Name: a murine model of ischemic retinal injury induced by transient bilateral common carotid artery occlusion
Uploaded: 2020-11-12T15:00:00
Description: Watch this Scientific Journal Video about A Murine Model of Ischemic Retinal Injury Induced by Transient Bilateral Common Carotid Artery Occlusion at JoVE.com

这项工作得到了教育、文化、体育、科学和技术部（MEXT）的科学研究援助赠款（18K09424，东芝库里原和20K18393对三和裕一郎）的支持。

这里描述的所有方法都得到了庆应大学医学院机构动物护理和使用委员会（IACUC）的批准。
1. 准备手术器械和动物
<ol><li>高压外科手术器械，并保持他们在70%的乙醇。在每一次新的外科手术之前，清洁手术器械小心使用70%的乙醇。</li>
	<li>在特定无病原体 （SPF） 室中准备雄性 BALB/cAJc1 小鼠（6 周大，26-28 公斤），以便在手术前、手术期间和手术后保持无菌状态。</li>
</ol><p cl...

在这项研究中，我们已经表明，tBCCAO，使用简单的缝合和夹子，可以诱导视网膜缺血和伴随视网膜功能障碍。此外，我们已证明，我们目前的协议，开发视网膜缺血小鼠模型是更容易和更快相比，其他以前的协议，开发视网膜缺血损伤模型2，3，7。
从解剖学上讲，左右脑动脉可以通过后沟通动脉（PCAs）连...

视网膜是一种神经感官组织，用于视觉功能。由于视觉功能需要大量的氧气，视网膜被称为身体中对氧气要求最高的组织之一。由于氧气通过血管输送，网膜易患血管疾病。各种血管疾病，如糖尿病视网膜病变和视网膜血管（静脉或动脉）闭塞，可诱发视网膜缺血。为了研究视网膜缺血的病理机制，认为有必要对视网膜缺血进行可重复和临床相关的实验模型。通过插入内脏丝进行中脑动脉闭塞（MCAO）是开发实验性脑缺血2、3体内啮齿动物模型的最普遍方法。由于眼动脉（OpA）接近MCA，MCAO模型也同时用于了解视网膜缺血的病理生理学4，5，6。为了诱导脑缺血和视网膜缺血，长丝通常通过普通胡萝卜动脉 （CCA） 或外部胡萝卜动脉 （ECA） 的切口插入。这些方法很难执行，需要很长时间才能完成手术（一只小鼠超过60分钟），并导致手术后结果的高变异性7。制定更好的模式以改善这些关切仍然很重要。
在这项研究中，我们只是使用短的瞬态双边CCA闭塞（tBCCAO）与针头和夹子诱导视网膜缺血在小鼠，并分析了视网膜缺血损伤的典型结果。在此视频中，我们将演示 tBCCAO 程序。

<table>
	<tbody>
		<tr>
			<td>Atipamezole hydrochloride</td>
			<td>Zenoaq</td>
			<td>Antisedan</td>
			<td>For anti-anesthesia</td>
		</tr>
		<tr>
			<td>Applied Biosystems 7500 Fast</td>
			<td>Applied Biosystems</td>
			<td>-</td>
			<td>For qPCR</td>
		</tr>
		<tr>
			<td>Butorphanol tartrate</td>
			<td>Meiji Seika Pharma</td>
			<td>Vetorphale</td>
			<td>For anesthesia</td>
		</tr>
		<tr>
			<td>BZ-II Analyzer</td>
			<td>KEYENCE</td>
			<td>-</td>
			<td>For an image merge</td>
		</tr>
		<tr>
			<td>BALB/cAJc1</td>
			<td>CLEA</td>
			<td>-</td>
			<td>Mouse strain</td>
		</tr>
		<tr>
			<td>&#946;-Actin (8H10D10) Mouse mAb</td>
			<td>CST</td>
			<td>3700</td>
			<td>For western blot</td>
		</tr>
		<tr>
			<td>Clamp Forcep</td>
			<td>World Precision Instruments</td>
			<td>WPI 500451</td>
			<td>For surgery</td>
		</tr>
		<tr>
			<td>Dumont forceps #5</td>
			<td>Fine Science Tools</td>
			<td>11251-10</td>
			<td>For surgery</td>
		</tr>
		<tr>
			<td>DAPI solution</td>
			<td>Dojindo</td>
			<td>340-07971</td>
			<td>For IHC</td>
		</tr>
		<tr>
			<td>Envisu SD-OCT system</td>
			<td>Leica</td>
			<td>R4310</td>
			<td>For OCT</td>
		</tr>
		<tr>
			<td>FITC-dextran</td>
			<td>Merk</td>
			<td>FD2000S</td>
			<td>For retinal blood perfusion</td>
		</tr>
		<tr>
			<td>Fluorescence microscope</td>
			<td>KEYENCE</td>
			<td>BZ-9000</td>
			<td>For fluorescence detection</td>
		</tr>
		<tr>
			<td>Gatifloxacin hydrate</td>
			<td>Senju Pharmaceutical</td>
			<td>Gachifuro</td>
			<td>For anti-bacterial infection</td>
		</tr>
		<tr>
			<td>GFAP Monoclonal Antibody (2.2B10)</td>
			<td>Thermo</td>
			<td>13-0300</td>
			<td>For IHC</td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Marukan</td>
			<td>RH-200</td>
			<td>For surgery</td>
		</tr>
		<tr>
			<td>HIF-1&#945; (D1S7W) XP Rabbit mAb</td>
			<td>CST</td>
			<td>36169</td>
			<td>For western blot</td>
		</tr>
		<tr>
			<td>ImageQuant LAS 4000 mini</td>
			<td>GE Healthcare</td>
			<td>-</td>
			<td>For chemiluminescence</td>
		</tr>
		<tr>
			<td>Midazolam</td>
			<td>Sandoz K.K</td>
			<td>SANDOZ</td>
			<td>For anesthesia</td>
		</tr>
		<tr>
			<td>Microtome Tissue-Tek TEC 6</td>
			<td>Sakura</td>
			<td>-</td>
			<td>For sectioning</td>
		</tr>
		<tr>
			<td>Medetomidine</td>
			<td>Orion Corporation</td>
			<td>Domitor</td>
			<td>For anesthesia</td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>Handaya</td>
			<td>HS-2307</td>
			<td>For surgery</td>
		</tr>
		<tr>
			<td>PuREC</td>
			<td>MAYO Corporation</td>
			<td>-</td>
			<td>For ERG</td>
		</tr>
		<tr>
			<td>Scissor</td>
			<td>Fine Science Tools</td>
			<td>91460-11</td>
			<td>For surgery</td>
		</tr>
		<tr>
			<td>Sodium hyaluronate</td>
			<td>Santen Pharmaceutical</td>
			<td>Hyalein</td>
			<td>For eye lubrication</td>
		</tr>
		<tr>
			<td>Tropicamide/Penylephrine hydrochloride</td>
			<td>Santen Pharmaceutical</td>
			<td>Mydrin-P</td>
			<td>For mydriasis</td>
		</tr>
		<tr>
			<td>6-0 silk suture</td>
			<td>Natsume</td>
			<td>E12-60N2</td>
			<td>For surgery</td>
		</tr>
	</tbody>
</table>

a murine model of ischemic retinal injury induced by transient bilateral common carotid artery occlusion

多种血管疾病，如糖尿病视网膜病变、视网膜静脉或动脉闭塞以及眼缺血综合征可导致视网膜缺血。为了研究视网膜缺血的病理机制，需要开发相关的实验模型。从解剖学上讲，主要的视网膜供血血管是眼动脉 （OpA）， OpA 起源于普通胡萝卜动脉 （CCA） 的内胡萝卜动脉。因此，CCA的中断可以有效地导致视网膜缺血。在这里，我们建立了视网膜缺血的鼠标模型，通过瞬态的双边常见胡萝卜素动脉闭塞（tBCCAO），用6-0丝缝合右CCA，并通过夹子暂时将左CCA遮挡2秒，并表明tBCCAO可以诱发急性视网膜缺血，导致视网膜功能障碍。目前的方法通过仅使用手术针头和夹子来减少对手术器械的依赖，缩短了闭塞时间，以尽量减少意外的动物死亡，这在中脑动脉闭塞小鼠模型中经常看到，并保持常见视网膜缺血结果的可重复性。该模型可用于研究小鼠缺血视网膜病变的病理生理学，并进一步用于体内药物筛选。

在FITC-dextran系统循环2分钟后，检查了假手术小鼠和tBCCAO手术小鼠左视网膜和右视网膜的视网膜血管（补充图1）。FITC-dextran在假手术小鼠的视网膜和tBCCAO操作的小鼠的左视网膜中完全可见，而在tBCCAO操作的小鼠的右视网膜中部分可检测到。
在tBCCAO之后，对眼睑下垂进行了检查（图2）。右眼显示温和（得分2;75%）和严重的眼睑（分数3和4：25%?...

Watch this Scientific Journal Video about A Murine Model of Ischemic Retinal Injury Induced by Transient Bilateral Common Carotid Artery Occlusion at JoVE.com

A Murine Model of Ischemic Retinal Injury Induced by Transient Bilateral Common Carotid Artery Occlusion

在这里，我们描述了一个鼠标模型的视网膜缺血通过瞬态的双边常见的胡萝卜动脉闭塞使用简单的缝合线和夹子。该模型可用于了解心血管异常引起的视网膜缺血的病理机制。

由瞬态双边常见胡萝卜动脉闭塞引起的缺血性视网膜损伤的 Murine 模型

Diverse vascular diseases such as diabetic retinopathy, occlusion of retinal veins or arteries and ocular ischemic syndrome can lead to retinal ischemia. ...

Research

JoVE Journal

Medicine

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

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

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

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 ...

Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat

内皮素-1诱导大脑中动脉闭塞模型缺血性中风激光多普勒血流仪在大鼠指导

Stroke is the number one cause of disability and third leading cause of death in the world, costing an estimated $70 billion in the United States in ...

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

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 ...

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

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

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, ...

Transient Middle Cerebral Artery Occlusion Model of Neonatal Stroke in P10 Rats

在P10大鼠新生儿中风的短暂大脑中动脉栓塞模型

A number of animal models have been used to study hypoxic-ischemic injury, traumatic injury, global hypoxia, or permanent ischemia in both the immature ...

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

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

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

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 ...