우리는이 모델에 작업 한 많은 학생들과 동료가 효율성을 개선하기 위해, 그 메커니즘을 이해 부탁드립니다. 특별 감사는 박사의에 기인한다. 메리 존슨 (메릴랜드 볼티모어 대학), Nitza 골덴 - 코헨 (Schneiderman 어린이 병원, Petah-티크, 이스라엘), 찰스 장 (아인슈타인 의과 대학, 브롱스, NY), 그리고 발레리 Touitou (원무 Salpetrie, 파리, 프랑스). 이 연구는 SLB에 RO1의 EY015304에 의해 부분적으로 투자되었다.

(, 볼티모어, MD, USA IACUC)이 프로토콜은 메릴랜드 기관 동물 관리 및 이용위원회의 대학에 의해 승인 1. 실험 셋업 <ol><li> 3mm 두께의 직경 플렉시 글라스의 명확한 광학 학년 원형 7mm에서 사용자 정의 설계 콘택트 렌즈를 확인합니다. 드릴 프레스와 원형 렌즈를 잘라. 내부 곡선을 만들기 위해 표준 드릴 비트를 사용하여, 최종적으로 초 미세 그릿 (3,000분의 1,000)의 콘택트 렌즈 용 광택제를 사용하여 외측 곡선 연마. </li><li> 사전에 pH 7.4의 인산염 완충 생리 식염수 (PBS)에 2.5 mM의 장미 벵골 (RB)를 준비, 필터는 최대 6 개월 동안 빛을 꽉 컨테이너에 0.45 미크론 주사기 필터 및 매장 -20 ° C에서 1 ml의 분취 량을 소독. 참고 : 같은 스프 라그 돌리 같은 흰둥이 이계 남성 동물의 사용은, 변형 따라 응답의 차이를 최소화 유도의 증가 용이성을 가능하게하고 발정 cycli 발생할 수있는 변동성을 감소NG 여성. </li><li> 설정 주파수는 네오디뮴 - 이트륨 알루미늄 가넷 532 nm의 레이저 광을 발생 (FD-YAG) 안과 의료 레이저를 두 배로. 표준 안과 레이저 어댑터를 사용하여 겐 - 스트 라이트 안과 슬릿 램프 레이저를 탑재합니다. 이 상업적으로 이용 가능한 장치는 동물의 눈과 레이저 스팟 응용 프로그램의 동시 시각화 할 수 있습니다. 의료 레이저는 적절한 레이저 유도로 같은 자리 크기를 사용하여 초점을 맞추고 자기 중심을위한 목표로 빔을 보유하고 있습니다. 다음과 같이 레이저 출력 매개 변수는 다음과 같습니다 <ol><li> 쥐 rNAION 유도의 경우 : 500 μm의 스폿 사이즈 / 50 mW의 레이저 파워 / 1,000 밀리 초 지속 시간 / 1,000 밀리 초 간격을 사용합니다. </li><li> 마우스 rNAION의 유도 : 작은 시신경 300 μm의 스폿 크기를 변경하고 래트 설정으로 다른 매개 변수는 동일하게 떠난다. </li><li> 일상적으로 레이저 출력을 보장하기 위해, 레이저 파워 미터를 사용한다. </li></ol></li></ol> 2. 실험 절차 <ol><li> 레이저 전원을 켭니다적절한 레이저 매개 변수를 설정합니다. 사용 전에 적어도 5 분 동안 레이저 따뜻한. </li><li> 케타민 / 자일 라진 및 RB 염료에 대한 적절한 투여 량을 결정하기 위해 동물을 단다. 80 ㎎ / ㎖ 케타민 4 ㎎ / ㎖의 자일 라진 1 ㎖ / kg의 혼합물을 복강 내 주사하여 동물을 마취. </li><li> 완전히 마취 될 때까지 가열 새장에 동물을 둡니다. 혐오 자극 (꼬리 또는 발가락 핀치)에 대한 응답을 확인합니다. 마취의 깊이 매 10 분 동안 동물을 확인합니다. </li><li> 1 % tropicamide과 동물의 동공을 팽창 및 0.5 % proparacaine와 눈 표면 마취. 이러한 긴 에반스와 같은 색소 동물을 사용하는 경우, 2.5 % neosynephrine 눈 방울 동공 팽창을 증가시킬 것이다. </li><li> 측면의 총구에 가까운 수염이보기를 막지 않도록 유도 할 잘라 가위를 사용합니다. </li><li> 1 % 메틸 셀룰로스 또는 맞춤형 콘택트 렌즈의 내부에 다른 안과 결합 방울 방울을 넣은 후 t에 렌즈를 적용쥐의 눈을 그가. </li><li> 슬릿 램프의 높이로 조정 플랫폼에서 동물을 놓습니다. 눈 슬릿 램프 및 레이저 광과 수직이되도록 45 ° 각도에서 동물의 머리를 설정한다. </li><li> 안과 세극등 통해 눈 시각화. 집중과 시각 시신경에 직접 중심으로 확인 조준 빔이 적당한 크기뿐만 아니라 확인하십시오. 맞춤 제작 어댑터 슬릿 램프의 눈 조각 중 하나에 장착 된 속도 - 고 ASA (2000 1200)와 디지털 카메라를 사용하여 망막 안저 사진. 주 : 맥락막 혈관을 볼 수있는 능력은 망막의 투명성을 보여준다. 이는 발생하면 해석을 혼동 할 수있는 이상, 망막 허혈을 검출 할 수 있기 위해서는 중요한 표시이다. rNAION 망막 손상에서 망막 허혈 결과가 망막 내층의 모든 셀에 영향을주는 반면, 절연 RGC 손실 결과 시신경 허혈이다. </li><li> 선택적으로, 상기 이미지 망막과유엔에 의한 눈을 평가하기 위해 스펙트럼 도메인 빛 간섭 단층 촬영기 (SD 간섭 단층 8)를 사용하여 시신경. 망막 (도 3c)를 통해 표면 (도 3b)과 (7)의 단면 검사 도중에 스캔. 는 SD 간섭 단층 촬영은 레이저 유도에 사용 된 것과 같은 콘택트 렌즈를 사용한다. </li><li> 1 ㎖ / 정맥 꼬리 정맥을 통해 kg RB를 주입하고, 30 초 후 레이저 파워를 활성화 기다린다. 이 시간 지연은 순환에 걸쳐 고르게 분포되도록 RB 수있다. 우리는 하나의 초 / 펄스에서 50 mW의 레이저 펄스를 사용합니다. 큰 에너지 (≥ 60 Mw)은 망막을 손상 시키거나 망막 혈관 허혈의 원인이 될 수 있습니다. 주의 : 있는지 확인하십시오 모든 사람이 길잃은 레이저 광이 조사의 눈에 들어가는 것을 방지하기 위해 적절한 차단 파장의 레이저 안전 필터링 안경을 보유하고 있습니다. 참고 : 염료를 순환에서 신속하게 제거되기 때문에 레이저 행정부가 IV RB 주입 후 신속하게 제공해야합니다. 긴 레이저 유도 더 심한 시신경 허혈. 빠른 연속 12 초 펄스 - 일반적으로, 동물은 7이 제공됩니다. 순환 염료 레이저 광의 접촉은 시신경 혈관 께 세극등 (도 4b)를 통해 볼 수있는 다운 황금 광선을 제공한다. 이는 염료를 주입 전신 혈류에 분배 하였다 증명한다. 글로우 모든 (그림 4A)에서 희미한 또는 없음 경우, 염료를 정맥 주입되지 않았습니다. 동물이 재 주입하기 전에 적어도 이틀 동안 복구해야하기 때문에이 경우, 즉시 두 번째 주사를주지 않는다. </li><li> 즉시 유도 후, 콘택트 렌즈를 제거한다. 덱사메타손과 안과 트리플 항생제 (Neosporin / 폴리 믹신 / 바시 트라 신) 연고와 두 눈을 커버하고, 전체 복구까지 가까운 관찰에서 하나의 수용 장에 37 ° C 온난화 패드에 동물을 배치합니다. <ol><li> 승 증류수로 콘택트 렌즈를 청소ater에는 나중에 사용하기 위해 비 마모성 청소 천으로 물기를 닦아. </li><li> 유도 후 2 일 안저 컬러 사진과 SD 간섭 단층 분석 (8) 모두에 의해 시신경 부종을 평가합니다. 주 : 시신경 부종의 정도를 비교하는 시신경 허혈의 심각도의 추정치를 제공한다. 이일에서, 시신경 디스크 이익률은 흐리게하고, 망막 정맥은 약간 반대측 (해제 유도) 눈에 비해 넓혀된다. 이 4 주에서 전위도 (ERG) 및 유발 전위 (VEP 11) 시각적 플래시를 수행 전기 생리학 분석을위한 유도을 게시 할 수 있습니다. </li></ol></li></ol>

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저자는 공개 아무것도 없어.

시신경 손상 (시신경의 호감 (12), 시신경 절개 (13), 및 PION 14)의 모델 번호가 있지만, rNAION 모델은 쥐 모두에 적용 할 인간이다. 그것은 더 밀접하게 NAION의 인간의 임상 상태를 닮았다. 이 조건은 장시간의 과정을 통해 점진적 전방 시신경 부종, 전방 시신경 구획 증후군, 초점 축삭 허혈, 고립 된 망막 신경절 세포 축삭 손상과 손실을 포함한다. 현재보고는 rNAION 유도하기위?...

Nonarteritic 전방 허혈성 시신경 병증 (NAION)은 시신경 (ON) (1)의 전방 부의 국소 허혈성 병소이다. NAION는기구 intraneural 부종을 초래 구획 증후군으로 생각된다. (50)이 세 이상의 사람들이 갑자기 시신경 관련 실명의 가장 흔한 원인이고, 시신경에서 축삭을 공급하는 모세 혈관의 압축을 야기 3. 온 실제로는 중추 신경계 (CNS) 계통이므로, 설치류 NAION (rNAION) 모델 절연 CNS 백질 스트로크에 응답 메커니즘을 연구하는데 사용될 수있다. rNAION 모델은 따라서 흰색 문제에 뇌졸중 관련 손상과 관련된 많은 문제를 해부에 유용 할 수있다. 백질 행정에서 다른 신경 전략과 제제를 평가하는데 사용될 수있다. 모델의 가장 매력적인 기능 중 하나가 있다는고통, 비 침습적 인 방법. 레이저 파워는 허혈성 손상의 다양한 정도를 생산하도록 조정될 수있다. 또 다른 특징은 누진 모세관 부전 제조 모세관 내피 세포를 손상하는 레이저 수퍼 옥사이드 라디칼에 의존한다는 것이다. 그것은 NAION 발생기구 현저 유사한 것으로 생각된다 부전이 진보적 종이다. 조사는 직접 모세관 응고가 발생하지 않는 것을 도시하지만, 적어도 두 가지 메커니즘을 작동되었다 : 과산화물 유도 된 죽음과 모세관의 일부 박리 내피 세포 (4) 및 활성화 된 B 세포의 NFkB (핵 인자 카파 경쇄 - 인핸서 간질 (5)로 세포막에 걸쳐 유체 수송 증가와 함께), 내피 나머지에 염증 규제 관련. 시신경 허혈에서 간질 유체의 축적 결과로 인한 시신경 모세관 및 압축 폐쇄. 개략적 픽처에 도시도 2. rNAION 모델은 모두 랫트 및 마우스 종 -6,7-에서 사용될 수 있고, 시신경의 완료되지만, 무통 파괴 망막 등을 온화한 병소에서 심각성 수준에서 변화 될 수있다 망막 중심 동맥 폐쇄 (CRAO) 등.

<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> 콘택트 렌즈 사용 가능 중앙 망막 시각화 (그림 1). 초점 레이저 스폿은 망막의 뒤쪽에있는 광학 디스크 (그림 2)를 조명한다. 정상적인 비 - 유도 망막 슬릿 램프 생체 현미경 (도 3a)에 의해 SD 간섭 단층 (도 3b 및도 3c)에 의해 촬상 나타낸다. 어떠한 염료 순환에 존재하지 않을 때, 레이저 유도시, 레이저 광 용기 디스크...

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 전방 허혈성 시신경 병증의 설치류 모델 (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 조회수.  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.

비디오: 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.

망막 신경절 세포의 생존을 공부 시신경의 호감 상해 Murine 모델

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

연구

신경과학

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.

비주얼 결함의 특성을 만성 안구 고혈압의 레이저 유도 마우스 모델

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

의학

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.

광 화학적으로 유도 된 후방 허혈성 시신경 병증의 쥐 모델

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.

레이저에 의한 맥락막 신생 혈관에 대한 마우스 모델

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.

녹내장 유도 절차<em&gt; 생체</em&gt; 쥐 모델 및 전체 마운트 망막 준비

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.

안압 상승을 통해 망막 허혈 - 재관류 손상의 마우스 모델

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.

쥐에 있는 부분 시 신경 Transection: 망막 신경 절 세포의 2 차 변성을 평가 하기 위해 새로운 수술 방식으로 설립 모델

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.

설치류에 있는 허혈성 망막 질병을 위한 산소 유도한 망막병증 모형

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.

설치류의 외과적으로 유도된 녹내장에 대한 Limbal Vascular Plexus의 전체 원형 소작

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.

과도 양측 공통 경동맥 폐색에 의해 유도된 허혈성 망막 손상의 뮤린 모델

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