Apr 15, 2000

In vivo gene transfer into choroidal neovascularization by the HVJ liposome method

Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie
T OtsujiY Kaneda

Abstract

To evaluate the efficacy of the HVJ liposome method for gene transfer in rats with experimentally induced choroidal neovascularization. Plasmid DNA containing the LacZ reporter gene, or fluorescein isothiocyanate (FITC)-labeled double-stranded phosphorothioate oligodeoxynucleotides (S-ODNs), was encapsulated in liposomes. The liposomes were coated with the envelope of inactivated hemagglutinating virus of Japan (HVJ). Intense laser burns were applied to the posterior pole of the retina of pigmented rats to induce choroidal neovascularization. Following photocoagulation, HVJ liposome suspension was injected into the vitreous. On days 3, 7, 14, and 28 after injection, the eyes were removed and fixed. The eyes injected with LacZ gene were reacted with X-gal, frozen, and cut into thin sections. The sections were examined for the expression of the LacZ gene by light microscopy. The enucleated eyes injected with double-stranded S-ODNs were frozen, cut into thin sections, and examined a confocal scanning laser microscope for FITC labeling. Eyes without injection of HVJ liposomes served as controls. Expression of LacZ genes (beta-galactosidase activity), or localization of FITC labeling, was observed mainly in the laser-induced choroid...Continue Reading

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Mentioned in this Paper

Establishment and Maintenance of Localization
Transfection
Entire Retina
5-bromo-4-chloro-3-indolyl beta-galactoside
Isothiocyanates
DNA, Viral
Glb1
Lac Gene
Fluorescein
Indirect Immunofluorescence

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