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Neurotransmission selectively regulates synapse formation in parallel circuits in vivo

Abstract

Activity is thought to guide the patterning of synaptic connections in the developing nervous system. Specifically, differences in the activity of converging inputs are thought to cause the elimination of synapses from less active inputs and increase connectivity with more active inputs1,2. Here we present findings that challenge the generality of this notion and offer a new view of the role of activity in synapse development. To imbalance neurotransmission from different sets of inputs in vivo, we generated transgenic mice in which ON but not OFF types of bipolar cells in the retina express tetanus toxin (TeNT). During development, retinal ganglion cells (RGCs) select between ON and OFF bipolar cell inputs (ON or OFF RGCs) or establish a similar number of synapses with both on separate dendritic arborizations (ON-OFF RGCs). In TeNT retinas, ON RGCs correctly selected the silenced ON bipolar cell inputs over the transmitting OFF bipolar cells, but were connected with them through fewer synapses at maturity. Time-lapse imaging revealed that this was caused by a reduced rate of synapse formation rather than an increase in synapse elimination. Similarly, TeNT-expressing ON bipolar cell axons generated fewer presynaptic active zones. The remaining active zones often recruited multiple, instead of single, synaptic ribbons. ON-OFF RGCs in TeNT mice maintained convergence of ON and OFF bipolar cells inputs and had fewer synapses on their ON arbor without changes to OFF arbor synapses. Our results reveal an unexpected and remarkably selective role for activity in circuit development in vivo, regulating synapse formation but not elimination, affecting synapse number but not dendritic or axonal patterning, and mediating independently the refinement of connections from parallel (ON and OFF) processing streams even where they converge onto the same postsynaptic cell.

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Figure 1: Selective blockade of glutamate release from retinal ON bipolar cells.
Figure 2: Silencing ON bipolar cells reduces synapse number on RGC dendrites in an input-specific manner without changes to laminar targeting or branching.
Figure 3: Axonal morphology is normal, but multiple ribbons accumulate at fewer synapses in TeNT-expressing ON bipolar cells.
Figure 4: Transmitter release regulates synapse formation but not elimination, causing a gradual divergence of synaptic development between wild-type and mGluR6-YFP/TeNT mice.

References

  1. 1

    Wong, R. O. L. & Lichtman, J. W. in Fundamental Neuroscience 2nd edn (eds Squire, L. R. et al.) Ch. 20 533–554 (Academic Press, 2002)

    Google Scholar 

  2. 2

    Miller, K. D. Synaptic economics: competition and cooperation in synaptic plasticity. Neuron 17, 371–374 (1996)

    MathSciNet  CAS  Article  Google Scholar 

  3. 3

    Masland, R. H. The fundamental plan of the retina. Nature Neurosci. 4, 877–886 (2001)

    CAS  Article  Google Scholar 

  4. 4

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

    Article  Google Scholar 

  5. 5

    Ueda, Y., Iwakabe, H., Masu, M., Suzuki, M. & Nakanishi, S. The mGluR6 5′ upstream transgene sequence directs a cell-specific and developmentally regulated expression in retinal rod and ON-type cone bipolar cells. J. Neurosci. 17, 3014–3023 (1997)

    CAS  Article  Google Scholar 

  6. 6

    Schiavo, G. et al. Tetanus and botulinum-B neurotoxins block neurotransmitter release by proteolytic cleavage of synaptobrevin. Nature 359, 832–835 (1992)

    ADS  CAS  Article  Google Scholar 

  7. 7

    Harms, K. J. & Craig, A. M. Synapse composition and organization following chronic activity blockade in cultured hippocampal neurons. J. Comp. Neurol. 490, 72–84 (2005)

    CAS  Article  Google Scholar 

  8. 8

    Chichilnisky, E. J. A simple white noise analysis of neuronal light responses. Network 12, 199–213 (2001)

    CAS  Article  Google Scholar 

  9. 9

    Maslim, J., Webster, M. & Stone, J. Stages in the structural differentiation of retinal ganglion cells. J. Comp. Neurol. 254, 382–402 (1986)

    CAS  Article  Google Scholar 

  10. 10

    Coombs, J. L., Van Der List, D. & Chalupa, L. M. Morphological properties of mouse retinal ganglion cells during postnatal development. J. Comp. Neurol. 503, 803–814 (2007)

    Article  Google Scholar 

  11. 11

    Wang, G. Y., Liets, L. C. & Chalupa, L. M. Unique functional properties of On and Off pathways in the developing mammalian retina. J. Neurosci. 21, 4310–4317 (2001)

    CAS  Article  Google Scholar 

  12. 12

    Nelson, R., Famiglietti, E. V. & Kolb, H. Intracellular staining reveals different levels of stratification for on- and off-center ganglion cells in cat retina. J. Neurophysiol. 41, 472–483 (1978)

    CAS  Article  Google Scholar 

  13. 13

    Morgan, J. L., Schubert, T. & Wong, R. O. L. Developmental patterning of glutamatergic synapses onto retinal ganglion cells. Neural Develop. 3, 8 (2008)

    Article  Google Scholar 

  14. 14

    Morgan, J. L., Dhingra, A., Vardi, N. & Wong, R. O. L. Axons and dendrites originate from neuroepithelial-like processes of retinal bipolar cells. Nature Neurosci. 9, 85–92 (2006)

    CAS  Article  Google Scholar 

  15. 15

    Ghosh, K. K., Bujan, S., Haverkamp, S., Feigenspan, A. & Wassle, H. Types of bipolar cells in the mouse retina. J. Comp. Neurol. 469, 70–82 (2004)

    Article  Google Scholar 

  16. 16

    Sterling, P. & Matthews, G. Structure and function of ribbon synapses. Trends Neurosci. 28, 20–29 (2005)

    CAS  Article  Google Scholar 

  17. 17

    Regus-Leidig, H., Tom Dieck, S., Specht, D., Meyer, L. & Brandstatter, J. H. Early steps in the assembly of photoreceptor ribbon synapses in the mouse retina: the involvement of precursor spheres. J. Comp. Neurol. 512, 814–824 (2009)

    Article  Google Scholar 

  18. 18

    Zhai, R. G. et al. Assembling the presynaptic active zone: a characterization of an active one precursor vesicle. Neuron 29, 131–143 (2001)

    CAS  Article  Google Scholar 

  19. 19

    Johnson, J. et al. Vesicular neurotransmitter transporter expression in developing postnatal rodent retina: GABA and glycine precede glutamate. J. Neurosci. 23, 518–529 (2003)

    CAS  Article  Google Scholar 

  20. 20

    Okabe, S., Miwa, A. & Okado, H. Spine formation and correlated assembly of presynaptic and postsynaptic molecules. J. Neurosci. 21, 6105–6114 (2001)

    CAS  Article  Google Scholar 

  21. 21

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

    CAS  Article  Google Scholar 

  22. 22

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

    ADS  CAS  Article  Google Scholar 

  23. 23

    Kasthuri, N. & Lichtman, J. W. The role of neuronal identity in synaptic competition. Nature 424, 426–430 (2003)

    ADS  CAS  Article  Google Scholar 

  24. 24

    Hua, J. Y., Smear, M. C., Baier, H. & Smith, S. J. Regulation of axon growth in vivo by activity-based competition. Nature 434, 1022–1026 (2005)

    ADS  CAS  Article  Google Scholar 

  25. 25

    Sanes, J. R. & Yamagata, M. Formation of lamina-specific synaptic connections. Curr. Opin. Neurobiol. 9, 79–87 (1999)

    CAS  Article  Google Scholar 

  26. 26

    Shaner, N. C. et al. Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nature Biotechnol. 22, 1567–1572 (2004)

    CAS  Article  Google Scholar 

  27. 27

    Marquardt, T. et al. Pax6 is required for the multipotent state of retinal progenitor cells. Cell 105, 43–55 (2001)

    CAS  Article  Google Scholar 

  28. 28

    Kerschensteiner, D. et al. Genetic control of circuit function: Vsx1 and Irx5 transcription factors regulate contrast adaptation in the mouse retina. J. Neurosci. 28, 2342–2352 (2008)

    CAS  Article  Google Scholar 

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Acknowledgements

We are grateful to J. Sanes and R. W. Burgess for TeNT–CFP, S. Naganishi for the mGluR6 promoter fragment, A. M. Craig for PSD95–CFP and R. Y. Tsien for tdTomato. We thank F. Soto, L. Godinho, A. Lewis, F. Dunn and T. Misgeld for comments on the manuscript. This work was supported by the National Institutes of Health (R.O.L.W., EY10699, J.L.M. T32 EY07031), the McDonnell Foundation at Washington University (R.O.L.W.), National Eye Institute core grant (E.D.P., EY01730) and the Deutsche Forschungsgemeinschaft (D.K., KE 1466/1-1).

Author Contributions D.K. and R.O.L.W. conceived the experiments. D.K. and R.M.L. generated transgenic constructs. D.K. performed and analysed patch-clamp and multi-electrode array recordings, and imaging experiments on fixed tissue. D.K. and J.L.M. performed and analysed live imaging experiments. D.K., E.D.P. and R.O.L.W. carried out the ultrastructural analysis. D.K. and R.O.L.W. wrote the paper.

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Correspondence to Daniel Kerschensteiner or Rachel O. L. Wong.

Supplementary information

Supplementary Information

This file contains a Supplementary Discussion, Supplementary Figures S1-S9 with Legends and Supplementary References. (PDF 1374 kb)

Supplementary Movie 1

This movie focuses through the representative RGC dendrite shown in Figure 2c expressing tdTomato (blue) and PSD95-CFP (red) in a P21 mGluR6-YFP/TeNT mouse (bipolar cell terminals in green). (MOV 2565 kb)

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Kerschensteiner, D., Morgan, J., Parker, E. et al. Neurotransmission selectively regulates synapse formation in parallel circuits in vivo. Nature 460, 1016–1020 (2009). https://doi.org/10.1038/nature08236

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