这项研究是由研究推广计划“START”，德国亚琛工业大学（标题：“的模具ArterialisationvenöserGefäße”START 690753）医学院的。我们要感谢她出色的编辑协助夫人琼Blümich的。

1。实验动物<ol><li>获得当地伦理委员会批准可用于所有的实验将要执行的，按照动物保护FELASA指引。 </li><li>使用雌性SD大鼠可用于购买任何商业饲养员（ 例如查尔斯河WIGA公司，Sulzfeld，德国），。我们使用了21只，分为三组各7只，术后3，6周和12周处死。在购买时，动物应该是12周龄，重250-300克。 </li><li>保持动物在气候控制的房间的环境温度为21°C，60％相对湿度，12小时免费获得食物和水的光暗周期。 </li></ol> 2。麻醉和手术的准备<ol><li>通过使用连接到麻醉机，设定为1升/分钟的氧气流速的有机玻璃箱中，诱导全身麻醉和异氟烷汽化器瓦特第i个5％的麻醉气体。 </li><li>维持麻醉尽快动物深感觉反射抑制在1.5％的异氟烷​​汽化器。 </li><li>修复大鼠在体保温板仰卧位。 </li><li>准备的操作区域的每个动物的脱毛霜，除去它的皮毛从钳口角向下柄sterni 2厘米以下。 </li><li>准备一个无菌外科领域关注手术的地区，该地区上的动物和外科医生的前面制备： </li><ul><li>清洁工作台上用一消毒。 </li><li>准备切口站点通过使用手术溶液擦洗的多个应用程序（ 例如，氯己定和70％异丙醇）。首先，应用洗必泰醇第二和，最后，溶液擦洗。 </li><li>穿一次性手术衣，手术手套，或者手套考试，此前已洗瓦特第i个外科手术消毒。 </li></ul><li>在操作过程中使用无菌显微仪器。 </li><li>使用手术显微镜（野生Heerbrugg M650，Gais的，瑞士），在10〜20倍的放大倍率。 </li></ol> 3。解剖学 图1描述了解剖老鼠的脖子上： 1A） 所在地前的右侧胸锁乳突肌切断。 1B） 所在地的右侧胸锁乳突肌切断后。 4。袖口准备剪切的长度为3毫米的袖口用手从一个塑料管（PORTEX，内径为0.75 mm，外径0.94毫米，史密斯医疗公司，英国）。这些翻边包括管状箍套主体，包括完整的卷的塑料管，和一个压脉袋的压脉袋的容积的一半组成的手柄。箍套主体和压脉袋手柄应同样离子克。用于操纵在手术过程中的压脉袋的压脉袋手柄是必不可少。袖带后滑过容器，迷你斗牛钳位同时放置的压脉袋的把手和容器上，以防止从出血，保持压脉袋的位置，并确保稳定。 5。袖套法<ol><li>滑袖该船只的。 </li><li>紧接其后，外翻容器，拉过来的外侧的压脉袋体，并用7-0丝线结扎固定。在这种方式中，压脉袋的​​管腔与内皮细胞覆盖。此后，滑静脉移植过的袖口的身体和一个新的7-0丝线结扎固定。为了防止血栓形成，它是必不可少的，移植静脉领带非常接近的袖带缘固定，以避免血液接触领带保持动脉到位。这是至关重要的，由于血流方向的远端吻合。然而，根据作者，它是不被认为出于同样的原因的近端吻合必不可少的。 </li></ol> 6。外科手术 6.1下游的右颈外静脉<ol><li>约2.5厘米长的中位数颈部切开直接以上的柄sterni。 </li><li>完全从周围组织中分离出1厘米长的段的右颈外静脉</li><li>用7-0丝线结扎静脉支流。 </li></ol> 6.2的右颈总动脉的下游<ol><li>准备胸锁乳突肌，在中间横切。 </li><li>准备右颈总动脉小心，确保不压缩的动脉或迷走神经。动员它的颈动脉分叉处，头臂干。 </li><li>随后，结扎两次，中间用7-0丝线缝合前横断。 </li><li>防滑的尼龙袖口consisting的身体并和常见的颈动脉，从而重要的是要确保后面的容器中（ 图2b）的压脉袋手柄的前端上的手柄（ 图2a）。 </li><li>钳位的带套囊的端部的前端与迷你牛头犬钳上（ 图2c），以确保稳定的压脉袋手柄完全相同常见的颈动脉。 </li><li>接着，直接切断动脉背后绑带，从而导致动脉残端应该只要作为压脉袋体。 </li><li>冲洗动脉管腔与肝素生理盐水溶液（20 IU肝素/毫升0.9％NaCl溶液中）。结扎环，除了在适当的位置以备后用。 </li><li>动脉残端拉起来，它像一个套在袖口身体滑倒。 </li><li>随后，拉的结扎环和结它紧紧地。 </li><li>为了防止动脉从它的袖子滑出，你可以实现一个绝招：掌握动脉壁缝合右手imultaneously与左手的钳子使结（ 图2e）（图2d）。 </li><li>重复同样的步骤，近端动脉残端（ 图2f）。 </li></ol> 6.3静脉移植定位<ol><li>消费税的1厘米长的段夹住颈外静脉</li><li>由于这两个手铐动脉静脉插入彼此相距甚远，正确的距离用3-0薇乔缝线在此过程中，你应该每一个的迷你牛头犬钳周围循环。 </li><li>纵横交错缝合。 </li><li>接着，在相反的方向上拉缝线，使铐动脉树桩的正确定位（ 图2g） </li><li>与肝素生理盐水溶液冲洗静脉段。 </li><li>滑在手铐的颈内动脉远端以前逆转静脉段的一端，将其固定在正确的位置tion与丝绸7-0结扎。 </li><li>在这里，重要的是要注意，静脉的圆周必须完全且均匀地把持。 </li><li>与肝素生理盐水溶液冲洗静脉段，和滑移近端颈残端静脉段的另一端。 </li><li>完成结扎（ 图2h）; </li><li>另一方面，第一个上的前端的颈动脉残端然后近端一个后取下微型斗牛夹具1，由此允许通畅的血流量和静脉移植物的动脉化（ 图2i）。手术被认为是成功的，如果血流量和脉动内观察到的接枝和颈总动脉（目测）。 </li></ol> 6.4。伤口愈合对于皮下连续缝合，用4-0薇乔。对于皮肤的连续缝合，用3-0薇乔。 7。恢复和术后管理&lt;/ P&gt; <ol><li>让动物从麻醉中恢复过来，下红外线变暖灯的平面纸床上用品。不要将经营的动物的动物控股的，直到所有的动物出现正常的。 </li><li>术后管理丁丙诺啡（Temgesic）的浓度为0.03-0.05毫克/公斤体重SC 2天，每8至12小时。丁丙诺啡可前给予异氟烷诱导。 </li><li>监测动物术后一般情况和体征的疾病或感染。 </li></ol> 8。血流测定术后的移植通畅证明了血液的流量测量，通过与血管周围的流量探头3毫米，（MA3PSB）的研究跨音速血流量测量系统（型号：TS420，跨音速系统欧洲BV，马斯特里赫特，荷兰）。该系统是基于多普勒原理。 9。代表性的成果由Faithfully保持上述的指示，手术的平均持续时间约为65分钟。提出了一个优秀的动物的身体状况和体重增加预计整个实验：在操作的起点的动物称重312±20.39克。手术后3周的时间点在牺牲的动物体重325.2±28.7克，在第6周333±39.9克的和在第12周362.22±29克。 无感染，观察到在操作的网站，开发没有疮。在预期的时间内，术后伤口完全愈合。脑梗塞或缺乏肌肉运动的神经控制没有任何迹象表明。的购买是很重要的一点，因为这些动物被操作在颈动脉和的两个颈外静脉之一除去，并用作自动移植。无过敏性反应，术后疼痛缓解治疗观察。 所有动物均提高警觉，社会整个实验。他们的生活习惯有关接收食物和水，在整个时间保持不变。 所有实验的存活率为100％的时间点牺牲。移植通畅率为60％，前10个动物的，因此82％，其余11只动物。总的移植物的通畅率为71％。测定动脉化的静脉移植物中的血流在时间点的牺牲为8±3毫升/分钟之间的三个牺牲时间点没有任何统计学上显着的差异。 <img src="/files/ftp_upload/4124/4124fig1.jpg" alt="图1" /> 图1。解剖老鼠的脖子。 1A） 所在地前的右侧胸锁乳突肌切断。 1B） 所在地的右侧胸锁乳突肌切断后。 e_content的“Fo：保持together.within”页面=“总是”&gt; <img src="/files/ftp_upload/4124/4124fig2.jpg" alt="图2" /> 图2。术显微照片：一）袖带组成的压脉袋体和袖口把手;二）袖带滑过常见的颈动脉;的前端）夹持的带套囊的远端常见的颈动脉残端;四）外翻在压脉袋的颈总动脉残端;五）固定外翻常见的颈动脉残端用绑带;六） 所在地后外翻的两个带套囊的远端和近端的常见的颈动脉树桩;克）纵横交错技术近似两个带套囊再灌注后的静脉插入移植的颈总动脉的树桩，H） 内脏静脉插入后，我） 所在地 。 <img src="/files/ftp_upload/4124/4124fig3.jpg" alt="图3" /> 图3描绘的拓扑示意图再灌注后的静脉插入移植。 <a href="http://www.jove.com/files/ftp_upload/4124/4124fig3large.jpg" target="_blank">点击此处查看大图</a> 。

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没有利益冲突的声明。

在颈总动脉插入颈外静脉移植是一个复杂的显微外科技术。视频帮助新手的外科医生在最短的时间内掌握这门技术，以较低的实验动物的浪费。 吸入性麻醉进行此操作是最好的选择，因为它避免了过深的麻醉阶段，可以很好的控制，并允许外科医生有足够的时间来操作。此外，大鼠是实验动物的练习这种复杂的外科手术技术，更容易，在鼠标相比，提供了一个合适的尺寸。 <...

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td>编织丝黑7-0 </td><td>爱惜康产品</td><td> 892100 </td><td></td></tr><tr><td>薇乔3-0 </td><td>爱惜康产品</td><td> J285G </td><td></td></tr><tr><td>薇乔4-0 </td><td>爱惜康产品</td><td> J284G </td><td></td></tr><tr><td> PORTEX非无菌尼龙管0.75毫米，0.94毫米的细口径聚乙烯（聚乙烯）管</td><td> PORTEX英国</td><td> 800/200/175/100 </td><td></td></tr><tr><td>异氟烷5％ </td><td></td><td></td><td>任何genericon </td></tr><tr><td>肝素</td><td></td><td></td><td>任何genericon </td></tr><tr><td> 0.9％的生理盐水</td><td></td><td&gt; </td><td>任何genericon </td></tr><tr><td>丁丙诺啡</td><td></td><td></td><td>任何genericon </td></tr></tbody></table>

training a sophisticated microsurgical technique: interposition of external jugular vein graft in the common carotid artery in rats

内膜增生是一个非常重要的动脉冠状动脉旁路移植术，外周动脉搭桥手术以及动静脉瘘，动脉静脉狭窄的主要原因。1-5实验过程中常见的颈动脉移植静脉插入使用袖扣技术已经应用在几个研究项目，研究新内膜增生的病因和治疗方案来解决这个问题。6-8袖带防止血管吻合口重构，并诱导移植物内的动荡，从而内膜增生的发展。 使用上腔静脉移植静脉动脉化实验的小动物模型是一个既定的9-11目前的协议，是指建立的颈静脉移植物插入技术首次描述了邹等人，以及其他。12-14 </sup然而，这些列举的小动物是复杂的协议。 为了简化该过程，并最大限度地减少所需的实验动物的数量，详细操作由视频训练协议。此视频帮助新手医生学习的袖口技术和移植静脉插入。由此，右颈外静脉接枝在袖带技术中常见的颈动脉21的雌性Sprague Dawley大鼠分为三个相等的组21，42和84天，分别处死。值得一提的是，必要的，因为没有供体动物进行自动移植。的存活率为100％的时间点牺牲。此外，移植通畅率为60％，前10个动物的82％，其余11只动物。在牺牲的时间的血流量为8±3毫升/分钟。总之，这种手术的协议大大简化，优化和规范这个组合licated程序。它给新手医生容易的，一步一步的指示，解释可能存在的缺陷，从而帮助他们快速获得专业知识，避免无谓牺牲的实验动物。

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

狭窄的动脉静脉内膜增生的首要原因。我们提出了一个新的协议，在SD大鼠颈总动脉，右颈外静脉用袖套法嫁接。的存活率为100％的时间点牺牲。

培养一个精密的显微外科技术在大鼠颈总动脉，颈外静脉移植：介入

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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Research

JoVE Journal

Medicine

20.3K 次观看.  University Hospital RWTH Aachen. 狭窄的动脉静脉内膜增生的首要原因。我们提出了一个新的协议，在SD大鼠颈总动脉，右颈外静脉用袖套法嫁接。的存活率为100％的时间点牺牲。

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

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

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model creating an artificial fistula between the common carotid artery (CCA) and the external jugular vein (EJV) has been widely described and proved technically feasible. This construct provokes a consistent cerebral venous hypertension (CVH), and therefore has helped studying the contribution of venous hypertension to formation, clinical symptoms, and prognosis of brain AVMs and dural AVFs. Equivalent mice models have been only scarcely described and have shown trouble with stenosis of the fistula. An established murine model would allow the study of not only pathophysiology but also potential genetic therapies for these cerebrovascular diseases.
We present a model of arteriovenous fistula that produces a durable intracranial venous hypertension in the mouse. Microsurgical anastomosis of the murine CCA and EJV can be difficult due to diminutive anatomy and frequently result in a non-patent fistula. In this step-by-step protocol we address all the important challenges encountered during this procedure. Avoiding excessive retraction of the vein during the exposure, using 11-0 sutures instead of 10-0, and making a carefully planned end-to-side anastomosis are some of the critical steps. Although this method requires advanced microsurgical skills and a longer learning curve that the equivalent in the rat, it can be consistently developed.
This novel model has been designed to integrate transgenic mouse techniques with a previously well-established experimental system that has proved useful to study brain AVMs and dural AVFs. By opening the possibility of using transgenic mice, a broader spectrum of valid models can be achieved and genetic treatments can also be tested. The experimental construct could also be further adapted to the study of other cerebrovascular diseases related with venous hypertension such as migraine, transient global amnesia, transient monocular blindness, etc.

We describe a method for creating a reliable model of cerebral venous hypertension in the adult mouse. This model has been widely described and tested in the rat. This new counterpart in the mice opens the possibility of using genetic modified animals and thereby broadens the applications of the model.

成年小鼠静脉高压型号：颈总动脉，以颈外静脉吻合。

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

科研

医学

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.

颈动脉和颈静脉导管在有意识大鼠模型进行血流动力学措施，输液和血液采样

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. ...

Carotid Artery Infusions for Pharmacokinetic and Pharmacodynamic Analysis of Taxanes in Mice

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study model. As the use of mouse models are often a vital step in preclinical drug discovery and drug development1-8, it is necessary to design a system to introduce drugs into mice in a uniform, reproducible manner. Ideally, the system should permit the collection of blood samples at regular intervals over a set time course. The ability to measure drug concentrations by mass-spectrometry, has allowed investigators to follow the changes in plasma drug levels over time in individual mice1, 9, 10. In this study, paclitaxel was introduced into transgenic mice as a continuous arterial infusion over three hours, while blood samples were simultaneously taken by retro-orbital bleeds at set time points. Carotid artery infusions are a potential alternative to jugular vein infusions, when factors such as mammary tumors or other obstructions make jugular infusions impractical. Using this technique, paclitaxel concentrations in plasma and tissue achieved similar levels as compared to jugular infusion. In this tutorial, we will demonstrate how to successfully catheterize the carotid artery by preparing an optimized catheter for the individual mouse model, then show how to insert and secure the catheter into the mouse carotid artery, thread the end of the catheter out through the back of the mouse&#8217;s neck, and hook the mouse to a pump to deliver a controlled rate of drug influx. Multiple low volume retro-orbital bleeds allow for analysis of plasma drug concentrations over time.

This method was developed with the goal of delivering a steady drug solution via the carotid artery, to assess the pharmacokinetics of novel drugs in mouse models.

颈动脉输液的紫杉烷类化合物在小鼠体内的药代动力学和药效学分析

When proposing the use of a drug, drug combination, or drug delivery into a novel system, one must assess the pharmacokinetics of the drug in the study ...

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.

执行永久远端大脑中与老年大鼠颈总动脉闭塞研究皮层缺血具有持续残疾

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

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. Research models for bypass graft failure are essential for a better understanding of pathobiological and pathophysiological processes during graft patency loss. Large animal models, such as pigs or sheep, resemble human anatomical structures but require special facilities and equipment. This video describes a rat vein interposition model to investigate vein graft patency loss. Rats are inexpensive and easy to handle. Compared to mouse models, the convenient size of rats permits better operability and enables a sufficient amount of material to be obtained for further diverse analysis. In brief, the inferior epigastric vein of a donor rat is harvested and used to replace a segment of the femoral artery. Anastomosis is conducted via single stitches and sealed with fibrin glue. Graft patency can be monitored non-invasively using duplex sonography. Myointimal hyperplasia, which is the main cause for graft patency loss, develops progressively over time and can be calculated from histological cross sections.

This video demonstrates a model to study the development of myointimal hyperplasia after venous interposition surgery in rats.

静脉介入型号：研究旁路移植通畅一个合适的模型

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

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.

体内兔颈总动脉内皮的基因转移

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.

颈动脉介入移植中颈静脉持久性转基因表达的家兔模型

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.

大鼠颈总动脉的相位对比磁共振成像

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

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.

颅内动脉闭塞允许经普通颈动脉修复再灌注的研究

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

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

大鼠胡萝卜动脉压力控制段气球损伤与腹腔ventit 治疗应用

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...