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Publication : Absence of functional and structural abnormalities associated with expression of EGFP in the retina.

First Author  Nour M Year  2004
Journal  Invest Ophthalmol Vis Sci Volume  45
Issue  1 Pages  15-22
PubMed ID  14691148 Mgi Jnum  J:87298
Mgi Id  MGI:2684433 Doi  10.1167/iovs.03-0663
Citation  Nour M, et al. (2004) Absence of functional and structural abnormalities associated with expression of EGFP in the retina. Invest Ophthalmol Vis Sci 45(1):15-22
abstractText  PURPOSE: The present study was undertaken to evaluate the effect of uniform EGFP expression on retinal morphology and function. METHODS: Electroretinography (ERG) was used to evaluate the recovery of scotopic a- and b-wave amplitudes after a single 137-cd.sec/m2 flash exposure. The cellular distribution of enhanced green fluorescent protein (EGFP) in the retina and its effect on retinal morphology were evaluated by fluorescence microscopy and histology, respectively. To evaluate its effect on retinal sensitivity to light, EGFP-expressing and control mice were exposed to constant light for 76 hours (3500 lux), and eyes were assessed functionally and structurally at 3 weeks after light exposure. RESULTS: Fluorescence microscopy showed a pronounced EGFP expression in the photoreceptor cell bodies and inner segments. ERG analysis revealed no significant differences in either a- or b-wave amplitudes or recovery between EGFP(+/-) and control mice under dark- or light-adapted conditions. Histologic assessment at as late as 4 months of age showed no difference in retinal morphology or photoreceptor nuclei count in EGFP(+/-) mice when compared with nontransgenic littermates. In addition, evaluation of animals, 3 weeks after constant light exposure, showed no difference between ERG amplitudes, recovery of the scotopic ERG response, or retinal morphology between EGFP(+/-) mice and control animals. CONCLUSIONS: Functional and morphologic evidence shows that long-term, high, uniform levels of EGFP expression have no deleterious effect on the mouse retina. This data demonstrates the safety of EGFP use as an indicator of viral transduction in retinal gene therapy.
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