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Spontaneous activity of opsin apoprotein is a cause of Leber congenital amaurosis

Nature Genetics volume 35, pages 158–164 (2003)Cite this article

Abstract

Mutations in Rpe65 disrupt synthesis of the opsin chromophore ligand 11-cis-retinal and cause Leber congenital amaurosis (LCA), a severe, early-onset retinal dystrophy. To test whether light-independent signaling by unliganded opsin causes the degeneration, we used Rpe65-null mice, a model of LCA. Dark-adapted Rpe65−/− mice behaved as if light adapted, exhibiting reduced circulating current, accelerated response turn-off, and diminished intracellular calcium. A genetic block of transducin signaling completely rescued degeneration irrespective of an elevated level of retinyl ester. These studies clearly show that activation of sensory transduction by unliganded opsin, and not the accumulation of retinyl esters, causes light-independent retinal degeneration in LCA. A similar mechanism may also be responsible for degeneration induced by vitamin A deprivation.

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Figure 1: Transduction and cyclic nucleotide–gated channel activity in rods.
Figure 2: Photoresponse amplitude and light sensitivity of Rpe65−/− rods are reduced.
Figure 4: Rpe65−/− rods have accelerated response kinetics.
Figure 3: Rods from Rpe65−/− mice show light saturation behavior and have functional cGMP channels.
Figure 5: Abrogation of transducin signaling protects Rpe65−/− mice from retinal degeneration, but dark-rearing does not.
Figure 6: Retinyl ester levels in wild-type (WT), Rpe65−/− and Rpe65−/− Gnat1−/− mice.
Figure 7: Free Ca2+ concentration in rods from wild-type (WT; blue), Rpe65−/− (red) and Rpe65−/− Gnat1−/− (black) mice.

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Acknowledgements

We thank M. Cilluffo for help with animal care, M. Fain for drawing Fig. 1, F. Celestin for assistance with histological sections, M. C. Cornwall and H. R. Matthews for assistance with early experiments and M. S. Obin for discussion and comments. Support for this research was provided by the US National Institutes of Health (to G.L.F., J.L. and Z.W.; Core Grants for Vision Research to the Jules Stein Eye Institute and Tufts Center for Vision Research) and the Massachusetts Lions Eye Research Fund (an institutional grant to the Department of Ophthalmology, New England Medical Center).

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Author notes

  1. Michael L Woodruff, Zhongyan Wang, Gordon L Fain and Janis Lem: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Physiological Science, University of California Los Angeles, Los Angeles, 90095, California, USA

    Michael L Woodruff & Gordon L Fain

  2. Molecular Cardiology Research Institute, Tufts-New England Medical Center, 750 Washington St. Box 5045, Boston, 02111, Massachusetts, USA

    Zhongyan Wang, Hae Yun Chung & Janis Lem

  3. Laboratory of Retinal Cell and Molecular Biology, National Eye Institute, National Institutes of Health, Bethesda, 20892, Maryland, USA

    T Michael Redmond

  4. Department of Ophthalmology, University of California Los Angeles, Los Angeles, 90095, California, USA

    Gordon L Fain

  5. Department of Ophthalmology, Program in Genetics, and Tufts Center for Vision Research, Tufts University School of Medicine, Boston, 02111, Massachusetts, USA

    Janis Lem

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Correspondence to Janis Lem.

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Woodruff, M., Wang, Z., Chung, H. et al. Spontaneous activity of opsin apoprotein is a cause of Leber congenital amaurosis. Nat Genet 35, 158–164 (2003). https://doi.org/10.1038/ng1246

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