Disruption of cell-cell junctions and induction of pathological cytokines in the retinal pigment epithelium of light-exposed mice

Toshio Narimatsu, Yoko Ozawa, Seiji Miyake, Shunsuke Kubota, Manabu Hirasawa, Norihiro Nagai, Shigeto Shimmura, Kazuo Tsubota

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

PURPOSE. To elucidate the influences of light exposure on the retinal pigment epithelium (RPE) in vivo that may be involved in the pathogenesis of AMD. METHODS. Six- to 7-week-old BALB/c mice were exposed to light at 2000 lux for 3 hours. Flatmount RPE samples were immunostained with anti-ZO-1 antibody for evaluating tight junction, anti-N-cadherin, and anti-b-catenin antibodies for adherens junction, and stained with phalloidin for actin cytoskeleton. The reactive oxygen species (ROS) level was measured using DCFH-DA; Rho-associated coiled-coil forming kinase (ROCK) activity was by ELISA. Cytokine expression was analyzed by real-time RT-PCR and/or ELISA in the RPE-choroid, and macrophage recruitment was by real-time RT-PCR and immunohistochemistry. Either an antioxidant, N-Acetyl-L-cysteine (NAC), or a ROCK inhibitor, Y-27632, were administered to analyze the roles of ROS and ROCK activation, respectively. RESULTS. Light exposure disrupted staining patterns of tight junctions, adherens junctions, and actin cytoskeleton in the RPE, where ROS was elevated. However, NAC treatment avoided the RPE changes, reducing ROS. ROCK activity increased after light exposure was suppressed by NAC, and the structural disruptions were suppressed by Y-27632. The levels of MCP-1, CCL11, and IL-6 increased after light exposure were suppressed by NAC. Light-induced MCP-1 and IL- 6 were suppressed by Y-27632. Macrophage recruitment after light exposure was also suppressed either by NAC or Y-27632. CONCLUSIONS. Light exposure induced ROS and Rho/ROCK activation, which caused disruption of cell-cell junctions (tight junctions and adherens junctions) and actin cytoskeleton, the RPE's barrier structure, and induced AMD-associated pathological changes in the RPE-choroid.

Original languageEnglish
Pages (from-to)4555-4562
Number of pages8
JournalInvestigative Ophthalmology and Visual Science
Volume54
Issue number7
DOIs
Publication statusPublished - 2013 Jul 15

Keywords

  • Age-related macular degeneration
  • Light damage
  • Oxidative damage
  • Retina
  • Retinal pigment epithelium

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

Fingerprint Dive into the research topics of 'Disruption of cell-cell junctions and induction of pathological cytokines in the retinal pigment epithelium of light-exposed mice'. Together they form a unique fingerprint.

  • Cite this