Name: a model of glaucoma induced by circumlimbal suture in rats and mice
Uploaded: 2018-10-05T14:00:00
Description: Watch this Scientific Journal Video about A Model of Glaucoma Induced by Circumlimbal Suture in Rats and Mice at JoVE.com

这项工作由澳大利亚国家卫生和医学研究理事会 (1046203)、澳大利亚研究理事会未来的奖学金 (FT130100338) 资助。

所有实验程序都是根据澳大利亚国家卫生和医学研究理事会制定的《澳大利亚保护和使用动物行为守则》进行的。伦理学的批准是从霍华德 Florey 研究所动物伦理委员会 (批准号 13-044-um 和 13-068-um 为老鼠和老鼠, 分别)。
1. 有意识大鼠的眼压测量
<ol><li>设置实验室回弹血压计到大鼠的设置。襁褓的老鼠在一块柔软的布里使动物平静下来。暴露头部和颈部。轻轻地握住躯干, 一?...

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circumlimbal 缝合是慢性眼高压症的新模型。除了有代表性的研究结果来源于16、18, 这一动物模型在最近的一些研究中被利用了15、23、24、25 ,26。以上这些报告的比较表明, 该方法产生可重复的结果, 包括眼压升高的幅度, 以及在模型?...

动物模型为实验研究青光眼发病机制的细胞过程提供了一个重要的平台, 并评估潜在的治疗干预措施。几种诱导模型的建立, 以产生持续眼压 (眼压) 升高, 最重要的危险因素, 青光眼。用于提高眼压的方法包括: 高渗盐水注射液在巩膜静脉1, 激光光凝的小梁网2或角膜缘静脉3, 和眼内注射液等物质鬼血红细胞4, 微球5,6和粘弹性剂7。每种方法都有其优点和局限性。
一个好的青光眼模型应该模仿疾病的过程, 以最小的并发症, 如创伤, 炎症和媒体混浊。这些并发症经常与用于诱发眼压升高的程序有关, 并会混淆结局的解释。例如, 即使在未引入异物的情况下, 前房穿刺也已被证明是造成外伤和炎症的原因, 而不是典型的青光眼改变8、9的代表性。除了避免炎症的重要性外, 保持光学清晰度有助于体内成像和电生理监测疾病进展。虽然目前尚不清楚这些并发症会对疾病调查有多大影响, 但在模型诱导过程中, 最好避免穿透眼球。circumlimbal 缝合方法避免了地球的穿透, 促进了体内视网膜结构和功能的纵向评估。更重要的是, 这个模型不同于以往的能力, 以返回眼压的基线值, 在必要时去除缝合。眼压正常化可能有助于研究可逆和不可逆转的神经节细胞损伤10,11,12,13,14的细胞和分子相关性。
本文着重介绍了模型归纳技术。该模型诱导的大鼠和小鼠视网膜损伤的特征可以在其他15、16、17、18、19中更详细地找到。

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			<td height="20" style="height:20px;width:157px;">normal saline</td>
			<td style="width:219px;">Baxter International Inc</td>
			<td style="width:135px;">AHB1323</td>
			<td style="width:365px;">Maintain corneal hydration during surgery</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:157px;">Chlorhexadine 0.5%</td>
			<td style="width:219px;">Orion Laboratories</td>
			<td style="width:135px;">27411, 80085</td>
			<td style="width:365px;">Disinfection of surgical instrument</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:157px;">Isoflurane 99.9%</td>
			<td style="width:219px;">Abbott Australasia Pty Ltd</td>
			<td style="width:135px;">CAS 26675-46-7</td>
			<td style="width:365px;">Proprietory Name: Isoflo(TM) Inhalation anaaesthetic. Pharmaceutical-grade inhalation anesthetic mixed with oxygen gas for suture procedure</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:157px;">ocular lubricant</td>
			<td style="width:219px;">Alcon Laboratories&#160;</td>
			<td style="width:135px;">1618611</td>
			<td style="width:365px;">Proprietory Name: Genteal, ocular lubricant to keep the other eye moist</td>
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			<td height="32" style="height:32px;width:157px;">Needle holder (microsurgery)</td>
			<td style="width:219px;">World Precision Instruments</td>
			<td style="width:135px;">555419NT</td>
			<td style="width:365px;">To hold needle during ocular surgery</td>
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			<td height="20" style="height:20px;width:157px;">Proxymetacaine 0.5%</td>
			<td style="width:219px;">Alcon Laboratories&#160;</td>
			<td style="width:135px;">CAS 5875-06-9</td>
			<td style="width:365px;">Topical ocular analgesia</td>
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			<td height="20" style="height:20px;width:157px;">Scissors (microsurgery)</td>
			<td style="width:219px;">World Precision Instruments</td>
			<td style="width:135px;">501232</td>
			<td style="width:365px;">To cut excessive suture stump during ligation</td>
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		<tr height="20">
			<td height="20" style="height:20px;width:157px;">Surgical drape</td>
			<td style="width:219px;">Vital Medical Supplies</td>
			<td style="width:135px;">GM29-612EE</td>
			<td style="width:365px;">Ensure sterile enviornment during surgery</td>
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			<td height="32" style="height:32px;width:157px;">Suture needle for rats (microsurgery)</td>
			<td style="width:219px;">Ninbo medical needles</td>
			<td style="width:135px;">151109</td>
			<td style="width:365px;">8-0 nylon suture attached with round needle, cutting edge 3/8, dual-needle, suture length 30cm</td>
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		<tr height="32">
			<td height="32" style="height:32px;width:157px;">Suture needle for mice (microsurgery)</td>
			<td style="width:219px;">Ninbo medical needles</td>
			<td style="width:135px;">160905</td>
			<td style="width:365px;">10-0 nylon suture attached with round needle, cutting edge 3/8, dual-needle, suture length 30cm</td>
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		<tr height="20">
			<td height="20" style="height:20px;width:157px;">Tweezers (microsurgery)</td>
			<td style="width:219px;">World Precision Instruments</td>
			<td style="width:135px;">500342</td>
			<td style="width:365px;">Manipulate tissues during ocular surgery</td>
		</tr>
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			<td height="20" style="height:20px;width:157px;">rebound tonometer</td>
			<td style="width:219px;">TONOLAB, iCare, Helsinki, Finland</td>
			<td style="width:135px;">TV02</td>
			<td style="width:365px;">for intraocular pressure monitoring</td>
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a model of glaucoma induced by circumlimbal suture in rats and mice

circumlimbal 缝合是一种以慢性抬高眼压 (眼压) 诱发啮齿动物实验性青光眼的技术, 是一种众所周知的青光眼危险因素。该协议说明了该技术在长埃文斯大鼠和 C57BL/6 小鼠的分步指导。在全身麻醉下, 在结膜、赤道周围和眼睛的后缘上应用 "钱包绳" 缝合。他的眼睛是一个未经处理的控制。在我们的研究期间, 这是一个8周的老鼠和12周的老鼠, 眼压保持升高, 在有意识的动物的反弹测量定期测量没有局部麻醉。在这两种类型中, 缝合眼睛显示视网膜电图的特点与优先内视网膜功能障碍一致。光学相干断层扫描显示视网膜神经纤维层有选择性细化。组织学的大鼠视网膜在横断面发现减少细胞密度在神经节细胞层, 但没有变化的其他细胞层。用神经节细胞特异标记 (RBPMS) 检测平板小鼠视网膜的染色, 证实神经节细胞丢失。circumlimbal 缝合术是一种简单、微创、经济有效的诱发眼内高压的方法, 可导致大鼠和小鼠神经节细胞损伤。

以下结果在老鼠18和老鼠16以前被报告了并且这里总结。circumlimbal 缝合在大鼠和小鼠中产生了类似的眼压升高模式 (图 2)。缝合术后立即发现一只小鼠眼压峰值, 高达 58.1 2.7 mmHg, 38.7, 2.2 mmHg。在大鼠中, 随着时间的推移, 眼压量逐渐减少 44, 为 6 mmHg 和 32, 2 毫米汞, 3 和24小时, 分别为15。在这最初的眼...

Watch this Scientific Journal Video about A Model of Glaucoma Induced by Circumlimbal Suture in Rats and Mice at JoVE.com

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

Circumlimbal 缝合法诱导大鼠和小鼠青光眼模型的建立

慢性眼高压是在大鼠和小鼠中应用 circumlimbal 缝合, 导致视网膜神经节细胞与青光眼的功能和结构恶化。

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

这种方法提供了一种可重复和微创的方式，以延长 IOP 的提升时间。这将有助于我们更好地了解青光眼。这种技术的一个优点是，IOP高程很容易逆转，使研究人员能够研究眼睛从慢性压力高程中恢复的能力。这种方法的视觉演示至关重要，因为单从阅读手稿中很难学习到张力结的苏特和绑紧结。为了提供精细的协助，我们建议一个新的这项技术，在显微镜下练习。首先用一块软布把清醒的老鼠擦去，使动物平静下来。一只手轻轻握住躯干，动物背靠着调查人员的胸部，露出头部和颈部。用另一只手将反弹吨位表放在大鼠眼附近，使内膜探针的尖端大约两到三毫米远，并垂直于角膜顶点。等待几秒钟，让老鼠平静下来，然后按一次测量按钮。观察 IOP 探头顶部轻轻击中角膜顶点一次，并听到反弹吨位计发出蜂鸣声。一两秒钟后，重复测量。总共进行 10 次测量，以得出时间点的平均 IOP 值。第五次读数后重置气压计。一旦您进行监控，在一天中的同一时间测量 IOP。在一元照明条件下，尽量减少 IOP 周期导致的变化。用无菌的手术窗帘盖住麻醉啮齿动物，使眼睛被缝合。接下来，在确保麻醉手术平面后，使用7-10尼龙缝合线，通过编织7-10缝合平行和2毫升后肢，对全球各地的球茎结膜进行包绳缝合。小心不要穿透他的sclera。手术过程中突然瞳孔扩张表明，性乳突可能已被穿透。用五到六个锚点将缝合固定在大鼠结节上。避免直接压缩的主要表皮静脉，通过线程缝合下结膜下在这些静脉的交叉。用打滑结来固定钱包串缝合线。然后打一个简单的结，并保持它松散。为了避免手术后 IOP 峰值过高，请提供辅助测量 IOP。如果发现 IOP 过高，则通过部分释放缝合线一端的张力来调整滑点。在达到30至60毫米汞的所需 IOP 后，系下第二个结，同时在缝合线端保持连续拉力。第二个结收紧后，修剪缝合的末端，以尽量减少任何异体感觉。在全身麻醉恢复期间监测动物。与小鼠一起工作时，将10-0尼龙缝合线放在边缘的一毫米后部。使用四到五个锚点将缝合线固定在鼠标的结节上。然后用绑一个滑点固定钱包串缝合线。打上一个简单的结并保持松动后，在助手的帮助下测量 IOP。为了保持长期 IOP 和视力，在紧固钱包串缝合时，确保结足够紧是十分紧密的，这一点很重要。在这里，我们演示如何将一个显著的结绑在模型眼上，以产生足够的张力。达到所需的 IOP 后，系下第二个结，同时在缝合线那一端保持连续拉力。在此图中，单个眼部高血压眼睛由红色符号表示，用灰色符号控制眼睛。圆环缝合在大鼠中升高了8周的眼内压力。IOP 在反向控制眼中保持不变。正顶点阈值反应，反映视网膜结节细胞健康，发现在红线指示的眼部高血压眼中降低;相对于黑线指示的控制眼。ERG a 和 b 波也小减少，可能分别反映光受体和双极细胞的轻微功能障碍。然而，最大的赤字是在积极的斯科托门槛响应中发现的。确认轻度慢性 IOP 升高后优先结节细胞功能障碍。在尝试此过程时，重要的是要记住用足够的内压来拧紧滑口。IOP长期未能上升的最常见原因是，随着时间的推移，结变得松动。按照这个程序，所有修改，如缝合去除，正常化眼内压力，可以回答其他问题，如常规结节细胞是否可以从维持的 IOP 高程恢复。涉及这种复苏的机制是什么？

Research

JoVE Journal

Medicine

The Spared Nerve Injury (SNI) Model of Induced Mechanical Allodynia in Mice

The Spared Nerve Injury SNI Model of Induced Mechanical Allodynia in Mice

幸免神经损伤（SNI）模型引起的机械触感小鼠

Peripheral neuropathic pain is a severe chronic pain condition which may result from trauma to sensory nerves in the peripheral nervous system. The spared ...

科研

医学

Cardiac Stress Test Induced by Dobutamine and Monitored by Cardiac Catheterization in Mice

多巴酚丁胺诱导的心脏压力测试和监控通过在小鼠心脏导管插入术

Dobutamine is a &beta;-adrenergic agonist with an affinity higher for receptor expressed in the heart (&beta;1) than for receptors expressed in the ...

Tibial Nerve Transection - A Standardized Model for Denervation-induced Skeletal Muscle Atrophy in Mice

胫骨神经切断术 - 一个标准化的模型失神经诱导小鼠骨骼肌

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. ...

Experimental Glaucoma Induced by Ocular Injection of Magnetic Microspheres

实验诱发青光眼通过磁性微球注射眼

Progress in understanding the pathophysiology, and providing novel treatments for glaucoma is dependent on good animal models of the disease. We present ...

Photothrombosis-induced Focal Ischemia as a Model of Spinal Cord Injury in Mice

Photothrombosis引起的局部脑缺血脊髓损伤的小鼠模型

Spinal cord injury (SCI) is a devastating clinical condition causing permanent changes in sensorimotor and autonomic functions of the spinal cord (SC) ...

A Magnetic Microbead Occlusion Model to Induce Ocular Hypertension-Dependent Glaucoma in Mice

A磁条微珠缺血模型，以诱导小鼠高眼压，青光眼相关

The use of rodent models of glaucoma has been essential to understand the molecular mechanisms that underlie the pathophysiology of this multifactorial ...

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

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

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 ...

Regenerative Therapy by Suprachoroidal Cell Autograft in Dry Age-related Macular Degeneration: Preliminary In Vivo Report

Regenerative Therapy by Suprachoroidal Cell Autograft in Dry Age-related Macular Degeneration: Preliminary In Vivo Report

脉络细胞自体移植治疗老年性黄斑变性的再生疗法: 初步体内报告

This study is aimed at examining whether a suprachoroidal graft of autologous cells can improve best corrected visual acuity (BCVA) and responses to ...

A Pleural Effusion Model in Rats by Intratracheal Instillation of Polyacrylate/Nanosilica

聚丙烯酸/纳米水的气管内灌注积液模型

Pleural effusion is a prevalent clinical finding of many pulmonary diseases. Having a useful animal pleural effusion model is very important to study ...

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

使用改性非缝合铐技术的宫颈主动脉移植模型

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. ...