Article | Published:

Wiring specificity in the direction-selectivity circuit of the retina

Nature volume 471, pages 183188 (10 March 2011) | Download Citation

Abstract

The proper connectivity between neurons is essential for the implementation of the algorithms used in neural computations, such as the detection of directed motion by the retina. The analysis of neuronal connectivity is possible with electron microscopy, but technological limitations have impeded the acquisition of high-resolution data on a large enough scale. Here we show, using serial block-face electron microscopy and two-photon calcium imaging, that the dendrites of mouse starburst amacrine cells make highly specific synapses with direction-selective ganglion cells depending on the ganglion cell’s preferred direction. Our findings indicate that a structural (wiring) asymmetry contributes to the computation of direction selectivity. The nature of this asymmetry supports some models of direction selectivity and rules out others. It also puts constraints on the developmental mechanisms behind the formation of synaptic connections. Our study demonstrates how otherwise intractable neurobiological questions can be addressed by combining functional imaging with the analysis of neuronal connectivity using large-scale electron microscopy.

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References

  1. 1.

    , & Retinal ganglion cells responding selectively to direction and speed of image motion in the rabbit. J. Physiol. (Lond.) 173, 377–407 (1964)

  2. 2.

    & The mechanism of directionally selective units in rabbit’s retina. J. Physiol. (Lond.) 178, 477–504 (1965)

  3. 3.

    & Diverse synaptic mechanisms generate direction selectivity in the rabbit retina. J. Neurosci. 22, 7712–7720 (2002)

  4. 4.

    , & Mechanisms and circuitry underlying directional selectivity in the retina. Nature 420, 411–414 (2002)

  5. 5.

    Synaptic organization of starburst amacrine cells in rabbit retina: analysis of serial thin sections by electron microscopy and graphic reconstruction. J. Comp. Neurol. 309, 40–70 (1991)

  6. 6.

    & The shape and arrangement of the cholinergic neurons in the rabbit retina. Proc. R. Soc. Lond. B 223, 101–119 (1984)

  7. 7.

    et al. A key role of starburst amacrine cells in originating retinal directional selectivity and optokinetic eye movement. Neuron 30, 771–780 (2001)

  8. 8.

    , & Co-release of acetylcholine and GABA by the starburst amacrine cells. J. Neurosci. 12, 1394–1408 (1992)

  9. 9.

    , & Directionally selective calcium signals in dendrites of starburst amacrine cells. Nature 418, 845–852 (2002)

  10. 10.

    & Starburst cells nondirectionally facilitate the responses of direction-selective retinal ganglion cells. J. Neurosci. 22, 10509–10513 (2002)

  11. 11.

    , & Is the input to a GABAergic or cholinergic synapse the sole asymmetry in rabbit’s retinal directional selectivity? Vis. Neurosci. 14, 39–54 (1997)

  12. 12.

    & New directions in retinal research. Trends Neurosci. 26, 379–385 (2003)

  13. 13.

    Cellular mechanisms for direction selectivity in the retina. Neuron 55, 179–186 (2007)

  14. 14.

    et al. Dendritic spikes amplify the synaptic signal to enhance detection of motion in a simulation of the direction-selective ganglion cell. PLOS Comput. Biol. 6, (2010)

  15. 15.

    The computation of directional selectivity in the retina occurs presynaptic to the ganglion cell. Nature Neurosci. 4, 176–183 (2001)

  16. 16.

    , , & Development of asymmetric inhibition underlying direction selectivity in the retina. Nature 469, 402–406 (2010)

  17. 17.

    , & Role of ACh-GABA co-transmission in detecting image motion and motion direction. Neuron 68, 1159–1172 (2010)

  18. 18.

    et al. Spatially asymmetric reorganization of inhibition establishes a motion-sensitive circuit. Nature 469, 407–410 (2010)

  19. 19.

    et al. Laminar circuit formation in the vertebrate retina. Prog. Brain Res. 147, 155–169 (2005)

  20. 20.

    A structural basis for omnidirectional connections between starburst amacrine cells and directionally selective ganglion cells in rabbit retina, with associated bipolar cells. Vis. Neurosci. 19, 145–162 (2002)

  21. 21.

    et al. Dendritic relationship between starburst amacrine cells and direction-selective ganglion cells in the rabbit retina. J. Physiol. (Lond.) 556, 11–17 (2004)

  22. 22.

    & Symmetric synaptic patterns between starburst amacrine cells and direction selective ganglion cells in the rabbit retina. J. Comp. Neurol. 508, 175–183 (2008)

  23. 23.

    , & Synaptic input to the on-off directionally selective ganglion cell in the rabbit retina. J. Comp. Neurol. 456, 267–278 (2003)

  24. 24.

    et al. The structure of the nervous system of the nematode Caenorhabditis elegans. Phil. Trans. R. Soc. Lond. 314, 1–340 (1986)

  25. 25.

    & Serial block-face scanning electron microscopy to reconstruct three-dimensional tissue nanostructure. PLoS Biol. 2, e329 (2004)

  26. 26.

    , & Dendritic architecture of ON-OFF direction-selective ganglion cells in the rabbit retina. Vision Res. 33, 579–608 (1993)

  27. 27.

    & Receptive fields and dendritic structure of directionally selective retinal ganglion cells. J. Neurosci. 14, 5267–5280 (1994)

  28. 28.

    & Optical recording of light-evoked calcium signals in the functionally intact retina. Proc. Natl Acad. Sci. USA 96, 7035–7040 (1999)

  29. 29.

    et al. Synaptic and extrasynaptic factors governing glutamatergic retinal waves. Neuron 62, 230–241 (2009)

  30. 30.

    et al. In vivo two-photon calcium imaging of neuronal networks. Proc. Natl Acad. Sci. USA 100, 7319–7324 (2003)

  31. 31.

    , & Two-photon laser scanning fluorescence microscopy. Science 248, 73–76 (1990)

  32. 32.

    et al. Eyecup scope–optical recordings of light stimulus-evoked fluorescence signals in the retina. Pflugers Arch. 457, 1393–1414 (2009)

  33. 33.

    & Direction-selective units in rabbit retina: distribution of preferred directions. Science 155, 841–842 (1967)

  34. 34.

    et al. Synaptic connections of starburst amacrine cells and localization of acetylcholine receptors in primate retinas. J. Comp. Neurol. 461, 76–90 (2003)

  35. 35.

    et al. Dendritic spread and functional coverage of starburst amacrine cells. J. Comp. Neurol. 505, 539–546 (2007)

  36. 36.

    et al. A dendrite-autonomous mechanism for direction selectivity in retinal starburst amacrine cells. PLoS Biol. 5, e185 (2007)

  37. 37.

    & The synaptic mechanism of direction selectivity in distal processes of starburst amacrine cells. Neuron 51, 787–799 (2006)

  38. 38.

    & Tetrodotoxin-resistant sodium channels contribute to directional responses in starburst amacrine cells. PLoS ONE 5, e12447 (2010)

  39. 39.

    , & The nondiscriminating zone of directionally selective retinal ganglion cells: comparison with dendritic structure and implications for mechanism. J. Neurosci. 19, 8049–8056 (1999)

  40. 40.

    & Effect of ON pathway blockade on directional selectivity in the rabbit retina. J. Neurophysiol. 73, 703–712 (1995)

  41. 41.

    , & Effects of picrotoxin and strychnine on rabbit retinal ganglion cells: lateral interactions for cells with more complex receptive fields. J. Physiol. (Lond.) 276, 277–298 (1978)

  42. 42.

    & GABA-like immunoreactivity in cholinergic amacrine cells of the rabbit retina. Brain Res. 438, 369–373 (1988)

  43. 43.

    , & Directional selectivity is formed at multiple levels by laterally offset inhibition in the rabbit retina. Neuron 46, 117–127 (2005)

  44. 44.

    et al. Identification of cholinoceptive glycinergic neurons in the mammalian retina. J. Comp. Neurol. 456, 167–175 (2003)

  45. 45.

    , & Expression of α7 nicotinic acetylcholine receptors by bipolar, amacrine, and ganglion cells of the rabbit retina. J. Histochem. Cytochem. 55, 461–476 (2007)

  46. 46.

    et al. Retrograde neuronal tracing with a deletion-mutant rabies virus. Nature Methods 4, 47–49 (2007)

  47. 47.

    et al. Fluorescence-based monitoring of in vivo neural activity using a circuit-tracing pseudorabies virus. PLoS ONE 4, e6923 (2009)

  48. 48.

    & Bulk electroporation and population calcium imaging in the adult mammalian retina. J. Neurophysiol (in the press)

  49. 49.

    et al. Toxicity assessment of intravitreal triamcinolone and bevacizumab in a retinal explant mouse model using two-photon microscopy. Invest. Ophthalmol. Vis. Sci. 50, 5880–5887 (2009)

  50. 50.

    An Extemporaneous Lead Citrate Stain for Electron Microscopy 148–149 (Proc. 25th Annu. EMSA Meeting, 1967)

  51. 51.

    & Biological specimen preparation for transmission electron microscopy. In Practical Methods in Electron Microscopy xxi (Princeton Univ. Press, 1998)

  52. 52.

    Use of Ferrocyanide-reduced osmium in electron microscopy 146 (Proc. 14th Annual Meeting Am. Soc. Cell Biol., 1971)

  53. 53.

    , & A new staining method (OTO) for enhancing contrast of lipid-containing membranes and droplets in osmium tetroxide-fixed tissue with osmiophilic thiocarbohydrazide (TCH). J. Cell Biol. 30, 424–432 (1966)

  54. 54.

    Lead asparate, an en bloc contrast stain particularly useful for ultrastructural enzymology. J. Histochem. Cytochem. 27, 1337–1342 (1979)

  55. 55.

    et al. User-guided 3D active contour segmentation of anatomical structures: significantly improved efficiency and reliability. Neuroimage 31, 1116–1128 (2006)

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Acknowledgements

We thank T. Euler for many useful discussions and help with the functional imaging experiments. We also thank H. Horstmann and S. Mikula for help with staining procedures; J. Kornfeld and F. Svara for programming KNOSSOS; J. Tritthardt for developing electronic circuits and M. Müller for help with the acquisition software; J. Hanne, H. Jakobi and H. Wissler for help with training tracers; M. Feller and Z. J. Zhou for discussion of their results; and J. Bollmann, A. Karpova and S. Seung for comments on the manuscript. We thank N. Abazova, E. Abs, A. Antunes, P. Bastians, M. Beining, J. Buhmann, F. Drawitsch, L. Ehm, F. Isensee, H. Jakobi, S. Kaspar, A. Khan, M. Kiapes, A. Klein, S. Laiouar, E. Möller, J. Trendel, P. Weber, K. Weiß, E. Wiegand and H. Wissler for the tracing work.

Author information

Affiliations

  1. Max Planck Institute for Medical Research, Department of Biomedical Optics, Heidelberg 69120, Germany

    • Kevin L. Briggman
    • , Moritz Helmstaedter
    •  & Winfried Denk

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Contributions

K.L.B. and W.D. designed the study; W.D. designed the microtome; K.L.B. performed the calcium imaging and SBEM experiments, K.L.B. and M.H. analysed data; K.L.B., M.H. and W.D. wrote the paper.

Competing interests

W.D. receives license income for SBEM technology.

Corresponding author

Correspondence to Kevin L. Briggman.

Supplementary information

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  1. 1.

    Supplementary Figures

    This file contains Supplementary Figures 1-7 with legends.

Zip files

  1. 1.

    Supplementary Image Stacks

    This zip file contains five small image stacks viewable in ImageJ corresponding to the examples in Figure 3. The centre of the stacks (64,64,64) coincides with the dendritic contact or proximity between a SAC and DSGC. Slice 64 in each stack contains colour-coded dots corresponding to the cell identities in Figure 3.

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DOI

https://doi.org/10.1038/nature09818

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