Article

Light-activated channels targeted to ON bipolar cells restore visual function in retinal degeneration

Received:
Accepted:
Published online:

Abstract

Genetically encoded optical neuromodulators create an opportunity for circuit-specific intervention in neurological diseases. One of the diseases most amenable to this approach is retinal degeneration, where the loss of photoreceptors leads to complete blindness. To restore photosensitivity, we genetically targeted a light-activated cation channel, channelrhodopsin-2, to second-order neurons, ON bipolar cells, of degenerated retinas in vivo in the Pde6brd1 (also known as rd1) mouse model. In the absence of 'classical' photoreceptors, we found that ON bipolar cells that were engineered to be photosensitive induced light-evoked spiking activity in ganglion cells. The rescue of light sensitivity was selective to the ON circuits that would naturally respond to increases in brightness. Despite degeneration of the outer retina, our intervention restored transient responses and center-surround organization of ganglion cells. The resulting signals were relayed to the visual cortex and were sufficient for the animals to successfully perform optomotor behavioral tasks.

  • Subscribe to Nature Neuroscience for full access:

    $59

    Subscribe

Additional access options:

Already a subscriber?  Log in  now or  Register  for online access.

References

  1. 1.

    et al. Gene therapy restores vision in a canine model of childhood blindness. Nat. Genet. 28, 92–95 (2001).

  2. 2.

    et al. Retinal repair by transplantation of photoreceptor precursors. Nature 444, 203–207 (2006).

  3. 3.

    , & Retinal prosthesis. Annu. Rev. Biomed. Eng. 7, 361–401 (2005).

  4. 4.

    et al. Ectopic expression of a microbial-type rhodopsin restores visual responses in mice with photoreceptor degeneration. Neuron 50, 23–33 (2006).

  5. 5.

    , , & Push-pull effect of surround illumination on excitatory and inhibitory inputs to mudpuppy retinal ganglion cells. J. Physiol. (Lond.) 388, 233–243 (1987).

  6. 6.

    & Receptive fields, binocular interaction and functional architecture in the cat's visual cortex. J. Physiol. (Lond.) 160, 106–154 (1962).

  7. 7.

    et al. Specific deficit of the ON response in visual transmission by targeted disruption of the mGluR6 gene. Cell 80, 757–765 (1995).

  8. 8.

    et al. Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proc. Natl. Acad. Sci. USA 100, 13940–13945 (2003).

  9. 9.

    , , , & Millisecond-timescale, genetically targeted optical control of neural activity. Nat. Neurosci. 8, 1263–1268 (2005).

  10. 10.

    , & The rd mouse story: seventy years of research on an animal model of inherited retinal degeneration. Prog. Retin. Eye Res. 13, 31–64 (1994).

  11. 11.

    & Electroporation and RNA interference in the rodent retina in vivo and in vitro. Proc. Natl. Acad. Sci. USA 101, 16–22 (2004).

  12. 12.

    Parallel processing in the mammalian retina. Nat. Rev. Neurosci. 5, 747–757 (2004).

  13. 13.

    , & The major cell populations of the mouse retina. J. Neurosci. 18, 8936–8946 (1998).

  14. 14.

    , & Concerted signaling by retinal ganglion cells. Science 270, 1207–1210 (1995).

  15. 15.

    & 2-amino-4-phosphonobutyric acid: a new pharmacological tool for retina research. Science 211, 182–185 (1981).

  16. 16.

    , & Phototransduction by retinal ganglion cells that set the circadian clock. Science 295, 1070–1073 (2002).

  17. 17.

    Discharge patterns and functional organization of mammalian retina. J. Neurophysiol. 16, 37–68 (1953).

  18. 18.

    et al. Intrinsic ON responses of the retinal OFF pathway are suppressed by the ON pathway. J. Neurosci. 26, 11857–11869 (2006).

  19. 19.

    & Vertical interactions across ten parallel, stacked representations in the mammalian retina. Nature 410, 583–587 (2001).

  20. 20.

    , & The visual physiology of the wild type mouse determined with pattern VEPs. Vision Res. 39, 3071–3081 (1999).

  21. 21.

    & The mouse light/dark box test. Eur. J. Pharmacol. 463, 55–65 (2003).

  22. 22.

    , & Defining stops in search pathways. J. Neurosci. Methods 60, 95–98 (1995).

  23. 23.

    et al. The optomotor response: a robust first-line visual screening method for mice. Vision Res. 45, 1439–1446 (2005).

  24. 24.

    , , , & Morphological and functional abnormalities in the inner retina of the rd/rd mouse. J. Neurosci. 22, 5492–5504 (2002).

  25. 25.

    , , , & Remodeling of second-order neurons in the retina of rd/rd mutant mice. Vision Res. 43, 867–877 (2003).

  26. 26.

    & Modifications of retinal neurons in a mouse model of retinitis pigmentosa. Proc. Natl. Acad. Sci. USA 97, 11020–11025 (2000).

  27. 27.

    et al. Neural reprogramming in retinal degeneration. Invest. Ophthalmol. Vis. Sci. 48, 3364–3371 (2007).

  28. 28.

    et al. Retinal remodeling triggered by photoreceptor degenerations. J. Comp. Neurol. 464, 1–16 (2003).

  29. 29.

    & Rod vision: pathways and processing in the mammalian retina. Prog. Retin. Eye Res. 20, 351–384 (2001).

  30. 30.

    , , & Retinal organization in the retinal degeneration 10 (rd10) mutant mouse: a morphological and ERG study. J. Comp. Neurol. 500, 222–238 (2007).

  31. 31.

    & Light-evoked responses of bipolar cells in a mammalian retina. J. Neurophysiol. 83, 1817–1829 (2000).

  32. 32.

    & Origin of transient and sustained responses in ganglion cells of the retina. J. Neurosci. 20, 7087–7095 (2000).

  33. 33.

    , & Stratum-by-stratum projection of light response attributes by retinal bipolar cells of Ambystoma. J. Physiol. (Lond.) 558, 249–262 (2004).

  34. 34.

    Bipolar cells use kainate and AMPA receptors to filter visual information into separate channels. Neuron 28, 847–856 (2000).

  35. 35.

    , & The intrinsic dynamics of retinal bipolar cells isolated from tiger salamander. Vis. Neurosci. 15, 425–438 (1998).

  36. 36.

    , & Heterogeneous expression of voltage-dependent Na+ and K+ channels in mammalian retinal bipolar cells. Vis. Neurosci. 22, 119–133 (2005).

  37. 37.

    TTX attenuates surround inhibition in rabbit retinal ganglion cells. Vis. Neurosci. 16, 285–290 (1999).

  38. 38.

    & Lateral inhibition in the inner retina is important for spatial tuning of ganglion cells. Nat. Neurosci. 1, 714–719 (1998).

  39. 39.

    & Spike-dependent GABA inputs to bipolar cell axon terminals contribute to lateral inhibition of retinal ganglion cells. J. Neurophysiol. 89, 2449–2458 (2003).

  40. 40.

    et al. Ganglion cell adaptability: does the coupling of horizontal cells play a role? PLoS ONE 3, e1714 (2008).

  41. 41.

    , & Behavioural visual acuity of wild-type and bcl2 transgenic mouse. Vision Res. 39, 569–574 (1999).

  42. 42.

    & Visual detection, pattern discrimination and visual acuity in 14 strains of mice. Genes Brain Behav. 5, 389–403 (2006).

  43. 43.

    , , & Rapid quantification of adult and developing mouse spatial vision using a virtual optomotor system. Invest. Ophthalmol. Vis. Sci. 45, 4611–4616 (2004).

  44. 44.

    , & Speed, spatial, and temporal tuning of rod and cone vision in mouse. J. Neurosci. 28, 189–198 (2008).

  45. 45.

    , , & Coordinated multivesicular release at a mammalian ribbon synapse. Nat. Neurosci. 7, 826–833 (2004).

  46. 46.

    , & Excitatory synaptic transmission in the inner retina: paired recordings of bipolar cells and neurons of the ganglion cell layer. J. Neurosci. 18, 4500–4510 (1998).

  47. 47.

    , , , & Light-activated ion channels for remote control of neuronal firing. Nat. Neurosci. 7, 1381–1386 (2004).

  48. 48.

    Will retinal implants restore vision? Science 295, 1022–1025 (2002).

  49. 49.

    & Multiple-color optical activation, silencing and desynchronization of neural activity, with single-spike temporal resolution. PLoS ONE 2, e299 (2007).

  50. 50.

    et al. Multimodal fast optical interrogation of neural circuitry. Nature 446, 633–639 (2007).

Download references

Acknowledgements

We thank B.G. Scherf, S. Djaffer, Y. Shimada and F. Ronay for technical assistance, K. Deisseroth for kindly providing us with the pLECYT lentiviral vector, and A. Lüthi, S. Picaud, M. Fendt, M. Stadler and C. Herry for their suggestions or help with the behavioral experiments. This study was supported by Friedrich Miescher Institute funds, an US Office of Naval Research Naval International Cooperative Opportunities in Science and Technology Program grant, a Marie Curie Excellence Grant, a Human Frontier Science Program Young Investigator grant (B.R.), a Marie Curie Postdoctoral Fellowship (D.B. and T.A.M.), a National Center for Competence in Research in Genetics fellowship (V.B.) and a Human Frontier Science Program Fellowship (G.B.A.)

Author information

Author notes

    • Gautam B Awatramani

    Present address: Department of Anatomy and Neurobiology, Dalhousie University, 5850 College Street, Halifax, NS B3H 1X5, Canada.

    • Pamela S Lagali
    • , David Balya
    •  & Gautam B Awatramani

    These authors contributed equally to this work.

Affiliations

  1. Neural Circuit Laboratories, Friedrich Miescher Institute for Biomedical Research, Maulbeerstrasse 66, CH-4058 Basel, Switzerland.

    • Pamela S Lagali
    • , David Balya
    • , Gautam B Awatramani
    • , Thomas A Münch
    • , Volker Busskamp
    •  & Botond Roska
  2. Department of Genetics and Howard Hughes Medical Institute, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, Massachusetts 02115, USA.

    • Douglas S Kim
    •  & Constance L Cepko

Authors

  1. Search for Pamela S Lagali in:

  2. Search for David Balya in:

  3. Search for Gautam B Awatramani in:

  4. Search for Thomas A Münch in:

  5. Search for Douglas S Kim in:

  6. Search for Volker Busskamp in:

  7. Search for Constance L Cepko in:

  8. Search for Botond Roska in:

Competing interests

Pamela S Lagali, David Balya, Thomas A Münch and Botond Roska have applied for a patent on the use of light-sensitive genes (WO 2008/022772).

Constance L Cepko, Douglas S Kim and Harvard University are submitting a patent application for the use of the Grm6 regulatory elements described in this publication.

Corresponding author

Correspondence to Botond Roska.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Figure 1 and Methods