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Hadas Stiebel-Kalish, Ehud Reich, Nir Rainy, Gad Vatine, Yael Nisgav, Anna Tovar, Yoav Gothilf, Michael Bach; Gucy2f Zebrafish Knockdown - A Model for GUCY2D-related Leber Congenital Amaurosis. Invest. Ophthalmol. Vis. Sci. 2012;53(14):763.
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In this study we will demonstrate the feasibility of using gene-targeting in the zebrafish as a model for the photoreceptor-specific guanylate cyclase (GUCY2D)-related LCA1, by reporting the visual phenotype and retinal histology resulting from Gucy2f-knockdown.
Gucy2f zebrafish LCA-orthologous cDNA was identified and isolated by PCR amplification. The expression pattern of Gucy2f was determined by whole-mount in-situ hybridization and its function was studied by gene knockdown utilizing two different morpholino-modified oligo (MO), one that blocks translation of Gucy2f and one that blocks splicing of Gucy2f. Visual function was assessed with an optomotor assay (moving grating of 0.16 cycles/degree) on 6 days post-fertilization larvae, and by analyzing changes in retinal histology.
Gucy2f knockdown resulted in significantly lower vision as measured by the optomotor response compared to uninjected zebrafish larvae and morpholino (MO)-controls. Histological changes in the Gucy2f-knockdown larvae included loss and shortening of cone and rod outer segments, retinal pigment epithelium thickening and loss of rod nuclei.
A zebrafish model of Gucy2f-related LCA1 displays early visual dysfunction and photoreceptor layer dystrophy. This study serves as proof of concept for the use of zebrafish as a simple, inexpensive model with excellent vision on which further study of LCA-related genes is possible.
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