Biz bu model üzerinde çalışmış birçok öğrenci ve arkadaşlarının kendi etkinliğini artırmak için, ve mekanizmalarını anlamak için teşekkür ederiz. Özel teşekkür Dr adlı kaynaklanmaktadır. Mary Johnson (Maryland Baltimore Üniversitesi), Nitza Goldenberg-Cohen (Schneiderman Çocuk Hastanesi, Petah-Tikva, İsrail), Charles Zhang (Einstein Tıp Koleji, Bronx, NY) ve Valerie TOUITOU (Hopital Salpetrie, Paris, Fransa). Bu çalışma SLB için RO1 EY015304 kısmen finanse edildi.

(; Baltimore, MD, ABD IACUC) Bu protokol Maryland Kurumsal Hayvan Bakım ve Kullanım Komitesi Üniversitesi tarafından onaylandı 1. Deney Seti-up <ol><li> 3 mm kalınlığında çapı Pleksiglas net bir optik dereceli dairesel 7 mm den özel olarak tasarlanmış kontakt lens olun. Bir matkap basın ile yuvarlak lensler kesti. İç eğri yapmak için standart bir matkap ucu kullanın ve nihayet ultra-ince kum (1000/3000) bir kontakt lens parlatıcı kullanarak dış ve iç eğrileri lehçe. </li><li> Önceden pH 7.4 fosfat tamponlu salin (PBS) içinde 2.5 mM Rose Bengal (RB) hazırlanması, filtre 6 aya kadar bir ışık geçirmez bir kap içinde bir 0.45 u&#39;luk şırınga filtre ve mağaza -20 ° C&#39;de 1 ml&#39;lik numuneler ile sterilize edin. NOT: Sprague Dawley olarak albino outbred erkek hayvanların kullanımı suşu bağımlı yanıt farklılıkları en aza indiren indüksiyon artan kolaylığı sağlayan ve östrus halkalandırma yapılmasından oluşabilir değişkenliği azaltırng kadın. </li><li> kurmak frekans Neodimyum-Alümoitri granat bir 532 nm lazer ışığı üretir (Fd-YAG) oftalmik medikal lazer, iki katına çıktı. Standart bir göz lazer adaptörü kullanarak bir Haag-Streit oftalmik yarık lamba lazer monte edin. Bu piyasada mevcut cihaz hayvanın göz ve lazer nokta uygulamasının eşzamanlı görselleştirme sağlar. tıbbi lazer de uygun lazer indüksiyon aynı nokta boyutunu kullanarak odaklama ve sentrasyonu için bir hedef ışın vardır. aşağıdaki gibi lazer güç parametreleri şunlardır: <ol><li> Sıçan rNAION indüksiyon için: 500 mikron spot büyüklüğü / 50 mW lazer güç / 1000 milisaniye süre / 1000 milisaniye aralığı kullanınız. </li><li> Fare rNAION indüksiyon için: küçük optik disk 300 mikron spot boyutunu değiştirmek ve sıçan ayarı gibi diğer parametreleri aynı bırakın. </li><li> Rutin lazer güç çıkışını sağlamak için bir lazer güç metre kullanın. </li></ol></li></ol> 2. Deneysel Prosedür <ol><li> lazer gücü açınve uygun lazer parametresini ayarlayın. Kullanmadan önce en az beş dakika süreyle lazer ısıtın. </li><li> ketamin / iksilazin ve RB boya için uygun dozu belirleyecektir hayvan tartılır. 80 mg / ml ketamin ve 4 mg / ml ksilazin 1 ml / kg karışımın peritoneal içi enjekte hayvansal anestezisi. </li><li> Tamamen anestezi kadar ısıtılmış kafeste hayvan bırakın. caydırıcı uyaranlara (kuyruk veya ayak tutam) için herhangi bir yanıt olup olmadığını kontrol edin. anestezi derinliği her 10 dk için hayvanları kontrol edin. </li><li> % 1 tropikamid ile hayvanın öğrencileri dilate ve% 0.5 proparakain ile gözün yüzeyini uyutmak. Böyle Uzun Evans gibi pigmentli hayvanlar, kullanılıyorsa,% 2.5 neosynephrine göz damlaları öğrenci dilatasyon artacaktır. </li><li> tarafında namlu yakın bıyıkları görünümü engelleyen uyarılan edilecek kesmek için makas kullanın. </li><li> % 1 metilselüloz veya ısmarlama kontakt lens içine diğer oftalmik bağlantı damla bir damla koymak ve daha sonra t üzerine lensi uygulamakSıçan gözü o. </li><li> biyomikroskopta yüksekliği ayarlanabilir bir platformda hayvan yerleştirin. Göz yarık lamba ve lazer ışınına dik olacak şekilde 45 ° &#39;lik açıyla hayvanın kafasını ayarlayın. </li><li> oftalmik yarık lamba ile göz gözünüzde canlandırın. odaklı ve görüntülenmiştir optik sinire doğrudan merkezli olarak emin amaçlayan kiriş sağ boyutu da olduğundan emin olun. Bir özel yapılmış adaptör ile yarık lamba göz parçalardan biri üzerine monte sürat - yüksek ASA (2.000 1.200) ile bir dijital kamerayı kullanarak retina fundus fotoğraf. NOT: koroid damarları görme yeteneği retinanın şeffaflığını ortaya koymaktadır. Bu ortaya çıkarsa yorumunu karıştırabilir ki, daha sonra retina iskemi tespit edebilmek için önemli bir işaretidir. rNAION retina hasarı retinal iskemi sonuçlar iç retina tabakaların tüm hücreler etki ederken, izole edilmiş RGC kaybına neden optik sinir iskemi vardır. </li><li> İsteğe bağlı olarak, daha fazla görüntü retinaun kaynaklı göz değerlendirmek için bir spektral domain-OCT (SD-OCT 8) kullanılarak optik sinir. Retina (Şekil 3C) vasıtasıyla yüze (Şekil 3B) ve 7 kesit taramalar en tarar. SD-OCT görüntüleme lazer indüksiyonu için kullanılan aynı kontakt lens kullanır. </li><li> 1 ml / kuyruk damarından üzerinden kg RB enjekte ve 30 saniye için, daha sonra lazer gücü aktif bekleyin. Bu gecikme süresi dolaşımı boyunca eşit dağıtmak için RB sağlar. Biz bir sn / darbe bir 50 mW lazer darbesi kullanın. Büyükşehir enerjiler (≥ 60 mW) retinaya zarar veya retinal vasküler iskemi neden olabilir. Dikkat: emin olun herkes kaçak lazer ışığı araştırıcı göze giren önlemek için uygun engelleme dalga boyunda lazer güvenlik filtreleme gözlük bir çift vardır. NOT: boya dolaşımdan hızla elimine edilir çünkü lazer idaresi IV RB enjeksiyonundan sonra hızla verilmelidir. uzun lazer indüksiyon, daha şiddetli optik sinir iskemi. Bir peş peşe 12 sn darbeleri - Genellikle, hayvanlar 7 verilmiştir. Dolaşan boya ile lazer ışığının temas optik sinir gemilerin biyomikroskopta (Şekil 4B) aracılığıyla görülebilir güzel bir altın bir parlaklık kazandırır. Bu boya sistemik enjekte ve kan dolaşımına dağıtıldı kanıtlıyor. Kızdırma tüm (Şekil 4A) de soluk veya hiçbiri değilse, boya damar içine enjekte değildi. hayvan reenjeksiyon önce en az iki gün boyunca kurtarmak gerekir çünkü bu durumda, hemen ikinci bir enjeksiyon vermeyin. </li><li> Hemen indüksiyon sonrası, kontakt lens çıkarın. deksametazon ile oftalmik üçlü antibiyotik (Neosporin / polimiksin / basitrasin) merhem her iki göz kapağı ve tam iyileşme kadar yakın gözlem altında tek bir yer kafes içinde 37 ° C ısınma pad üzerinde hayvan yerleştirin. <ol><li> w distile ile kontakt lens temizlemebırakmayın ileride kullanmak üzere aşındırıcı olmayan bir temizlik bezi ile kurulayın ve. </li><li> Indüksiyon iki gün sonra fundus renkli fotoğraf ve SD-OCT analizi 8 hem optik sinir ödemi değerlendirmek. NOT: Optik sinir ödemi derecesini karşılaştırılması optik sinir iskemi şiddetinin bir tahmin verir. iki gün, optik sinir diski marjı bulanık ve retina damarları biraz kontralateral (un-kaynaklı) göz ile karşılaştırıldığında, dilate edilir. Iki ve dört hafta elektroretinografi (ERG) ve uyarılmış potansiyel (VEP 11) görsel flaş gerçekleştirin elektrofizyoloji analizi için indüksiyon sonrası. </li></ol></li></ol>

<ol>
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Yazarlar ifşa hiçbir şey yok.

Optik sinir hasarı (optik sinir ezilme 12, optik sinirin 13 ve PION 14) modelleri bir dizi varken, rNAION modeli, sıçan ve fareler hem de uyarlanabilir insancıl olduğunu. Daha yakın NAION insan klinik durumunu andırmaktadır. Bu durum uzun bir süre boyunca progresif ön optik sinir ödemi, anterior optik sinir kompartman sendromu, fokal akson iskemi, izole retina ganglion hücre akson hasarı ve kaybı içerir. Mevcut rapor, rNAION indüksiyon için uygun adımları sağlar indü...

Nonarteritik anterior iskemik optik nöropati (NAİON) optik sinir (ON) 1 ön kısmının fokal iskemik lezyonlardır. NAİON mekanizması intranöral ödem sonuçlanan bir kompartman sendromu olduğuna inanılmaktadır. 50 2 yaşın üzerindeki kişilerde ani optik sinir bağlı görme kaybının en sık nedenidir ve optik sinir içinde aksonlar temin kılcal damarların sıkışmasına neden olmaktadır 3.. AÇIK aslında bir merkezi sinir sistemi (MSS) yolu olduğundan, kemirgen NAİON (rNAION) modeli izole MSS beyaz cevher vuruş mekanizmaları ve yanıtları incelemek için kullanılabilir. rNAION modeli bu nedenle beyaz cevher inme ile ilgili hasar ile ilişkili pek çok sorun diseksiyon yararlı olabilir. Beyaz madde darbesi esnasmda farklı nöro-koruyucu strateji ve maddeleri değerlendirmek için de kullanılabilir. Modelin en çekici özelliklerinden biri olmasıdırağrısız, non-invaziv prosedür. lazer gücü iskemik hasar çeşitli seviyesi sağlamak için ayarlanabilir. Bir başka özelliği de ilerici bir kılcal disfonksiyonu üreten kılcal endotel hasar lazer kaynaklı süperoksit radikalleri dayanıyor olmasıdır. Bu NAION neden mekanizmasına oldukça benzer olduğuna inanılan bu disfonksiyon ve ilerici ödem olduğunu. Araştırma doğrudan kılcal pıhtılaşma neden olmadığını göstermiştir, ancak en az iki mekanizma ile çalışır etti: süperoksit bağlı ölüm ve kılcal bazı sıyırma endotelial hücreler 4, ve aktive edilmiş B hücreleri NFkB (Nükleer Faktör kappa hafif zincir-güçlendirici interstisyuma 5 içine hücre zarları arasında sıvı geçişini arttı ile), endotel kalan kadar inflamatuar düzenleme ilişkili. optik sinir başı iskemi interstisyel sıvı birikimi sonucu yol açtığı optik sinir kılcal damarların ve sıkıştırma kapatılması. Şematik resim gösterilirŞekil 2. RNAION model hem sıçan ve fare türleri 6,7 kullanılabilen ve optik sinirin tam ama ağrısız yıkım ve retina, örneğin hafif bir lezyondan, şiddeti seviyesinde değişebilir santral retinal arter tıkanıklığı (SRAT) olarak.

<table><tbody><tr><td>50 mW 532 nm laser</td><td>Iridex</td><td></td><td>Standard Ophthalmic Laser</td></tr><tr><td>0 - 100 mW 532 nm laser</td><td>Laserglow technologies</td><td></td><td>Substitute for iridex</td></tr><tr><td>Laser slit lamp adapter</td><td>Iridex</td><td></td><td>SMA coupled adapter for laser output</td></tr><tr><td>Coherent Fieldmate laser meter with thermopile sensor</td><td>Coherent</td><td></td><td>others also appropriate</td></tr><tr><td>Ophthalmic Examing Slit lamp biomicroscope</td><td>Various</td><td></td><td>Haag-Streit is the best; cheaper versions available on ebay</td></tr><tr><td>Rose Bengal</td><td>Sigma</td><td>330000-1G</td><td>Photoinducing agent</td></tr><tr><td>Fundus Contact lens or glass cover slip</td><td>custom/Cantor and Nissel (UK)</td><td></td><td>Custom designed planoconvex plastic lens for eye exam and induction</td></tr><tr><td>Tropicamide 1%</td><td></td><td></td><td></td></tr><tr><td>Tamiya polishing compound</td><td>Tamiya, INC</td><td>87068</td><td>polishing contact lens</td></tr><tr><td>2.5% Hypromellose (Goniovisc)/1% Methycellulose</td><td>HUB Pharmaceuticals</td><td></td><td>contact lens coupling agent</td></tr><tr><td>2.5% Neosynephrine Ophthalmic drops</td><td>Alcon labs</td><td></td><td>pupil dilating agent</td></tr><tr><td>Tropicamide 1%</td><td>Alcon labs</td><td></td><td>pupil dilating agent</td></tr><tr><td>0.5% Proparacaine</td><td>Alcon labs</td><td></td><td>topical Anesthetic</td></tr><tr><td>30 G fused needle insulin syringe</td><td>Various</td><td>Various</td><td>for intravenous injection of rose bengal</td></tr><tr><td>Ophthamic Antibiotic ointment with dexamethasone added (Triple antibiotic ointment)</td><td>Various</td><td>Various</td><td>Apply after induciton to minimize corneal scarring</td></tr><tr><td>Heidelberg Corporation Spectral domain-Optical Coherence Tomograph</td><td>Heidelberg Corporation</td><td></td><td>For Optical coherence measurements baseline and post-induction; not essential for induction</td></tr></tbody></table>

the rodent model of nonarteritic anterior ischemic optic neuropathy (rnaion)

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their lifetime. NAION results in optic nerve edema, selective loss of the retinal ganglion cell neurons (RGCs) and atrophy of the optic nerve. A rodent model of NAION that expresses most NAION features and sequelae has been developed, which is applicable to both rats and mice. This model utilizes a focal laser application of 532 nm wavelength to illuminate a photoactive dye, Rose Bengal (RB), to cause capillary damage and leakage at the targeted anterior optic nerve (the laminar region). After rNAION induction, there is an early optic nerve ischemia, optic nerve edema, and intraneural inflammation, followed by selective RGC and axonal loss. Since the optic nerve is a CNS white matter tract, the rNAION model is applicable to mechanistic studies of selective white matter ischemia, as well as neuroprotective analyses and short and long-term mechanisms of glial and neuronal response to ischemia.

<html> Kontakt lens özellikli santral retina görselleştirme (Şekil 1). Fokal lazer nokta retinanın arkasında optik disk (Şekil 2) yanar. Normal un kaynaklı retina yarık lamba biyo-mikroskop (Şekil 3A) tarafından ve SD-OCT (Şekil 3B ve 3C) tarafından görüntülendi gösterilmiştir. Hiçbir boya maddesinin dolaşımda mevcut olduğu zaman lazer İndüksiyon sırasında, lazer ışığı kabı ve disk floresan...

Watch this Scientific Journal Video about The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy rNAION at JoVE.com

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy rNAION

The following report describes how to replicate the rodent model of nonarteritic anterior ischemic optic neuropathy (rNAION), using the appropriate dye, contact lens and laser parameters. We also reveal the appropriate steps for evaluating the rNAION lesion in vivo.

Nonarteritik Anterior İskemik Optik Nöropati Kemirgen Modeli (rNAION)

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their ...

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Medicine

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy (rNAION)

8.9K Görüntüleme.  University of Maryland-Baltimore School of Medicine. The following report describes how to replicate the rodent model of nonarteritic anterior ischemic optic neuropathy (rNAION), using the appropriate dye, contact lens and laser parameters. We also reveal the appropriate steps for evaluating the rNAION lesion in vivo.

Video: The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy rNAION

An Optic Nerve Crush Injury Murine Model to Study Retinal Ganglion Cell Survival

Injury to the optic nerve can lead to axonal degeneration, followed by a gradual death of retinal ganglion cells (RGCs), which results in irreversible vision loss. Examples of such diseases in human include traumatic optic neuropathy and optic nerve degeneration in glaucoma. It is characterized by typical changes in the optic nerve head, progressive optic nerve degeneration, and loss of retinal ganglion cells, if uncontrolled, leading to vision loss and blindness.
The optic nerve crush (ONC) injury mouse model is an important experimental disease model for traumatic optic neuropathy, glaucoma, etc. In this model, the crush injury to the optic nerve leads to gradual retinal ganglion cells apoptosis. This disease model can be used to study the general processes and mechanisms of neuronal death and survival, which is essential for the development of therapeutic measures. In addition, pharmacological and molecular approaches can be used in this model to identify and test potential therapeutic reagents to treat different types of optic neuropathy.
Here, we provide a step by step demonstration of (I) Baseline retrograde labeling of retinal ganglion cells (RGCs) at day 1, (II) Optic nerve crush injury at day 4, (III) Harvest the retinae and analyze RGC survival at day 11, and (IV) Representative result.

This protocol shows how to retrogradely label retinal ganglion cells, and how to subsequently make an optic nerve crush injury in order to analyze retinal ganglion cell survival and apoptosis. It is an experimental disease model for different types of optic neuropathy, including glaucoma.

Retina Ganglion Hücre Survival Eğitim Bir Optik Sinir Crush Yaralanma murin Modeli

Injury to the optic nerve can lead to axonal degeneration, followed by a gradual death of retinal ganglion cells (RGCs), which results in irreversible ...

Nörobilim

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.

Görme Kusurları karakterize Kronik Oküler Hipertansiyon bir Lazer-kaynaklı Fare Modeli

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

Tıp

Rat Model of Photochemically-Induced Posterior Ischemic Optic Neuropathy

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history have&#160;remained poorly understood. Recently, we developed a reliable, reproducible animal model of PION and tested the treatment effect of some neurotrophic factors in this model1. The purpose of this video is to demonstrate our photochemically induced model of posterior ischemic optic neuropathy, and to evaluate its effects with retrograde labeling of retinal ganglion cells. Following surgical exposure of the posterior optic nerve, a photosensitizing dye, erythrosin B, is intravenously injected and a laser beam is focused onto the optic nerve surface. Photochemical interaction of erythrosin B and the laser during irradiation damages the vascular endothelium, prompting microvascular occlusion mediated by platelet thrombosis and edematous compression. The resulting ischemic injury yields a gradual but pronounced retinal ganglion cell dieback, owing to a loss of axonal input - a remote, injury-induced and clinically relevant outcome. Thus, this model provides a novel platform to study the pathophysiologic course of PION,&#160;and can be further optimized for testing therapeutic approaches for optic neuropathies as well as other CNS ischemic diseases.

The goal of this protocol is to photochemically induce ischemic injury to the posterior optic nerve in rat. This model is critical to studies of the pathophysiology of posterior ischemic optic neuropathy, and therapeutic approaches for this and other optic neuropathies, as well as of other CNS ischemic diseases.

Fotokimyasal Bağlı Posterior İskemik Optik Nöropati Rat Modeli

Posterior Ischemic optic neuropathy (PION) is a sight-devastating disease in clinical practice. However, its pathogenesis and natural history ...

A Mouse Model for Laser-induced Choroidal Neovascularization

The mouse laser-induced choroidal neovascularization (CNV) model has been a crucial mainstay model for neovascular age-related macular degeneration (AMD) research. By administering targeted laser injury to the RPE and Bruch&#8217;s membrane, the procedure induces angiogenesis, modeling the hallmark pathology observed in neovascular AMD. 
First developed in non-human primates, the laser-induced CNV model has come to be implemented into many other species, the most recent of which being the mouse. Mouse experiments are advantageously more cost-effective, experiments can be executed on a much faster timeline, and they allow the use of various transgenic models. The miniature size of the mouse eye, however, poses a particular challenge when performing the procedure. Manipulation of the eye to visualize the retina requires practice of fine dexterity skills as well as simultaneous hand-eye-foot coordination to operate the laser. However, once mastered, the model can be applied to study many aspects of neovascular AMD such as molecular mechanisms, the effect of genetic manipulations, and drug treatment effects. 
The laser-induced CNV model, though useful, is not a perfect model of the disease. The wild-type mouse eye is otherwise healthy, and the chorio-retinal environment does not mimic the pathologic changes in human AMD. Furthermore, injury-induced angiogenesis does not reflect the same pathways as angiogenesis occurring in an age-related and chronic disease state as in AMD.
Despite its shortcomings, the laser-induced CNV model is one of the best methods currently available to study the debilitating pathology of neovascular AMD. Its implementation has led to a deeper understanding of the pathogenesis of AMD, as well as contributing to the development of many of the AMD therapies currently available.

Here, we present the mouse laser-induced choroidal neovascularization (CNV) protocol, an experimental model that re-creates the vascular hallmarks of neovascular age-related macular degeneration (AMD). Once mastered, it can reliably and effectively induce CNV as a model system to test various experimental measures.

Lazer kaynaklı Koroid Neovaskülarizasyonlarında A Fare Modeli

The mouse laser-induced choroidal neovascularization (CNV) model has been a crucial mainstay model for neovascular age-related macular degeneration (AMD) ...

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.

Glokom uyaran bir de Prosedürü<em&gt; İn Vivo</em&gt; Rat Modeli ve Tüm montaj Retina Hazırlık

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

Retinal ischemia-reperfusion (I/R) is a pathophysiological process contributing to cellular damage in multiple ocular conditions, including glaucoma, diabetic retinopathy, and retinal vascular occlusions. Rodent models of I/R injury are providing significant insights into mechanisms and treatment strategies for human I/R injury, especially with regard to neurodegenerative damage in the retinal neurovascular unit. Presented here is a protocol for inducing retinal I/R injury in mice through elevation of intraocular pressure (IOP). In this protocol, the ocular anterior chamber is cannulated with a needle, through which flows the drip of an elevated saline reservoir. Using this drip to raise IOP above systolic arterial blood pressure, a practitioner temporarily halts inner retinal blood flow (ischemia). When circulation is reinstated (reperfusion) by removal of the cannula, severe cellular damage ensues, resulting ultimately in retinal neurodegeneration. Recent studies demonstrate inflammation, vascular permeability, and capillary degeneration as additional elements of this model. Compared to alternative retinal I/R methodologies, such as retinal arterial ligation, retinal I/R injury by elevated IOP offers advantages in its&#160;anatomical specificity, experimental tractability, and technical accessibility, presenting itself as a valuable tool for examining neuronal pathogenesis and therapy in the retinal neurovascular unit.

This article describes a procedure for inducing retinal ischemia-reperfusion injury by elevated intraocular pressure in mice. Retinal ischemia-reperfusion injury by elevated intraocular pressure serves to model human pathologies characterized by compromised oxygen and nutrient delivery in the retina, enabling researchers to examine potential cellular mechanisms and treatments for human diseases of the retinal neurovascular unit.

Göz İçi Basıncı Yükselmesi ile Retina İskemi-Reperfüzyon Hasarı Fare Modeli

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

Partial Optic Nerve Transection in Rats: A Model Established with a New Operative Approach to Assess Secondary Degeneration of Retinal Ganglion Cells

Previous studies have shown that the secondary degeneration of retinal ganglion cells (RGCs) occurs commonly in glaucoma. Partial optic nerve transection is considered a useful and reproducible model. Compared with other optic nerve injury models used commonly for assessing secondary degeneration, e.g. complete optic nerve transection and optic nerve crush models, the partial optic nerve transection model is superior as it distinguishes primary from secondary degeneration in situ. Therefore, it serves as an excellent tool for evaluating secondary degeneration. This study describes a novel operative approach of partial optic nerve transection by directly accessing the area of the retrobulbar optic nerve through the orbital lateral wall of the eyeball. Moreover, we present a newly designed, low cost surgical instrument to assist with transection. As demonstrated by the representative results in distinguishing the boundary of primary and secondary injury areas, the new approach and instrument ensures high efficiency and stability of the model by providing adequate space for surgical operation. This in turn makes it easy to separate the meningeal sheath and ophthalmic vessels from the optic nerve before transection. An additional benefit is that this space-saving operative approach improves the investigators&#39; ability to administer drugs, carriers, or selective RGC tracers to the stump of the partially transected optic nerve, allowing the exploration of mechanisms behind secondary injury in RGCs, in a new way.

Secondary degeneration of retinal ganglion cells (RGCs) occurs commonly in glaucoma. This study describes an innovative operative approach for partial optic nerve transection. The use of this space-saving operative approach extends the model's application range, and allows exploration of secondary injury mechanisms in RGCs in a new way.

Kısmi optik sinir transeksiyon Sıçanlarda: ikincil dejenerasyonu retina Ganglion hücrelerinin değerlendirmek için yeni bir operatif yaklaşım ile kurulan bir Model

Previous studies have shown that the secondary degeneration of retinal ganglion cells (RGCs) occurs commonly in glaucoma. Partial optic nerve transection ...

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.

Kemirgenlerde İskemik Retina Hastalıkları için Oksijen Kaynaklı Retinopati Modeli

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.

Kemirgenlerde Cerrahi Olarak İndüklenen Glokom için Limbal Vasküler Pleksusun Tam Daire Koterizasyonu

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

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.

Geçici Bilateral Ortak Karotid Arter Tıkanması sonucu İndüklenen İskemik Retina Yaralanmasının Murine Modeli

Diverse vascular diseases such as diabetic retinopathy, occlusion of retinal veins or arteries and ocular ischemic syndrome can lead to retinal ischemia. ...