（; EJD EY022383和EY022683）和Core补助（P30EY001765），成像和显微镜核心模块这项工作是由美国国立卫生研究院的研究资助。

伦理学声明:所有程序均按照由约翰霍普金斯大学机构动物护理和使用委员会规定的准则进行。 注意:在拍摄过程中使用的小鼠是从杰克逊C57BL / 6小鼠，虽然也可以使用其他啮齿动物品系或物种。当使用其他菌株或物种，应注意麻醉剂量和伤害时间表可能会有所不同。它适应I / R的条件，以适应应变，物种和实验的变化是重要的。 1.准备麻醉鸡尾酒<ol><li>结合1.25毫升氯胺酮，0.625毫升赛拉嗪，0.375毫升乙酰丙嗪，和22.75毫升磷酸盐缓冲盐水在50毫升离心管中。 注意:对于手稿的其余部分，该解决方案将被称为鸡尾酒。 </li><li>过滤到鸡尾酒用60毫升注射器和0.20微米的过滤器的新的无菌50ml离心管中。标签和日期这个新涂是。 </li><li>完全包住鸡尾酒管在铝箔以防止麻醉剂的光诱导降解。 注:鸡尾酒可在室温下保存和再使用直至其最早的过期的成分的到期日。 </li></ol> 2.准备麻醉助推器<ol><li>在50ml离心管中，结合4毫升氯胺酮和16毫升磷酸盐缓冲盐水。 注:该解决方案将作为今后助推器被称为。 </li><li>过滤成助推器用60毫升注射器和0.20微米的过滤器的新的无菌50ml离心管中。标签和日期这一新管。 注:助力器可以在室温下保存和再使用直至其最早的过期的成分的到期日。 </li></ol> 3.准备手术套房<ol><li>设定18℃，21℃之间的室温。 </li><li>打开手术表，并调整其表面热到解释第1条ST温度。 </li><li>覆盖手术底衬所有的工作表面。 </li><li>排列在手术台的空笼或其他容器温暖。 </li><li>安排一个规模上台面的耳朵恶搞。 </li></ol> 4.准备肝素钠的平衡盐溶液<ol><li>将0.5ml肝素钠到500ml四瓶平衡盐溶液（HBSS）中的。 </li><li>将prepierced Y形管插入瓶中0.1％肝素钠的主要集的尖底。 </li></ol> 5.设置输液架<ol><li>挂在输液架上延伸0​​.1％肝素钠的瓶子，并扣打开prepierced Y形管的主要集空气过滤器盖。 </li><li>提升0.1％瓶肝素钠到163厘米（120毫米汞柱）。测量从桌面到肝素钠滴的峰的高度。 </li><li>通过手动弹prepierced Y形管的主要集中移除在静脉输液管的气泡。 </l我&gt; </ol> 6.设置肝素钠滴灌<ol><li> prepierced Y形管的主要组连接到五阀歧管。 </li><li>连接30号½使用露儿男性对男性露儿管接头英寸针到五端口歧管。 </li><li>将每个30号½英寸针到30号管自己的10寸段。 </li><li>使用止血，打破新的30口径，½英寸针针尖和他们的钝端插入30号管。针尖无菌消毒或将是必要的步骤8.3。 </li><li>使用磁带，安排与针，使得连接到内口管被定位在连接到外端口的管的顶部的30号管。这种安排将防止前房插管过程中管缠绕。 </li><li>打开0.1％肝素钠流至五个阀歧管和每个单独的端口。 <ol><li>确保0.1％肝素钠是为每个端口强烈流动。如果一个端口弱流动，更换端口或无菌注射器与空气清除。 </li><li>允许0.1％肝素钠流动为2 - 3分钟以从30号管和5-阀歧管除去气泡。 </li></ol></li><li>关闭于5阀组的所有端口。 </li></ol> 7.准备手术的小鼠<ol><li>把老鼠的外科手术室。供水瓶每个网箱以防止手术过程中动物脱水。 </li><li>记录每只小鼠的体重。 </li><li>注射每只小鼠腹腔注射每克体重0.02毫升鸡尾酒。 </li><li>标记和记录各小鼠的数目。 </li><li>将所有的老鼠进入上手术台的空容器。允许所有的老鼠5-10分钟，以达到深度麻醉由无踏板戒断反应到脚趾捏的证实。 </li><li>对于每个小鼠，施用一滴托到每个眼睛的瞳孔扩张和短期润滑。 </li><li>对于每个小鼠，施用丙美卡因一滴到每个眼的局部麻醉和短期润滑。 </li><li>安排在麻醉的顺序小鼠，使得第一鼠标被麻醉将是第一个鼠标被插管。 </li><li>允许约2分钟的眼药水才能生效。 </li><li>由磁带上紧紧拉制备胶带的直4英寸件。预留每鼠一块胶带。 注意:如果不紧密地拉在磁带上会导致磁带的卷曲。 </li></ol> 8.导管插入前房<ol><li>安排手术显微镜下的第一个鼠标和聚焦显微镜的首选角膜。 注:可以插管在任的眼睛进行。右手主导的外科医生可能会发现我吨最容易导管插入左眼，而左侧优势的外科医生可能更喜欢的权利。 </li><li>打开第一个0.1％肝素钠端口上的五阀组。 </li><li>在手术显微镜，用一双镊子轻轻proptose眼球。大约中途插入30号套管针进入前房的小带纤维和角膜的顶点之间。 <ol><li>小心避免划伤或刺破虹膜，晶状体，或内角膜表面。 </li><li>避免穿透角膜第二次。 </li><li>使用温和的扭转运动，克服了套管和角膜之间的摩擦，在所述前房深深插入插管。 </li></ol></li><li>使用胶带的条带的30号管件固定到该表。为了尽量减少套管插入运动，按30号管对桌面同时达到磁带。 </li><li>记录手术的开始时间。 </li><li>使用显微镜，verify表示无泄漏是明显的。如果泄漏存在，流体的运动将是眼睛附近可见。 </li><li>通过观察，该I / R眼睛比对侧眼大目视确认眼部腹胀。一起，由于没有泄漏和眼腹胀的存在表明眼压的一个成功的高程。 </li><li>应用羟丙甲纤维素到双眼。羟丙甲纤维素起到润滑角膜和密封中微小。根据需要（大约每30分钟）持续润滑再涂羟丙甲纤维素。 </li><li>重复步骤8，直到所有的动物已经被插管。 </li></ol> 9.监控麻醉<ol><li>使用脚趾捏，晶须，或尾部的视觉观察的视觉观察，以确认每个小鼠保持麻醉。如果鼠标演示踏板戒断反应，晶须抽搐，或尾部的运动，立即进行步骤9.2。 </li><li>如果一只老鼠开始在手术过程中，电梯唤醒尾到腹膜内注入0.05毫升增压到下腹部从鼠标后面。允许1 - 2分钟，助推器生效。 </li><li>如果鼠标需要额外的镇静，必要时重复步骤9。 </li></ol> 10.前房取出套管<ol><li>对于每个动物，90分钟后已经过去了，轻轻拉动从前房插管。 </li><li>从手术表Untape鼠标，注意不要扰乱相邻动物的套管管道。 </li><li>润滑双眼具有润滑果冻。 </li><li>随后的套管被删除，安排在手术台上的空容器中的小鼠从手术中恢复。不要关闭手术台的热量。 </li><li>允许，至少，2 - 3小时于加热手术台复苏。经常观察，直到他们充分从麻醉中恢复的老鼠。 </li></ol> 11.清洁式uipment <ol><li>用消毒酒精擦拭插管。 注意:为消毒或灭菌的其它方法，如高压灭菌，可以被取代。 </li><li>用60毫升注射器填充有空气排出从5阀组0.1％肝素钠，30号针头，30号管和套管。 </li><li>用60毫升注射器充满蒸馏水冲洗5阀组，30号针头，30号管和套管。 </li><li>用60毫升注射器充满空气从5端口歧管，30号针头，30号管和插管排出的蒸馏水。 </li><li>消毒插管和冲洗管道设备后，存放该设备的再利用。 </li><li>商店，放弃或关闭所有其它设备。留在手术台的热量。 </li></ol> 12.返回所有小鼠饲养笼<ol><li>后小鼠从手术唤醒（后2 - 3小时），每个动物返回其家笼。每个笼子提供凝胶的食品。返回所有笼子到其指定的房间。 </li><li>关掉手术台的热量。丢弃所有垃圾，并擦拭手术套件。 </li></ol> 13.执行视网膜评估<ol><li>适当时，收集组织学分析或暗适应小鼠视网膜电图记录视网膜。 </li></ol>

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A., Schoen, T., Kuwabara, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+ischemia+induces+cell+mitosis+in+the+adult+rat+retina.">Experimental ischemia induces cell mitosis in the adult rat retina.</a> Invest Ophthalmol Vis Sci. 29 (7), 1050-1055 (1988).</li><li>Honjo, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Statin+inhibits+leukocyte-endothelial+interaction+and+prevents+neuronal+death+induced+by+ischemia-reperfusion+injury+in+the+rat+retina.">Statin inhibits leukocyte-endothelial interaction and prevents neuronal death induced by ischemia-reperfusion injury in the rat retina.</a> Arch Ophthalmol. 120 (12), 1707-1713 (2002).</li><li>Otori, Y., et al. <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+c-fos+and+c-jun+mRNA+following+transient+retinal+ischemia:+an+approach+using+ligation+of+the+retinal+central+artery+in+the+rat.">Expression of c-fos and c-jun mRNA following transient retinal ischemia: an approach using ligation of the retinal central artery in the rat.</a> Surv Ophthalmol. 42, 96-104 (1997).</li><li>Liu, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epac2-deficiency+leads+to+more+severe+retinal+swelling,+glial+reactivity+and+oxidative+stress+in+transient+middle+cerebral+artery+occlusion+induced+ischemic+retinopathy.">Epac2-deficiency leads to more severe retinal swelling, glial reactivity and oxidative stress in transient middle cerebral artery occlusion induced ischemic retinopathy.</a> Sci China Life Sci. 58 (6), 521-530 (2015).</li><li>Zhang, Y., Zhang, Z., Yan, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simvastatin+inhibits+ischemia/reperfusion+injury-induced+apoptosis+of+retinal+cells+via+downregulation+of+the+tumor+necrosis+factor-alpha/nuclear+factor-kappaB+pathway.">Simvastatin inhibits ischemia/reperfusion injury-induced apoptosis of retinal cells via downregulation of the tumor necrosis factor-alpha/nuclear factor-kappaB pathway.</a> Int J Mol Med. , (2015).</li><li>Li, B., Pang, I. H., Barnes, G., McLaughlin, M., Holt, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+method+and+device+to+induce+transient+retinal+ischemia+in+the+rat.">A new method and device to induce transient retinal ischemia in the rat.</a> Curr Eye Res. 24 (6), 458-464 (2002).</li></ol>

The authors have no disclosures.

视网膜我/ R损伤的高眼压已经证明了其在模拟细胞损伤和功能障碍，神经退行性疾病特别是在啮齿动物的视网膜神经血管单位效用。这个程序提供了一个强大的控制组织在技术成熟度而言很方便。它已经在这方面和其他I / R损伤模型，增加的压力和缺血的持续时间可能会增加损伤程度24被注意到。出于这个原因，一些从业者选择使用缺血性压力和持续时间与此处4,6-10,12提出不同。?...

视网膜缺血再灌注（I / R）表征许多人视网膜病变，包括青光眼，糖尿病性视网膜病和视网膜血管闭塞1。在视网膜我/ R，血流量减少（缺血）在视网膜血管造成视网膜过敏的氧气和其他营养物质，沉淀严重的氧化和炎症损伤的状态时，循环后来又恢复（再灌注）2。视网膜神经似乎特别容易受到这些变化，患有视网膜神经变性的也许就是我/ R引起的损伤最鲜明的特点。这里介绍的是在小鼠模型视网膜I / R损伤的协议。这项技术使研究人员能够检查视网膜神经血管单元的人类疾病的潜在机制和治疗策略。 通过寻求理解手术贫血3的神经退行性后果外科医生于1952年首创，罗登ŧ视网膜的I /高眼压（IOP）R于1991年重新建立缺血损伤后4标准化神经退行性端点的目的。用盐水贮存提高眼压以上收缩压的滴，这些研究表明，加压眼部插管足以中止视网膜循环，并由此引发的神经元变性。使用视网膜我/ R高眼压最近的努力已开始拟订底层I / R引起的视网膜神经性变性5-12机制。多组报道额外的病理改变包括炎症13,14，血管通透性15,16，和毛细血管变性14,17。总之，这些研究已经更加普遍建立高眼压视网膜I / R损伤视网膜神经血管疾病的模型。 表征I / R损伤的机制是对VA的研究必不可少scular疾病。视网膜我/ R损伤的高眼压是众多缺氧性损伤模型，包括I / R损伤肺18，19心脏，脑20，21肝脏，肾脏22，和小肠23中的一个。这些模型已经在推进我们的血管疾病的认识及临床补救措施至关重要。通过扩展I / R进程眼部组织，视网膜我/ R损伤的高眼压的调查有助于绘制的这些相关条件，更全面的了解。 在临床视网膜神经退行性疾病，视网膜我/ R损伤的高眼压密切相应提出了兴趣探索缺血的发病机制研究的宝贵工具。本文描述的协议是有针对性的，听话的，并且可以访问。它是通过在神经元变性的端点，例如视网膜神经元的定量，视网膜厚度的测量，和电ř补充井视网膜神经元功能的ECORDING。这种模式已经证明了其在促进神经血管查询工具，它显示了在收入中的视觉医学研究的一个基础性协议状态的承诺。

<table><tbody><tr><td>Heparin Sodium Injection, USP</td><td>Abraxis Pharmaceutical Products</td><td></td><td>1,000 USP/ml</td></tr><tr><td>BSS Sterile Irrigating Solution</td><td>Alcon Laboratories, Inc.</td><td>9007754-0212</td><td>500 ml</td></tr><tr><td>SC-2 kg Digital Pocket Scale</td><td>American Weigh Scales, Inc.</td><td>SC-2 kg</td><td></td></tr><tr><td>Tropicamide Ophthalmic Solution USP 1%</td><td>Bausch + Lomb</td><td></td><td>1% (10 mg/ml)</td></tr><tr><td>Proparacaine Hydrochloride Ophthalmic Solution USP, 0.5%</td><td>Bausch + Lomb</td><td></td><td>0.5% (5 mg/ml)</td></tr><tr><td>INTRAMEDIC Polyethylene Tubing</td><td>Becton Dickinson and Company</td><td>427400</td><td>Inner diameter: 427400</td></tr><tr><td>30 G 1/2 PrecisionGlide Needles</td><td>Benton Dickinson and Company</td><td>305106</td><td></td></tr><tr><td>BC 1 ml TB Syringe, Slim Tip with Intradermal Bevel Needle, 26 G x 3/8</td><td>Benton Dickinson and Company</td><td>309625</td><td></td></tr><tr><td>BD 60 ml Syringe Luer-Lok Tip</td><td>Benton Dickinson and Company</td><td>309653</td><td></td></tr><tr><td>Zeiss OPMI Visu 200/S8 Microscope</td><td>Carl Zeiss AG</td><td>000000-1179-101</td><td></td></tr><tr><td>Sterile Syringe Filter</td><td>Corning Inc.</td><td>CLS431224</td><td>0.20 &amp;micro;m</td></tr><tr><td>Durasorb Underpads</td><td>Covidien</td><td>1038</td><td>23 x 24 inches</td></tr><tr><td>Alcohol Prep</td><td>Covidien</td><td>6818</td><td>2 Ply, Medium</td></tr><tr><td>Student Dumont #5 Forceps</td><td>Fine Science Tools</td><td>91150-20</td><td></td></tr><tr><td>Hartman Hemostats</td><td>Fine Science Tools</td><td>13002-10</td><td></td></tr><tr><td>Primary Set, Macrobore, Prepierced Y-Site, 80 Inch</td><td>Hospira</td><td>12672-28</td><td></td></tr><tr><td>Phosphate Buffered Saline pH 7.4 (1x)</td><td>Invitrogen</td><td>10010-049</td><td>500 ml</td></tr><tr><td>Distilled water</td><td>Invitrogen</td><td>15230-204</td><td>500 ml</td></tr><tr><td>C57BL/6J Mice</td><td>The Jackson Laboratory</td><td>664</td><td></td></tr><tr><td>AnaSed Injection: Xylazine Sterile Solution</td><td>LLOYD, Inc.</td><td></td><td>20 mg/ml</td></tr><tr><td>Lubricating Jelly, Water Soluble Bacteriostatic</td><td>MediChoice</td><td></td><td>3-Gram Packet</td></tr><tr><td>NAMIC Angiographic Pressure Monitoring Manifold</td><td>Navilyst Medical, Inc.</td><td>70039355</td><td>5-Valve Manifold with Seven Female Ports</td></tr><tr><td>Goniosoft, Hypromellose 2.5% Ophthalmic Demulcent Solution: Hydroxypropyl Methylcellulose</td><td>OCuSOFT, Inc.</td><td></td><td>2.5% (25 mg/ml)</td></tr><tr><td>Ketaset CIII: Ketamine Hydrochloride</td><td>Pfizer, Inc.</td><td></td><td>100 mg/ml</td></tr><tr><td>Trans-Pal I.V. Stand&amp;nbsp;</td><td>Pryor Products</td><td>372</td><td>Furnished with a home-constructed 60-cm stainless steel extension</td></tr><tr><td>Acepromazine: Acepromazine Maleate Injection, USP</td><td>Vet One</td><td></td><td>10 mg/ml</td></tr><tr><td>V-Top Surgery Table/Adjustable Hydraulic</td><td>VSSI</td><td>100-4041-21</td><td></td></tr><tr><td>Tube Fitting Luer Male to Luer Male</td><td>Warner Instruments</td><td>64-1579</td><td></td></tr></tbody></table>

a mouse model of retinal ischemia-reperfusion injury through elevation of intraocular pressure

视网膜缺血再灌注（I / R）是有助于在多个眼部病症，包括青光眼，糖尿病性视网膜病和视网膜血管阻塞的细胞损伤病理生理过程。 I / R损伤的啮齿动物模型中提供显著见解为人类I / R损伤的机制和治疗策略，特别是关于在视网膜神经血管单元神经变性损伤。这里介绍的是通过眼内压（IOP）升高诱导小鼠视网膜I / R损伤的协议。在这个协议中，眼部前房插管用针，通过该流动升高​​盐水储层的滴注。使用该滴提高眼压上述收缩动脉血压，从业者暂时停止内视网膜血流（缺血）。当循环被除去套管的恢复（再灌注），严重的细胞损伤随之而来，在视网膜的神经变性，最终导致。最近螺柱IES表明炎症，血管通透性和毛细管变性作为该模型的另外的元件。相比于替代视网膜I / R方法，如视网膜动脉结扎术，视网膜我/ R损伤的高眼压有优势在其解剖的特殊性，实验可追踪性和技术辅助，呈现自己作为一个有价值的工具，用于检查神经元病机及治疗视网膜神经血管单元。

由高眼压视网膜的I / R的神经变性的影响通常进行评价使用两个标准的方法。神经核的NeuN免疫标记已经揭示以下I / R损伤（ 图1）显著神经元细胞的损失。简单地说，眼睛剜7天I / R固定后，多聚甲醛，与神经细胞的NeuN标记标记和全装。图像用共焦显微镜拍摄的，并标有的NeuN细胞通过计数11定量。减少在神经节细胞层的神经元计数指示I / R诱导的细胞死?...

Watch this Scientific Journal Video about A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure at JoVE.com

A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure

本文介绍了在小鼠高眼压诱导视网膜缺血再灌注损伤的过程。视网膜缺血再灌注损伤的高眼压用于人类疾病视网膜特点是妥协的氧气和营养物质的交付模式，使研究人员能够检查视网膜神经血管单元的人类疾病的潜在细胞机制和治疗。

视网膜缺血再灌注损伤的小鼠模型通过眼压升高

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, ...

a-mouse-model-retinal-ischemia-reperfusion-injury-through-elevation

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

14.8K 次观看.  Johns Hopkins University. 本文介绍了在小鼠高眼压诱导视网膜缺血再灌注损伤的过程。视网膜缺血再灌注损伤的高眼压用于人类疾病视网膜特点是妥协的氧气和营养物质的交付模式，使研究人员能够检查视网膜神经血管单元的人类疾病的潜在细胞机制和治疗。 

视频: A Mouse Model of Retinal Ischemia-Reperfusion Injury Through Elevation of Intraocular Pressure

Laser-Induced Chronic Ocular Hypertension Model on SD Rats

Glaucoma is one of the major causes of blindness in the world. Elevated intraocular pressure is a major risk factor. Laser photocoagulation induced ocular hypertension is one of the well established animal models. This video demonstrates how to induce ocular hypertension by Argon laser photocoagulation in rat.

Glaucoma is one of the major causes of blindness in the world. Elevated intraocular pressure is a major risk factor. Laser photocoagulation induced ocular hypertension is one of the well established animal models. This video demonstrates how to induce ocular hypertension by Argon laser photocoagulation in rat.

激光诱导SD大鼠慢性高眼压高压模型

Glaucoma is one of the major causes of blindness in the world. Elevated intraocular pressure is a major risk factor. Laser photocoagulation induced ocular ...

科研

生物学

A Laser-induced Mouse Model of Chronic Ocular Hypertension to Characterize Visual Defects

Glaucoma, frequently associated with elevated intraocular pressure (IOP), is one of the leading causes of blindness. We sought to establish a mouse model of ocular hypertension to mimic human high-tension glaucoma. Here laser illumination is applied to the corneal limbus to photocoagulate the aqueous outflow, inducing angle closure. The changes of IOP are monitored using a rebound tonometer before and after the laser treatment. An optomotor behavioral test is used to measure corresponding changes in visual capacity. The representative result from one mouse which developed sustained IOP elevation after laser illumination is shown. A decreased visual acuity and contrast sensitivity is observed in this ocular hypertensive mouse. Together, our study introduces a valuable model system to investigate neuronal degeneration and the underlying molecular mechanisms in glaucomatous mice.

Chronic ocular hypertension is induced using laser photocoagulation of the trabecular meshwork in mouse eyes. The intraocular pressure (IOP) is elevated for several months after laser treatment. The decrease of visual acuity and contrast sensitivity of experimental animals are monitored using the optomotor test.

激光诱导的慢性高眼压表征视觉缺陷的小鼠模型

Glaucoma, frequently associated with elevated intraocular pressure (IOP), is one of the leading causes of blindness. We sought to establish a mouse model ...

医学

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the brain for visual perception. Damage to RGCs and their axons leads to vision loss and/or blindness. Although the specific cause of glaucoma is unknown, the primary risk factor for the disease is an elevated intraocular pressure. Glaucoma-inducing procedures in animal models are a valuable tool to researchers studying the mechanism of RGC death. Such information can lead to the development of effective neuroprotective treatments that could aid in the prevention of vision loss. The protocol in this paper describes a method of inducing glaucoma - like conditions in an in vivo rat model where 50 &#181;l of 2 M hypertonic saline is injected into the episcleral venous plexus. Blanching of the vessels indicates successful injection. This procedure causes loss of RGCs to simulate glaucoma. One month following injection, animals are sacrificed and eyes are removed. Next, the cornea, lens, and vitreous are removed to make an eyecup. The retina is then peeled from the back of the eye and pinned onto sylgard dishes using cactus needles. At this point, neurons in the retina can be stained for analysis. Results from this lab show that approximately 25% of RGCs are lost within one month of the procedure when compared to internal controls. This procedure allows for quantitative analysis of retinal ganglion cell death in an in vivo rat glaucoma model.

Glaucoma-inducing Procedure in an In Vivo Rat Model and Whole-mount Retina Preparation

Glaucoma is characterized by damage to retinal ganglion cells. Inducing glaucoma in animal models can provide insight into the study of this disease. Here, we outline a procedure that induces loss of RGCs in an in vivo rat model and demonstrates the preparation of whole-mount retinas for analysis.

青光眼诱导在程序<em&gt;在体内</em&gt;大鼠模型和全安装视网膜准备

Glaucoma is a disease of the central nervous system affecting retinal ganglion cells (RGCs). RGC axons making up the optic nerve carry visual input to the ...

A Model of Glaucoma Induced by Circumlimbal Suture in Rats and Mice

The circumlimbal suture is a technique for inducing experimental glaucoma in rodents by chronically elevating intraocular pressure (IOP), a well-known risk factor for glaucoma. This protocol demonstrates a step-by-step guide on this technique in Long Evans rats and C57BL/6 mice. Under general anesthesia, a "purse-string" suture is applied on the conjunctiva, around the equator and behind the limbus of the eye. The fellow eye serves as an untreated control. Over the duration of our study, which was a period of 8 weeks for rats and 12 weeks for mice, IOP remained elevated, as measured regularly by rebound tonometry in conscious animals without topical anesthesia. In both species, the sutured eyes showed electroretinogram features consistent with preferential inner retinal dysfunction. Optical coherence tomography showed selective thinning of the retinal nerve fiber layer. Histology of the rat retina in cross-section found reduced cell density in the ganglion cell layer, but no change in other cellular layers. Staining of flat-mounted mouse retinae with a ganglion cell specific marker (RBPMS) confirmed ganglion cell loss. The circumlimbal suture is a simple, minimally invasive and cost-effective way to induce ocular hypertension that leads to ganglion cell injury in both rats and mice.

Chronic ocular hypertension is induced by applying a circumlimbal suture in rats and mice, leading to functional and structural deterioration of the retinal ganglion cells consistent with glaucoma.

Circumlimbal 缝合法诱导大鼠和小鼠青光眼模型的建立

The circumlimbal suture is a technique for inducing experimental glaucoma in rodents by chronically elevating intraocular pressure (IOP), a well-known ...

A Reversible Silicon Oil-Induced Ocular Hypertension Model in Mice

Elevated intraocular pressure (IOP) is a well-documented risk factor for glaucoma. Here we describe a novel, effective method for consistently inducing stable IOP elevation in mice that mimics the post-operative complication of using silicone oil (SO) as a tamponade agent in human vitreoretinal surgery. In this protocol, SO is injected into the anterior chamber of the mouse eye to block the pupil and prevent inflow of aqueous humor. The posterior chamber accumulates aqueous humor and this in turn increases the IOP of the posterior segment. A single SO injection produces reliable, sufficient, and stable IOP elevation, which induces significant glaucomatous neurodegeneration. This model is a true replicate of secondary glaucoma in the eye clinic. To further mimic the clinical setting, SO can be removed from the anterior chamber to reopen the drainage pathway and allow inflow of aqueous humor, which is drained through the trabecular meshwork (TM) at the angle of the anterior chamber. Because IOP quickly returns to normal, the model can be used to test the effect of lowering IOP on glaucomatous retinal ganglion cells. This method is straightforward, does not require special equipment or repeat procedures, closely simulates clinical situations, and may be applicable to diverse animal species. However, minor modifications may be required.

Here, we present a protocol to induce ocular hypertension and glaucomatous neurodegeneration in mouse eyes by intracameral injection of silicone oil and the procedure for silicone oil removal from the anterior chamber to return elevated intraocular pressure to normal.

小鼠中可逆硅油诱发眼部高血压模型

Elevated intraocular pressure (IOP) is a well-documented risk factor for glaucoma. Here we describe a novel, effective method for consistently inducing ...

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 ...

Full-Circle Cauterization of Limbal Vascular Plexus for Surgically Induced Glaucoma in Rodents

Glaucoma, the second leading cause of blindness worldwide, is a heterogeneous group of ocular disorders characterized by structural damage to the optic nerve and retinal ganglion cell (RGC) degeneration, resulting in visual dysfunction by interrupting the transmission of visual information from the eye to the brain. Elevated intraocular pressure is the most important risk factor; thus, several models of ocular hypertension have been developed in rodents by either genetic or experimental approaches to investigate the causes and effects of the disease. Among those, some limitations have been reported such as surgical invasiveness, inadequate functional assessment, requirement of extensive training, and highly variable extension of retinal damage. The present work characterizes a simple, low-cost, and efficient method to induce ocular hypertension in rodents, based on low-temperature, full-circle cauterization of the limbal vascular plexus, a major component of aqueous humor drainage. The new model provides a technically easy, noninvasive, and reproducible subacute ocular hypertension, associated with progressive RGC and optic nerve degeneration, and a unique post-operative clinical recovery rate that allows in vivo functional studies by both electrophysiological and behavioral methods.

The goal of this protocol is to characterize a novel model of glaucomatous neurodegeneration based on 360&#176; thermic cauterization of limbal vascular plexus, inducing subacute ocular hypertension.

角膜缘血管丛全圆烧灼治疗啮齿动物手术诱发的青光眼

Glaucoma, the second leading cause of blindness worldwide, is a heterogeneous group of ocular disorders characterized by structural damage to the optic ...

神经科学

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

Diverse vascular diseases such as diabetic retinopathy, occlusion of retinal veins or arteries and ocular ischemic syndrome can lead to retinal ischemia. To investigate pathological mechanisms of retinal ischemia, relevant experimental models need to be developed. Anatomically, a main retinal blood supplying vessel is the ophthalmic artery (OpA) and OpA originates from the internal carotid artery of the common carotid artery (CCA). Thus, disruption of CCA could effectively cause retinal ischemia. Here, we established a mouse model of retinal ischemia by transient bilateral common carotid artery occlusion (tBCCAO) to tie the right CCA with 6-0 silk sutures and to occlude the left CCA transiently for 2 seconds via a clamp, and showed that tBCCAO could induce acute retinal ischemia leading to retinal dysfunction. The current method reduces reliance on surgical instruments by only using surgical needles and a clamp, shortens occlusion time to minimize unexpected animal death, which is often seen in mouse models of middle cerebral artery occlusion, and maintains reproducibility of common retinal ischemic findings. The model can be utilized to investigate the pathophysiology of ischemic retinopathies in mice and further can be used for in vivo drug screening.

Here, we describe a mouse model of retinal ischemia by transient bilateral common carotid artery occlusion using simple sutures and a clamp. This model can be useful for understanding the pathological mechanisms of retinal ischemia caused by cardiovascular abnormalities.

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. ...

Monitoring Dynamic Growth of Retinal Vessels in Oxygen-Induced Retinopathy Mouse Model

Oxygen-induced retinopathy (OIR) is widely used to study abnormal vessel growth in ischemic retinal diseases, including retinopathy of prematurity (ROP), proliferative diabetic retinopathy (PDR), and retinal vein occlusion (RVO). Most OIR studies observe retinal neovascularization at specific time points; however, the dynamic vessel growth in live mice along a time course, which is essential for understanding the OIR-related vessel diseases, has been understudied. Here, we describe a step-by-step protocol for the induction of the OIR mouse model, highlighting the potential pitfalls, and providing an improved method to quickly quantify areas of vaso-obliteration (VO) and neovascularization (NV) using immunofluorescence staining. More importantly, we monitored vessel regrowth in live mice from P15 to P25 by performing fluorescein fundus angiography (FFA) in the OIR mouse model. The application of FFA to the OIR mouse model allows us to observe the remodeling process during vessel regrowth.

This protocol describes a detailed method for the preparation and immunofluorescence staining of mice retinal flat mounts and analysis. The use of fluorescein fundus angiography (FFA) for mice pups and image processing are described in detail as well.

监测氧诱导视网膜病变小鼠模型中视网膜血管的动态生长

Oxygen-induced retinopathy (OIR) is widely used to study abnormal vessel growth in ischemic retinal diseases, including retinopathy of prematurity (ROP), ...

<meta charset="utf8"></head><body>用于研究视网膜病变的视网膜缺血再灌注损伤小鼠模型

Induction of Retinal Ischemia-Reperfusion Injury in a Mouse Eye Model

Ischemia-reperfusion injuries are known to cause a range of retinal pathologies, including diabetic retinopathy, glaucoma, retinal vascular occlusions, and other vaso-occlusive conditions. This manuscript presents a method for inducing ischemia-reperfusion injury in a mouse model. The method utilized anterior chamber cannulation attached to a saline reservoir, generating hydrostatic pressure to raise the intraocular pressure to 90-100 mmHg. This method effectively caused constriction of retinal capillaries to induce retinal ischemia. At the end of the ischemic period (60 min), the intraocular pressure was normalized (&#8804;20 mmHg) before removing the cannula from the anterior chamber to initiate reperfusion. Days after the ischemia/reperfusion procedure, the eyes were collected and sectioned for histological staining. The histopathology of the retinal sections was scored by evaluating eight parameters of retinal injury: folds, hemorrhage, deformation, cell loss in the ganglion cell, inner nuclear, outer nuclear, and photoreceptor layers, and damage to retinal pigment epithelial cells. This method provided a reproducible model to study the mechanisms and pathology of retinal ischemia/reperfusion injury. In addition, this model can facilitate the discovery of potential therapeutic targets to treat retinal ischemia/reperfusion injury, advancing the study of retinal pathologies and improving patient outcomes.

<meta charset="utf8"></head><body>小鼠眼模型中视网膜缺血再灌注损伤的诱导

This protocol models retinal ischemia-reperfusion injury in a mouse eye by inducing retinal ischemia via anterior chamber cannulation and intraocular pressure elevation, followed by intraocular pressure normalization to initiate the reperfusion.

小鼠眼模型中视网膜缺血再灌注损伤的诱导

Ischemia-reperfusion injuries are known to cause a range of retinal pathologies, including diabetic retinopathy, glaucoma, retinal vascular occlusions, ...