The absence of cyclic nucleotide-gated (CNG) channels in cone photoreceptor outer segments leads to achromatopsia, a severely disabling disease associated with the complete lack of cone photoreceptor function. In a common form, loss of the CNGA3 subunit disrupts visual transduction in cones and causes progressive degeneration. Here, we show that adeno-associated viral vector-mediated gene replacement therapy added the lacking sensual quality, cone-mediated vision, in the CNGA3−/− mouse model of the human disease. The functional rescue of cone vision was assessed at different sites along the visual pathway. In particular, we show electrophysiologically that treated CNGA3−/− mice became able to generate cone-mediated responses and to transfer these signals to bipolar and finally ganglion cells. In support, we found morphologically that expression of CNGA3 delayed cone cell death. Finally, we show in a behavioral test that treated mice acquired photopic vision suggesting that achromatopsia patients may as well benefit from gene replacement therapy.
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We thank Peter Humphries (Trinity College Dublin) for providing Rho−/− mice, James M. Wilson (Univ Pennsylvania) and Alberto Auricchio (TIGEM) for the gift of AAV plasmids. This work was supported by the Deutsche Forschungsgemeinschaft (Se837/6-1, Se837/7-1, and Bi484/4-1), the German Ministry of Education and Research (BMBF 0314106), the European Union (EU HEALTH-F2-2008-200234), and the Max Planck Society.
Akimoto M, Filippova E, Gage PJ et al (2004) Transgenic mice expressing Cre-recombinase specifically in M- or S-cone photoreceptors. Invest Ophthalmol Vis Sci 45:42–47PubMedCrossRefGoogle Scholar
Auricchio A, Hildinger M, O’Connor E et al (2001) Isolation of highly infectious and pure adeno-associated virus type 2 vectors with a single-step gravity-flow column. Hum Gene Ther 12:71–76PubMedCrossRefGoogle Scholar
Biel M, Seeliger M, Pfeifer A et al (1999) Selective loss of cone function in mice lacking the cyclic nucleotide-gated channel CNG3. Proc Natl Acad Sci U S A 96:7553–7557PubMedCrossRefGoogle Scholar
Fischer MD, Huber G, Beck SC et al (2009) Noninvasive, in vivo assessment of mouse retinal structure using optical coherence tomography. PLoS One 4:e7507PubMedCrossRefGoogle Scholar
Grieger JC, Choi VW, Samulski RJ (2006) Production and characterization of adeno-associated viral vectors. Nat Protoc 1:1412–1428PubMedCrossRefGoogle Scholar
Kohl S, Marx T, Giddings I et al (1998) Total colourblindness is caused by mutations in the gene encoding the alpha-subunit of the cone photoreceptor cGMP-gated cation channel. Nat Genet 19:257–259PubMedCrossRefGoogle Scholar
Kohl S, Varsanyi B, Antunes GA et al (2005) CNGB3 mutations account for 50% of all cases with autosomal recessive achromatopsia. Eur J Hum Genet 13:302–308PubMedCrossRefGoogle Scholar
Michalakis S, Geiger H, Haverkamp S et al (2005) Impaired opsin targeting and cone photoreceptor migration in the retina of mice lacking the cyclic nucleotide-gated channel CNGA3. Invest Ophthalmol Vis Sci 46:1516–1524PubMedCrossRefGoogle Scholar
Petrs-Silva H, Dinculescu A, Li Q et al (2009) High-efficiency transduction of the mouse retina by tyrosine-mutant AAV serotype vectors. Mol Ther 17:463–471PubMedCrossRefGoogle Scholar
Seeliger MW, Grimm C, Stahlberg F et al (2001) New views on RPE65 deficiency: the rod system is the source of vision in a mouse model of Leber congenital amaurosis. Nat Genet 29:70–74PubMedCrossRefGoogle Scholar
Seeliger MW, Beck SC, Pereyra-Munoz N et al (2005) In vivo confocal imaging of the retina in animal models using scanning laser ophthalmoscopy. Vis Res 45:3512–3519PubMedCrossRefGoogle Scholar
Tanimoto N, Muehlfriedel RL, Fischer MD et al (2009) Vision tests in the mouse: Functional phenotyping with electroretinography. Front Biosci 14:2730–2737PubMedCrossRefGoogle Scholar