该项目由美国国立卫生研究院T32 GM008440到埃斯彭·沃克，R01 NS27713威廉L.Young，P01 NS44155威廉L.Young和华缩，R21 NS070153华SU和美国心脏协会AHA 10GRNT3130004到华苏部分支持。医生安娜·罗德里格斯·埃尔南德斯从"OBRA社会的La Caixa银行"支持赠款

1.准备鼠标<ol><li>诱发全身麻醉的小鼠异氟醚气。注0.15毫升腹腔brupenorphine的疼痛管理。在开始之前，检查麻醉水平通过刺老鼠的爪子令人满意。 </li><li>把鼠标在背卧与四肢固定胶带。除去颈部的头发和用剪刀上胸部。通过皮下注射，辖0.2-0.4毫升0.9％盐水中，以保持鼠标的外科手术过程中水合。 </li><li>准备手术视野下了严格的消毒方法。的皮肤切口的面积应为90％的醇进行清洗。 </li></ol> 2.解剖颈总动脉和颈外静脉<ol><li>使整个鼠标的下颈部区域的水平中线颈切口。深化伤口后，用在提升唾液腺及颈部软组织raction缝合（ 图1）。暴露右侧颈外静脉（EJV）横向到胸锁乳突肌（SCM）。这一步应该在显微镜下进行，因为过度牵拉过静脉可能会损坏，并诱导其血栓形成。 </li><li>小心地从锁骨到颅底解剖沿其课程的权利合资企业。通常电双极钳，凝聚和分裂的任何分支机构，准备了充足的长度为后来的临时安置夹子吻合。 </li><li>外侧到气管和内侧到SCM，探索颈总动脉（CCA）。它应该从锁骨小心接触到刚超过其分叉成外部和内部颈动脉。在此步骤期间，应注意再次给予，以避免过度牵引到合资企业可能购买损害吻合的通畅。 </li></ol> 3.准备吻合<ol><li>结扎CCA与10-0尹恩惠LON刚好接近其分叉。接着，应用一个临时剪辑在近端CCA尽可能靠近锁骨越好。 </li><li>一旦流已经被中断，横切动脉正下方分叉结扎并用盐水灌溉它洗出任何残留的血液的管腔内。避免双极电凝在这个步骤中，由于热损伤动脉壁可以把将来吻合处于危险之中。 </li><li>为了提高合资企业的边缘的可见性，内侧壁标有沿计划venotomy的过程中一个蓝色标记笔。一旦合资企业被标记，用10-0缝线结扎前端作为尾尽可能和应用上的近端颅尽可能临时血管夹。 </li><li>用细地下30针0.5毫升注射器，使最初的光圈在合资企业的标志区，并立即灌溉管腔用生理盐水，避免血栓形成。接下来，延长与microscissors的venotomy直到该长度是CCA约2-3倍的直径。避免剧烈拉伸，注意保持锋利，整洁的前沿。 </li><li>近似​​CCA的结束合资企业。使侧切切口CCA的施主端与直径调整到长度与venotomy大小（ 图2）。 </li></ol> 4.终端侧吻合<ol><li>用11-0尼龙单丝缝合的CCA到EJV端侧吻合。缝合内侧壁吻合的craneo脚方向是初始步骤。每一针应放在从外到内的静脉壁的第一（ 图3），并从内到外的动脉壁（ 图4）下一个。这将保持结上的吻合血管的时刻（ 图5）的外表面。任一间断或连续的缝合线都可以使用，但所有的连续的缝合线应紧固作为最终步骤。 </li><li> ØNCE内侧墙已被缝合，从尾重复该过程颅与侧壁。现在每一个线迹应放在从外到内的动脉壁的第一和从内到外的静脉壁旁边。生理盐水冲洗将有助于在手术过程中保持在任何时候都可见吻合的管腔。 </li><li>在完成吻合的所有步骤之后，第一个和下一个从动脉除去从静脉临时剪辑。动脉血液会流入合资企业没有或很少渗出从吻合（见视频VH模式和VH模式2）。最小的出血应该停止不使用棉花的压缩比吻合。这必须避免，以防止微妙静脉血栓形成。 </li><li>一旦搏动流经吻合确认并没有明显的出血观察，灌溉手术区域用盐水和用6-0尼龙缝线闭合颈切口。最后，管理0.15毫升更多intrap的eritoneal brupenorphine术后疼痛管理和0.2-0.4毫升皮下0.9％盐水的手术过程中，以补充任何血液损失。 </li></ol>

<ol>
	<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>

实验程序，实验动物批准了加州大学的机构动物护理和使用委员会，旧金山分校（UCSF）。 作者有没有潜在的利益有关此过程中使用的药物和物质的冲突。

持续脑静脉高血压已密切相关的更严重的临床表现及预后差的患者硬脑膜动静脉瘘和脑动静脉畸形3。 CVH这些作用已被广泛研究在大鼠模型1,2,8。在鼠标的等效模型将允许使用转基因动物这将最终允许参与对静脉高压的发病机制及其与硬脑膜动静脉瘘，脑动静脉畸形的关系分子途径的分析。 这里，我们报告用于通过终端到侧颈颈静脉瘘创建在成年小鼠脑静脉高...

脑静脉高压动物模型已被证明是脑动静脉畸形和动静脉瘘1-7的病理生理的认识的重要工具。最广泛使用的是通过颈总动脉（CCA）和颈外静脉（合资企业），这引发了一致脑静脉高血压（CVH）在大鼠1,8-10之间的人工瘘创建的大鼠模型。相当于小鼠模型，通过打开使用转基因小鼠品系不同的可能性，将允许不仅病理生理学，但也是潜在的基因疗法，这些脑血管疾病的进一步研究。此外，实验结构还可以进一步适应于与静脉压增高，如偏头痛，短暂性失忆，瞬态单眼失明等 11然而，以前的尝试相关的其他脑血管疾病的研究，构建这些小鼠模型ħAVE展示与瘘管通畅的困难，由于身材矮小的解剖5,12。在这里，我们描述一步一步的协议，用于小鼠CCA和合资企业的转化为长期的专利瘘和鼠标耐用静脉高压的成功吻合。

<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.

脑动静脉畸形和动静脉瘘的病理生理的认识有了提高得益于动物模型。大鼠模型创建颈总动脉（CCA）和外部颈静脉（合资企业）之间的人工瘘已被广泛描述和证明了在技术上是可行的。该构建激起一个一致脑静脉高血压（CVH），因此，已帮助学习静脉高压，以形成，临床症状，和脑动静脉畸形和硬脑膜动静脉瘘的预后的贡献。相当于小鼠模型已经只有很少描述和证明与瘘管狭窄的麻烦。一个既定的小鼠模型将允许不仅病理生理学的研究，但也是潜在的基因疗法，这些脑血管疾病。 我们提出的动静脉瘘的模型，产生一个持久的颅内静脉高​​血压的鼠标。显微外科吻合术Øf显示鼠CCA和合资企业可以是困难的，因为小型的解剖学和经常导致非专利瘘。在该步骤中的分步协议中，我们解决所有在此过程中所遇到的重大挑战。在曝光过程中避免了静脉过度内陷，用11-0缝线，而不是10-0，并作出精心策划端侧吻合是一些关键步骤。虽然这种方法需要先进的显微技术和其等效在大鼠，可以一致地开发了一种更长的学习曲线。 这种新颖的模型已被设计为转基因小鼠技术结合起来已经证明，研究脑动静脉畸形和硬脑膜动静脉瘘有用一个预先既定的实验系统。通过打开使用的转基因小鼠的可能性，有效模型的一个更广泛，可以实现和遗传治疗也可进行测试。该实验的构建体也可以被进一步适配以OT的研究她静脉高压，如偏头痛，短暂性失忆，瞬态单眼失明等相关的脑血管疾病

该模型的一个成功的结果是一个专利动静脉瘘诱导静脉高压在小鼠大脑。为了验证该模型，我们最初测定小鼠的矢状窦的颅内静脉压力在2，3和4周后手术。 6种不同的小鼠被分配到每一个时间组。窦的压力为8.8±1.2毫米汞柱的组中测得的两个星期后手术。在手术后第3周测量的6只小鼠中，鼻窦压为4.7±1.4毫米汞柱。最后，将6只小鼠测定4周手术具有3.9±0.6毫米汞柱（ 图6）一窦压力后。术...

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.

我们描述了在成年小鼠产生的脑静脉高血压可靠的模型的方法。该模型已被广泛描述和试验的大鼠。在小鼠中这个新的对应打开利用遗传修饰的动物的可能性，从而拓宽了该模型的应用程序。

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 ...

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

Research

JoVE Journal

Medicine

14.0K 次观看.  University of California, San Francisco. 我们描述了在成年小鼠产生的脑静脉高血压可靠的模型的方法。该模型已被广泛描述和试验的大鼠。在小鼠中这个新的对应打开利用遗传修饰的动物的可能性，从而拓宽了该模型的应用程序。 

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

A Model of Disturbed Flow-Induced Atherosclerosis in Mouse Carotid Artery by Partial Ligation and a Simple Method of RNA Isolation from Carotid Endothelium

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified version of carotid partial ligation methods [1,2] to show that it acutely induces low and oscillatory flow conditions, two key characteristics of disturbed flow, in the mouse common carotid artery. Using this model, we have provided direct evidence that disturbed flow indeed leads to rapid and robust atherosclerosis development in Apolipoprotein E knockout mouse [3]. We also developed a method of endothelial RNA preparation with high purity from the mouse carotid intima [3]. Using this mouse model and method, we found that partial ligation causes endothelial dysfunction in a week, followed by robust and rapid atheroma formation in two weeks in a hyperlipidemic mouse model along with features of complex lesion formation such as intraplaque neovascularization by four weeks. This rapid in vivo model and the endothelial RNA preparation method could be used to determine molecular mechanisms underlying flow-dependent regulation of vascular biology and diseases. Also, it could be used to test various therapeutic interventions targeting endothelial dysfunction and atherosclerosis in considerably reduced study duration.

This describes a partial carotid ligation surgery, which causes disturbed flow conditions and subsequent atherosclerosis development (in two weeks) with intraplaque neo-vascularization (in four weeks) in the mouse common carotid artery. We also describe a novel method of RNA isolation from the carotid intima, providing high purity endothelial RNA.

一个型号的不安流致动脉粥样硬化小鼠颈动脉部分结扎术和颈动脉内皮细胞的RNA分离的简单方法

Despite the well-known close association, direct evidence linking disturbed flow to atherogenesis has been lacking. We have recently used a modified ...

科研

医学

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.

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 ...

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.

双侧颈总动脉闭塞足够的预处理刺激诱导局灶性脑缺血早期缺血耐受

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.

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. ...

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.

三氯化铁诱发血栓形成小鼠模型上的颈动脉和肠系膜血管

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.

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, ...

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 ...

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, we have translated a vascular injury model from a small animal into a large animal (canine), with slight modifications that can be used for pre-clinical screening of prophylactic and thrombolytic agents. In addition to the surgical methods, the modified protocol describes the step-by-step methods to assess carotid artery canalization by angiography, detailed instructions to process both the brain and carotid artery for histological analysis to verify carotid canalization and cerebral hemorrhage, and specific parameters to complete an assessment of downstream thromboembolic events by utilizing magnetic resonance imaging (MRI). In addition, specific procedural changes from the previously well-established small animal model necessary to translate into a large animal (canine) vascular injury are discussed.

Here, we present the modifications necessary to a well characterized and commonly used small animal ferric chloride-induced (FeCl3) carotid artery injury model for use in a large animal vascular injury model. The resulting model can be utilized for pre-clinical trial assessment of both prophylactic and thrombolytic pharmacological and mechanical interventions.

氯化铁诱导的犬颈动脉血栓形成: 一种大的血管损伤动物模型

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, ...