本文所载的工作已经得到了青光眼研究由美国健康援助基金会（XL），威廉＆玛丽格雷夫特别学术研究奖，以防止失明（XL），博士道格拉斯·约翰逊奖伊利诺伊防盲协会（HC）和美国国立卫生研究院：授予R01EY019034（XL）。

程序 C57BL/6J小鼠（杰克逊实验室的Bar Harbor，ME）提出在西北大学的动物护理设施。按照协议，由西北大学机构动物护理和使用委员会，符合使用动物在神经科学的研究由美国国立卫生研究院的指引批准用于所有动物。 1。激光光凝从先前公布的5-7协议的程序修改光凝。 <ol><li>麻醉40-60日龄鼠标，腹腔内注射氯胺酮（100毫克/公斤，管家施恩动物健康，OH）和甲苯噻嗪（10毫克/千克，劳埃德公司，雪兰爱荷华，IA）。 </li><li>实验动物的右眼的瞳孔扩张，局部治疗与一个或两个滴1％硫酸阿托品溶液（爱尔康实验室公司，德克萨斯州沃思堡）。 </li><li>瞳孔散大，后压平了内六角圆柱腔，以提高激光诱导6。插入玻璃微裂隙灯下前间隙（SL-3E，拓普康，奥克兰，新泽西州）锐利尖端（世界精密仪器公司，萨拉索塔，佛罗里达州），前房液排出。 </li><li>抑制鼠标在一个的塑料锥持有布伦特里科学公司（MA）和自制的平台上捆绑起来（ 见图 1a）。按住鼠标限制器和公开右眼裂隙灯背后的光源的鼠标。麻醉鼠标对准右眼裂隙灯下。 </li><li>用双手按住鼠标限制器，适用于激光照射到角膜缘使用氩激光（终极2000SE，相干，圣克拉拉，CA）。提供约80-100激光点（514纳米，100毫瓦，50毫秒的脉冲，和200μm的光斑）垂直的圆周周围的小梁。 C57BL / 6小鼠虹膜色素作为任意的p的一个障碍otential杂散能量7。 </li><li>灌输外用0.5％莫西沙星（爱尔康实验室公司，沃斯堡，德克萨斯州）眼球表面的消毒激光治疗的面积和0.5％的丙美卡因（博士伦，纽约州罗切斯特市），以减轻疼痛。 </li><li>动物保持一个加热垫（新光产品公司，佛罗里达州博卡拉顿）恢复了大约一个小时，直到它完全清醒。 </li><li>左眼是未经处理的作为对照。 </li></ol> 2。眼压测量<ol><li>将清醒的鼠标成筒装入塑料锥持有人，然后克制它的平台上（ 见图 2a）。 </li><li>允许五到十分钟，让鼠标适应持有人的位置。接近反弹眼压（TonoLab，殖民地医疗用品，新罕布什尔州Franconia）老鼠的眼睛，直到探头尖端是2-3毫米，从表面角膜14。 </li><li>按下测量键，让探头尖击中中心表面轻轻角膜。连续三组六相同的眼眼压测量采集和眼的眼压的平均值。未处理的对照眼总是首先测量测量激光治疗眼睛得到一个基准值。 </li></ol> 3。 Optomotor测试视力及对比敏感度测试14,15。个别老鼠的两只眼睛进行检查，分别由反转的漂流的光栅方向， 即沿顺时针漂流光栅是用来识别的视觉功能的左眼和用于右眼16沿反时针的漂移光栅。每个测试大约需要15分钟，由两个独立观察家重复。 <ol><li>将鼠标，让鼠标上自由移动的高架平台，四周由四个电脑显示器（ 图3A-B）。 </li><li>设置了监视器，让他们显示水平漂流正弦的光栅作为视觉刺激，平均亮度为39 cd /米2。光栅的移动方向上应交替 之间连续的顺时针和逆时针。 </li><li>分析动物的运动。漂流光栅音乐会动物的动作被认为是“积极的”视觉刺激后，在15秒内，然后逐渐增加。最高的响应被定义为引发视觉刺激动物的视力17。 </li><li>检查对比敏感度在预先选定的三个空间频率：0.075，0.16，和0.3个周期，每度（CPD）。为每只眼睛的对比度阈值被定义为，在预先固定的频率引出的视觉反应的最低对比度。对比敏感度的阈值17的倒数。 </li></ol>

<ol>
	<li>Gupta, N., Yucel, Y. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glaucoma+as+a+neurodegenerative+disease.">Glaucoma as a neurodegenerative disease.</a> Curr. Opin. Ophthalmol. 18, 110-114 (2007).</li><li>Quigley, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuronal+death+in+glaucoma.">Neuronal death in glaucoma.</a> Prog. Retin. Eye Res. 18, 39-57 (1999).</li><li>McKinnon, S. J., Schlamp, C. L., Nickells, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+retinal+ganglion+cell+death+and+glaucoma.">Mouse models of retinal ganglion cell death and glaucoma.</a> Experimental Eye Research. 88, 816-824 (2009).</li><li>Pang, I. H., Clark, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rodent+models+for+glaucoma+retinopathy+and+optic+neuropathy.">Rodent models for glaucoma retinopathy and optic neuropathy.</a> J. Glaucoma. 16, 483-505 (2007).</li><li>Levkovitch-Verbin, H., 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=Translimbal+laser+photocoagulation+to+the+trabecular+meshwork+as+a+model+of+glaucoma+in+rats.">Translimbal laser photocoagulation to the trabecular meshwork as a model of glaucoma in rats.</a> Investigative Ophthalmology &amp; Visual Science. 43, 402-410 (2002).</li><li>Aihara, M., Lindsey, J. D., Weinreb, R. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+mouse+ocular+hypertension:+establishment+of+the+model.">Experimental mouse ocular hypertension: establishment of the model.</a> Investigative Ophthalmology &amp; Visual Science. 44, 4314-4320 (2003).</li><li>Grozdanic, S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser-induced+mouse+model+of+chronic+ocular+hypertension.">Laser-induced mouse model of chronic ocular hypertension.</a> Investigative ophthalmology &amp; visual science. 44, 4337-4346 (2003).</li><li>Sappington, R. M., Carlson, B. J., Crish, S. D., Calkins, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+microbead+occlusion+model:+a+paradigm+for+induced+ocular+hypertension+in+rats+and+mice.">The microbead occlusion model: a paradigm for induced ocular hypertension in rats and mice.</a> Investigative ophthalmology &amp; visual science. 51, 207-216 (2010).</li><li>Ding, C., Wang, P., Tian, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+general+anesthetics+on+IOP+in+elevated+IOP+mouse+model.">Effect of general anesthetics on IOP in elevated IOP mouse model.</a> Experimental Eye Research. 92, 512-520 (2011).</li><li>Kalesnykas, G., 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=Retinal+ganglion+cell+morphology+after+optic+nerve+crush+and+experimental+glaucoma.">Retinal ganglion cell morphology after optic nerve crush and experimental glaucoma.</a> Investigative Ophthalmology &amp; Visual Science. 53, 3847-3857 (2012).</li><li>Shareef, S. R., Garcia-Valenzuela, E., Salierno, A., Walsh, J., Sharma, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chronic+ocular+hypertension+following+episcleral+venous+occlusion+in+rats.">Chronic ocular hypertension following episcleral venous occlusion in rats.</a> Experimental Eye Research. 61, 379-382 (1995).</li><li>Chiu, K., Chang, R., So, K. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser-induced+chronic+ocular+hypertension+model+on+SD+rats.">Laser-induced chronic ocular hypertension model on SD rats.</a> J. Vis. Exp. (10), e549 (2007).</li><li>Fu, C. T., Sretavan, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser-induced+ocular+hypertension+in+albino+CD-1+mice.">Laser-induced ocular hypertension in albino CD-1 mice.</a> Investigative Ophthalmology &amp; Visual Science. 51, 980-990 (2010).</li><li>Rangarajan, K. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+visual+deficits+in+aging+DBA/2J+mice+by+two+behavioral+assays.">Detection of visual deficits in aging DBA/2J mice by two behavioral assays.</a> Curr. Eye Res. 36, 481-491 (2011).</li><li>Wang, L., 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=Direction-specific+disruption+of+subcortical+visual+behavior+and+receptive+fields+in+mice+lacking+the+beta2+subunit+of+nicotinic+acetylcholine+receptor.">Direction-specific disruption of subcortical visual behavior and receptive fields in mice lacking the beta2 subunit of nicotinic acetylcholine receptor.</a> J. Neurosci. 29, 12909-12918 (2009).</li><li>Douglas, R. 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=Independent+visual+threshold+measurements+in+the+two+eyes+of+freely+moving+rats+and+mice+using+a+virtual-reality+optokinetic+system.">Independent visual threshold measurements in the two eyes of freely moving rats and mice using a virtual-reality optokinetic system.</a> Visual Neuroscience. 22, 677-684 (2005).</li><li>Prusky, G. T., Alam, N. M., Beekman, S., Douglas, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+quantification+of+adult+and+developing+mouse+spatial+vision+using+a+virtual+optomotor+system.">Rapid quantification of adult and developing mouse spatial vision using a virtual optomotor system.</a> Investigative Ophthalmology &amp; Visual Science. 45, 4611-4616 (2004).</li><li>Morrison, J. C., 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=A+rat+model+of+chronic+pressure-induced+optic+nerve+damage.">A rat model of chronic pressure-induced optic nerve damage.</a> Experimental Eye Research. 64, 85-96 (1997).</li><li>Cone, F. E., 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+effects+of+anesthesia,+mouse+strain+and+age+on+intraocular+pressure+and+an+improved+murine+model+of+experimental+glaucoma.">The effects of anesthesia, mouse strain and age on intraocular pressure and an improved murine model of experimental glaucoma.</a> Experimental Eye Research. 99, 27-35 (2012).</li><li>Liu, X., 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=Brain-derived+neurotrophic+factor+and+TrkB+modulate+visual+experience-dependent+refinement+of+neuronal+pathways+in+retina.">Brain-derived neurotrophic factor and TrkB modulate visual experience-dependent refinement of neuronal pathways in retina.</a> J. Neurosci. 27, 7256-7267 (2007).</li><li>Liu, X., 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=Regulation+of+neonatal+development+of+retinal+ganglion+cell+dendrites+by+neurotrophin-3+overexpression.">Regulation of neonatal development of retinal ganglion cell dendrites by neurotrophin-3 overexpression.</a> The Journal of Comparative Neurology. 514, 449-458 (2009).</li><li>Sun, W., Li, N., He, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-scale+morphological+survey+of+mouse+retinal+ganglion+cells.">Large-scale morphological survey of mouse retinal ganglion cells.</a> The Journal of Comparative Neurology. 451, 115-126 (2002).</li><li>Feng, L., 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=Sustained+Ocular+Hypertension+Induces+Dendritic+Degeneration+of+Mouse+Retinal+Ganglion+Cells+that+Depends+on+Cell-type+and+Location.">Sustained Ocular Hypertension Induces Dendritic Degeneration of Mouse Retinal Ganglion Cells that Depends on Cell-type and Location.</a> Investigative Ophthalmology &amp; Visual Science. , (2013).</li></ol>

作者宣称，他们有没有竞争经济利益。 作者是美国西北大学的全职雇员。 作者没有收到拨款，在这篇文章中所使用的试剂和仪器的公司，生产提供。

我们报告以上，持续的高眼压可诱发小鼠眼睛的激光照射。盐水注射模型18和静脉烧灼模型的11两者都需要广泛的显微技能相比，激光照射是相对简单和容易执行的。通常情况下，我们就可以进行激光照射4-6小鼠在2-3小时。实现持续的高眼压的关键步骤是前房压扁之前，激光和激光照射的参数。排水便于目标激光的小梁区和减少伤害到附近的睫状体和血管6的前房中的流体?...

<table border="1">
 <tbody>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Reagent</td>
 </tr>
 <tr>
 <td>moxifloxacin</td>
 <td>Alcon Labs, Inc.</td>
 <td>NDC 0065-4013-03</td>
 <td>0.5 %, Rx only</td>
 </tr>
 <tr>
 <td>Proparacaine Hydrochloride</td>
 <td>Bausch &amp; Lomb</td>
 <td>NDC 24208-730-06</td>
 <td>0.5 %, Rx only</td>
 </tr>
 <tr>
 <td>Ophthalmic Solution USP</td>
 <td>Bausch &amp; Lomb</td>
 <td>NDC 24208-730-06</td>
 <td>.5 %, Rx only</td>
 </tr>
 <tr>
 <td>ketamine</td>
 <td>Butler Schein Animal Health</td>
 <td>NDC 11695-0550-1</td>
 <td>100 mg / kg </td>
 </tr>
 <tr>
 <td>xylazine</td>
 <td>LLOYD Inc. of Iowa</td>
 <td>NADA 139-236</td>
 <td>10 mg / kg </td>
 </tr>
 <tr>
 <td>atropine sulfate solution</td>
 <td>Alcon Labs, Inc.</td>
 <td>NDC 61314-303-02</td>
 <td>1 %, Rx only</td>
 </tr>
 <tr>
 <td></td>
 <td></td>
 <td></td>
 <td>Equipment </td>
 </tr>
 <tr>
 <td>Slit Lamp, TOPCON </td>
 <td>Visual Systems Inc</td>
 <td>SL-3E</td>
 <td>powered by PS-30A</td>
 </tr>
 <tr>
 <td>OptoMotry 1.8.0 virtual</td>
 <td>CerebralMechanics Inc.</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>opto-kinetic testing system</td>
 <td>CerebralMechanics Inc.</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Tonometer, TonoLab, for mice</td>
 <td>Colonial Medical Supply</td>
 <td></td>
 <td></td>
 </tr>
 <tr>
 <td>Heating pad</td>
 <td>Sunbeam Products Inc</td>
 <td>722-810</td>
 <td></td>
 </tr>
 <tr>
 <td>Argon laser </td>
 <td>Coherent Inc</td>
 <td>Ultima 2000SE</td>
 <td></td>
 </tr>
 <tr>
 <td>DECAPICONE Plastic cone holder </td>
 <td>Braintree Sci Inc.</td>
 <td>MDC-200</td>
 <td>for mouse</td>
 </tr>
 </tbody>
 </table>

a laser-induced mouse model of chronic ocular hypertension to characterize visual defects

经常伴有高眼压（IOP），青光眼，失明的主要原因之一。我们试图建立高眼压症的小鼠模型来模拟人类高眼压性青光眼。这里激光照射施加到角膜缘photocoagulate水外流，诱导闭角。使用激光治疗之前和之后的反弹式眼压计的眼压的变化进行监测。是用于测量相应的变化在视觉能力一个optomotor的行为测试。代表性的结果显示从一个鼠标开发激光照射后持续高眼压。下降的视力，对比敏感度观察在高眼压鼠标。总之，我们的研究引入了一个有价值的模型系统研究青光眼的小鼠神经元变性的分子机制。

中所描述的程序，目的是激光照明在角膜缘的区域photocoagulate水外流的小梁，诱导闭角型青光眼（ 图1）。大多数光刻眼睛表现出没有明显的物理伤害，色素脱落或感染，与以前的研究结果相一致6。当一小群​​小鼠（小于5％所有的光刻动物）展出，如的瘪眼球，严重的白内障，显着的色素支队，或出血严重损害的体征，我们立刻处死他们。约有30％的光刻眼睛轻微的角膜疤?...

Watch this Scientific Journal Video about A Laser-induced Mouse Model of Chronic Ocular Hypertension to Characterize Visual Defects at JoVE.com

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

慢性高眼压诱导使用激光光凝小梁在小鼠眼睛。激光治疗后的几个月内升高的眼压（IOP）。减少实验动物的视力和对比敏感度使用optomotor测试监测。

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 ...

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12.9K 次观看.  Northwestern University. 慢性高眼压诱导使用激光光凝小梁在小鼠眼睛。激光治疗后的几个月内升高的眼压（IOP）。减少实验动物的视力和对比敏感度使用optomotor测试监测。