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Summary

Abstract

Introduction

Protocol

Representative Results

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Medicine

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

Published: November 20th, 2016

DOI:

10.3791/54504

1Department of Ophthalmology and Visual Sciences, University of Maryland-Baltimore School of Medicine, 2Anatomy and Neurobiology, 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.

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.

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the anterior portion of the optic nerve (ON)1. NAION is the most common cause of sudden optic nerve related vision loss in individuals over the age of 502. The mechanism is believed to be a compartment syndrome that results in intraneural edema, and causes compression of the capillaries supplying the axons within the optic nerve3.

Since the ON is actually a central nervous system (CNS) tract, the rodent NAION (rNAION) model can be used to study the mechanisms and responses to isolated CNS white matter strokes. The rNAION mo....

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This protocol was approved by the University of Maryland Institutional Animal Care and Utilization Committee (IACUC; Baltimore, MD, USA)

1. Experimental Set-up

  1. Make a custom designed contact lens from a clear optical grade circular 7 mm in diameter Plexiglas, of 3 mm thickness. Cut the circular lenses with a drill press. Use a standard drill bit to make the inner curve, and finally polish the outer and inner curves using a contact lens polisher of ultrafine grit (1000/3000).
  2. .......

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The contact lens enabled central retina visualization (Figure 1). The focal laser spot illuminates the optic disc at the back of the retina (Figure 2). The normal un-induced retina is shown imaged by slit lamp bio-microscope (Figure 3A) and by SD-OCT (Figure 3B and 3C). During laser induction, when no dye is present in the circulation, laser light does not result in vessel and disk fluorescence (Figure 4A.......

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While there are a number of models of optic nerve damage (optic nerve crush12, optic nerve transection13, and PION14), the rNAION model is humane, adaptable to both rats and mice. It more closely resembles the human clinical condition of NAION. This condition includes progressive anterior optic nerve edema, an anterior optic nerve compartment syndrome, focal axonal ischemia, isolated retinal ganglion cell axonal damage and loss over a prolonged time course. The current report gives the ap.......

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We thank the many students and fellows who have worked on this model to improve its effectiveness, and to understand its mechanisms. Special thanks are due to Dr's. Mary Johnson (University of Maryland-Baltimore), Nitza Goldenberg-Cohen (Schneiderman Childrens Hospital, Petah-Tikva, Israel), Charles Zhang (Einstein Medical College, Bronx, NY), and Valerie Touitou (Hopital Salpetrie, Paris, France). This study was funded in part by RO1 EY015304 to SLB.

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Name Company Catalog Number Comments
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

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