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R. Y. Choi, A. Coughlin, G. Jones, A. S. Viczian, G. A. Engbretson, M. E. Zuber; Targeted Rod Photoreceptor Ablation in Transgenic Xenopus Laevis. Invest. Ophthalmol. Vis. Sci. 2009;50(13):4461.
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© ARVO (1962-2015); The Authors (2016-present)
Rod photoreceptor death is a common feature of blinding diseases such as retinitis pigmentosa. We have developed a transgenic approach to specifically ablate rod photoreceptors in the retina of Xenopus laevis in order to mimic rod cell loss.
Our method utilizes the E.coli enzyme nitroreductase, and its ability to reduce the antibiotic metronidazole to a DNA cross-linker. In theory, metronidazole treatment will kill any cell expressing nitroreductase via DNA damage. We have generated a transgenic line of frogs driving nitroreductase expression under the control of the rod specific rhodopsin promoter.
Metronidazole treated transgenic tadpoles show a dramatic loss in rod photoreceptors when compared with wild type and untreated transgenic animals. Rod cell ablation was metronidazole dose and time dependent. Using a behavioral assay we also show that vision loss is correlated with rod photoreceptor cell death. Cone photoreceptor loss followed rod cell death in treated animals; suggesting that retinal changes may mimic those observed in rod-cone dystrophies. Additional changes in the retina following rod ablation will also be described.
These results suggest this cell ablation technique may be a useful tool for studying the mechanisms driving retinal degenerations and potentially retinal regeneration.
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