The influence of visible light exposure on cultured RGC-5 cells.
OBJECTIVE
To determine the effects of visible light on normal or metabolically compromised cultured rat RGC-5 cells.


METHODS
Cultured RGC-5 cells were exposed to different durations as well as intensities of optical radiation, filtered to exclude wavelengths below 400 nm. Some cells were also subjected to metabolic compromise by depriving them of serum (serum deprivation; SD). Treated cells were assayed for cell viability using the 3,(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) reduction assay, for DNA breakdown by terminal deoxynucleotidyl transferase (TdT)-mediated d-UTP-linked nick end labeling (TUNEL), apoptotic protein activation by immunoblotting, and the production of reactive oxygen species (ROS) with dihydroethidium. A subset of cells was treated with 100 pM rotenone as an alternative means to induce metabolic stress; this was to determine that the influence of light on compromised cells was not specific to serum-deprivation alone.


RESULTS
Exposure to the light for 48 h activated both caspase-3 and Bcl-associated X-protein (Bax) in cultured RGC-5 cells. Furthermore, light (1000 or 4000 lux), SD, and rotenone caused minor but significant decreases in cellular MTT reduction. SD and light also led to cellular DNA breakdown, although only light caused ROS production. Light (48 h) significantly exacerbated the effect of SD on MTT reduction and DNA cleavage. Furthermore, the antioxidant, trolox, significantly blunted the detrimental influence of light on cell viability, increase in TUNEL-positive cells, and the generation of ROS.


CONCLUSIONS
Exposure to light was slightly, but significantly, harmful to healthy RGC-5 cells alone, but was much more toxic to those cells that were energetically compromised. Continuous light exposure can therefore detrimentally affect metabolically stressed RGC-5 cells. This may have implications for some ocular retinopathies such as glaucoma.
Wood
John P M
JP
Nuffield Laboratory of Ophthalmology, University of Oxford, Oxford, United Kingdom.
Lascaratos
Gerassimos
G
Bron
Anthony J
AJ
Osborne
Neville N
NN
D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects / Q000528:radiation effects; D002478:Cells, Cultured; D002839:Chromans / Q000494:pharmacology; D004249:DNA Damage; D004996:Ethidium / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D020287:In Situ Nick-End Labeling; D008027:Light; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism; D012165:Retinal Ganglion Cells / Q000166:cytology / Q000187:drug effects / Q000528:radiation effects*; D012402:Rotenone / Q000494:pharmacology; D044967:Serum; D013194:Staining and Labeling
D051017:Apoptosis Regulatory Proteins; D002839:Chromans; D017382:Reactive Oxygen Species; D012402:Rotenone; C067883:dihydroethidium; D004996:Ethidium; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid