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Open Access Research article

Expression of netrin-1 receptors in retina of oxygen-induced retinopathy in mice

Dan Liu1, Si-Qi Xiong1, Lei Shang2, Xiao-feng Tian1, Jing Yang1 and Xiao-Bo Xia1*

Author Affiliations

1 Department of Ophthalmology, Xiangya Hospital, Central South University, Changsha, Hunan 410008, China

2 Department of Anatomy and Neurobiology, School of Basic Medical Sciences, Central South University, Changsha, Hunan 410013, China

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BMC Ophthalmology 2014, 14:102  doi:10.1186/1471-2415-14-102

Published: 22 August 2014

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.

Keywords:
Netrin-1; UNC5B; Neogenin; Retinal neovascularization