title: Expression of netrin-1 receptors in retina of oxygen-induced retinopathy in mice.
abstract: BACKGROUND
  Netrin-1 has been reported to promote retinal neovascularization in oxygen-induced retinopathy (OIR). However, netrin-1 receptors, which may mediate netrin-1 action during retinal neovascularization, have not been characterized. In this study, we investigated netrin-1 receptor subtype expression and associated changes in the retinas of mice with OIR.
  
  
  METHODS
  C57BL/6J mice were exposed to 75±2% oxygen for 5 days and then returned to normal air to induce retinal neovascularization. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot were used to examine the expression of netrin-1 receptor subtypes in the mouse retinas. Double staining of netrin-1 receptor subtypes and isolectin B4 was used to determine the location of the netrin-1 receptor subtypes in the retinas. Inhibition of retinal neovascularization was achieved by UNC5B shRNA plasmid intravitreal injection. Retinal neovascularization was examined by fluorescein angiography and quantification of preretinal neovascular nuclei in retinal sections.
  
  
  RESULTS
  RT-PCR results showed that, except for UNC5A, netrin-1 receptor subtypes UNC5B, UNC5C, UNC5D, DCC, neogenin, and A2b were all expressed in the retinas of OIR mice 17 days after birth. Western blots showed that only UNC5B expression was significantly increased on that day, and immunofluorescence results showed that only UNC5B and neogenin were expressed in retinal vessels. Treatment of OIR mice with the UNC5B shRNA plasmid dramatically reduced neovascular tufts and neovascular outgrowth into the inner limiting membrane.
  
  
  CONCLUSIONS
  UNC5B may promote retinal neovascularization in OIR mice.
authors.forename: Dan
authors.initials: D
authors.lastname: Liu
authors.forename: Si-Qi
authors.initials: SQ
authors.lastname: Xiong
authors.forename: Lei
authors.initials: L
authors.lastname: Shang
authors.forename: Xiao-feng
authors.initials: XF
authors.lastname: Tian
authors.forename: Jing
authors.initials: J
authors.lastname: Yang
authors.affiliation: Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, Hunan 410008, China. xiaxiaobo2013@163.com.
authors.forename: Xiao-Bo
authors.initials: XB
authors.lastname: Xia
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D004195:Disease Models, Animal; D005260:Female; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D008810:Mice, Inbred C57BL; D000075386:Netrin Receptors; D010100:Oxygen / Q000633:toxicity; D012333:RNA, Messenger / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D011956:Receptors, Cell Surface / Q000096:biosynthesis / Q000235:genetics*; D012164:Retinal Diseases / Q000139:chemically induced / Q000235:genetics* / Q000378:metabolism
chemical_list: D000075386:Netrin Receptors; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; C000617308:Unc5A protein, mouse; D010100:Oxygen