Quantification of retinal transneuronal degeneration in human glaucoma: a novel Multiphoton-DAPI approach

Lei, Yuan, Garrahan, Nigel John, Hermann, Boris ORCID: https://orcid.org/0000-0001-8658-485X, Becker, David L., Hernandez, M. Rosario, Boulton, Michael E. and Morgan, James Edwards ORCID: https://orcid.org/0000-0002-8920-1065 2008. Quantification of retinal transneuronal degeneration in human glaucoma: a novel Multiphoton-DAPI approach. Investigative Ophthalmology and Visual Science 49 (5) , pp. 1940-1945. 10.1167/iovs.07-0735

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Abstract

purpose. Glaucoma is presumed to result in the selective loss of retinal ganglion cells. In many neural systems, this loss would initiate a cascade of transneuronal degeneration. The quantification of changes in neuronal populations in the middle layers of the retina can be difficult with conventional histologic techniques. A method was developed based on multiphoton imaging of 4′,6′-diamino-2-phenylindole (DAPI)–stained tissue to quantify neuron loss in postmortem human glaucomatous retinas. methods. Retinas from normal and glaucomatous eyes fixed in 4% paraformaldehyde were incubated at 4°C overnight in DAPI solution. DAPI-labeled neurons at different levels of the retina were imaged by multiphoton confocal microscopy. Algorithms were developed for the automated identification of neurons in the retinal ganglion cell layer (RGCL), inner nucleus layer (INL), and outer nuclear layer (ONL). results. In glaucomatous retinas, the mean density of RGCs within 4 mm eccentricity was reduced by approximately 45%, with the greatest RGC loss occurring in a region that corresponds to the central 6° to 14° of vision. Significant neuron loss in the INL and ONL was also seen at 2 to 4 mm and 2 to 3 mm eccentricities, respectively. The ratios of neuron densities in the INL and ONL relative to the RGCL (INL/RGC and ONL/RGC, respectively) were found to increase significantly at 3 to 4 mm eccentricity. conclusions. The data confirm that the greatest neuronal loss occurs in the RGCL in human glaucoma. Neuronal loss was also observed in the outer retinal layers (INL and ONL) that correlated spatially with changes in the RGCL. Further work is necessary to confirm whether these changes arise from transneuronal degeneration.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Optometry and Vision Sciences Medicine Systems Immunity Research Institute (SIURI)
Subjects:	R Medicine > RE Ophthalmology
Additional Information:	Presented at the annual meeting of the Association for Research in Vision and Ophthalmology, Fort Lauderdale, Florida, May 2007. Supported by Research into Ageing Grant 127. Confirmation received by publisher on 21 February 2014 that publisher's pdf can be self-archived 6 months after publication.
Publisher:	Association for Research in Vision and Ophthalmology
ISSN:	0146-0404
Date of First Compliant Deposit:	30 March 2016
Last Modified:	07 May 2023 17:28
URI:	https://orca.cardiff.ac.uk/id/eprint/19712

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