Role of the Fas-signaling pathway in photoreceptor neuroprotection.
OBJECTIVE
To determine whether inhibiting the Fas proapoptosis pathway will result in increased photoreceptor survival after separation of the retina from the retinal pigment epithelium (RPE).


METHODS
Retina/RPE separation was induced in rat and mouse eyes by the subretinal injection of hyaluronic acid, 1%. Fas-pathway signaling was inhibited by the concomitant injection of a Fas receptor-neutralizing antibody, small inhibitory RNA against the Fas-receptor transcript (siFAS), or the use of the Fas-receptor defective mouse strain LPR. Indices of photoreceptor death included terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) staining, cell counts, and retinal thickness measurements. Retinas were immunostained with antibodies against rhodopsin and cone opsin to evaluate rod and cone photopigment production, respectively.


RESULTS
Inhibition of Fas signaling using Fas receptor-neutralizing antibody, siFas, or LPR mice resulted in a significant reduction in the number of TUNEL-positive photoreceptor cells as well as in a significant preservation of outer nuclear layer cell counts and thickness as compared with retina/RPE separation in eyes with intact Fas signaling. Fas-pathway inhibition resulted in preservation of both rhodopsin- and cone opsin-positive cells.


CONCLUSIONS
Inhibition of the Fas proapoptosis pathway results in significant photoreceptor preservation after retinal separation from the RPE.


CONCLUSIONS
Fas-pathway inhibition might serve as a novel mechanism for preserving photoreceptor cells during retinal disease.
Zacks
David N
DN
Retina Service, Kellog Eye Center, University of Michigan Medical School, Department of Ophthalmology and Visual Sciences, Ann Arbor, MI 48105, USA.
Boehlke
Christopher
C
Richards
Ayo-Lynn
AL
Zheng
Qiong-Duan
QD
D000818:Animals; D019138:Antibodies, Blocking / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D002452:Cell Count; D002470:Cell Survival / Q000502:physiology; D019084:Fluorescent Antibody Technique, Indirect; D015151:Immunoblotting; D020287:In Situ Nick-End Labeling; D051379:Mice; D008810:Mice, Inbred C57BL; D019463:Mice, Inbred MRL lpr; D020419:Photoreceptor Cells, Vertebrate / Q000502:physiology*; D012313:RNA / Q000302:isolation & purification; D034741:RNA, Small Interfering / Q000494:pharmacology; D051381:Rats; D011914:Rats, Inbred BN; D012163:Retinal Detachment / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012243:Rhodopsin / Q000378:metabolism; D017299:Rod Opsins / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D019014:fas Receptor / Q000037:antagonists & inhibitors / Q000502:physiology*
D019138:Antibodies, Blocking; C491899:Fas protein, mouse; C491898:Fas protein, rat; D034741:RNA, Small Interfering; D017299:Rod Opsins; D019014:fas Receptor; D012313:RNA; D012243:Rhodopsin