我们感谢Eleut埃尔南德斯畜牧业，加布Gaidosh显微镜的专业知识，以及奎Tran和镇扬赵视频编辑。该研究由美国国家眼科研究所授予R01-EY022129到JLG和P30补助EY022589到加州大学圣地亚哥分校和EY014801到UM;美国心脏协会，詹姆斯和Esther国王基金会，复旦大学研究生院（编号2010033）的博士研究生交流计划资金，以及无限制的赠款防盲研究公司

所有动物的程序批准了加州大学圣地亚哥和迈阿密体制动物护理和使用委员会（IACUC）大学等按照ARVO声明对动物的眼科和视觉研究中的应用进行。所有试剂和外科手术使用的仪器是不育的。 1.麻醉和准备鼠外科手术<ol><li>前程序，将大鼠按照体重麻醉腹膜内注射氯胺酮（60毫克/千克）和赛拉嗪（8毫克/千克）的。麻醉足够的深度应到脚趾捏刺激了否定的答复来确定。 </li><li>麻醉后，将舌头向前，以防止窒息和应用润滑软膏双眼，以防止手术中角膜干燥。 </li><li>剃使用理发推子的手术部位和擦拭区域三次，用10％providone碘洗涤剂溶液和70％的乙醇。 </li><li>悬垂吨无菌区域内，他的动物。无菌手套和手术器械的过程中生存手术中使用。重新消毒使用的动物之间的热珠灭菌仪器的提示。 </li></ol> 2.手术方法<ol><li> PION感应 <ol><li> 股静脉插管 <ol><li>制备并清洗手术部位。剃使用理发推子的右大腿内侧和擦拭区域三次，每次用10％providone碘洗涤剂溶液和70％的乙醇。 </li><li>准备管。切聚乙烯管道的40厘米长度（PE 10）灭菌70％的乙醇。冲洗管道用盐水并将其连接到一个1ml注射器含有2％藻红乙染料预先测定溶液（1微升/毫克，得到20毫克/公斤体重的剂量）。装入注射器插入一个脚踏开关控输注泵设定为600微升/分钟的速率。 </li><li>使用15号刀片，做一个小的水平切口处右大腿的基础。剪切和传播里面的膜，清洁用无菌棉签的区域。 </li><li>分离钳肌肉直到股静脉的分支是可见的。鞘周围动脉，静脉和神经。捏，向上拉出此套钳（精尖杜蒙钳），切一小切口（2-4毫米通常就足够了）的三角形楔Vannas弹簧剪刀基地附近。展开切割是必要的。 </li><li>分离静脉和动脉与显微手术钩平行于静脉的方向上的钝。要小心，不要损坏精致膜，静脉分支。然后，轻轻提起静脉和它从底层结缔组织分开。 </li><li>获得无针尼龙缝合线，并把它旁边的股静脉。使用显微外科钩，提升静脉，并通过细尖镊子远侧区域的下方。抓住缝线的一端，并把它下面的静脉。紧紧结扎远端静脉。通过以类似的方式的第二缝合线下的近端静脉和使一个松散结。 </li><li>做一个小切口在靠近远端结扎Vannas弹簧剪刀静脉。如果必要，展开与细尖镊子孔。有些血液可能会泄漏通过晋级。清洁冷，无菌BSS和无菌棉签手术区。 </li><li>在切割的边缘保持静脉壁，导尿容器使用一针固定器制备的盐水冲洗管道。拧紧静脉周围和油管近结。然后，通过将其直接连接到所述远端的缝合锚的管道。 </li><li>检查导管的质量按下脚踏开关注入生理盐水，&lt;1 m1为充分。确保管道通畅，没有泄漏。暂时关闭缝合切口，以保护导管和组织。 </li></ol></li><li> 曝光视神经 <ol><li> PreparE中的手术部位通过擦拭预先刮净区域上面的左眼三次，每次用10％providone碘洗涤剂溶液和70％的乙醇。 </li><li>使沿皮肤2-3毫米的眼睛后面用第15号刀片的切口。捏住并解除与锯齿钳的结缔组织，并做一个小切口，Vannas弹簧剪刀。这个小切口通常为约5毫米的长度，但可以更长，以提供在训练外科医生更大的曝光。通过继续沿眼眶骨的上缘的结缔组织直言不讳地剖析，同时注意避免破坏血管。清洁用棉签手术区。 </li><li>通过结膜向下解剖直到上直肌是可见的。捏并剖析通过肌肉;肌肉会从深被解放的轨道内。现在，周围组织可用于在眼睛的缩回和仰角，以帮助为便于可视化。 </li><li>回覆道皮肤和结缔组织的瓣横向和向下，并保持在适当位置与缝合和止血钳。这将向前和向外转动眼睛，以揭示所述含脂肪鞘围绕视神经。 </li><li>小心地将一对尖锐的镊子，扩大平行于视神经分开周围鞘的结缔组织。请勿触摸视神经与镊子的尖头。 </li><li>视神经和周围鞘的5毫米长现在应该是可见的。微血管上的套表面的网络环绕视神经。这些将激光照射过程中有的放矢。 </li></ol></li><li> 静脉注射赤藓红B和激光照射 <ol><li>同时经营的激光照射装置从绿色激光保护自己戴上橙色护目镜在任何时候。打开激光，打开快门，调整峰值和平均值鲍威激光器的RS作为必要的。关闭快门。 </li><li>定位在激光照射装置的老鼠。为了确保适当的光束的位置，弱对准光束是通过钻在关闭快门叶片的直径为100微米的孔空间滤波的激光产生。再暴露有一对细尖镊子的视神经，和对准光束定位到3之间毫米和视神经头后面4毫米眶内视神经。 </li><li>通过激活输液泵的注入2％赤藓红B中的溶液。它可循环几秒钟，而外科医生增加一小滴的BSS的滋润视神经的表面上。 </li><li>验证瞄准光束的位置，然后单击脚踏开关来启动辐射。橙色安全过滤器，这是添加到显微镜的光路，将立即触发随后快门的一个1秒的延迟后开口。 </li><li>照射为9视神经0秒与135MW的一个峰值功率和18mW功率由光束斩波旋转在250赫兹以15％的占空比产生的平均功率（这可以减少热效应）。黄色荧光，通过安全过滤器可视化为亮橙色，将被从视神经的上表面发射的，并足以确保光束照射神经对称。 </li><li>在辐射后，橙色的安全过滤器将自动打开。微出血可以在某些情况下可以观察到。 </li><li>减轻对额外眼肌牵引和眼睛返回到中立位置。关闭切口间断缝合。然后撤出导管和打结股静脉严密，防止泄漏;关闭间断缝合。应用抗生素软膏，以两个切口。检查眼底验证视网膜中央静脉和动脉的血管完整性。 </li></ol></li></ol></li><li> 逆行标记。视网膜神经节细胞（RGC的）。注：为了评估RGC存活，用荧光金（FG）逆行标记应PION前一星期内完成。该方法中详细朱庇特协议819 13中描述 。 <ol><li>简而言之：麻醉与氯胺酮（60毫克/公斤）和甲苯噻嗪（8毫克/千克）的动物和剃光头。 </li><li>手术擦洗切口部位，使整个头部正中切口，暴露颅骨。 </li><li>钻透颅骨双侧孔（直径2×2毫米）0.5毫米无论从矢状缝。 </li><li>小心吸用真空泵即在于在上丘（SC）的背表面上的脑的内容。然后，将一小块明胶海绵用4％FG浸泡到SC的表面。 </li><li>关闭缝合切口和爱护使用标准的术后护理的动物。 </li></ol></li><li> 术后护理和安乐死 <ol><li>手术后，将动物在一个单独的干净笼子上的再循环水加热垫的顶部，直到动物进行回收。 </li><li>手术后止痛药（盐酸丁丙诺啡，0.01毫克/千克），应每天给予两次，连续三天，以减少不适。 </li><li>鼠应分开存放，并观察，直到他们能够保持胸骨斜卧，恢复足够的意识。 </li><li>回收和身体健康的迹象每天监测至少5天的手术后，或直到拆线和外科手术部位，为准最新的适当愈合。 </li><li>根据调查感兴趣的手术后安乐死的动物通过灌注4％PFA在科学上适当的时间点。 </li></ol></li></ol>

<ol>
	<li>Wang, Y., Brown, D. P., Duan, Y., Kong, W., Watson, B. D., Goldberg, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+rodent+model+of+posterior+ischemic+optic+neuropathy..">A novel rodent model of posterior ischemic optic neuropathy..</a> JAMA Ophthalmology. 131, 194-204 (2013).</li><li>Rucker, J. C., Biousse, V., Newman, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ischemic+optic+neuropathies.">Ischemic optic neuropathies.</a> Curr Opin Neurol. 17, 27-35 (2004).</li><li>Hayreh, S. 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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-arteritic+anterior+ischemic+optic+neuropathy:+role+of+systemic+corticosteroid+therapy..">Non-arteritic anterior ischemic optic neuropathy: role of systemic corticosteroid therapy..</a> Graefe's archive for clinical and experimental ophthalmology. 246, 1029-1046 (2008).</li><li>Prado, R., Dietrich, W. D., Watson, B. D., Ginsberg, M. D., Green, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photochemically+Induced+Graded+Spinal-Cord+Infarction+-+Behavioral,+Electrophysiological,+and+Morphological+Correlates..">Photochemically Induced Graded Spinal-Cord Infarction - Behavioral, Electrophysiological, and Morphological Correlates..</a> J Neurosurg. 67, 745-753 (1987).</li><li>Dietrich, W. D., Busto, R., Watson, B. D., Scheinberg, P., Ginsberg, M. 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The authors have nothing to disclose.

Here we describe in detail a method for inducing PION in a rat model. The most critical part of the protocol is the exposure and irradiation of the optic nerve &#8211; to expose the nerve as long as possible while avoiding damage caused by the sharp fine tip forceps or from stretching. In rats, the ophthalmic artery enters the optic nerve &#8804;1 mm from the optic nerve head. Therefore, irradiation of the optic nerve 3-4 mm away from the optic nerve head should only result in ischemia of the capillaries feeding the nerv...

患者50岁，缺血性视神经病变（ION）是最常见的类型的急性视神经病变2。前（AION）或后（π介子）3：条件可根据具体的受影响的血液供给和临床上介绍的源呈现为一个两种亚型的。虽然AION的发病机理和过程已被广泛研究4-7，PION一直由于其低流行，变呈现，不明确的诊断标准，缺乏动物模型知之甚少。此外，没有任何的治疗已被证明有效地防止或从AION或π介子反向视力丧失。因此，π介子可重复的和可靠的动物模型是很有价值的研究在体内的疾病过程和神经保护和轴突再生测试新的治疗方案。 化学诱发缺血性损伤导致vasog微血管ENIC水肿和血栓形成有效地创建区域组织缺血8-12。注射到血管循环后，光敏染料赤藓红B最终产生在激活活性单线氧分子靶血管激光照射。单重态氧直接peroxidizes血管内皮，刺激血小板粘附/聚集并导致闭塞性血栓的形成。缺血性损伤蔓延到邻近地区和微血管压迫因血管性水肿进一步加剧。此协议的总体目标是光化学诱发缺血的球后视神经镜像造成PION的损害。 据我们所知，这是在后视神经1缺血性损伤的第一模型。由于这种模式产生缺血，同时避免身体创伤，对后部缺血性视神经病变的生理过程是更好的模仿和学习。此外，这种模式提供了一个新的平台，为候选人为治疗视神经病变等中枢神经系统缺血性疾病的筛查。在这里，股静脉置管，视神经接触，静脉注射赤藓红的B和激光照射大鼠模型PION了详细的方案进行了说明。

<table>
	<tbody>
		<tr>
			<td>532-nm Nd:YAG laser&#160;</td>
			<td>Laser glow</td>
			<td>LRS-532-KM-200-3</td>
			<td></td>
		</tr>
		<tr>
			<td>Beam chopper</td>
			<td>custom-made</td>
			<td>custom-made</td>
			<td></td>
		</tr>
		<tr>
			<td>Mechanical shutter and corresponding shutter drive timer</td>
			<td>AAM Vincent Associates</td>
			<td>SD-10</td>
			<td></td>
		</tr>
		<tr>
			<td>25-cm focal length spherical lens</td>
			<td>CVI/Mellles-Griot</td>
			<td>01 LPX 293</td>
			<td>plano-convexBK7 glass lens with HEBBARTM antireflection coating</td>
		</tr>
		<tr>
			<td>Erythrosin B&#160;</td>
			<td>MP Biomedicals</td>
			<td>190449</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorogold</td>
			<td>Fluorochrome,LLC</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Gelfoam</td>
			<td>Cardinal Health</td>
			<td>CAH1203421</td>
			<td></td>
		</tr>
		<tr>
			<td>Polyethylene tubing (PE10)</td>
			<td>BD Intramedic</td>
			<td>427400</td>
			<td></td>
		</tr>
		<tr>
			<td>No. 10 Blade</td>
			<td>Miltex</td>
			<td>4-110</td>
			<td></td>
		</tr>
		<tr>
			<td>Fine Forceps</td>
			<td>F.S.T.</td>
			<td>91150-20</td>
			<td>DUMONT #5 RUSTLESS NON-MAGNETIC</td>
		</tr>
		<tr>
			<td>Forceps with Teeth</td>
			<td>F.S.T.</td>
			<td>11153-10</td>
			<td>Germany stainless</td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>F.S.T.</td>
			<td>18025-10&#160;</td>
			<td>Germany stainless</td>
		</tr>
		<tr>
			<td>Vannas spring scissors</td>
			<td>F.S.T.</td>
			<td>2-220&#160;</td>
			<td>JJECK Stainless</td>
		</tr>
		<tr>
			<td>Polyglactin suture</td>
			<td>Ethicon</td>
			<td>J488G</td>
			<td>7-0 suture</td>
		</tr>
		<tr>
			<td>hemostat</td>
			<td>F.S.T.</td>
			<td>12075-12&#160;</td>
			<td>Germany stainless</td>
		</tr>
	</tbody>
</table>

rat model of photochemically-induced posterior ischemic optic neuropathy

后部缺血性视神经病变（PION）是一见倾心，破坏性疾病的临床实践。然而，其发病机制与自然史仍然知之甚少。最近，我们开发PION的一个可靠的，可再现的动物模型和测试在这个模型1的一些神经营养因子的治疗效果。该视频的目的是表明我们光化学诱导的后部缺血性视神经病变模型，并评估其与视网膜神经节细胞的逆行标记作用。以下后视神经，光敏染料，赤藓红乙手术暴露，静脉注射和将激光束聚焦到视神经表面。赤藓红B和激光的期间，照射损伤血管内皮，促使微血管闭塞血小板血栓和水肿压缩介导的光化学相互作用。由此产生的缺血性损伤产生一个渐进的，但pronounc编视网膜神经节细胞枯死，由于亏损轴索输入 - 远程，损伤诱发和临床相关的结果。因此，这种模型提供了一种新颖的平台来研究PION的病理生理过程中，而且可用于测试治疗方法为视神经病变以及其它CNS缺血性疾病进一步优化。

诱导这种技术将所得的缺血性损伤的产生视网膜神经节细胞轴突缺血损伤后逐渐但明显的死亡。这类似于在人类疾病观察到临床上相关的结果。 FG逆行标记用于PION后量化RGC存活。相同的方法被用来验证一个模型的创建成功以及评估不同治疗方案的效果。 图1示出 FG阳性细胞的代表性共聚焦图像在视网膜平面坐骑控制（图1A），假治疗（激光只/无赤藓红B， 图1B），</s...

Watch this Scientific Journal Video about Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy at JoVE.com

Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy

该协议的目的是光化学诱导缺血性损伤至大鼠后视神经。这个模型是至关重要的后部缺血性视神经病变的病理生理的研究，以及治疗方法，这和其他视神经病变，以及其它CNS缺血性疾病。

光化学诱导后部缺血性视神经病变的大鼠模型

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history ...

rat-model-photochemically-induced-posterior-ischemic-optic

Research

JoVE Journal

Medicine

8.5K 次观看.  University of Miami Miller School of Medicine. 该协议的目的是光化学诱导缺血性损伤至大鼠后视神经。这个模型是至关重要的后部缺血性视神经病变的病理生理的研究，以及治疗方法，这和其他视神经病变，以及其它CNS缺血性疾病。 

视频: Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy

Creation of Reversible Cholestatic Rat Model

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Cholestasis can cause various physiological changes and affect the nutritional status and surgical outcomes. Study of the pathophysiological changes occurring in the liver and other organs is of importance. Various studies have been done in cholestatic rat models.
We used a reversible cholestatic rat model in our recent study looking at the role of methylprednisolone in the ischemia reperfusion injury. Various techniques for creation of a reversible cholestatic model have been described. Creation of a reversible cholestatic rat model can be challenging in view of the smaller size and unique hepatopancreatobiliary anatomy in rats. This video article demonstrates the creation of a reversible cholestatic model.
This model can be used in various studies, such as looking at the changes in nutritional, physiological, pathological, histological and immunological changes in the gastrointestinal tract. This model can also be used to see the effects of cholestasis and various therapeutic interventions on major hepatic surgeries.

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Creation of a reversible cholestatic rat model can be challenging in view of the smaller size and unique hepatopancreatobiliary anatomy in rats. This video article demonstrates the creation of a reversible cholestatic model.

可逆的胆汁淤积大鼠模型的建立

Cholestasis is a clinical condition commonly encountered by both surgeons and gastroenterologists. Cholestasis can cause various physiological changes and ...

科研

医学

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine their efficacy and safety.
Gene therapy using viral vectors is one of the most promising approaches for treating cardiac diseases. Viral delivery of various different genes by changing the carrier gene has immeasurable therapeutic potential.
In this video, the full process of an animal model of heart failure creation followed by gene transfer is presented using a swine model. First, myocardial infarction is created by occluding the proximal left anterior descending coronary artery. Heart remodeling results in chronic heart failure. Unique to our model is a fairly large scar which truly reflects patients with severe heart failure who require aggressive therapy for positive outcomes. After myocardial infarct creation and development of scar tissue, an intracoronary injection of virus is demonstrated with simultaneous nitroglycerine infusion. Our injection method provides simple and efficient gene transfer with enhanced gene expression. This combination of a myocardial infarct swine model with intracoronary virus delivery has proven to be a consistent and reproducible methodology, which helps not only to test the effect of individual gene, but also compare the efficacy of many genes as therapeutic candidates.

A method of gene transfer for the treatment of ischemic heart failure is described using a swine model of myocardial infarction. Our simple and reproducible method enables us to readily evaluate the efficacy of various gene transfers with a very simple and reproducible way.

缺血性心力衰竭在临床前模型的转基因

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine ...

A Simplified Technique for Producing an Ischemic Wound Model

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work well in non-ischemic wounds may not work in human diabetic wounds because vasculopathy is one major factor that hinders healing of these wounds. We published an article in 2007 describing a rabbit ear ischemic wound model created by a minimally invasive surgical technique. Since then, we have further simplified the procedure for easier operation. On one ear, three small skin incisions were made on the vascular pedicles, 1-2 cm from the ear base. The central artery was ligated and cut along with the nerve. The whole cranial bundle was cut and ligated, leaving only the caudal branch intact. A circumferential subcutaneous tunnel was made through the incisions, to cut subcutaneous tissues, muscles, nerves, and small vessels. The other ear was used as a non-ischemic control. Four wounds were made on the ventral side of each ear. This technique produces 4 ischemic wounds and 4 non-ischemic wounds in one animal for paired comparisons. After surgery, the ischemic ear was cool and cyanotic, and showed reduced movement and a lack of pulse in the ear artery. Skin temperature of the ischemic ear was 1-10 &deg;C lower than that on the normal ear and this difference was maintained for more than one month. Ear tissue high-energy phosphate contents were lower in the ischemic ear than the control ear. Wound healing times were longer in the ischemic ear than in the non-ischemic ear when the same treatment was used. The technique has now been used on more than 80 rabbits in which 23 were diabetic (diabetes time ranging from 2 weeks to 2 years). No single rabbit has developed any surgical complications such as bleeding, infection, or rupture in the skin incisions. The model has many advantages, such as little skin disruption, longer ischemic time, and higher success rate, when compared to many other models. It can be safely used in animals with reduced resistance, and can also be modified to meet different testing requirements.

We have developed a minimally invasive technique to create a rabbit ischemic ear wound model by dividing the central artery and nerve and the cranial neurovascular bundle. A subcutaneous tunnel then cuts all subcutaneous tissues. This procedure causes minimal skin disruption and can be safely used in diabetic animals.

一个生产缺血性损伤模型的简化技术

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work ...

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

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 20091, 2. Several models of cerebral ischemia have been developed to mimic the human condition of stroke. It has been suggested that up to 80% of all strokes result from ischemic damage in the middle cerebral artery (MCA) area3. In the early 1990s, endothelin-1 (ET-1) 4 was used to induce ischemia by applying it directly adjacent to the surface of the MCA after craniotomy. Later, this model was modified 5 by using a stereotaxic injection of ET-1 adjacent to the MCA to produce focal cerebral ischemia. The main advantages of this model include the ability to perform the procedure quickly, the ability to control artery constriction by altering the dose of ET-1 delivered, no need to manipulate the extracranial vessels supplying blood to the brain as well as gradual reperfusion rates that more closely mimics the reperfusion in humans5-7. On the other hand, the ET-1 model has disadvantages that include the need for a craniotomy, as well as higher variability in stroke volume8. This variability can be reduced with the use of laser Doppler flowmetry (LDF) to verify cerebral ischemia during ET-1 infusion. Factors that affect stroke variability include precision of infusion and the batch of the ET-1 used6. Another important consideration is that although reperfusion is a common occurrence in human stroke, the duration of occlusion for ET-1 induced MCAO may not closely mimic that of human stroke where many patients have partial reperfusion over a period of hours to days following occlusion9, 10. This protocol will describe in detail the ET-1 induced MCAO model for ischemic stroke in rats. It will also draw attention to special considerations and potential drawbacks throughout the procedure.

Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats. In addition, important considerations, advantages, and shortcomings of this model are discussed.

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 ...

Angiogenesis in the Ischemic Rat Lung

The adult lung is perfused by both the systemic bronchial artery and the entire venous return flowing through the pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that responds to a need for enhanced lung perfusion and shows robust neovascularization. Pulmonary vascular ischemia induced by pulmonary artery obstruction has been shown to result in rapid systemic arterial angiogenesis in man as well as in several animal models. Although the histologic assessment of the time course of bronchial artery proliferation in rats was carefully described by Weibel 1, mechanisms responsible for this organized growth of new vessels are not clear. We provide surgical details of inducing left pulmonary artery ischemia in the rat that leads to bronchial neovascularization. Quantification of the extent of angiogenesis presents an additional challenge due to the presence of the two vascular beds within the lung. Methods to determine functional angiogenesis based on labeled microsphere injections are provided.

The lung is perfused by both the systemic bronchial artery and pulmonary arteries. In most lung pathologies, it is the smaller systemic vasculature that shows robust neovascularization. Cessation of pulmonary blood flow promotes brisk bronchial angiogenesis. We provide surgical details of inducing left pulmonary artery ischemia that promotes bronchial neovascularization.

在缺血大鼠肺的血管生成

The  adult lung is perfused by both the systemic bronchial artery and the entire  venous return flowing through the pulmonary arteries. In most lung ...

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the world&#8217;s 2.9 million neonatal deaths. Animal models of HIE have contributed to the understanding of the pathophysiology in HIE, and have highlighted the dynamic process that occur in brain injury due to perinatal asphyxia. Thus, animal studies have suggested a time-window for post-insult treatment strategies. Hypothermia has been tested as a treatment for HIE in pdiglet models and subsequently proven effective in clinical trials. Variations of the model have been applied in the study of adjunctive neuroprotective methods and piglet studies of xenon and melatonin have led to clinical phase I and II trials1,2. The piglet HIE model is further used for neonatal resuscitation- and hemodynamic studies as well as in investigations of cerebral hypoxia on a cellular level. However, it is a technically challenging model and variations in the protocol may result in either too mild or too severe brain injury. In this article, we demonstrate the technical procedures necessary for establishing a stable piglet model of neonatal HIE. First, the newborn piglet (&#60; 24 hr old, median weight 1500 g) is anesthetized, intubated, and monitored in a setup comparable to that found in a neonatal intensive care unit. Global hypoxia-ischemia is induced by lowering the inspiratory oxygen fraction to achieve global hypoxia, ischemia through hypotension and a flat trace amplitude integrated EEG (aEEG) indicative of cerebral hypoxia. Survival is promoted by adjusting oxygenation according to the aEEG response and blood pressure. Brain injury is quantified by histopathology and magnetic resonance imaging after 72 hr.

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

新生儿缺氧缺血性脑病的小猪模型

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the ...

Demonstration of the Rat Ischemic Skin Wound Model

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and unrelieved pressure due to immobility. Animal models have been developed that attempt to mimic these conditions for the purpose of furthering our understanding of the complexity of chronic wounds. The model described herein is a rat ischemic skin flap model that permits a prolonged reduction of blood flow resulting in wounds that become ischemic and resemble a chronic wound phenotype (reduced vascularization, increased inflammation and delayed wound closure). It consists of a bipedicled dorsal flap with 2 ischemic wounds placed centrally and 2 non-ischemic wounds lateral to the flap as controls. A novel addition to this ischemic skin flap model is the placement of a silicone sheet beneath the flap that functions as a barrier and a splint to prevent revascularization and reduce contraction as the wounds heal. Despite the debate of using rats for wound healing studies due to their quite distinct anatomic and physiologic differences compared to humans (i.e., the presence of a panniculus carnosus muscle, short life-span, increased number of hair follicles, and their ability to heal infected wounds) the modifications employed in this model make it a valuable alternative to previously developed ischemic skin flap models.

The rat, due to its size, availability, and rather docile behavior, has been utilized as a research model for many years. The goal of this protocol is to utilize the rat as an ischemic skin wound healing model to provide valuable insight into the pathophysiology of chronic wounds.

在大鼠缺血性皮肤创伤模型的演示

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and ...

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy (rNAION)

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their lifetime. NAION results in optic nerve edema, selective loss of the retinal ganglion cell neurons (RGCs) and atrophy of the optic nerve. A rodent model of NAION that expresses most NAION features and sequelae has been developed, which is applicable to both rats and mice. This model utilizes a focal laser application of 532 nm wavelength to illuminate a photoactive dye, Rose Bengal (RB), to cause capillary damage and leakage at the targeted anterior optic nerve (the laminar region). After rNAION induction, there is an early optic nerve ischemia, optic nerve edema, and intraneural inflammation, followed by selective RGC and axonal loss. Since the optic nerve is a CNS white matter tract, the rNAION model is applicable to mechanistic studies of selective white matter ischemia, as well as neuroprotective analyses and short and long-term mechanisms of glial and neuronal response to ischemia.

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy rNAION

The following report describes how to replicate the rodent model of nonarteritic anterior ischemic optic neuropathy (rNAION), using the appropriate dye, contact lens and laser parameters. We also reveal the appropriate steps for evaluating the rNAION lesion in vivo.

非动脉炎性前部缺血性视神经病变的啮齿动物模型（rNAION）

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their ...

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.

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Oxygen-Induced Retinopathy Model for Ischemic Retinal Diseases in Rodents

One of the commonly used models for ischemic retinopathies is the oxygen-induced retinopathy (OIR) model. Here we describe detailed protocols for the OIR model induction and its readouts in both mice and rats. Retinal neovascularization is induced in OIR by exposing rodent pups either to hyperoxia (mice) or alternating levels of hyperoxia and hypoxia (rats). The primary readouts of these models are the size of neovascular (NV) and avascular (AVA) areas in the retina. This preclinical in vivo model can be used to evaluate the efficacy of potential anti-angiogenic drugs or to address the role of specific genes in the retinal angiogenesis by using genetically manipulated animals. The model has some strain and vendor specific variation in the OIR induction which should be taken into consideration when designing the experiments.

Oxygen-induced retinopathy (OIR) can be used to model ischemic retinal diseases such as retinopathy of prematurity and proliferative diabetic retinopathy and to serve as a model for proof-of-concept studies in evaluating antiangiogenic drugs for neovascular diseases. OIR induces robust and reproducible neovascularization in the retina that can be quantified.

罗登斯缺血性视网膜疾病的氧诱导视网膜病变模型

One of the commonly used models for ischemic retinopathies is the oxygen-induced retinopathy (OIR) model. Here we describe detailed protocols for the OIR ...