title: Proteomic changes in the photoreceptor outer segment upon intense light exposure.
abstract: Acute light-induced photoreceptor degeneration has been studied in experimental animals as a model for photoreceptor cell loss in human retinal degenerative diseases. Light absorption by rhodopsin in rod photoreceptor outer segments (OS) induces oxidative stress and initiates apoptotic cell death. However, the molecular events that induce oxidative stress and initiate the apoptotic cascade remain poorly understood. To better understand the molecular mechanisms of light-induced photoreceptor cell death, we studied the proteomic changes in OS upon intense light exposure by using a proteolytic (18)O labeling method. Of 171 proteins identified, the relative abundance of 98 proteins in light-exposed and unexposed OS was determined. The quantities of 11 proteins were found to differ by more than 2-fold between light-exposed OS and those remaining in darkness. Among the 11 proteins, 8 were phototransduction proteins and 7 of these were altered such that the efficiency of phototransduction would be reduced or quenched during light exposure. In contrast, the amount of OS rhodopsin kinase was reduced by 2-fold after light exposure, suggesting attenuation in the mechanism of quenching phototransduction. Liquid chromatography multiple reaction monitoring (LC-MRM) was performed to confirm this reduction in the quantity of rhodopsin kinase. As revealed by immunofluorescence microscopy, this reduction of rhodopsin kinase is not a result of protein translocation from the outer to the inner segment. Collectively, our findings suggest that the absolute quantity of rhodopsin kinase in rod photoreceptors is reduced upon light stimulation and that this reduction may be a contributing factor to light-induced photoreceptor cell death. This report provides new insights into the proteomic changes in the OS upon intense light exposure and creates a foundation for understanding the mechanisms of light-induced photoreceptor cell death.
authors.affiliation: Case Center for Proteomics and Bioinformatics, Case Western Reserve University, Cleveland, Ohio 44106, USA.
authors.forename: Dagmar
authors.initials: D
authors.lastname: Hajkova
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Imanishi
authors.forename: Vikram
authors.initials: V
authors.lastname: Palamalai
authors.forename: K C Sekhar
authors.initials: KC
authors.lastname: Rao
authors.forename: Chao
authors.initials: C
authors.lastname: Yuan
authors.forename: Quanhu
authors.initials: Q
authors.lastname: Sheng
authors.forename: Haixu
authors.initials: H
authors.lastname: Tang
authors.forename: Rong
authors.initials: R
authors.lastname: Zeng
authors.forename: Ruth M
authors.initials: RM
authors.lastname: Darrow
authors.forename: Daniel T
authors.initials: DT
authors.lastname: Organisciak
authors.forename: Masaru
authors.initials: M
authors.lastname: Miyagi
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D008027:Light; D008297:Male; D018384:Oxidative Stress; D020419:Photoreceptor Cells, Vertebrate / Q000737:chemistry*; D040901:Proteomics; D051381:Rats; D017207:Rats, Sprague-Dawley