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Impressum
Comparing functional, immunological and histochemical methods for visualization of laser-induced choroidal neovascularization (CNV)
Peters S., Luther T. T., Jordan J., Schraermeyer U.
University of Cologne, Joseph-Stelzmann-Str. 9, D-50931 Köln
Objective: Choroidal neovascularization (CNV) play an important
role in pathogenesis of age-related macular degeneration. The aim of this
study was to compare different methods for the detection of laser-induced
CNV in a rat model.
Methods: Laserspots (Argon, blue-green, 150 mW, 100 ms, 100 µm)
were applied to the fundus of Long Evans rats in a circle around the optic
disc. 10 days after laser coagulation 3 different methods were used to
detect CNV in the laser spots: 1) After intravenous injection of high
molecular fluorescein isothiocyanate dextran flat mounts consisting of
choroid and RPE or of choroid, RPE and retina were examined by fluorescent
microscopy. 2) Flat mounts consisting of choroid and RPE were labeled
with the endothelial cell antibody PECAM-1 and assessed by fluorescent
microscopy. 3) The alkaline phosphatase of endothelial cells was shown
by histochemistry for light microscopical examination.
Results: At the rim of the laser spots we found circularly intravasal
dextran accumulation (method 1) and vessel-forming endothelial cells in
the same area (method 2,3). In the center of the scars there was a diffuse
accumulation of dextran (method 1) and a diffuse arrangement of non vessel-forming
endothelial cells (method 2).
Conclusion: Apart from the classical techniques (fluorescein angiography,
histology) CNV can reliably be represented by 1) vascular perfusion of
high molecular fluorescein dextran, which shows the intravasal lumen as
well as leakage; 2) immunohistochemical labeling of proliferated vascular
endothelial cells by endothelial cell antibodies and 3) histochemical
representation of alkaline phosphatase in endothelial cells. There is
a good correlation between the different methods, because all methods
show not leaking neovascularization at the border of the laser scars but
diffuse leakage and flat endothelial cells, which do not form vessels,
in the center of the lesion.