Bu proje kısmen William L.Young, William L.Young ve Hua Su, Hua SU R21 NS070153 için P01 NS44155 için Espen Walker NIH T32 GM008440, R01 NS27713 tarafından Hua Su Amerikan Kalp Derneği AHA 10GRNT3130004 tarafından desteklenmektedir. Dr Ana Rodríguez-Hernández &quot;Obra Sosyal La Caixa&quot; bir hibe ile desteklenmektedir

1. Fare Hazırlanması <ol><li> Izofluran gazla fare genel anestezi neden olur. Ağrı yönetimi için intraperitoneal brupenorphine 0.15 ml enjekte edilir. Anestezi seviyesi farenin pençeleri iğneleyici tatmin edici olup olmadığını Devam etmeden önce, kontrol edin. </li><li> Yapışkan bant ile sabit dört ekstremite ile sırt yatma fare koyun. Boyun saç ve makas ile üst göğüs çıkarın. Deri altı enjeksiyon yoluyla, cerrahi işlem sırasında nemli fare tutmak için% 0.9 tuzlu 0.2-0.4 ml yönetmek. </li><li> Sıkı bir steril yöntemle aşağıdaki operatif alan hazırlayın. cilt kesi yeri% 90 alkol ile temizlenmelidir. </li></ol> 2. Ortak Karotis Arter ve Dış Boyun Damar Kesme <ol><li> Fare alt boyun alanı boyunca yatay bir orta hat servikal kesi olun. Yarayı derinleşen sonra en kullanarak tükürük bezleri ve servikal yumuşak doku yükseltmesinereaksiyona karşı sütür (Şekil 1). Sternoklaydomastoid kas (SCM) lateral sağ eksternal juguler ven (EJV) Açığa. Ven üzerinde aşırı çekiş zarar ve trombozu neden olabilir çünkü bu adım, mikroskop altında yapılmalıdır. </li><li> Dikkatle kafa tabanına klavikula onun kurs boyunca sağ EJV teşrih. Genellikle elektrik bipolar forseps, katılaştırır ve daha sonra geçici klip yerleştirme ve anastomoz için yeterli uzunluğa hazırlamak için herhangi bir dalları bölün. </li><li> SCM trakea ve medial lateral, karotis arter (CCA) keşfetmek. Dikkatle sadece dış ve iç karotid arterlerin ayrılmasından ötesinde klavikula gelen açık olmalıdır. Bu adım sırasında, dikkat sonra anastomoz açıklığını tehlikeye atabilecek EJV aşırı çekiş önlemek için tekrar verilmelidir. </li></ol> 3. anastomoz hazırlanması <ol><li> 10-0 Ny ile CCA ligeonun çatallanma boylam sadece yakın. Sonraki mümkün olduğunca klavikula kadar yakın proksimal CCA üzerinde geçici bir klip uygulayın. </li><li> Akış kesildi sonra, hemen çatallanma ligature altında arter transect ve lümen içinde kalan kanı yıkamak için tuzlu su ile sulamak. Arter duvarına termal yaralanma tehlikeye gelecekteki anastomoz koyabilirsiniz çünkü, bu aşamada bipolar koagülasyon kaçının. </li><li> EJV kenarlarında görünürlüğünü artırmak amacıyla, medial duvar planlanan venotomi seyri boyunca mavi işaretleme kalemi ile işaretlenir. EJV işaretlenmiş sonra, mümkün olduğunca kaudal olarak uzak ucunu bağlayan ve mümkün olduğunca kranial gibi yakın ucuna geçici vasküler klibi uygulamak için 10-0 sütür kullanın. </li><li> Ince 30 G iğne ile 0.5 ml şırınga ile EJV belirgin alana bir başlangıç ​​yapmak ve diyafram hemen trombüs oluşumunu önlemek için tuzlu su ile lümen sulamak. Daha sonra, microscissors venotomi uzatmakuzunluğu CCA yaklaşık 2-3 kat çapı kadar. Germe şiddet kaçının ve keskin ve düzgün kesme kenarı tutmak için dikkat. </li><li> EJV için CCA ucunu yaklaşmaktadır. CCA donör sonunda bir yan kesim kesi venotomi boyutuna uzunluğu (Şekil 2) çapını ayarlamak için emin olun. </li></ol> 4. sonu-yan anastomoz <ol><li> CCA kadar EJV uç-yan anastomoz için 11-0 tek filamentli naylon dikiş kullanın. Bir craneo-kaudal yönde anastomoz medial duvarını dikilmesi ilk adımdır. Her dikiş venöz duvarında dışarıdan birinci (Şekil 3) yerleştirilmiş ve aşağıdaki iç-dış atardamar duvarında (Şekil 4) ile ilgili olmalıdır. Bu, her zaman (Şekil 5) anastomoz damarların dış yüzeyi üzerindeki düğüm devam edecektir. Kesilebilir veya sürekli dikiş kullanılabilir, ancak tüm sürekli dikiş son adım olarak sıkılmalıdır ya. </li><li> Çmedial duvar sütüre edilmiştir nce, yan duvar ile kranyal kaudal prosedürü tekrarlayın. Şimdi her dikiş dışında-arteriyel duvarın birinci ve sonraki venöz duvarın iç-dışarı konulmalıdır. Tuzlu sulama işlemi sırasında her zaman görünür anastomoz lümen tutmaya yardımcı olacaktır. </li><li> Anastomoz tüm adımları tamamladıktan sonra, ilk ve sonraki arter ven geçici klibi silmek. arteriyel kan anastomoz hiçbir ya da çok az sızma ile EJV akmaya (videolar, VH modeli ve VH modeli 2 bakınız). az kanama anastomoz üzerine pamuk sıkıştırma kullanmadan durdurmak gerekir. Bu hassas ven trombozu önlemek için kaçınılmalıdır. </li><li> Pulsatil anastomoz akmasına sonra onaylanır ve hiçbir belirgin kanama görülmektedir, tuzlu su ile cerrahi alan sulamak ve 6-0 naylon sütür ile servikal kesi kapatın. Son olarak, intrap 0.15 ml daha yönetmekpostoperatif ağrı yönetimi ve derialtı% 0.9 tuzlu 0.2-0.4 ml eritoneal brupenorphine ameliyat sırasında herhangi bir kan kaybını doldurmak için. </li></ol>

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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. 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Laboratuar hayvanlarında deneysel prosedürler Kaliforniya Üniversitesi Kurumsal Hayvan Bakım ve Kullanım Kurulu, San Francisco (UCSF) tarafından onaylanmıştır. Yazarlar bu prosedürde kullanılan ilaçlar ve malzemelere ilişkin ilgi hiçbir potansiyel çatışmaları var.

Sürekli serebral venöz hipertansiyon yakından daha ağır klinik bulguları ve dura fistüller ve beyin AVM&#39;leri 3 olan hastalarda kötü prognoz ile ilişkili olmuştur. KVH Bu etkiler yaygın sıçan modellerinde 1,2,8 incelenmiştir. Fare eşdeğer bir model sonuçta venöz hipertansiyon patogenezinde ve dura AVF ve beyin AVM ile ilişkisi üzerinde yer alan moleküler yollar analizini sağlayacak genetiği değiştirilmiş hayvanların kullanımına olanak sağlayacak. <p class="jove...

Serebral venöz hipertansiyon hayvan modelleri beyin arteriovenöz malformasyonlar ve arteriovenöz fistül 1-7 patofizyolojisinde anlamada önemli bir araç olduğu kanıtlanmıştır. En yaygın olarak kullanılan karotis arter (CCA) ve sıçan 1,8-10 tutarlı serebral venöz hipertansiyon (CVH) kışkırtır eksternal juguler ven (EJV) arasında yapay bir fistül aracılığıyla oluşturulan sıçan modelidir. Eşdeğer fareler modelleri, farklı transgenik fareler suşları kullanılarak olasılığını açarak, sadece patofizyoloji aynı zamanda bu serebrovasküler hastalıklar için potansiyel genetik tedaviler üzerinde daha fazla çalışma izin verecek. Bundan başka, deneysel yapı ayrıca bu fareler modelleri h oluşturmak için, migren, venöz hipertansiyon, geçici global amnezi, geçici monoküler körlük, vb 11 Bununla birlikte, önceki girişimlerde ile ilgili diğer serebrovasküler hastalıkların çalışma uyarlanabilirave nedeniyle küçücük anatomi 5,12 için fistül açıklığı zorlukları gösterdi. Burada, biz uzun vadeli bir patent fistül ve fare dayanıklı venöz hipertansiyon çevirir Murin CCA ve EJV başarılı bir anastomoz için bizim adım adım protokol açıklar.

<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.

Beyin arteriovenöz malformasyonlar ve arteriovenöz fistül patofizyoloji anlayışı hayvan modellerinde sayesinde geliştirdi. Karotis arter (CCA) ve eksternal juguler ven (EJV) arasında yapay bir fistül oluşturarak bir sıçan modeli yaygın açıklanan ve teknik açıdan kanıtlanmıştır. Bu yapı tutarlı bir serebral venöz hipertansiyon (CVH) kışkırtır ve bu nedenle oluşumu, klinik belirtiler ve beyin AVM&#39;leri ve dura AVF&#39;lerin prognozu venöz hipertansiyon katkısını okuyan yardımcı oldu. Eşdeğer fareler modelleri güçlükle tarif edilmiştir ve fistül darlığı sorun göstermiştir. Bir kurulan fare modeli bu serebrovasküler hastalıklar için sadece patofizyoloji çalışma değil, aynı zamanda potansiyel genetik tedaviler sağlayacak. Biz fare dayanıklı intrakranial hipertansiyon venöz üreten arteriyovenöz fistül bir model sunuyoruz. Mikrocerrahi anastomoz omurin f CCA ve EJV bağlı küçültme anatomisine zor olabilir ve sıklıkla bir patent dışı fistül neden olur. Bu adım adım protokolde bu işlem sırasında karşılaşılan tüm önemli zorlukları. 11-0 sütür yerine 10-0 kullanarak, pozlama sırasında ven aşırı çekilmesini kaçınmak, ve dikkatlice planlanmış uç-yan anastomoz yaparak kritik adımlardan bazıları. Bu yöntem, gelişmiş mikrocerrahi beceri ve sıçan eşdeğer, sürekli geliştirilebilir bir uzun bir öğrenme eğrisi gerektirir rağmen. Bu yeni model, beyin AVM&#39;u ve dura AVF incelemek için yararlı olduğu kanıtlanmıştır önceden köklü deneysel sistemi ile transgenik fare tekniklerini entegre şekilde dizayn edilmiştir. Transgenik fareler kullanılarak olasılığını açarak, geçerli bir model, daha geniş bir spektrum elde edilebilir ve genetik tedaviler de test edilebilir. Deneysel yapı ayrıca ot çalışma uyarlanabilirMigren gibi venöz hipertansiyon, geçici global amnezi, geçici monoküler körlük, vb ilgili onu serebrovasküler hastalıklar

Modelin başarılı bir sonuç kemirgen beyin venöz hipertansiyon neden olur patent arteriyovenöz fistül olduğunu. Modeli doğrulamak için başlangıçta ameliyat sonrası 2, 3 ve 4 hafta farelerin sagital sinüs intrakranial venöz basınç ölçüldü. 6 farklı fareler, her zaman grubuna ayrıldı. sinüs basıncı grubunda 8.8 ± 1.2 mmHg ameliyattan iki hafta sonra ölçülmüştür. 3 hafta ameliyat sonrası ölçülen 6 farelerde, sinüs basıncı 4.7 ± 1.4 mmHg idi. Cerrahi 3.9 ± 0.6 mm Hg (Şekil...

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.

Biz yetişkin fare serebral venöz hipertansiyon güvenilir bir model oluşturmak için bir yöntem açıklanmaktadır. Bu model yaygın fare tarif edilir ve test edilmiştir. Farelerde bu yeni mevkidaşı genetik modifiye hayvanları kullanma imkanı açar ve böylece modelin uygulamalarını genişletmektedir.

Yetişkin Fare Venöz Hipertansiyon Modeli: Dış Boyun Ven Anastomoza Ortak Karotis Arter.

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 Görüntüleme.  University of California, San Francisco.  Biz yetişkin fare serebral venöz hipertansiyon güvenilir bir model oluşturmak için bir yöntem açıklanmaktadır. Bu model yaygın fare tarif edilir ve test edilmiştir. Farelerde bu yeni mevkidaşı genetik modifiye hayvanları kullanma imkanı açar ve böylece modelin uygulamalarını genişletmektedir. 

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.

Sıçanlarda Common Karotid Arter interpozisyon Harici Juguler Ven Greft: Gelişmiş Mikrocerrahi Tekniği Eğitimi

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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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.

Odak Serebral İskemi Erken İskemik Tolerans tetikleyebilecek bir Yeterli Önkoşullama Uyarıcı olarak bilateral Ortak Karotis Arter Tıkanıklığı

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.

Karotis Arter ve Ven Juguler kateterizasyon bir Bilinçli Rat Modelinde, İnfüzyonları ve Kan Örnekleme hemodinamik Tedbirleri Gerçekleştirme

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.

Ferric Klorür kaynaklı Karotis Arter ve Mezenter Gemi Tromboz Fare Modeli

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.

Yaşlı Sıçanlarda Ortak Karotis Arter Tıkanıklığı ile Kalıcı Distal Orta Serebral Sahne Kortikal İskemi Sürekli Engelli Eğitim için

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.

Vivo Gen transferi için tavşan ortak Karotid arter endotel

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.

Tavşan manken dayanıklı Transgene ifade juguler ven ortak karotid arter interpozisyonu Greftler için

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.

Faz kontrast manyetik rezonans görüntüleme sıçan ortak karotis arter

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.

İzometrik Contractility ölçüm tel Myography kullanarak fare mezenterik arter

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.

Orta serebral arter tıkanıklığı ortak karotid arter onarım ile reperfüzyon farelerde izin

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