Bystander effects elicited by single-cell photo-oxidative blue-light stimulation in retinal pigment epithelium cell networks

Abstract

'Bystander effect' refers to the induction of biological effects in cells not directly targeted. The retinal pigment epithelium consists of hexagonal cells, forming a monolayer interconnected by gap junctions (GJs). Oxidative stress initiated in an individual cell by photostimulation (488 nm) triggered changes in reactive oxygen species (ROS), Ca²⁺ and mitochondrial membrane potential (ψ_m). The Ca²⁺ signal was transmitted to neighboring cells slowly and non-uniformly; the ROS signal spread fast and radially. Increased Ca²⁺ levels were associated with a loss in ψ_m. GJ blockers prevented the spreading of the Ca²⁺, but not the ROS-related signal. The GJ-mediated Ca²⁺ wave was associated with cell death by 24 h, requiring endoplasmic reticulum-mitochondria Ca²⁺ transfer. Ensuing cell death was correlated with baseline Ca²⁺ levels, and baseline Ca²⁺ levels were correlated with pigmentation. Hence, local oxidative stress in a donor cell can trigger changes in certain connected recipient cells, a signal that required GJ communication and an ROS-Ca²⁺ dual-hit. Finally, damage apparently occurred in susceptible cells, which correlated with baseline Ca²⁺ levels.