Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Axons and dendrites originate from neuroepithelial-like processes of retinal bipolar cells

Nature Neuroscience volume 9, pages 85–92 (2006)Cite this article

Abstract

The cellular mechanisms underlying axogenesis and dendritogenesis are not completely understood. The axons and dendrites of retinal bipolar cells, which contact their synaptic partners within specific laminae in the inner and outer retina, provide a good system for exploring these issues. Using transgenic mice expressing enhanced green fluorescent protein (GFP) in a subset of bipolar cells, we determined that axonal and dendritic arbors of these interneurons develop directly from apical and basal processes attached to the outer and inner limiting membranes, respectively. Selective stabilization of processes contributed to stratification of axonal and dendritic arbors within the appropriate synaptic layer. This unusual mode of axogenesis and dendritogenesis from neuroepithelial-like processes may act to preserve neighbor-neighbor relationships in synaptic wiring between the outer and inner retina.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: GFP is expressed by ON cone bipolar cells and rod bipolar cells in the Grm6-GFP transgenic mouse.
Figure 2: GFP expression reveals bipolar cells with neuroepithelial-like processes during development.
Figure 3: Bipolar cell axons originate from the basal process.
Figure 4: The axons of P5 ON bipolar cells branch throughout their length within the IPL, but branches are preferentially stabilized in the ON sublamina.
Figure 5: Dendrites originate from the apical process, at or proximal to the OPL.
Figure 6: Time-lapse imaging reveals dynamics of dendritic outgrowth and apical process retraction.

Similar content being viewed by others

References

  1. Goslin, K. & Banker, G. Experimental observations on the development of polarity by hippocampal neurons in culture. J. Cell Biol. 108, 1507–1516 (1989).

    Article  CAS  Google Scholar 

  2. Craig, A.M. & Banker, G. Neuronal polarity. Annu. Rev. Neurosci. 17, 267–310 (1994).

    Article  CAS  Google Scholar 

  3. Shi, S.H., Jan, L.Y. & Jan, Y.N. Hippocampal neuronal polarity specified by spatially localized mPar3/mPar6 and PI 3-kinase activity. Cell 112, 63–75 (2003).

    Article  CAS  Google Scholar 

  4. Hatanaka, Y. & Murakami, F. In vitro analysis of the origin, migratory behavior, and maturation of cortical pyramidal cells. J. Comp. Neurol. 454, 1–14 (2002).

    Article  Google Scholar 

  5. Morest, D.K. & Silver, J. Precursors of neurons, neuroglia, and ependymal cells in the CNS: what are they? Where are they from? How do they get where they are going? Glia 43, 6–18 (2003).

    Article  Google Scholar 

  6. Snow, R.L. & Robson, J.A. Migration and differentiation of neurons in the retina and optic tectum of the chick. Exp. Neurol. 134, 13–24 (1995).

    Article  CAS  Google Scholar 

  7. Nakajima, Y. et al. Molecular characterization of a novel retinal metabotropic glutamate receptor mGluR6 with a high agonist selectivity for L-2-amino-4-phosphonobutyrate. J. Biol. Chem. 268, 11868–11873 (1993).

    CAS  PubMed  Google Scholar 

  8. Nomura, A. et al. Developmentally regulated postsynaptic localization of a metabotropic glutamate receptor in rat rod bipolar cells. Cell 77, 361–369 (1994).

    Article  CAS  Google Scholar 

  9. Vardi, N. & Morigiwa, K. ON cone bipolar cells in rat express the metabotropic receptor mGluR6. Vis. Neurosci. 14, 789–794 (1997).

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  11. Greferath, U., Grunert, U. & Wassle, H. Rod bipolar cells in the mammalian retina show protein kinase C-like immunoreactivity. J. Comp. Neurol. 301, 433–442 (1990).

    Article  CAS  Google Scholar 

  12. Claes, E. et al. Morphological characterization of the retina of the CNGA3−/−Rho−/− mutant mouse lacking functional cones and rods. Invest. Ophthalmol. Vis. Sci. 45, 2039–2048 (2004).

    Article  Google Scholar 

  13. Tehrani, A., Wheeler-Schilling, T.H. & Guenther, E. Coexpression patterns of mGluR mRNAs in rat retinal ganglion cells: a single-cell RT-PCR study. Invest. Ophthalmol. Vis. Sci. 41, 314–319 (2000).

    CAS  PubMed  Google Scholar 

  14. Cepko, C.L., Austin, C.P., Yang, X., Alexiades, M. & Ezzeddine, D. Cell fate determination in the vertebrate retina. Proc. Natl. Acad. Sci. USA 93, 589–595 (1996).

    Article  CAS  Google Scholar 

  15. Lin, B. & Masland, R.H. Synaptic contacts between an identified type of ON cone bipolar cell and ganglion cells in the mouse retina. Eur. J. Neurosci. 21, 1257–1270 (2005).

    Article  Google Scholar 

  16. Wong, G.T., Ruiz-Avila, L. & Margolskee, R.F. Directing gene expression to gustducin-positive taste receptor cells. J. Neurosci. 19, 5802–5809 (1999).

    Article  CAS  Google Scholar 

  17. Bishop, D.L., Misgeld, T., Walsh, M.K., Gan, W.B. & Lichtman, J.W. Axon branch removal at developing synapses by axosome shedding. Neuron 44, 651–661 (2004).

    Article  CAS  Google Scholar 

  18. Dotti, C.G., Sullivan, C.A. & Banker, G.A. The establishment of polarity by hippocampal neurons in culture. J. Neurosci. 8, 1454–1468 (1988).

    Article  CAS  Google Scholar 

  19. Goslin, K. & Banker, G. Rapid changes in the distribution of GAP-43 correlate with the expression of neuronal polarity during normal development and under experimental conditions. J. Cell Biol. 110, 1319–1331 (1990).

    Article  CAS  Google Scholar 

  20. Snow, R.L. & Robson, J.A. Ganglion cell neurogenesis, migration and early differentiation in the chick retina. Neuroscience 58, 399–409 (1994).

    Article  CAS  Google Scholar 

  21. de Anda, F.C. et al. Centrosome localization determines neuronal polarity. Nature 436, 704–708 (2005).

    Article  Google Scholar 

  22. Quesada, A., Prada, F., Armengol, J.A. & Genis-Galvez, J.M. Early morphological differentiation of the bipolar neurons in the chick retina. A Golgi analysis. Anat. Histol. Embryol. 10, 328–341 (1981).

    Article  CAS  Google Scholar 

  23. Quesada, A. & Genis-Galvez, J.M. Morphological and structural study of Landolt's club in the chick retina. J. Morphol. 184, 205–214 (1985).

    Article  CAS  Google Scholar 

  24. Rowan, S. & Cepko, C.L. Genetic analysis of the homeodomain transcription factor Chx10 in the retina using a novel multifunctional BAC transgenic mouse reporter. Dev. Biol. 271, 388–402 (2004).

    Article  CAS  Google Scholar 

  25. Cajal, R.Y. Studies on Vertebrate Neurogenesis (Thomas, Springfield, Illinois, 1960).

    Google Scholar 

  26. Kirby, M.A. & Steineke, T.C. Morphogenesis of retinal ganglion cells: a model of dendritic, mosaic, and foveal development. Perspect. Dev. Neurobiol. 3, 177–194 (1996).

    CAS  PubMed  Google Scholar 

  27. Hatanaka, Y., Hisanaga, S., Heizmann, C.W. & Murakami, F. Distinct migratory behavior of early- and late-born neurons derived from the cortical ventricular zone. J. Comp. Neurol. 479, 1–14 (2004).

    Article  Google Scholar 

  28. Freed, M.A., Smith, R.G. & Sterling, P. Computational model of the on-alpha ganglion cell receptive field based on bipolar cell circuitry. Proc. Natl. Acad. Sci. USA 89, 236–240 (1992).

    Article  CAS  Google Scholar 

  29. Cohen, E. & Sterling, P. Microcircuitry related to the receptive field center of the on-beta ganglion cell. J. Neurophysiol. 65, 352–359 (1991).

    Article  CAS  Google Scholar 

  30. Cohen, E. & Sterling, P. Demonstration of cell types among cone bipolar neurons of cat retina. Phil. Trans. R. Soc.B 330, 305–321 (1990).

    Article  CAS  Google Scholar 

  31. Niell, C.M., Meyer, M.P. & Smith, S.J. In vivo imaging of synapse formation on a growing dendritic arbor. Nat. Neurosci. 7, 254–260 (2004).

    Article  CAS  Google Scholar 

  32. Stacy, R.C. & Wong, R.O. Developmental relationship between cholinergic amacrine cell processes and ganglion cell dendrites of the mouse retina. J. Comp. Neurol. 456, 154–166 (2003).

    Article  Google Scholar 

  33. Kay, J.N. et al. Transient requirement for ganglion cells during assembly of retinal synaptic layers. Development 131, 1331–1342 (2004).

    Article  CAS  Google Scholar 

  34. Gunhan-Agar, E., Kahn, D. & Chalupa, L.M. Segregation of on and off bipolar cell axonal arbors in the absence of retinal ganglion cells. J. Neurosci. 20, 306–314 (2000).

    Article  CAS  Google Scholar 

  35. Higgins, D., Burack, M., Lein, P. & Banker, G. Mechanisms of neuronal polarity. Curr. Opin. Neurobiol. 7, 599–604 (1997).

    Article  CAS  Google Scholar 

  36. Rice, D.S. et al. The reelin pathway modulates the structure and function of retinal synaptic circuitry. Neuron 31, 929–941 (2001).

    Article  CAS  Google Scholar 

  37. Vardi, N., Duvoisin, R., Wu, G. & Sterling, P. Localization of mGluR6 to dendrites of ON bipolar cells in primate retina. J. Comp. Neurol. 423, 402–412 (2000).

    Article  CAS  Google Scholar 

  38. Demas, J., Eglen, S.J. & Wong, R.O. Developmental loss of synchronous spontaneous activity in the mouse retina is independent of visual experience. J. Neurosci. 23, 2851–2860 (2003).

    Article  CAS  Google Scholar 

  39. Lohmann, C. et al. Imaging the developing retina. in Imaging in Neuroscience and Development. (eds. Yuste, R. & Konnerth, A.) 159–170 (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 2005).

    Google Scholar 

Download references

Acknowledgements

We thank R.F. Margolskee and R.H. Masland for providing the GUS8.4-GFP mice. This work is supported by grants from the US National Institutes of Health (EY10699 to R.O.L.W. and EY11105 to N.V.) and the Bakewell and Alafi Neuroimaging Laboratories.

Author information

Author notes

  1. Josh L Morgan and Anuradha Dhingra: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Anatomy and Neurobiology, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, 63110, Missouri, USA

    Josh L Morgan & Rachel O L Wong

  2. Department of Neuroscience, University of Pennsylvania School of Medicine, 123 Anatomy/Chemistry Building, Philadelphia, 19104–6058, Pennsylvania, USA

    Anuradha Dhingra & Noga Vardi

Authors
  1. Josh L Morgan
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Anuradha Dhingra
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Noga Vardi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Rachel O L Wong
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Rachel O L Wong.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Video 1

3D rotation of a confocal stack from a P3 Grm6-GFP retinal cross-section. In this region, a GFP positive bipolar axon terminal can be distinguished from the relatively dimmer IPL processes of neighboring amacrine cells and RGCs upon rotation of the image stack. For example, a GFP-labeled process (arrow 2) that appears close to the bipolar cell axon (arrow 1) in the initial frame, clearly moves apart from the axon in subsequent frames when the stack is rotated. Here, we have also highlighted the bipolar axon and its fine terminals in white using the 3D segmentation program in Amira, to provide a high contrast view of the axon. Inner nuclear layer (INL), inner plexiform layer (IPL), ganglion cell layer (GCL). (MOV 3156 kb)

Supplementary Video 2

Time-lapse showing process motility in some axonal arbors of P6 bipolar cells in the Grm6-GFP mouse retina (confocal image stacks, 7 time frames, 30 min between frames). Note extensions and retractions that are particularly evident in the bright axon. (MOV 1373 kb)

Supplementary Video 3

3D confocal reconstruction of P10 Grm6-GFP retina showing extensions of bipolar cell axons into the ganglion cell layer (GCL), some of which retracted during the recording period. The boundaries of the inner plexiform layer (IPL) and inner nuclear layer (INL) are provided. In the first segment of the movie, the image stack is cropped and rotated in 3D to provide a clearer view of the morphology of the processes within the GCL. In the second segment, the retraction of an axonal terminal (middle of the field of view) in the GCL is apparent in a zoomed up view of this region (7 time points, 20 min between frames). (MOV 2941 kb)

Supplementary Video 4

Movie showing process extension and retraction from the apical process of the cell shown in Fig. 6b (7 frames, 20 min between frames). Outer nuclear layer (ONL), outer plexiform layer (OPL); inner nuclear layer (INL). (MOV 376 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Morgan, J., Dhingra, A., Vardi, N. et al. Axons and dendrites originate from neuroepithelial-like processes of retinal bipolar cells. Nat Neurosci 9, 85–92 (2006). https://doi.org/10.1038/nn1615

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nn1615

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing