Purchase this article with an account.
Wenfei Wu, Kevin Joseph Donaldson, Kristie Ling Liao, Isabelle Gefke, Jeffrey H Boatright, Hans E Grossniklaus, John M Nickerson; Novel in and ex vivo methods to reveal abnormal RPE morphology and microglial cell migration following light-induced retinal damage. Invest. Ophthalmol. Vis. Sci. 2019;60(9):2382. doi: https://doi.org/.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Light-induced retinal degeneration (LIRD) in rodents is a model of retinal degeneration, accompanied by abnormal RPE morphology and recruitment of IBA-positive microglia/macrophages into the subretinal space. However, this process has not been systematically described morphologically, especially in pigmented mice. We used novel in and ex vivo methods to reveal RPE dysmorphology and microglial cell migration following light-induced retinal damage.
We used efficient LED technology to establish a LIRD model in adult C57BL/6J mice. Funduscopy, SD-OCT and fundus autofluorescence (AF) scanning were performed at multiple time points (0–30days) after sufficient light exposure. RPE flatmounts were then prepared at each time point correspondingly, stained with ZO-1, alpha-catenin and IBA1. To verify condition of microglial cells in the RPE layer at the level of individual cells, in vivo cSLO and funduscopic images were then co-registered with post-mortem flatmounts. Additionally, we used transgenic mouse lines expressing RPE-specific fluorescent protein to track individual RPE cells and patterns dynamically following light-induced damage.
RPE dysmorphology and a substantial recruitment of activated, amoeboid microglial cells into the RPE layer were detected in the superior region of retina at multiple time points within 21 days after light damage, while the changes in the inferior were relatively moderate. This reaction is more voilent in younger mice than in older groups. However, abnormal RPE morphology and remodeling persisted for up to 30 days. Visible discoloration patterns in color funduscopy and cSLO images corresponded with unique, individual, microglial cells in RPE flatmounts.
Here, we highlight the success of utilizing a novel technique that allows for simple, effective and repeated visualization of RPE and microglial cells following light-induced damage in pigmented mice, which makes the LIRD model more closely aligned with human physiology and more efficient to offer clues to mechanisms that underpin retinal disease. Registration between AF images and RPE flatmounts revealed that in vivo imaging corresponded with our ex vivo method to measure incursion of microglial cells, indicating an effective approach to longitudinally monitoring migration of microglial cells and morphological changes of RPE cells caused by light damage.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
This PDF is available to Subscribers Only