title: RNA interference of GADD153 protects photoreceptors from endoplasmic reticulum stress-mediated apoptosis after retinal detachment.
abstract: BACKGROUND
  Apoptosis of photoreceptors plays a critical role in the vision loss caused by retinal detachment (RD). Pharmacologic inhibition of photoreceptor cell death may prevent RD. This study investigated the role of GADD153 that participates in endoplasmic reticulum (ER) stress-mediated apoptosis of photoreceptor cells after RD.
  
  
  METHODS
  Retinal detachment was created in Wistar rats by subretinal injection of hyaluronic acid. The rats were then randomly divided into four groups: normal control group, RD group, GADD153 RNAi group and vehicle group. RNA interference of GADD153 was performed using short hairpin RNA (shRNA). Expressions of GADD153 mRNA and protein were examined by RT-PCR and Western blotting analysis, respectively. GADD153 protein distribution in the retinal cells was observed using immunofluorescence confocal laser scanning microscopy. Apoptosis of retinal cells was determined by TdT-mediated fluorescein-16-dUTP nick-end labeling (TUNEL) assay.
  
  
  RESULTS
  Lentivirus GADD153 shRNA with the most effective silencing effect was chosen for in vivo animal study and was successfully delivered into the retinal tissues. GADD153 mRNA and protein expressions in GADD153 RNAi group were significantly lower than those in the RD group. Silencing of GADD153 by RNAi protected photoreceptors from ER stress-induced apoptosis.
  
  
  CONCLUSIONS
  ER stress-mediated pathway is involved in photoreceptor cell apoptosis after RD. GADD153 is a key regulatory molecule regulating ER-stress pathways and plays a crucial role in the apoptosis of photoreceptor cells after RD.
authors.affiliation: Department of Ophthalmology, Shanghai First People's Hospital, School of Medicine, Shanghai JiaoTong University, Eye Research Institute of Shanghai JiaoTong University, Shanghai, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Zhu
authors.forename: Jin
authors.initials: J
authors.lastname: Qian
authors.forename: Wenqiu
authors.initials: W
authors.lastname: Wang
authors.forename: Quan
authors.initials: Q
authors.lastname: Yan
authors.forename: Ying
authors.initials: Y
authors.lastname: Xu
authors.forename: Yuan
authors.initials: Y
authors.lastname: Jiang
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Fengqing
authors.initials: F
authors.lastname: Lu
authors.forename: Weiting
authors.initials: W
authors.lastname: Hu
authors.forename: Xi
authors.initials: X
authors.lastname: Zhang
authors.forename: Fenghua
authors.initials: F
authors.lastname: Wang
authors.forename: Xiaodong
authors.initials: X
authors.lastname: Sun
mesh_terms: D000818:Animals; D017209:Apoptosis; D059865:Endoplasmic Reticulum Stress / Q000235:genetics*; D015870:Gene Expression; D005822:Genetic Vectors; D006820:Hyaluronic Acid; D020287:In Situ Nick-End Labeling; D056965:Injections, Intraocular; D016086:Lentivirus; D008297:Male; D020419:Photoreceptor Cells, Vertebrate / Q000378:metabolism* / Q000473:pathology; D034622:RNA Interference; D012333:RNA, Messenger; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use*; D051381:Rats; D017208:Rats, Wistar; D012160:Retina / Q000378:metabolism* / Q000473:pathology; D012163:Retinal Detachment / Q000139:chemically induced / Q000235:genetics / Q000473:pathology / Q000517:prevention & control*; D015398:Signal Transduction; D051742:Transcription Factor CHOP / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism
chemical_list: C497251:Ddit3 protein, rat; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D051742:Transcription Factor CHOP; D006820:Hyaluronic Acid
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