Name: assessment of vascular regeneration in the cns using the mouse retina
Uploaded: 2014-06-23T18:45:00
Description: Watch this Scientific Journal Video about Assessment of Vascular Regeneration in the CNS Using the Mouse Retina at JoVE.com

PS持有加拿大研究主席在视网膜细胞生物学和爱尔康研究院新研究者奖。这项工作是由健康研究加拿大学院（221478）的资金支持，加拿大糖尿病协会（OG-3-11-3329-PS）的加拿大自然科学和工程研究理事会（418637）和基金会战斗失明加拿大。技术支持也由送货线路德RECHERCHE恩桑特德拉视觉魁北克提供。

伦理声明：所有动物实验坚持以研究协会在视觉与眼科在眼科用动物和视觉研究和动物保健的加拿大委员会（ARVO）语句建立的动物护理指引。 1。氧诱导视网膜病变（OIR） <ol><li>鼠标幼仔为P0出生记录日期。 </li><li>在进入O 2，记录动物的全部重量，以确保有足够的体重范围。注意：对于C57BL / 6小鼠在P17，体重应5和7.5 g的为最大NV 32系列 。为了?...

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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue-specific+selection+for+different+mtDNA+genotypes+in+heteroplasmic+mice.">Tissue-specific selection for different mtDNA genotypes in heteroplasmic mice.</a> Nat Genet. 16, 93-95 (1997).</li><li>Mattapallil, M. 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=The+Rd8+mutation+of+the+Crb1+gene+is+present+in+vendor+lines+of+C57BL/6N+mice+and+embryonic+stem+cells,+and+confounds+ocular+induced+mutant+phenotypes.">The Rd8 mutation of the Crb1 gene is present in vendor lines of C57BL/6N mice and embryonic stem cells, and confounds ocular induced mutant phenotypes.</a> Investigative ophthalmolog., &amp; visual science. 53, 2921-2927 (2012).</li></ol>

什么是刺激在缺血性神经组织中新血管的健康成长最有效的方法是什么？它是治疗有效的干预和加速自然发生的血管再生？在神经缺血性疾病，如缺血性视网膜病或中风，血管退化与减少神经元功能35-38相关联。因此，应对早期损伤，疾病的直接/早段期间复原区域微循环可能证明是有益的。在眼部背景下，实验范式是加速缺血性视网膜血管重建减少病理性新生血管形成21-25，34，?...

整个中枢神经系统发育，神经，免疫细胞和血管建立显着耦合网络，以确保有足够的组织灌注，并允许感官信息1-5传输。血管系统，导致组织氧合不足和代谢受到影响供给和故障日益被视为一个重要因素神经退行性疾病6的发病机制。血管漏失和神经血管单元的大脑内的劣化，例如，与血管性痴呆，脑7的白质血管病变和阿尔茨海默氏病与小动脉和小血管8的狭窄相关联。此外，受损的血管屏障功能被认为有助于多发性硬化症9和肌萎缩性侧索硬化症10。 直接的相关性，以在本协议中所述，致盲的视网膜模型的疾病，如糖 ​​尿病性视网膜病变11和早产儿12的视网膜病变，13的特征是早期血管退化的相位。对神经血管视网膜随后的缺血性应激触发过度和病理血管生成中的第二阶段中可能源于作为一种代偿性反应重新死刑犯氧和能量供应14-16。一个有吸引力的策略来克服缺血性应激是中央到疾病进展是恢复功能性血管网络特别是在神经视网膜（ 图2和3）的局部缺血区。挑起控制血管生成的响应可能会遇到如反直觉对于其中的抗血管生成治疗，如抗的VEGF被认为是适于处理的条件。然而，证据表明这种方法的有效性安装。例如，加强ischem“生理状”血管再生IC视网膜病变已经通过引进内皮前体细胞17，抑制缪勒细胞表达VEGF诱导的其他血管生成因子18，注射髓系祖细胞19，抑制NADPH氧化酶诱导的细胞凋亡20，增加饮食中ω-3多不饱和脂肪酸下调的优雅展现酸摄入21，具有色氨酸tRNA合成酶22的羧基末端片段和VEGF或FGF-2的保护胶质细胞23的直接给药治疗。此外，我们已经表明，调节神经古典的指导线索，如脑信号或导蛋白在缺血性视网膜病变加速视网膜内的良性血管的血管再生，从而减少病理性血管生成24，25。直接的临床意义，几个上述动物研究提供的证据表明，促进血管再生成期间视网膜病的早期缺血阶段可以显著减轻威胁视力的视网膜前新生血管形成19，23，24，26，有可能通过对缺血性负担的减轻。 制定刺激的功能性血管再生治疗策略仍然是血管生物学家一个显著的挑战。在这里，我们描述了一个实验系统，它采用的氧诱导视网膜病变（OIR）小鼠模型，探讨策略，视网膜内调节血管再生。 Smith 等人开发的。于1994年27，这种模式可作为人类增生性视网膜病变的代理，由露出P7鼠标幼仔至75％的O 2，直到P12和随后重新引进幼犬室内环境氧张力（ 图中1）。这一模式松散模仿一个场景：一个早产儿通风与O 2。鼠标幼仔高氧暴露引发视网膜毛细血管和微血管病变，并产生血管闭塞的可重复的区域（VO）通常在评估从O 2的出口在P12，虽然最大VO面积达到48小时（P9）后接触到O 2 28。在小鼠中，股骨头缺血性VO区自发地重新在一周的过程中下面再介绍了室内空气，并最终VO区域被完全重新血管化（ 图2）。重新引入到经受OIR小鼠的室内空气也引发视网膜前新生血管形成（NV）（最多为P17），它通常评估，以确定抗血管生成治疗模式的疗效。在其最纯粹的形式，OIR模型提供了一个高度可重复的，可量化的工具来评估氧 ​​诱导的血管退化，并确定破坏性预视网膜新生血管形成29-31的程度。 30,31被彻底说明。在这里，我们提供了一个简单的一步一步的过程由药物化合物，有意治疗，病毒载体进行调查生理血运重建的调节神经视网膜内或研究的候选基因在转基因或基因敲除小鼠的影响。

<table border="0" cellpadding="0" cellspacing="0" width="1224">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:639px;">C57Bl/6 mice ((Other strains may be used; angiogenic response varies from one strain to the other)</td>
			<td style="width:181px;"></td>
			<td style="width:175px;"></td>
			<td style="width:231px;"></td>
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		<tr height="38">
			<td height="38" style="height:38px;width:639px;">CD1 nursing mothers</td>
			<td style="width:181px;">Vendor of choice</td>
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			<td style="width:181px;">World Precision Instruments</td>
			<td style="width:175px;">14192</td>
			<td></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:639px;">Dissecting Scissors straight</td>
			<td style="width:181px;">World Precision Instruments</td>
			<td style="width:175px;">14393</td>
			<td></td>
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			<td height="21" style="height:21px;width:639px;">Vannas Eye Scissors</td>
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			<td style="width:175px;">72-8483</td>
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			<td height="42" style="height:42px;width:639px;">Iris Forceps, curved, serrated</td>
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			<td style="width:175px;">15915</td>
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			<td height="21" style="height:21px;width:639px;">Brushes 362R size 0</td>
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			<td height="42" style="height:42px;width:639px;">Dumont Forceps #3; straight</td>
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			<td style="width:175px;">500338</td>
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			<td height="21" style="height:21px;width:639px;">Rhodamine Griffonia (Bandeiraea) Simplicifolia Lectin I</td>
			<td style="width:181px;">Vector Laboratories, Inc</td>
			<td style="width:175px;">RL-1102</td>
			<td></td>
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			<td height="21" style="height:21px;width:639px;">Microscope slides</td>
			<td style="width:181px;">VWR</td>
			<td style="width:175px;">16004-368</td>
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			<td height="42" style="height:42px;width:639px;">Fluoromount G</td>
			<td style="width:181px;">Electron Microscopy Sciences</td>
			<td style="width:175px;">17984-25</td>
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			<td height="21" style="height:21px;width:639px;">Zeiss Axio Observer Z1 Inverted Phase and Fluorescence Microscope</td>
			<td style="width:181px;">Zeiss</td>
			<td style="width:175px;"></td>
			<td></td>
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			<td height="42" rowspan="2" style="height:42px;width:639px;">Leica MZ9.5 Stereomicroscope</td>
			<td rowspan="2" style="width:181px;">Leica</td>
			<td rowspan="2" style="width:175px;"></td>
			<td></td>
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</table>

assessment of vascular regeneration in the cns using the mouse retina

啮齿动物视网膜也许是最方便的哺乳动物系统中，中枢神经系统（CNS）内神经血管调查的相互作用。它被越来越多地认识到，一些神经变性疾病如阿尔茨海默氏症，多发性硬化症，与血管妥协肌萎缩性侧索硬化症本元素。此外，失明在儿科和工作年龄人口（早产儿视网膜病变和糖尿病性视网膜病变，分别）的最突出的原因是其特征在于，生理血管再生长的血管退化和故障。本技术文件的目的是提供一个详细的协议来研究中枢神经系统血管再生的视网膜。该方法可以用来阐明导致血管生长的故障缺血性损伤后的分子机制。此外，潜在的治疗方式，以加速和恢复健康的血管丛可以探讨。结果obtaineD使用所描述的方法可以提供的治疗途径，缺血性视网膜病变，如糖尿病或早产儿，可能有利于中枢神经系统的其他血管疾病。

的OIR模型被广泛用于研究氧诱导血管变性和局部缺血引起的病理性新生血管在视网膜中和一直在为眼部疾病27，29，30目前使用的抗血管生成治疗的发展。利用该模型得到的结果可以大致推断缺血性视网膜病变，如增殖性糖尿病视网膜病变和30早产儿视网膜病变。 在这里，我们提出一个替代使用这个模型来研究血管再生。我们将描述一个策略的例子来再生，这?...

Watch this Scientific Journal Video about Assessment of Vascular Regeneration in the CNS Using the Mouse Retina at JoVE.com

Assessment of Vascular Regeneration in the CNS Using the Mouse Retina

啮齿动物视网膜长期以来被认为是可访问的窗口到大脑。在此技术论文中，我们提供了一个协议，它采用的氧诱导视网膜病变研究，导致血管再生的失败缺血性损伤后的中枢神经系统内的机制的小鼠模型。所描述的系统也可以利用，以探讨策略，视网膜和中枢神经系统内促进功能性血管再生。

血管再生在中枢神经系统中使用的小鼠视网膜评估

The rodent retina is perhaps the most accessible mammalian system in which to investigate neurovascular interplay within the central nervous system (CNS). ...

Research

JoVE Journal

Neuroscience

Quantifying the Activity of cis-Regulatory Elements in the Mouse Retina by Explant Electroporation

Quantifying the Activity of cis-Regulatory Elements in the Mouse Retina by Explant Electroporation

量化的活动顺</em在小鼠视网膜>调控元件外植体电穿孔

Transcription factors within cellular gene networks control the spatiotemporal pattern and levels of expression of their target genes by binding to ...

科研

神经科学

In vivo Electroporation of Developing Mouse Retina

In vivo Electroporation of Developing Mouse Retina

在发展中国家小鼠视网膜体内电穿孔

The functional characterization of genes expressed during mammalian retinal development remains a significant challenge.  Gene targeting to generate ...

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

谷氨酸和GABA快速微离子导入：一个有用​​的工具，调查突触整合

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of ...

The Corneal Micropocket Assay: A Model of Angiogenesis in the Mouse Eye

角膜微囊含量：血管生成在小鼠眼模型

The mouse corneal micropocket assay is a robust and quantitative in vivo assay for evaluating angiogenesis. By using standardized slow-release pellets ...

Imaging Ca2+ Dynamics in Cone Photoreceptor Axon Terminals of the Mouse Retina

Imaging Ca2+ Dynamics in Cone Photoreceptor Axon Terminals of the Mouse Retina

成像钙<sup&gt; 2+</sup&gt;在小鼠视网膜锥光感受器轴突终末动态

Retinal cone photoreceptors (cones) serve daylight vision and are the basis of color discrimination. They are subject to degeneration, often leading to ...

Using the Electroretinogram to Assess Function in the Rodent Retina and the Protective Effects of Remote Limb Ischemic Preconditioning

使用电生理评估功能，在啮齿动物的视网膜和远程肢体缺血预适应的保护作用

The ERG is the sum of all retinal activity. The ERG is usually recorded from the cornea, which acts as an antenna that collects and sums signals from the ...

Mouse Microsurgery Infusion Technique for Targeted Substance Delivery into the CNS via the Internal Carotid Artery

Mouse Microsurgery Infusion Technique for Targeted Substance Delivery into the CNS via the Internal Carotid Artery

鼠标显微注水技术进行有针对性的物质传递到中枢神经系统<i&gt;通过</i&gt;的颈内动脉

Animal models of central nervous system (CNS) diseases and, consequently, blood-brain barrier disruption diseases, require the delivery of exogenous ...

Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion

Investigating Mammalian Axon Regeneration: In Vivo Electroporation of Adult Mouse Dorsal Root Ganglion

哺乳动物轴突再生: 成年小鼠背根神经节体内电穿孔的研究

Electroporation is an essential non-viral gene transfection approach to introduce DNA plasmids or small RNA molecules into cells. A sensory neuron in the ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

间刺激间隔对啮齿类动物时间处理评估的力量

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in ...

Transpupillary Two-Photon In Vivo Imaging of the Mouse Retina

小鼠视网膜的经瞳双光子体内成像

The retina transforms light signals from the environment into electrical signals that are propagated to the brain. Diseases of the retina are prevalent ...