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Leah C. Byrne, Deniz Dalkara, Emmanuelle Clerin, Steven K. Fisher, Jose A. Sahel, Jr., Thierry D. Leveillard, John G. Flannery; AAV-Mediated Delivery of RdCVF and RdCVFL in a Mouse Model of Retinal Degeneration. Invest. Ophthalmol. Vis. Sci. 2012;53(14):1907.
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© ARVO (1962-2015); The Authors (2016-present)
Rod-derived cone viability factor (RdCVF), a truncated thioredoxin-like protein, is secreted and has been shown to have a protective effect on the survival of cones in rodent models of rod-cone dystrophy, while RdCVFL, a full length alternative splicing product encoded by the same gene, contains a thioredoxin fold and may be involved in oxidative signaling. RdCVFL has been shown to interact with the microtubule binding protein Tau, and in vitro to protect against phosphorylation of Tau through oxidative stress. Here we evaluate the effects of AAV-mediated delivery of RdCVF and RdCVFL in the rd10 mouse model of retinal degeneration.
AAV vectors were used to deliver RdCVF or RdCVFL in neonatal rd10 mice via tail vein injections or via intravitreal injections at P14. A CAG promoter drove ubiquitous expression of RdCVF or RdCVFL while a rho promoter restricted expression of RdCVFL to photoreceptors. A viral 2A peptide was used to simultaneously allow expression of a fluorescent reporter gene. Electroretinograms were recorded to evaluate rescue of rod and cone-mediated responses. The density of cones was quantified to determine the effect of expression on cone survival. Western blotting was used to observe the effects of expression of RdCVFL on levels of Tau phosphorylation.
Injection of AAV-RdCVF and RdCVFL resulted in high levels of protein expression. Delivery of RdCVF mediated functional rescue of cone photoreceptors while delivery of RdCVFL resulted in a delay in the loss of the rod-driven scotopic ERG, indicating its involvement in protecting rods.
Nxnl1 is a bifunctional gene encoding two alternative splicing products. AAV-mediated delivery of these two products, RdCVF and RdCVFL, mediates high levels of protein expression and rescue of photoreceptors in the retina. RdCVF promotes survival of cones while the expression of RdCVFL delays rod degeneration. The synergistic effect of co-expression of these two proteins is being evaluated.
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