Kenji Suda1, Congrong Guo1, Akio Oishi1, Shinya Ikeda1, Akiyoshi Uemura2, Nagahisa Yoshimura1. 1. Department of Ophthalmology and Visual Sciences, Kyoto University Graduate School of Medicine, Kyoto, Japan. 2. Department of Retinal Vascular Biology, Nagoya City University Graduate School of Medicine, Nagoya, Japan.
Abstract
PURPOSE: We investigated the effects of semaphorin 3E (Sema3E)/PlexinD1 signaling in the development of choroidal neovascularization (CNV) and explored the therapeutic potential of the pathway. METHODS: We used a laser-induced CNV model in the mouse. The expression of Sema3E and PlexinD1 was evaluated with immunohistochemistry, real-time RT-PCR, and Western blotting. After the intravitreal injection of Sema3E, the CNV size was measured on immunostained flat mounts. The CNV size was compared between inducible PlexinD1 homozygous mice and heterozygous knockout mice. RESULTS: The presence of PlexinD1 was confirmed immunohistochemically in the endothelial cells of CNV. The expression of PlexinD1 mRNA was elevated after laser photocoagulation with its peak expression at day 5 (1.72 times, compared to baseline; P < 0.01). The intravitreal injection of recombinant Sema3E decreased the CNV size (16,320.9 ± 8316.0 μm(2) in the control eyes versus 9487.4 ± 4424.1 μm(2) in the eyes administered 0.1-μg Sema3E, P < 0.01). This inhibitory effect of Sema3E in the formation of CNV was canceled in the inducible PlexinD1 knockout mice (24,920.8 ± 24,684.8 μm(2) in homozygous knockout mice and 13,152.6 ± 7973.0 μm(2) in heterozygous knockout mice, P = 0.014). CONCLUSIONS: These results indicate that Sema3E-PlexinD1 signaling is involved in the development of CNV. Stimulation of the pathway has therapeutic potential for CNV. Further studies are needed to evaluate the effects for clinical applications. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.
PURPOSE: We investigated the effects of semaphorin 3E (Sema3E)/PlexinD1 signaling in the development of choroidal neovascularization (CNV) and explored the therapeutic potential of the pathway. METHODS: We used a laser-induced CNV model in the mouse. The expression of Sema3E and PlexinD1 was evaluated with immunohistochemistry, real-time RT-PCR, and Western blotting. After the intravitreal injection of Sema3E, the CNV size was measured on immunostained flat mounts. The CNV size was compared between inducible PlexinD1 homozygous mice and heterozygous knockout mice. RESULTS: The presence of PlexinD1 was confirmed immunohistochemically in the endothelial cells of CNV. The expression of PlexinD1 mRNA was elevated after laser photocoagulation with its peak expression at day 5 (1.72 times, compared to baseline; P < 0.01). The intravitreal injection of recombinant Sema3E decreased the CNV size (16,320.9 ± 8316.0 μm(2) in the control eyes versus 9487.4 ± 4424.1 μm(2) in the eyes administered 0.1-μg Sema3E, P < 0.01). This inhibitory effect of Sema3E in the formation of CNV was canceled in the inducible PlexinD1 knockout mice (24,920.8 ± 24,684.8 μm(2) in homozygous knockout mice and 13,152.6 ± 7973.0 μm(2) in heterozygous knockout mice, P = 0.014). CONCLUSIONS: These results indicate that Sema3E-PlexinD1 signaling is involved in the development of CNV. Stimulation of the pathway has therapeutic potential for CNV. Further studies are needed to evaluate the effects for clinical applications. Copyright 2014 The Association for Research in Vision and Ophthalmology, Inc.