Article | Published:

The scaffold protein, Homer1b/c, regulates axon pathfinding in the central nervous system in vivo

Nature Neuroscience volume 4, pages 499506 (2001) | Download Citation

Subjects

Abstract

Homer proteins are a family of multidomain cytosolic proteins that have been postulated to serve as scaffold proteins that affect responses to extracellular signals by regulating protein–protein interactions. We tested whether Homer proteins are involved in axon pathfinding in vivo, by expressing both wild-type and mutant isoforms of Homer in Xenopus optic tectal neurons. Time-lapse imaging demonstrated that interfering with the ability of endogenous Homer to form protein–protein interactions resulted in axon pathfinding errors at stereotypical choice points. These data demonstrate a function for scaffold proteins such as Homer in axon guidance. Homer may facilitate signal transduction from cell-surface receptors to intracellular proteins that govern the establishment of axon trajectories.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.

from$8.99

All prices are NET prices.

References

  1. 1.

    Mechanisms and molecules that control growth cone guidance. Annu. Rev. Neurosci. 19, 341–377 (1996).

  2. 2.

    & From membrane to cytoskeleton: enabling a connection. Neuron 22, 419–422 (1999).

  3. 3.

    , & Cytoskeletal reorganization underlying growth cone motility. Curr. Opin. Neurobiol. 4, 640–647 (1994).

  4. 4.

    & Modulation of ion channels: a “current” view of AKAPs. Neuron 23, 423–426 (1999).

  5. 5.

    et al. Coordination of three signaling enzymes by AKAP79, a mammalian scaffold protein. Science 271, 1589–1592 (1996).

  6. 6.

    & Glutamate receptor anchoring proteins and the molecular organization of excitatory synapses. Ann. NY Acad. Sci. 868, 483–493 (1999).

  7. 7.

    & Regulation of growth cone behavior by calcium. J. Neurosci. 11, 891–899 (1991).

  8. 8.

    , , , & Calcium signalling in the guidance of nerve growth by netrin-1. Nature 403, 93–98 (2000).

  9. 9.

    Turning of nerve growth cones induced by localized increases in intracellular calcium ions. Nature 403, 89–93 (2000).

  10. 10.

    , & Calcium and chemotropic turning of nerve growth cones. Perspect. Dev. Neurobiol. 4, 205–213 (1996).

  11. 11.

    & Cytoskeletal events in growth cone steering. Curr. Opin. Neurobiol. 4, 43–48 (1994).

  12. 12.

    et al. Homer: a protein that selectively binds metabotropic glutamate receptors. Nature 386, 284–288 (1997).

  13. 13.

    , , , & vesl, a gene encoding VASP/Ena family related protein, is upregulated during seizure, long-term potentiation and synaptogenesis. FEBS Lett. 412, 183–189 (1997).

  14. 14.

    , , & A set of genes expressed in response to light in the adult cerebral cortex and regulated during development. Proc. Natl. Acad. Sci. USA 93, 2048–2053 (1996).

  15. 15.

    , , , & Involvement of unique leucine-zipper motif of PSD-Zip45 (Homer 1c/vesl-1L) in group 1 metabotropic glutamate receptor clustering. Proc. Natl. Acad. Sci. USA 96, 13801–13806 (1999).

  16. 16.

    et al. Cupidin, an isoform of Homer/Vesl, interacts with the actin cytoskeleton and activated rho family small GTPases and is expressed in developing mouse cerebellar granule cells. J. Neurosci. 19, 8389–8400 (1999).

  17. 17.

    et al. Homer regulates the association of group 1 metabotropic glutamate receptors with multivalent complexes of homer-related, synaptic proteins. Neuron 21, 707–716 (1998).

  18. 18.

    et al. Homer binds a novel proline-rich motif and links group 1 metabotropic glutamate receptors with IP3 receptors. Neuron 21, 717–726 (1998).

  19. 19.

    et al. Coupling of mGluR/Homer and PSD-95 complexes by the Shank family of postsynaptic density proteins. Neuron 23, 583–592 (1999).

  20. 20.

    , , & Homer-1c/vesl-1L modulates the cell surface targeting of metabotropic glutamate receptor type 1a: Evidence for an anchoring function. Mol. Cell. Neurosci. 15, 36–50 (2000).

  21. 21.

    , & Co-expression of metabotropic glutamate receptor type 1alpha with homer-1a/Vesl-1S increases the cell surface expression of the receptor. Biochem. J. 795–803 (1999).

  22. 22.

    et al. Homer 1b regulates the trafficking of group I metabotropic glutamate receptors. J. Biol. Chem. 274, 25953–25957 (1999).

  23. 23.

    et al. Novel members of the Vesl/Homer family of PDZ proteins that bind metabotropic glutamate receptors. J. Biol. Chem. 273, 23969–23975 (1998).

  24. 24.

    , , , & Homer proteins regulate coupling of group I metabotropic glutamate receptors to N-type calcium and M-type potassium channels. J. Neurosci. 20, 7238–7245 (2000).

  25. 25.

    et al. Structure of the Homer EVH1 domain-peptide complex reveals a new twist in polyproline recognition. Neuron 26, 143–154 (2000).

  26. 26.

    , , & Dendritic dynamics in vivo change during neuronal maturation. J. Neurosci. 19, 4472–4483 (1999).

  27. 27.

    , , , & Single-cell electroporation for in vivo neuronal gene expression. Neuron 29, 571–581 (2001).

  28. 28.

    , & Small GTPases in axon outgrowth. Perspect. Dev. Neurobiol. 4, 199–204 (1996).

  29. 29.

    , & Genetic analysis of axon guidance and mapping in the zebrafish. Trends Neurosci. 20, 3–8 (1997).

  30. 30.

    & Developmental regulation of mossy fiber afferent interactions with target granule cells. Dev. Biol. 195, 75–87 (1998).

  31. 31.

    & Error correction during guidance of pioneer axons in the leg of the cockroach embryo. Roux's Arch. Dev. Biol. 205, 476–485 (1996).

  32. 32.

    , , , & Control of dendritic field formation in Drosophila: the roles of Flamingo and competition between homologous neurons. Neuron 28, 91–101 (2000).

  33. 33.

    , & Dishevelled is a component of the frizzled signaling pathway in Drosophila. Development 121, 4095–4102 (1995).

  34. 34.

    & A single frizzled protein has a dual function in tissue polarity. Development 120, 1883–1893 (1994).

  35. 35.

    , , & Blockade of endogenous ligands of trkB inhibits formation of ocular dominance columns. Neuron 19, 63–76 (1997).

  36. 36.

    , & Inhibition of ocular dominance column formation by infusion of NT-4/5 or BDNF. Science 267, 1662–1666 (1995).

  37. 37.

    & In vivo regulation of axon extension and pathfinding by growth-cone calcium transients. Nature 397, 350–355 (1999).

  38. 38.

    , , , & Regulation of nerve growth mediated by inositol 1,4,5-trisphosphate receptors in growth cones. Science 282, 1705–1708 (1998).

  39. 39.

    Signal transduction molecules at the glutamatergic postsynaptic membrane. Brain Res. Brain Res. Rev. 26, 243–257 (1998).

  40. 40.

    , & in DNA Cloning: A Practical Approach (ed. Glover, D. M.) 191–210 (IRL, Oxford, 1985).

  41. 41.

    , , & Infection of frog neurons with Vaccinia Virus permits in vivo expression of foreign proteins. Neuron 14, 681–684 (1995).

  42. 42.

    Dendritic arbor development and synaptogenesis. Curr. Opin. Neurobiol. 11, 118–126 (2001).

  43. 43.

    , , , & in Imaging: A Laboratory Manual (eds. Yuste, R., Lanni, F. & Konnerth, A.) 13.1–13.12 (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 1999).

Download references

Acknowledgements

We thank K. Bronson, N. Dawkins and B. Burbach for technical help, E. Ruthazer, P. O'Brien, B. Burbach and K. Svoboda for building the two-photon microscope, E. Ruthazer for help with the two-photon image acquisition and analysis, J.C. Tu and B. Xiao for constructs, and members of the Cline lab for discussions. A. Demetriades helped with experiments through the Undergraduate Research Program at CSHL. Supported by the NIH (H.T.C., K.H. and P.W.), the Eppley Foundation (H.T.C.) and the Helen Hoffritz Fund (H.T.C.).

Author information

Author notes

    • Lisa Foa
    •  & Indrani Rajan

    The first two authors contributed equally to this work

Affiliations

  1. Cold Spring Harbor Laboratory, Beckman Building, 1 Bungtown Road, Cold Spring Harbor, New York 11724, USA

    • Lisa Foa
    • , Indrani Rajan
    • , Kurt Haas
    • , Gang-Yi Wu
    •  & Hollis Cline
  2. Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA

    • Paul Brakeman
    •  & Paul Worley

Authors

  1. Search for Lisa Foa in:

  2. Search for Indrani Rajan in:

  3. Search for Kurt Haas in:

  4. Search for Gang-Yi Wu in:

  5. Search for Paul Brakeman in:

  6. Search for Paul Worley in:

  7. Search for Hollis Cline in:

Corresponding author

Correspondence to Hollis Cline.

About this article

Publication history

Received

Accepted

Published

DOI

https://doi.org/10.1038/87447