabbit common carotid arteries and an arteriotomy to deliver a transgene-expressing viral vector into the arterial lumen. A short incubation period of the vector in the lumen, with subsequent aspiration of the lumen contents, is sufficient to achieve efficient and durable expression of the transgene in the endothelium, with no detectable transduction or expression outside of the isolated arterial segment. The method allows assessment of the biological activities of transgene products both in normal arteries and in models of human vascular disease, while avoiding systemic effects that could be caused either by targeting gene delivery to other sites (e.g. the liver) or by the alternative approach of delivering genetic constructs to the endothelium by germ line transgenesis. Application of the method is limited by the need for a skilled surgeon and anesthetist, a well-equipped operating room, the costs of purchasing and housing rabbits, and the need for expertise in gene-transfer vector construction and use. Results obtained with this method include: transgene-related alterations in arterial structure, cellularity, extracellular matrix, or vasomotor function; increases or reductions in arterial inflammation; alterations in vascular cell apoptosis; and progression, retardation, or regression of diseases such as intimal hyperplasia or atherosclerosis. The method also allows measurement of the ability of native and synthetic DNA regulatory sequences to alter transgene expression in endothelial cells, providing results that include: levels of transgene mRNA, levels of transgene protein, and levels of transgene enzymatic activity.

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

This method is to introduce a transgene into the endothelium of rabbit carotid arteries. Introduction of the transgene allows the assessment of the biological role of the transgene product either in normal arteries or disease models. The method is also useful for measuring activity of DNA regulatory sequences.

In Vivo Gentransfer in die Kaninchen gemeinsame Halsschlagader Endothel

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves ...

A Rabbit Model of Durable Transgene Expression in Jugular Vein to Common Carotid Artery Interposition Grafts

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, intimal hyperplasia, and atherosclerosis. The goal of this article is to demonstrate a method for placing bilateral venous interposition grafts in a rabbit, then transducing the grafts with a gene transfer vector that achieves durable transgene expression. The method allows the investigation of the biological roles of genes and their protein products in normal vein graft homeostasis. It also allows the testing of transgenes for the activities that could prevent vein graft failure, e.g., whether the expression of a transgene prevents the neointimal growth, reduces the vascular inflammation, or reduces atherosclerosis in rabbits fed with a high-fat diet. During an initial survival surgery, the segments of right and left external jugular vein are excised and placed bilaterally as reversed end-to-side common carotid artery interposition grafts. During a second survival surgery, performed 28 days later, each of the grafts is isolated from the circulation with vascular clips and the lumens are filled (via an arteriotomy) with a solution containing a helper-dependent adenoviral (HDAd) vector. After a 20-min incubation, the vector solution is aspirated, the arteriotomy is repaired, and flow is restored. The veins are harvested at time points dictated by individual experimental protocols. The 28-day delay between the graft placement and the transduction is necessary to ensure the adaptation of the vein graft to the arterial circulation. This adaptation avoids rapid loss of transgene expression that occurs in vein grafts transduced before or immediately after grafting. The method is unique in its ability to achieve durable, stable transgene expression in grafted veins. Compared to other large animal vein graft models, rabbits have advantages of low cost and easy handling. Compared to rodent vein graft models, rabbits have larger and easier-to-manipulate blood vessels that provide abundant tissue for analysis.

This method describes the placement of interposition vein grafts in rabbits, the transduction of the grafts, and the achievement of durable transgene expression. This allows the investigation of physiological and pathological roles of transgenes and their protein products in grafted veins, and testing of gene therapies for vein graft disease.

Ein Kaninchen-Modell der dauerhafte Transgene Ausdruck in Halsschlagader, gemeinsame Halsschlagader Interposition Grafts

Vein graft bypass surgery is a common treatment for occlusive arterial disease; however, long-term success is limited by graft failure due to thrombosis, ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

Phase Kontrast Magnetresonanz-Bildgebung in der Ratte gemeinsame Halsschlagader

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

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.

Messung der isometrischen Kontraktilität der Maus mesenterialen Arterie mit Draht Myographie

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

Middle Cerebral Artery Occlusion Allowing Reperfusion via Common Carotid Artery Repair in Mice

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue plasminogen activator (tPA) as an approved drug treatment to target ischemic strokes. Current research in the field of ischemic stroke focuses on better understanding the pathophysiology of stroke, to develop and investigate novel pharmaceutical targets. Reliable experimental stroke models are crucial for the progression of potential treatments. The middle cerebral artery occlusion (MCAO) model is clinically relevant and the most frequently used surgical model of ischemic stroke in rodents. However, the outcomes of this model, such as lesion volume, are associated with high levels of variability, particularly in mice. The alternative MCAO model described here allows the reperfusion of the common carotid artery (CCA) and the increased perfusion of the middle cerebral artery (MCA) territory, using a tissue pad with fibrinogen-based sealant to repair the vessel, and the improved welfare of the mice by avoiding external carotid artery (ECA) ligation. This reduces the reliance on the Circle of Willis, which is known to be highly anatomically variable in mice. Representative data show that using this alternative surgical approach decreases the variability in lesion volumes between the traditional MCAO approach and the alternative approach described here.

Intraluminal filament occlusion of the middle cerebral artery is the most frequently used in vivo model of experimental stroke in rodents. An alternative surgical approach to allow common carotid artery repair is performed here, which allows the reperfusion of the common carotid artery and a full reperfusion to the middle cerebral artery territory.

Mittlere zerebrale Arterie Okklusion so dass Reperfusion über gemeinsame Halsschlagader Reparatur bei Mäusen

The ischemic stroke is a major cause of adult long-term disability and death worldwide. The current treatments available are limited, with only tissue ...