Vascular endothelial growth factor expression in choroidal neovascularization in rats.
BACKGROUND
The pathogenesis of choroidal neovascularization is largely unknown. We investigated vascular endothelial growth factor (VEGF) expression in laser-induced choroidal neovascularization (CNV) in rats.


METHODS
Intense krypton laser photocoagulation was applied to the posterior poles of the eyes of pigmented rats to induce CNV, which was confirmed by fluorescein angiography and histopathology. The eyeballs were enucleated 1, 3, 7, 14 and 28 days after laser photocoagulation. Cryostat sections were prepared for immunofluorescence staining using anti-VEGF and macrophage marker (ED1) antibodies. The posterior segments of eyeballs pooled from photocoagulated and control rats were submitted for immunoprecipitation and immunoblotting by the anti-VEGF antibody, and reverse transcriptase-polymerase chain reaction (RT-PCR) amplification of VEGF mRNA.


RESULTS
Very weak immunoreactivity for anti-VEGF antibody was found in the ganglion cell layer, inner nuclear layer, and retinal pigment epithelium (RPE) in the normal retina. In the development of CNV, strong positive staining for anti-VEGF antibody was found in photocoagulated areas in the subretinal space and choroid. Double immunofluorescence staining showed that many cells in lasered lesions were positive both for anti-VEGF and macrophage marker ED1 antibody staining in the early stage of this model. Immunoblots showed a positive band for the VEGF molecule in treated but not control animals. RT-PCR results demonstrated upregulation of VEGF transcripts in the CNV model compared with normal animals.


CONCLUSIONS
Our findings showed the upregulation of VEGF expression in experimentally induced CNV, where it may be involved in promoting choroidal angiogenesis. Macrophages may be one of the main sources of VEGF in the early stage of the disease.
Yi
X
X
Department of Ophthalmology, Kansai Medical University, Osaka, Japan.
Ogata
N
N
Komada
M
M
Yamamoto
C
C
Takahashi
K
K
Omori
K
K
Uyama
M
M
D000818:Animals; D000906:Antibodies / Q000032:analysis; D002829:Choroid / Q000098:blood supply*; D004195:Disease Models, Animal; D016228:Endothelial Growth Factors / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D005451:Fluorescein Angiography; D019084:Fluorescent Antibody Technique, Indirect; D005654:Fundus Oculi; D015151:Immunoblotting; D017075:Laser Coagulation / Q000009:adverse effects; D008222:Lymphokines / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D008264:Macrophages / Q000378:metabolism; D008297:Male; D009389:Neovascularization, Pathologic / Q000175:diagnosis / Q000209:etiology / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D011914:Rats, Inbred BN; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors
D000906:Antibodies; D016228:Endothelial Growth Factors; D008222:Lymphokines; D012333:RNA, Messenger; D042461:Vascular Endothelial Growth Factor A; D042442:Vascular Endothelial Growth Factors