The work contained in this paper has been supported by the Dr. Douglas H. Johnson Award for Glaucoma Research from the American Health Assistance Foundation (XL), the William &amp; Mary Greve Special Scholar Award from the Research to Prevent Blindness (XL), the Illinois Society for the Prevention of Blindness (HC) and NIH grant R01EY019034 (XL).

Procedures
C57BL/6J mice (Jackson Laboratory, Bar Harbor, ME) are raised at Northwestern University's Animal Care Facility. All animals are used in accordance with protocols approved by Northwestern University Institutional Animal Care and Use Committee and conformed to the guidelines on the Use of Animals in Neuroscience Research from the NIH.
1. Laser Photocoagulation
The procedure of laser photocoagulation is modified from previously publis...

<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>

The authors declare that they have no competing financial interests.
	The authors are full-time employees of Northwestern University.
	The authors received NO funding that was provided by companies which produce reagents and instruments used in this article.

We report above that sustained ocular hypertension can be induced by laser illumination in mouse eyes. Compared to the saline injection model 18 and the vein cautery model 11 both of which require extensive microsurgical skills, the laser illumination is relatively simple and easy to perform. Usually we can perform the laser illumination for 4-6 mice in 2-3 hr. The critical steps to achieve sustained IOP elevation are the anterior chamber flattening before laser and the parameters for laser illumina...

<table border="1">
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 <td>Ophthalmic Solution USP</td>
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 <td>NDC 24208-730-06</td>
 <td>.5 %, Rx only</td>
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 <td>ketamine</td>
 <td>Butler Schein Animal Health</td>
 <td>NDC 11695-0550-1</td>
 <td>100 mg / kg </td>
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 <td>xylazine</td>
 <td>LLOYD Inc. of Iowa</td>
 <td>NADA 139-236</td>
 <td>10 mg / kg </td>
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 <td>1 %, Rx only</td>
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 <td>Slit Lamp, TOPCON </td>
 <td>Visual Systems Inc</td>
 <td>SL-3E</td>
 <td>powered by PS-30A</td>
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 <td>Tonometer, TonoLab, for mice</td>
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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.

As described in the Procedures, laser illumination is aimed at the trabecular meshwork in the limbal region to photocoagulate the aqueous outflow, inducing angle closure (Figure 1). Most lasered eyes exhibited no significant physical damage, pigment detachment or infection, consistent with previous findings 6. When a small group of mice (less than 5% of all lasered animals) exhibited physical signs of severe damage such as deflated eye balls, severe cataract, significant pigment detachment, or...

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

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.

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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12.8K ビュー.  Northwestern University. 慢性高眼圧症は、マウスの目に小柱網のレーザー光凝固を使用して誘導される。眼圧（IOP）は、レーザー治療後数ヶ月上昇する。視力と実験動物のコントラスト感度の低下は微細運動テストを使用して監視している。