Den gnaver Model of Nonarteritic anterior iskæmisk opticusneuropati (rNAION)
Summary
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.
Abstract
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.
Introduction
Nonarteritic anterior iskæmisk optisk neuropati (NAION) er et knudepunkt iskæmisk læsion af den forreste del af synsnerven (ON) 1. NAION er den mest almindelige årsag til pludselig synsnerven relateret synstab hos personer over 50 år 2. Mekanismen antages at være en kompartment syndrom, der resulterer i intraneural ødem, og forårsager kompression af kapillærerne leverer axonerne i synsnerven 3.
Da ON er faktisk et centralnervesystem (CNS) tarmkanalen, kan gnaveren NAION (rNAION) model anvendes til at studere de mekanismer og reaktioner på isolerede CNS hvide substans slagtilfælde. Den rNAION model kan derfor være egnede i dissekere mange problemer forbundet med slagtilfælde relateret skade på hvid substans. Det kan bruges til at vurdere forskellige neuroprotektive strategier og agenter i hvid substans slagtilfælde.
Et af de mest attraktive træk ved modellen er, at det eren smertefri, noninvasive procedure. Lasereffekten kan justeres til at producere forskellige grad af iskæmisk skade. Et andet træk er, at den bygger på laser induceret superoxidradikaler at beskadige det kapillære endotel, der producerer en progressiv kapillær dysfunktion. Det er denne dysfunktion og progressiv ødem, der menes at være bemærkelsesværdigt lig den mekanisme, der forårsager NAION. Forskning har vist, at det ikke forårsager direkte kapillær koagulation, men virker gennem mindst to mekanismer: superoxid induceret død og fjernelse af noget af den kapillære endotelceller 4, og NFkB (nuklear faktor kappa-let kæde-enhanceren af aktiverede B-celler ) associeret inflammatorisk op regulering resterende endotel, med øget væske transport over cellemembraner i interstitium fem. Lukningen af synsnerven kapillærer og kompression forårsaget af interstitiel væske ophobning resultat i synsnerven hoved iskæmi. En skematisk billede er vist iFigur 2. Den rNAION modellen kan anvendes i både rotte- og muse arter 6,7, og kan varieres i niveauet af dens sværhedsgrad, fra en mild læsion til en fuldstændig, men smertefri ødelæggelse af synsnerven og nethinden, såsom som central retinal arterieokklusion (CRAO).
Protocol
Representative Results
Discussion
Mens der er en række modeller af synsnervebeskadigelse (synsnerven crush 12, synsnerven transektion 13, og PION 14), hvor rNAION model er human, kan tilpasses til både rotter og mus. Det ligner mere den humane kliniske tilstand NAION. Denne betingelse omfatter progressiv anterior synsnerve ødem, en forreste synsnerve kompartment syndrom, fokal axonal iskæmi, isoleret retinal ganglion celle axonal skade og tab over en længere tidsforløb. Den aktuelle rapport giver de relevante trin …
Disclosures
The authors have nothing to disclose.
Acknowledgements
Vi takker de mange studerende og stipendiater, der har arbejdet på denne model til at forbedre dens effektivitet, og for at forstå dens mekanismer. Særlige tak til dr s. Mary Johnson (University of Maryland-Baltimore), Nitza Goldenberg-Cohen (Schneiderman Childrens Hospital, Petah-Tikva, Israel), Charles Zhang (Einstein Medical College, Bronx, NY), og Valerie Touitou (Hopital Salpetrie, Paris, Frankrig). Denne undersøgelse blev finansieret delvist af RO1 EY015304 til SLB.
Materials
| 50mW 532nm laser | Iridex | Standard Ophthalmic Laser | |
| 0-100mW 532nm laser | Laserglow technologies | Substitute for iridex | |
| Laser slit lamp adapter | Iridex | SMA coupled adapter for laser output | |
| Cpherent Fieldmate laser meter with thermopile sensor | Coherent | others also appropriate | |
| Ophthalmic Examing Slit lamp biomicroscope | VArious | Haag-Streit is the best; cheaper versions available on ebay | |
| Rose Bengal | Sigma | 330000-1G | Photoinducing agent |
| Fundus Contact lens or glass cover slip | custom/Cantor and Nissel (UK) | Custom designed planoconvex plastic lens for eye exam and induction | |
| Tropicamide 1% | |||
| Tamiya polishing compound | Tamiya, INC | 87068 | polishing contact lens |
| 2.5% Hypromellose (Goniovisc)/1% Methycellulose | HUB Pharmaceuticals | contact lens coupling agent | |
| 2.5% Neosynephrine Ophthalmic drops | Alcon labs | pupil dilating agent | |
| Tropicamide 1% | Alcon labs | pupil dilating agent | |
| 0.5% Proparacaine | Alcon labs | topical Anesthetic | |
| 30ga fused needle insulin syringe | Various | Various | for intravenous injection of rose bengal |
| Ophthamic Antibiotic ointment with dexamethasone added (Triple antibiotic ointment) | Various | Various | Apply after induciton to minimize corneal scarring |
| Heidelberg Corporation Spectral domain-Optical Coherence Tomograph | Heidelberg Corportion | For Optical coherence measurements baseline and post-induction; not essential for induction |
References
- Banik, R. Nonarteritic Anterior Ischemic Optic Neuropathy: An Update on Demographics, Clinical Presentation, Pathophysiology, Animal Models, Prognosis, and Treatment. J. Clin. Experimental Ophthalmol. 10, 1-5 (2013).
- IONDT study group. Characteristics of patients with nonarteritic anterior ischemic optic neuropathy eligible for the Ischemic Optic Neuropathy Decompression Trial. Arch Ophthalmol. 114, 1366-1374 (1996).
- Tesser, R. A., Niendorf, E. R., Levin, L. A. The morphology of an infarct in nonarteritic anterior ischemic optic neuropathy. Ophthalmology. 110, 2031-2035 (2003).
- Bernstein, S. L., Johnson, M. A., Miller, N. R. Nonarteritic anterior ischemic optic neuropathy (NAION) and its experimental models. Prog Retin Eye Res. 30, 167-187 (2011).
- Nicholson, J. D., et al. PGJ2 Provides Prolonged CNS Stroke Protection by Reducing White Matter Edema. PLoS One. 7 (12), (2012).
- Goldenberg-Cohen, N., et al. Oligodendrocyte Dysfunction Following Induction of Experimental Anterior Optic Nerve Ischemia. Invest Ophthalmol Vis Sci. 46, 2716-2725 (2005).
- Bernstein, S. L., Guo, Y., Kelman, S. E., Flower, R. W., Johnson, M. A. Functional and cellular responses in a novel rodent model of anterior ischemic optic neuropathy. Invest Ophthalmol Vis Sci. 44, 4153-4162 (2003).
- Berger, A., et al. Spectral-Domain Optical Coherence Tomography of the Rodent Eye: Highlighting Layers of the Outer Retina Using Signal Averaging and Comparison with Histology. PLoS One. 9 (5), (2014).
- Huang, T. L., et al. Protective effects of systemic treatment with methylprednisolone in a rodent model of non-arteritic anterior ischemic optic neuropathy (rNAION). Exp Eye Res. 131, 69-76 (2015).
- Mantopoulos, D., et al. An Experimental Model of Optic Nerve Head Injury. Invest Ophthalmol Vis Sci. 57, 6222 (2014).
- You, Y., et al. Visual Evoked Potential Recording in a Rat Model of Experimental Optic Nerve Demyelination. J. Vis. Exp. (101), e52934 (2015).
- Templeton, J. P., Geisert, E. E. A practical approach to optic nerve crush in the mouse. Mol Vis. 18, 2147-2152 (2012).
- Magharious, M. M., D’Onofrio, P. M., Koeberle, P. D. Optic Nerve Transection: A Model of Adult Neuron Apoptosis in the Central Nervous System. J Vis Exp. (51), e2241 (2011).
- Wang, Y., et al. A Novel Rodent Model of Posterior Ischemic Optic Neuropathy. JAMA Ophthalmol. 131 (2), 194-204 (2013).
- Huang, T. L., Chang, C. H., Chang, S. W., Lin, K. H., Tsai, R. K. Efficacy of Intravitreal Injections of Antivascular Endothelial Growth Factor Agents in a Rat Model of Anterior Ischemic Optic Neuropathy. IOVS. 56, 2290-2296 (2015).
