July 2019
Volume 60, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2019
Novel in and ex vivo methods to reveal abnormal RPE morphology and microglial cell migration following light-induced retinal damage
Author Affiliations & Notes
  • Wenfei Wu
    Ophthalmology, Emory University, Atlanta, Georgia, United States
    Ophthalmology, The First Affiliated Hospital of Medical School of Xi'an Jiaotong University, Xi'an, Shaanxi, China
  • Kevin Joseph Donaldson
    Neuroscience Institute, Georgia State University, Georgia, United States
    Ophthalmology, Emory University, Atlanta, Georgia, United States
  • Kristie Ling Liao
    Ophthalmology, Emory University, Atlanta, Georgia, United States
  • Isabelle Gefke
    Ophthalmology, Emory University, Atlanta, Georgia, United States
  • Jeffrey H Boatright
    Ophthalmology, Emory University, Atlanta, Georgia, United States
    Atlanta VAMC Center for Visual & Neurocognitive Rehabilitation, Atlanta, Georgia, United States
  • Hans E Grossniklaus
    Ophthalmology, Emory University, Atlanta, Georgia, United States
  • John M Nickerson
    Ophthalmology, Emory University, Atlanta, Georgia, United States
  • Footnotes
    Commercial Relationships   Wenfei Wu, None; Kevin Donaldson, None; Kristie Liao, None; Isabelle Gefke, None; Jeffrey Boatright, None; Hans Grossniklaus, None; John Nickerson, None
  • Footnotes
    Support  NIH R01EY028450, R01EY021592, P30EY006360, R01EY028859; Abraham and Phyllis Katz Foundation; VA RR&D I01RX002806 and I21RX001924; VA RR&D C9246C (Atlanta Veterans Administration Center of Excellence in Vision and Neurocognitive Rehabilitation); and an unrestricted grant to the Department of Ophthalmology at Emory University from Research to Prevent Blindness, Inc.
Investigative Ophthalmology & Visual Science July 2019, Vol.60, 2382. doi:https://doi.org/
  • Views
  • Share
  • Tools
    • Alerts
      ×
      This feature is available to authenticated users only.
      Sign In or Create an Account ×
    • Get Citation

      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)

      ×
  • Supplements
Abstract

Purpose : 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.

Methods : 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.

Results : 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.

Conclusions : 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

Sign in or purchase a subscription to access this content. ×

You must be signed into an individual account to use this feature.

×