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Regulation of axon growth in vivo by activity-based competition

Nature volume 434, pages 1022–1026 (2005)Cite this article

Abstract

The formation of functional neural networks requires precise regulation of the growth and branching of the terminal arbors of axons, processes known to be influenced by early network electrical activity1,2,3. Here we show that a rule of activity-based competition between neighbouring axons appears to govern the growth and branching of retinal ganglion cell (RGC) axon arbors in the developing optic tectum of zebrafish. Mosaic expression of an exogenous potassium channel or a dominant-negative SNARE protein was used to suppress electrical or neurosecretory activity in subsets of RGC axons. Imaging in vivo showed that these forms of activity suppression strongly inhibit both net growth and the formation of new branches by individually transfected RGC axon arbors. The inhibition is relieved when the activity of nearby ‘competing’ RGC axons is also suppressed. These results therefore identify a new form of activity-based competition rule that might be a key regulator of axon growth and branch initiation.

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Figure 1: Kir2.1 overexpression inhibits zebrafish neuronal calcium spiking; expression in single RGCs inhibits axon arbor growth.
Figure 2: VAMPm overexpression suppresses presynaptic vesicle function in zebrafish neurons; expression in single RGCs inhibits axon arbor growth.
Figure 3: Transfection of multiple neighbouring RGC axons with Kir2.1 or VAMPm mitigates their growth-suppressing effects compared with transfection of a single axon.
Figure 4: Selective activity suppression inhibits axon motility and branch formation rate with little effect on branch stability.

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References

  1. Sanes, J. R. & Lichtman, J. W. Development of the vertebrate neuromuscular junction. Annu. Rev. Neurosci. 22, 389–442 (1999)

    Article  CAS  Google Scholar 

  2. Katz, L. C. & Shatz, C. J. Synaptic activity and the construction of cortical circuits. Science 274, 1133–1138 (1996)

    Article  ADS  CAS  Google Scholar 

  3. Zhang, L. I. & Poo, M. M. Electrical activity and development of neural circuits. Nature Neurosci. 4 (Suppl), 1207–1214 (2001)

    Article  CAS  Google Scholar 

  4. Buffelli, M. et al. Genetic evidence that relative synaptic efficacy biases the outcome of synaptic competition. Nature 424, 430–434 (2003)

    Article  ADS  CAS  Google Scholar 

  5. Lichtman, J. W. & Balice-Gordon, R. J. Understanding synaptic competition in theory and in practice. J. Neurobiol. 21, 99–106 (1990)

    Article  CAS  Google Scholar 

  6. Shatz, C. J. & Stryker, M. P. Ocular dominance in layer IV of the cat's visual cortex and the effects of monocular deprivation. J. Physiol. (Lond.) 281, 267–283 (1978)

    Article  CAS  Google Scholar 

  7. Stryker, M. P. & Harris, W. A. Binocular impulse blockade prevents the formation of ocular dominance columns in cat visual cortex. J. Neurosci. 6, 2117–2133 (1986)

    Article  CAS  Google Scholar 

  8. Antonini, A. & Stryker, M. P. Rapid remodeling of axonal arbors in the visual cortex. Science 260, 1819–1821 (1993)

    Article  ADS  CAS  Google Scholar 

  9. Yu, C. R. et al. Spontaneous neural activity is required for the establishment and maintenance of the olfactory sensory map. Neuron 42, 553–566 (2004)

    Article  CAS  Google Scholar 

  10. Zhao, H. & Reed, R. R. X inactivation of the OCNC1 channel gene reveals a role for activity-dependent competition in the olfactory system. Cell 104, 651–660 (2001)

    Article  CAS  Google Scholar 

  11. Ruthazer, E. S., Akerman, C. J. & Cline, H. T. Control of axon branch dynamics by correlated activity in vivo . Science 301, 66–70 (2003)

    Article  ADS  CAS  Google Scholar 

  12. Schmidt, J. T., Buzzard, M., Borress, R. & Dhillon, S. MK801 increases retinotectal arbor size in developing zebrafish without affecting kinetics of branch elimination and addition. J. Neurobiol. 42, 303–314 (2000)

    Article  CAS  Google Scholar 

  13. Schmidt, J. T., Fleming, M. R. & Leu, B. Presynaptic protein kinase C controls maturation and branch dynamics of developing retinotectal arbors: possible role in activity-driven sharpening. J. Neurobiol. 58, 328–340 (2004)

    Article  CAS  Google Scholar 

  14. Gnuegge, L., Schmid, S. & Neuhauss, S. C. Analysis of the activity-deprived zebrafish mutant macho reveals an essential requirement of neuronal activity for the development of a fine-grained visuotopic map. J. Neurosci. 21, 3542–3548 (2001)

    Article  CAS  Google Scholar 

  15. Johnson, F. A., Dawson, A. J. & Meyer, R. L. Activity-dependent refinement in the goldfish retinotectal system is mediated by the dynamic regulation of processes withdrawal: an in vivo imaging study. J. Comp. Neurol. 406, 548–562 (1999)

    Article  CAS  Google Scholar 

  16. Burrone, J., O'Byrne, M. & Murthy, V. N. Multiple forms of synaptic plasticity triggered by selective suppression of activity in individual neurons. Nature 420, 414–418 (2002)

    Article  ADS  CAS  Google Scholar 

  17. Drapeau, P. et al. Development of the locomotor network in zebrafish. Prog. Neurobiol. 68, 85–111 (2002)

    Article  CAS  Google Scholar 

  18. Koster, R. W. & Fraser, S. E. Tracing transgene expression in living zebrafish embryos. Dev. Biol. 233, 329–346 (2001)

    Article  CAS  Google Scholar 

  19. Sorensen, J. B. et al. The SNARE protein SNAP-25 is linked to fast calcium triggering of exocytosis. Proc. Natl Acad. Sci. USA 99, 1627–1632 (2002)

    Article  ADS  CAS  Google Scholar 

  20. Li, W., Ono, F. & Brehm, P. Optical measurements of presynaptic release in mutant zebrafish lacking postsynaptic receptors. J. Neurosci. 23, 10467–10474 (2003)

    Article  CAS  Google Scholar 

  21. Scales, S. J. et al. SNAREs contribute to the specificity of membrane fusion. Neuron 26, 457–464 (2000)

    Article  CAS  Google Scholar 

  22. Hua, J. Y. & Smith, S. J. Neural activity and the dynamics of central nervous system development. Nature Neurosci. 7, 327–332 (2004)

    Article  CAS  Google Scholar 

  23. Cogen, J. & Cohen-Cory, S. Nitric oxide modulates retinal ganglion cell axon arbor remodeling in vivo . J. Neurobiol. 45, 120–133 (2000)

    Article  CAS  Google Scholar 

  24. Cohen-Cory, S. & Fraser, S. E. Effects of brain-derived neurotrophic factor on optic axon branching and remodelling in vivo . Nature 378, 192–196 (1995)

    Article  ADS  CAS  Google Scholar 

  25. Haas, K., Jensen, K., Sin, W. C., Foa, L. & Cline, H. T. Targeted electroporation in Xenopus tadpoles in vivo—from single cells to the entire brain. Differentiation 70, 148–154 (2002)

    Article  CAS  Google Scholar 

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Acknowledgements

We thank T. R. Clandinin, R. W. Tsien, R. W. Aldrich, L. Luo and the Smith laboratory for comments on the manuscript, and C. M. Niell for developing the Matlab routines used in image analysis. We thank T. Roeser for isolating the brn3c promoter. The US National Institutes of Health and the Vincent Coates Foundation provided financial support. J.Y.H. was supported by a Stanford Graduate Fellowship and a Coates Foundation Fellowship. M.C.S. was supported by a predoctoral fellowship from the American Heart Association.

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Authors and Affiliations

  1. Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, 94305, USA

    Jackie Yuanyuan Hua & Stephen J. Smith

  2. Neurosciences Program, Stanford University, Stanford, California, 94305, USA

    Jackie Yuanyuan Hua

  3. Department of Physiology, Program in Neuroscience, University of California, San Francisco, 1550 4th Street, San Francisco, California, 94143-2722, USA

    Matthew C. Smear & Herwig Baier

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Correspondence to Jackie Yuanyuan Hua.

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Supplementary information

Supplementary Figure S1

This document contains Supplementary Figure S1 and accompanying legend. The figure shows that Kir2.1 and VAMP-GFP coexpress with high efficiency. (DOC 406 kb)

Supplementary Movie S1

This movie shows the growth of a VAMP-GFP expressing RGC axon arbor. Images were collected at two-minute intervals for 48 minutes. Scale bars represent 5µm in all movies. Auto-fluorescence of the skin is seen in the upper left corner of this movie and in some of the other movies presented (QuickTime; 5MB). (MOV 517 kb)

Supplementary Movie S2

This movie shows the growth of a Kir2.1 and VAMP-GFP expressing RGC axon arbor. Images were collected at two-minute intervals for 48 minutes. The axon shows less exploratory behaviour compared with control axons expressing VAMP-GFP (Supplementary movie 1). (MOV 514 kb)

Supplementary Movie S3

This movie shows the growth of a VAMPm expressing RGC axon arbor. Images were collected at two-minute intervals for 48 minutes. The axon shows less exploratory behaviour compared with control axons expressing VAMP-GFP (Supplementary movie 1). (MOV 707 kb)

Supplementary Movie S4

This movie shows the growth of a VAMP-GFP expressing RGC axon arbor. Images were collected at 20-minute intervals for 8 hours. In each of this movie and Supplementary movie S5-6, a new branch formed during the imaging period and lasted beyond 4 hours is pointed out by arrowhead in the last time point of imaging. (MOV 700 kb)

Supplementary Movie 5

This movie shows the growth of a Kir2.1 and VAMP-GFP expressing RGC axon arbor. Images were collected at 20-minute intervals for 8 hours. The axon is less exploratory compared with control (Supplementary movie S5), but new branch stability is not significantly affected. A new branch that lasted for more than 4 hours, and was likely stabilized, is pointed out by arrowhead in the last time point of this movie. (MOV 1534 kb)

Supplementary Movie S6

This movie shows the growth of a VAMPm expressing RGC axon arbor. Images were collected at 20-minute intervals for 8 hours. The axon is less exploratory compared to control (Supplementary movie S5), but new branch stability is not significantly affected. (MOV 591 kb)

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Hua, J., Smear, M., Baier, H. et al. Regulation of axon growth in vivo by activity-based competition. Nature 434, 1022–1026 (2005). https://doi.org/10.1038/nature03409

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