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(Investigative Ophthalmology and Visual Science. 2001;42:1172-1178.)
© 2001 by The Association for Research in Vision and Ophthalmology, Inc.

Colocalization of Neuropilin-1 and Flk-1 in Retinal Neovascularization in a Mouse Model of Retinopathy

Hidenori Ishihama1,2, Masaharu Ohbayashi3, Nobuyuki Kurosawa1, Takashi Kitsukawa4, Onrai Matsuura1,5, Yozo Miyake2 and Takashi Muramatsu1

1 From the Departments of Biochemistry, 2 Ophthalmology, and 5 Pediatrics, Nagoya University School of Medicine; 3 Photon Medical Research Center, Hamamatsu University School of Medicine; and the 4 Laboratory of Speciation Mechanisms 1, National Institute for Basic Biology, Okazaki, Japan.

PURPOSE. To investigate the mechanisms of the development of retinal neovascularization, the localizations of vascular endothelial (VEGF) receptors Flk-1 and neuropilin (NP)-1 mRNAs were examined.

METHODS. The model of retinopathy of prematurity (ROP) was produced by ischemia-induced ocular neovascularization, by exposing postnatal day-7 mice to 75% oxygen for 5 days and then returning them to room air for 5 days. Retinal neovascularization was visualized by injection of fluorescein-dextran. Expression of Flk-1 and NP-1 mRNAs were examined by in situ hybridization with flatmount and serial sections of the retina. The localization of NP-1 was also confirmed by immunohistochemistry. Blood vessel patterns were characterized by immunohistochemical localization of von Willebrand factor (vWF).

RESULTS. Flatmount in situ hybridization showed intense expression of NP-1 and Flk-1 mRNAs colocalized in the area of neovascularization. In situ hybridization of serial sections of the retina revealed that expression of Flk-1 and NP-1 was restricted to neovascularized vessels of the retina from ROP mice.

CONCLUSIONS. The restricted expression of Flk-1 and NP-1 on neovascularized vessels suggests that these molecules may play important roles in retinal neovascularization. This is the first report of the colocalization of NP-1 and Flk-1 on neovascularized vessels of the retina from ROP mice.




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