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.
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.
Authors: A Suzuki; K Obi; T Urabe; H Hayakawa; M Yamada; S Kaneko; M Onodera; Y Mizuno; H Mochizuki Journal: J Neurochem Date: 2002-08 Impact factor: 5.372
Authors: Ann C Morris; Eric H Schroeter; Joseph Bilotta; Rachel O L Wong; James M Fadool Journal: Invest Ophthalmol Vis Sci Date: 2005-12 Impact factor: 4.799
Authors: Daniel M Lipinski; Mohammed Yusuf; Alun R Barnard; Christopher Damant; Peter Charbel Issa; Mandeep S Singh; Edward Lee; Wayne L Davies; Emanuela V Volpi; Robert E MacLaren Journal: Invest Ophthalmol Vis Sci Date: 2011-08-22 Impact factor: 4.799
Authors: Alicia A Brunet; Paula I Fuller-Carter; Annie L Miller; Valentina Voigt; Sophia Vasiliou; Rabab Rashwan; David M Hunt; Livia S Carvalho Journal: Transl Vis Sci Technol Date: 2020-08-18 Impact factor: 3.283