Questo progetto è in parte sostenuto da NIH T32 GM008440 a Espen Walker, R01 NS27713 a William L.Young, P01 NS44155 a William L.Young e Hua Su, R21 NS070153 a Hua SU e dalla American Heart Association AHA 10GRNT3130004 a Hua Su. Dr Ana Rodríguez-Hernández è sostenuta da una sovvenzione &quot;Obra Social La Caixa&quot;

1. Preparare il mouse <ol><li> Indurre l&#39;anestesia generale nel topo con gas isoflurano. Iniettare 0,15 ml di brupenorphine intraperitoneale per la gestione del dolore. Prima di procedere, verificare che il livello di anestesia è soddisfacente pungendo zampe del topo. </li><li> Ponga il mouse in decubito dorsale con i quattro arti fissati dal nastro adesivo. Rimuovere il pelo del collo e la parte superiore del torace con le forbici. Via iniezione sottocutanea, somministrare 0,2-0,4 ml di soluzione salina allo 0,9% per mantenere il mouse idratato durante la procedura chirurgica. </li><li> Preparare il campo operatorio seguendo un metodo rigoroso sterili. L&#39;area della incisione cutanea deve essere pulito con il 90% di alcol. </li></ol> 2. dissezione della carotide comune e la Vena giugulare esterna <ol><li> Fare un linea mediana incisione orizzontale cervicale in tutta l&#39;area del collo inferiore del mouse. Dopo approfondire la ferita, elevare le ghiandole salivari e dei tessuti molli del collo dell&#39;utero utilizzando araction sutura (Figura 1). Esporre la giusta vena giugulare esterna (EJV) lateralmente al muscolo sternocleidomastoideo (SCM). Questa operazione deve essere eseguita al microscopio, poiché trazione eccessiva sopra la vena potrebbe danneggiarlo e indurre il trombosi. </li><li> Sezionare con cura il EJV destra lungo il suo corso dalla clavicola alla base del cranio. Di solito pinze bipolari elettrici, coagulano e si dividono i rami di preparare una lunghezza adeguata per il successivo posizionamento di clip temporanea e anastomosi. </li><li> Laterale alla trachea e mediale alla SCM, esplorare l&#39;arteria carotide comune (CCA). Si deve essere attentamente esposta dalla clavicola a poco oltre la sua biforcazione nelle arterie carotidi interne ed esterne. Durante questa fase, l&#39;attenzione deve essere nuovamente somministrato per evitare un&#39;eccessiva trazione sul EJV che potrebbe successivamente compromettere la pervietà dell&#39;anastomosi. </li></ol> 3. Preparare l&#39;anastomosi <ol><li> Legare il CCA con 10-0 Nylon solo prossimale alla sua biforcazione. Successivamente, applicare una clip temporaneo del prossimale CCA più vicino alla clavicola possibile. </li><li> Una volta che il flusso è stato interrotto, transetto l&#39;arteria appena sotto la legatura biforcazione ed irrigare con soluzione fisiologica per lavare il sangue residuo all&#39;interno del lume. Evitare di coagulazione bipolare in questa fase, in quanto danno termico alla parete dell&#39;arteria potrebbe mettere il futuro anastomosi in pericolo. </li><li> Al fine di migliorare la visibilità dei bordi della EJV, la parete mediale è contrassegnato con un pennarello blu lungo il corso del venotomia prevista. Una volta che il EJV è contrassegnata, utilizzare un 10-0 sutura per legare l&#39;estremità distale come caudale possibile e applicare una clip vascolare temporanea sull&#39;estremità prossimale craniale possibile. </li><li> Con un bel 30 ago G e 0,5 ml siringa, effettuare un&#39;apertura iniziale tramite marcata zona della EJV e immediatamente irrigare il lume con soluzione fisiologica per evitare la formazione di trombi. Avanti, estendere il venotomia con i microscissorsfinché la lunghezza è di circa 2-3 volte il diametro del CCA. Evitare violenti stretching e prestare attenzione a mantenere un filo di lama tagliente e pulito. </li><li> Approssimi la fine del CCA al EJV. Effettuare una incisione laterale tagliata alla fine donatore del CCA per regolare il diametro alla lunghezza alle dimensioni venotomia (Figura 2). </li></ol> 4. end-to-side anastomosi <ol><li> Utilizzare un nylon sutura monofilamento 11-0 per l&#39;anastomosi CCA-to-EJV end-to-side. Suturando la parete mediale della anastomosi in una direzione craniofacciale caudale è il passo iniziale. Ogni punto deve essere collocato al di fuori-nella parete venosa prima (figura 3) e da dentro-fuori della parete arteriosa (Figura 4) successiva. Ciò manterrà il nodo sulla superficie esterna dei vasi anastomotiche in ogni momento (Figura 5). Interrotta o continui suture possono essere usati, ma tutti suture continue devono essere serrati come fase finale. </li><li> Once parete mediale è stato suturato, ripetere la procedura dal caudale a cranico con la parete laterale. Ora ogni punto deve essere collocato al di fuori-nella parete arteriosa prima e dall&#39;interno verso la parete venosa successivo. Saline irrigazione contribuirà a mantenere il lume della anastomosi visibile in qualsiasi momento durante la procedura. </li><li> Dopo aver terminato tutte le fasi della anastomosi, rimuovere la clip temporanea dalla vena prima e dalla arteria successivo. Il sangue arterioso scende nella EJV con nessuna o poca che trasuda dalla anastomosi (vedi video modello VH e modello VH 2). Il sanguinamento minimo dovrebbe smettere senza utilizzare la compressione cotone sopra l&#39;anastomosi. Questo deve essere evitata per evitare la trombosi della vena delicata. </li><li> Una volta che il flusso pulsatile attraverso l&#39;anastomosi è confermato e nessun sanguinamento apparente si osserva, irrigare il campo chirurgico con soluzione salina e chiudere l&#39;incisione cervicale con una sutura 6-0 nylon. Infine, amministrare 0,15 ml più di intrapbrupenorphine eritoneal per la gestione del dolore postoperatorio e 0,2-0,4 ml di sottocutanea 0,9% soluzione salina per ricostituire la perdita di sangue durante l&#39;intervento chirurgico. </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>

Le procedure sperimentali con animali da laboratorio sono stati approvati dalla cura e l&#39;uso degli animali Comitato Istituzionale dell&#39;Università della California, San Francisco (UCSF). Gli autori non hanno potenziali conflitti di interesse relativi ai farmaci e materiali utilizzati in questa procedura.

Ipertensione venosa cerebrale prolungata è stata strettamente correlata con più gravi manifestazioni cliniche e prognosi sfavorevole nei pazienti con FAV durale e MAV cerebrali 3. Questi effetti di CVH sono stati ampiamente studiati in modelli di ratto 1,2,8. Un modello equivalente nel topo dovrebbe consentire l&#39;uso di animali geneticamente modificati, che finirebbe per consentire l&#39;analisi dei meccanismi molecolari coinvolti nella patogenesi di ipertensione venosa e la sua relazione con ...

I modelli animali di ipertensione venosa cerebrale hanno dimostrato di essere uno strumento fondamentale per la comprensione della fisiopatologia del cervello malformazioni artero-venose e fistole arterovenose 1-7. Il più diffuso è il modello di ratto creata attraverso una fistola artificiale tra l&#39;arteria carotide comune (CCA) e la vena giugulare esterna (EJV), che provoca una ipertensione venosa cerebrale coerente (CVH) nel ratto 1,8-10. Modelli di topi equivalenti, aprendo la possibilità di utilizzare diversi ceppi di topi transgenici, consentirebbe ulteriori studi sulla fisiopatologia non solo, ma anche potenziali terapie genetiche per queste malattie cerebrovascolari. Inoltre, il costrutto sperimentale potrebbe anche essere ulteriormente adattata allo studio di altre malattie cerebrovascolari sono collegati con ipertensione venosa come emicrania, amnesia globale transitoria, cecità monoculare transitoria, ecc 11 Tuttavia, i tentativi precedenti per costruire tali modelli topi have dimostrato le difficoltà con pervietà della fistola grazie all&#39;anatomia diminutivo 5,12. Qui, descriviamo nostro protocollo passo-passo per una anastomosi successo della murino CCA e EJV che si traduce in una fistola brevetto a lungo termine e una ipertensione venosa durevole nel topo.

<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.

La comprensione della fisiopatologia del cervello malformazioni artero-venose e fistole arterovenose è migliorata grazie a modelli animali. Un modello di ratto creare una fistola artificiale tra l&#39;arteria carotide comune (CCA) e la vena giugulare esterna (EJV) è stato ampiamente descritto ed dimostrato tecnicamente fattibile. Questo costrutto provoca una consistente ipertensione venosa cerebrale (CVH), e quindi ha contribuito a studiare il contributo di ipertensione venosa alla formazione, i sintomi clinici, e la prognosi di MAV cerebrali e FAV durale. Modelli di topi equivalenti sono stati solo poco descritti e hanno mostrato problemi con stenosi della fistola. Un modello murino stabilito permetterebbe lo studio non solo fisiopatologia, ma anche potenziali terapie genetiche per queste malattie cerebrovascolari. Presentiamo un modello di fistola artero che produce una ipertensione venosa intracranica durevole nel topo. Microsurgical anastomosi of murino CCA e EJV possono essere difficili a causa di anatomia diminutivo e spesso provocare una fistola non-brevetto. In questo protocollo step-by-step affrontiamo tutte le importanti sfide incontrate durante questa procedura. Evitare eccessiva retrazione della vena durante l&#39;esposizione, utilizzando 11-0 suture invece di 10-0, e facendo un attentamente pianificato anastomosi end-to-side sono alcuni dei passaggi critici. Anche se questo metodo richiede abilità microchirurgia avanzata ed una curva di apprendimento più che l&#39;equivalente nel ratto, può essere sviluppato in modo coerente. Questo modello romanzo è stato progettato per integrare le tecniche di topi transgenici con un sistema sperimentale già ben consolidata che si è dimostrato utile per studiare MAV cerebrali e FAV durale. Aprendo la possibilità di utilizzare topi transgenici, un più ampio spettro di modelli validi può essere raggiunta e trattamenti genetici può essere testato. Il costrutto sperimentale potrebbe anche essere ulteriormente adattata allo studio di otle sue malattie cerebrovascolari legati con ipertensione venosa, come l&#39;emicrania, amnesia globale transitoria, transitoria cecità monoculare, etc.

Un esito positivo del modello è una fistola artero-venosa brevetto che induce l&#39;ipertensione venosa nel cervello murino. Per convalidare il modello che abbiamo inizialmente misurato la pressione venosa intracranica nel seno sagittale dei topi a 2, 3 e 4 settimane dopo l&#39;intervento chirurgico. 6 diversi topi sono stati assegnati a ogni gruppo il tempo. La pressione del seno era 8.8 ± 1.2 mmHg nel gruppo misurate due settimane dopo l&#39;intervento chirurgico. Nei 6 topi misurati 3 settimane dopo l&#39;intervent...

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.

Noi descriviamo un metodo per creare un modello affidabile di ipertensione venosa cerebrale nel topo adulto. Questo modello è stato ampiamente descritto ed testati nel ratto. Questo nuovo omologo nei topi apre la possibilità di utilizzare animali geneticamente modificate e amplia così le applicazioni del modello.

Topo adulto venosa Ipertensione Modello: carotide comune di External Vena giugulare anastomosi.

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

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Research

JoVE Journal

Medicine

14.0K Views.  University of California, San Francisco.  Noi descriviamo un metodo per creare un modello affidabile di ipertensione venosa cerebrale nel topo adulto. Questo modello è stato ampiamente descritto ed testati nel ratto. Questo nuovo omologo nei topi apre la possibilità di utilizzare animali geneticamente modificate e amplia così le applicazioni del modello. 

Video: 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.

La formazione di una sofisticata tecnica microchirurgica: Interposizione della vena giugulare esterna innesto nella carotide comune 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 ...

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Medicina

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 occlusione comune Arteria carotide come uno stimolo precondizionamento in grado di indurre tolleranza ischemica precoce di focale Ischemia Cerebrale

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.

Cateterismo dell&#39;arteria carotide e giugulare Vena di eseguire misure emodinamiche, Infusi e campionamento del sangue in un modello di ratto cosciente

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.

Cloruro ferrico-indotta Trombosi modello murino sulla carotide e Mesentere 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.

Esecuzione permanente distale Medio cerebrale con arteria carotide comune di occlusione in ratti anziani per studiare corticale Ischemia con disabilità sostenuta

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 Trasferimento del gene per l'endotelio dell'arteria carotica comune di coniglio

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.

Un modello del coniglio di espressione del Transgene durevole nella vena giugulare di innesti di interposizione dell'arteria carotica comune

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.

Fase di contrasto risonanza magnetica nell'arteria carotica comune ratto

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.

Misurazione di contrattilità isometrica dell'arteria mesenterica Mouse con filo Myography

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.

Occlusione dell'arteria cerebrale centrale consentendo di riperfusione tramite riparazione dell'arteria carotica comune nei topi

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