Name: the rodent model of nonarteritic anterior ischemic optic neuropathy (rnaion)
Uploaded: 2016-11-20T13:00:00.000+00:00
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Description: Watch this Scientific Journal Video about The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy rNAION at JoVE.com

私たちは、その有効性を改善するために、そのメカニズムを理解するために、このモデルに働いている多くの学生や仲間に感謝します。特別な感謝は、博士のによるものです。メアリー・ジョンソン（メリーランド州バルチモアの大学）、Nitzaゴールデン・コーエン（シュナイダーマン小児病院、Petah-Tikva、イスラエル）、チャールズチャン（アインシュタイン医科大学、ブロンクス、NY）、およびヴァレリーTouitou（Hopital Salpetrie、パリ、フランス）。この研究は、SLBにRO1 EY015304によって部分的に資金を供給されました。

このプロトコルは、メリーランド大学の施設内動物管理及び利用委員会（ボルチモア、MD、USA IACUC）によって承認されました 1.実験の設定<ol><li> 3ミリメートルの厚さの直径プレキシグラスで明確な光学グレード円形7ミリメートル、からカスタム設計されたコンタクトレンズを作ります。ドリルプレスの円形レンズをカットします。内側の曲線を作るために、標準のドリルビットを使用し、最終的には超微細グリット（3000分の1000）のコンタクトレンズ研磨機を使用して、外側と内側のカーブを磨きます。 </li><li>事前にpH7.4のリン酸緩衝生理食塩水（PBS）に2.5 mMのローズベンガル（RB）を準備し、フィルタまでの6ヶ月間、光を通さない容器に-20℃で0.45ミクロンのシリンジフィルターと店舗1ミリリットルのアリコートで滅菌します。 注：このようなスプラーグドーリーなどのアルビノの非近交系雄の動物の使用は、ひずみ依存性応答の違いを最小限に誘導の増加使いやすさを可能にし、発情cycliで発生する可能性が変動性を減少させますngの女性。 </li><li>周波数を設定しネオジムイットリウム・アルミニウム・ガーネット（FD-YAG）532 nmのレーザー光を発生する眼科医療レーザーを、倍増しました。標準的な眼科用レーザーアダプタを使用して、ハーグ・シュトライト眼科スリットランプ上にレーザをマウントします。この市販の装置は、動物の眼とレーザスポットアプリケーションの同時可視化を可能にします。医療レーザーは、レーザー誘導と同じスポットサイズを使用して焦点を当て、適切および中心のための照準ビームを持っています。次のようにレーザパワーパラメータは次のとおりです。 <ol><li>ラットrNAIONの誘導のために：500μmのスポットサイズ/ 50mWのレーザパワー/千ミリ秒の持続時間/千ミリ秒間隔を使用します。 </li><li>マウスrNAION誘導のため：小さい視神経乳頭のために300μmのスポットサイズを変更し、ラットの設定と同じ他のパラメータを残します。 </li><li>日常的にレーザー出力を保証するために、レーザーパワーメーターを使用。 </li></ol></li></ol> 2.実験方法<ol><li>レーザーパワーをオンにしますそして、適切なレーザーパラメータを設定します。使用前に少なくとも5分間レーザーを温めます。 </li><li>ケタミン/キシラジンおよびRB色素に対する適切な用量を決定するために動物を秤量します。 80 mg / mlでケタミンおよび4 mg / mlとキシラジンを1ml / kgの混合物の腹腔内注射によって動物を麻酔。 </li><li>完全に麻酔になるまで加熱されたケージに動物を残します。嫌悪刺激（尾や足指のピンチ）への応答がないか確認してください。麻酔の深さごとに10分のために動物をチェックしてください。 </li><li> 1％トロピカミドで動物の瞳孔を拡張し、0.5％のプロパラカインと眼の表面を麻酔。このようなロング・エヴァンスなどの着色された動物を、使用している場合は、2.5％neosynephrine眼滴が瞳孔拡張が増加します。 </li><li>ビューをブロックしないようするように誘導される側の銃口に近いウィスカーをカットするはさみを使用してください。 </li><li> 1％メチルセルロースまたはカスタムメイドコンタクトレンズの内側に他の眼科用カップリングドロップのドロップを入れて、[トンの上にレンズを適用ラットの目を彼。 </li><li>スリットランプの高さに調節プラットフォーム上で動物を置きます。眼は、スリットランプ、レーザービームに対して垂直になるように45°の角度で、動物の頭を設定します。 </li><li>眼科スリットランプを通して目を可視化します。照準ビームは、右のサイズだけでなく、集中して可視化視神経に直接中心にあることを確認します。特注のアダプタを使用してスリットランプの目の作品の一つに搭載された高速 - 高ASA（2000 1200）でデジタルカメラを使用して眼底を撮影。 注：脈絡膜血管を見る能力が網膜の透明性を明らかにする。これが発生した場合の解釈を混乱することができ、後に網膜虚血を検出することができるようにするために重要な兆候です。 rNAIONは、網膜損傷における網膜虚血の結果は、内網膜層の細胞のすべてに影響を与えながら、孤立したRGCの損失をもたらす視神経虚血、です。 </li><li>必要に応じて、さらに画像網膜と非誘導の目を評価するために、スペクトルドメイン光コヒーレンス断層撮影（SD-10月8日）を使用して視神経。網膜（ 図3C）を介して、 面 （ 図3B）、ならびに7断面スキャンエンスキャンします。 SD-OCT画像化は、レーザー誘導のために使用される同一のコンタクトレンズを使用します。 </li><li> 1ml /尾静脈を介してキロRBを注入し、30秒間、次いで、レーザーパワーをアクティブに待機。この遅延時間は、循環全体に均等に分配するためにRBを可能にします。私たちは、1秒/パルスで50mWのレーザーパルスを使用しています。グレーターエネルギー（≥60 MW）は、網膜を損傷したり、網膜血管虚血を引き起こす可能性があります。 注意：誰もが研究者の目に入る迷光レーザ光を防止するために、適切な遮断波長のレーザー安全フィルタリング眼鏡を持っていることを確認します。 注：染料は循環から迅速に除去されるので、レーザーの管理は、IV RB注射後急速に与えられなければなりません。長いレーザー誘導、より深刻な視神経虚血。立て続けに12秒のパルス - 一般に、動物を7が与えられます。循環色素レーザ光の接触は、視神経血管にスリットランプ（ 図4B）を通して見ることができる美しい金色の輝きを与えます。これは、染料が全身に注入し、血流中に分布していたことを証明します。グローはかすかな、または全くなし（ 図4A）である場合、色素が静脈内に注射されませんでした。動物が再注入する前に、少なくとも2日間回復する必要がありますので、この場合には、すぐに二回目の注射を与えることはありません。 </li><li>すぐに誘導した後、コンタクトレンズを取り外してください。デキサメタゾンと眼科トリプル抗生物質（ネオスポリン/ポリミキシン/バシトラシン）軟膏で両目を覆い、完全に回復するまで近い観察下で、単一の収容されたケージ内の37℃の加温パッドの上に動物を配置します。 <ol><li>蒸留ワットでコンタクトレンズを清掃してくださいATERは、将来の使用のための非研磨洗浄布で水分を拭き取ります。 </li><li>誘導後の二日は、眼底のカラー写真とSD-OCT分析8の両方によって視神経浮腫を評価します。 注：視神経浮腫の程度を比較するとは、視神経虚血の重症度の推定を与えます。 2日目に、視神経ディスクマージンがぼやけて、網膜静脈はやや反対側（非誘導）目と比較して、拡張されています。電図（ERG）を実行し、2と4週目に視覚誘発電位（VEP 11）を点滅電気生理学解析のための誘導を投稿してください。 </li></ol></li></ol>

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視神経損傷（視神経挫滅12、視神経離断13、及びパイオン14）のモデルの数がありますが、rNAIONモデルは、ラットとマウスの両方に適応人道的です。これは、より密接NAIONのヒト臨床症状に似ています。この条件は、プログレッシブ前方視神経浮腫、前視神経コンパートメント症候群、焦点軸索虚血、長時間経過にわたって孤立した網膜神経節細胞の軸索損傷および損?...

Nonarteritic前部虚血性視神経症（NAION）が1（ON）視神経の前方部分の焦点虚血性病変です。 NAIONは、機構が神経内の浮腫につながるコンパートメント症候群であると考えられる。50 2歳以上の個人の突然の視神経関連失明の最も一般的な原因であり、視神経内の軸索を供給する毛細血管の圧迫を引き起こします3。 ONは、実際に、中枢神経系（CNS）管であることから、齧歯類NAION（rNAION）モデルは、単離されたCNS白質ストロークの機構および応答を研究するために使用することができます。 rNAIONモデルは、したがって、白質に脳卒中に関連する損傷に関連した多くの問題を解剖するのに有用であり得ます。白質行程で異なる神経保護戦略と薬剤を評価するために使用することができます。 モデルの中で最も魅力的な特徴の一つは、それがあるということです無痛、非侵襲的な手順。レーザー出力は、虚血性損傷の種々の程度を生成するように調整することができます。別の特徴は、進行性の毛細管機能不全を生成、毛細血管内皮を損傷するレーザー誘起スーパーオキシドラジカルに依存することです。それはNAIONを引き起こすメカニズムと非常に類似していると考えられているこの機能障害および進行性の浮腫です。研究は、それが直接の毛細管凝固を起こさないことを示したが、少なくとも二つのメカニズムを介して動作している：毛細血管内皮細胞の一部4のスーパーオキシド誘導死とストリッピング、およびNFkBの（核因子カッパ軽鎖-エンハンサー活性化B細胞の間質5に細胞膜を横切る流体輸送を増加して）、内皮の残りにまで炎症レギュレーションに関連。視神経乳頭虚血における間質液の蓄積結果に起因する視神経の毛細血管および圧縮の閉鎖。模式画像がに示されています図2。rNAIONモデルは両方ラットおよびマウス種6,7で使用することができ、軽度病変から視神経と網膜の完全な、しかし痛み破壊する、その重症度のレベルに変化させることができる、そのような網膜中心動脈閉塞（CRAO）など。

<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>コンタクトレンズ有効中心網膜の視覚化（ 図1）。焦点レーザスポットは、網膜の奥の視神経乳頭（ 図2）を照射します。通常の非誘導網膜は細隙灯生体顕微鏡（ 図3A）によって、およびSD-OCT（ 図3Bおよび3C）によって撮像示されています。全く色素が循環中に存在しない場合、レーザー誘導中、レーザ光は、容?...

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.

Nonarteritic前部虚血性視神経症の齧歯類モデル（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 ビュー.  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.

ビデオ: 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.

網膜神経節細胞の生存を研究するために視神経挫滅モデルマウス

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

神経科学

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.

ビジュアル欠陥を特徴づけるために慢性高眼圧症のレーザー誘起マウスモデル

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

医学

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.

光化学誘起後方虚血性視神経障害のラットモデル

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.

レーザ誘起脈絡膜血管新生のためのマウスモデル

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.

緑内障誘導の手順<em&gt;インビボ</em&gt;ラットモデルと全体のマウント網膜準備

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.

眼圧の上昇を通じて、網膜虚血再灌流障害のマウスモデル

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.

ラットの視神経の部分断裂: 網膜神経節細胞の二次変性を評価する新しい手術アプローチとの確立モデル

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.

げっ歯類における虚血性網膜症の酸素誘導網膜症モデル

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.

げっ歯類の外科的緑内障に対する辺縁血管神経叢の全周焼灼

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.

一過性両側性頸動脈閉塞による虚血性筋炎損傷のマウスモデル

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