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:

GRIP1 controls dendrite morphogenesis by regulating EphB receptor trafficking

Nature Neuroscience volume 8, pages 906–915 (2005)Cite this article

Abstract

The function of the multi-PDZ domain scaffold protein GRIP1 (glutamate receptor interacting protein 1) in neurons is unclear. To explore the function of GRIP1 in hippocampal neurons, we used RNA interference (RNAi) to knock down the expression of GRIP1. Knockdown of GRIP1 by small interfering RNA (siRNA) in cultured hippocampal neurons caused a loss of dendrites, associated with mislocalization of the GRIP-interacting proteins GluR2 (AMPA receptor subunit), EphB2 (receptor tyrosine kinase) and KIF5 (also known as kinesin 1; microtubule motor). The loss of dendrites by GRIP1-siRNA was rescued by overexpression of the extracellular domain of EphB2, and was phenocopied by overexpression of the intracellular domain of EphB2 and extracellular application of ephrinB-Fc fusion proteins. Neurons from EphB1-EphB2-EphB3 triple knockout mice showed abnormal dendrite morphogenesis. Disruption of the KIF5-GRIP1 interaction inhibited EphB2 trafficking and strongly impaired dendritic growth. These results indicate an important role for GRIP1 in dendrite morphogenesis by serving as an adaptor protein for kinesin-dependent transport of EphB receptors to dendrites.

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: RNAi knockdown of GRIP1 impairs dendrite morphogenesis of hippocampal neurons.
Figure 2: GRIP1 is essential for the maintenance of dendrites in mature hippocampal neurons.
Figure 3: GRIP1 constructs containing PDZ456 and L2 rescue the GRIP1-siRNA phenotype.
Figure 4: Effect of GRIP1 knockdown on subcellular distribution of GluR2 and EphB2.
Figure 5: Extracellular domain of EphB2 rescues GRIP1 siRNA phenotype.
Figure 6: Abnormal dendrite morphology in hippocampal neurons deficient for EphB1, EphB2 and EphB3.
Figure 7: EphB2 accumulates in the Golgi region in the absence of GRIP1.
Figure 8: KIF5/GRIP interaction required for surface expression of GluR2 and EphB2 and dendrite morphogenesis.

Similar content being viewed by others

References

  1. Hausser, M., Spruston, N. & Stuart, G.J. Diversity and dynamics of dendritic signaling. Science 290, 739–744 (2000).

    Article  CAS  PubMed  Google Scholar 

  2. 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  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  5. Scott, E.K. & Luo, L. How do dendrites take their shape? Nat. Neurosci. 4, 359–365 (2001).

    Article  CAS  PubMed  Google Scholar 

  6. Wong, R.O. & Ghosh, A. Activity-dependent regulation of dendritic growth and patterning. Nat. Rev. Neurosci. 3, 803–812 (2002).

    Article  CAS  PubMed  Google Scholar 

  7. Miller, F.D. & Kaplan, D.R. Signaling mechanisms underlying dendrite formation. Curr. Opin. Neurobiol. 13, 391–398 (2003).

    Article  CAS  PubMed  Google Scholar 

  8. Jan, Y.N. & Jan, L.Y. The control of dendrite development. Neuron 40, 229–242 (2003).

    Article  CAS  PubMed  Google Scholar 

  9. Van Aelst, L. & Cline, H.T. Rho GTPases and activity-dependent dendrite development. Curr. Opin. Neurobiol. 14, 297–304 (2004).

    Article  CAS  PubMed  Google Scholar 

  10. Liu, Z., Steward, R. & Luo, L. Drosophila Lis1 is required for neuroblast proliferation, dendritic elaboration and axonal transport. Nat. Cell Biol. 2, 776–783 (2000).

    Article  CAS  PubMed  Google Scholar 

  11. Yu, W. et al. Depletion of a microtubule-associated motor protein induces the loss of dendritic identity. J. Neurosci. 20, 5782–5791 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Dong, H. et al. GRIP: a synaptic PDZ domain-containing protein that interacts with AMPA receptors. Nature 386, 279–284 (1997).

    Article  CAS  PubMed  Google Scholar 

  13. Wyszynski, M. et al. Association of AMPA receptors with a subset of glutamate receptor-interacting protein in vivo . J. Neurosci. 19, 6528–6537 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Srivastava, S. et al. Novel anchorage of GluR2/3 to the postsynaptic density by the AMPA receptor-binding protein ABP. Neuron 21, 581–591 (1998).

    Article  CAS  PubMed  Google Scholar 

  15. DeSouza, S., Fu, J., States, B.A. & Ziff, E.B. Differential palmitoylation directs the AMPA receptor-binding protein ABP to spines or to intracellular clusters. J. Neurosci. 22, 3493–3503 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Dong, H. et al. Characterization of the glutamate receptor-interacting proteins GRIP1 and GRIP2. J. Neurosci. 19, 6930–6941 (1999).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Bruckner, K. et al. EphrinB ligands recruit GRIP family PDZ adaptor proteins into raft membrane microdomains. Neuron 22, 511–524 (1999).

    Article  CAS  PubMed  Google Scholar 

  18. Lin, D., Gish, G.D., Songyang, Z. & Pawson, T. The carboxyl terminus of B class ephrins constitutes a PDZ domain binding motif. J. Biol. Chem. 274, 3726–3733 (1999).

    Article  CAS  PubMed  Google Scholar 

  19. Torres, R. et al. PDZ proteins bind, cluster, and synaptically colocalize with Eph receptors and their ephrin ligands. Neuron 21, 1453–1463 (1998).

    Article  CAS  PubMed  Google Scholar 

  20. Wyszynski, M. et al. Interaction between GRIP and liprin-α/SYD2 is required for AMPA receptor targeting. Neuron 34, 39–52 (2002).

    Article  CAS  PubMed  Google Scholar 

  21. Ye, B. et al. GRASP-1: a neuronal RasGEF associated with the AMPA receptor/GRIP complex. Neuron 26, 603–617 (2000).

    Article  CAS  PubMed  Google Scholar 

  22. Takamiya, K. et al. A direct functional link between the multi-PDZ domain protein GRIP1 and the Fraser syndrome protein Fras1. Nat. Genet. 36, 172–177 (2004).

    Article  CAS  PubMed  Google Scholar 

  23. Setou, M. et al. Glutamate-receptor-interacting protein GRIP1 directly steers kinesin to dendrites. Nature 417, 83–87 (2002).

    Article  CAS  PubMed  Google Scholar 

  24. Bladt, F., Tafuri, A., Gelkop, S., Langille, L. & Pawson, T. Epidermolysis bullosa and embryonic lethality in mice lacking the multi-PDZ domain protein GRIP1. Proc. Natl. Acad. Sci. USA 99, 6816–6821 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Hering, H. & Sheng, M. Activity-dependent redistribution and essential role of cortactin in dendritic spine morphogenesis. J. Neurosci. 23, 11759–11769 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Grunwald, I.C. et al. Kinase-independent requirement of EphB2 receptors in hippocampal synaptic plasticity. Neuron 32, 1027–1040 (2001).

    Article  CAS  PubMed  Google Scholar 

  27. Dalva, M.B. et al. EphB receptors interact with NMDA receptors and regulate excitatory synapse formation. Cell 103, 945–956 (2000).

    Article  CAS  PubMed  Google Scholar 

  28. Takasu, M.A., Dalva, M.B., Zigmond, R.E. & Greenberg, M.E. Modulation of NMDA receptor-dependent calcium influx and gene expression through EphB receptors. Science 295, 491–495 (2002).

    Article  CAS  PubMed  Google Scholar 

  29. Henderson, J.T. et al. The receptor tyrosine kinase EphB2 regulates NMDA-dependent synaptic function. Neuron 32, 1041–1056 (2001).

    Article  CAS  PubMed  Google Scholar 

  30. Henkemeyer, M., Itkis, O.S., Ngo, M., Hickmott, P.W. & Ethell, I.M. Multiple EphB receptor tyrosine kinases shape dendritic spines in the hippocampus. J. Cell Biol. 163, 1313–1326 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Henkemeyer, M. et al. Nuk controls pathfinding of commissural axons in the mammalian central nervous system. Cell 86, 35–46 (1996).

    Article  CAS  PubMed  Google Scholar 

  32. Birgbauer, E., Cowan, C.A., Sretavan, D.W. & Henkemeyer, M. Kinase independent function of EphB receptors in retinal axon pathfinding to the optic disc from dorsal but not ventral retina. Development 127, 1231–1241 (2000).

    CAS  PubMed  Google Scholar 

  33. Bredt, D.S. & Nicoll, R.A. AMPA receptor trafficking at excitatory synapses. Neuron 40, 361–379 (2003).

    Article  CAS  PubMed  Google Scholar 

  34. Song, I. & Huganir, R.L. Regulation of AMPA receptors during synaptic plasticity. Trends Neurosci. 25, 578–588 (2002).

    Article  CAS  PubMed  Google Scholar 

  35. Kim, E. & Sheng, M. PDZ domain proteins of synapses. Nat. Rev. Neurosci. 5, 771–781 (2004).

    Article  CAS  PubMed  Google Scholar 

  36. Burette, A., Wyszynski, M., Valtschanoff, J.G., Sheng, M. & Weinberg, R.J. Characterization of glutamate receptor interacting protein-immunopositive neurons in cerebellum and cerebral cortex of the albino rat. J. Comp. Neurol. 411, 601–612 (1999).

    Article  CAS  PubMed  Google Scholar 

  37. Kanai, Y. et al. KIF5C, a novel neuronal kinesin enriched in motor neurons. J. Neurosci. 20, 6374–6384 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Ferreira, A., Niclas, J., Vale, R.D., Banker, G. & Kosik, K.S. Suppression of kinesin expression in cultured hippocampal neurons using antisense oligonucleotides. J. Cell Biol. 117, 595–606 (1992).

    Article  CAS  PubMed  Google Scholar 

  39. Horton, A.C. & Ehlers, M.D. Dual modes of endoplasmic reticulum-to-Golgi transport in dendrites revealed by live-cell imaging. J. Neurosci. 23, 6188–6199 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Krijnse-Locker, J., Parton, R.G., Fuller, S.D., Griffiths, G. & Dotti, C.G. The organization of the endoplasmic reticulum and the intermediate compartment in cultured rat hippocampal neurons. Mol. Biol. Cell 6, 1315–1332 (1995).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Baas, P.W., Deitch, J.S., Black, M.M. & Banker, G.A. Polarity orientation of microtubules in hippocampal neurons: uniformity in the axon and nonuniformity in the dendrite. Proc. Natl. Acad. Sci. USA 85, 8335–8339 (1988).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Goldstein, L.S. & Yang, Z. Microtubule-based transport systems in neurons: the roles of kinesins and dyneins. Annu. Rev. Neurosci. 23, 39–71 (2000).

    Article  CAS  PubMed  Google Scholar 

  43. Lin, S.X., Gundersen, G.G. & Maxfield, F.R. Export from pericentriolar endocytic recycling compartment to cell surface depends on stable, detyrosinated (glu) microtubules and kinesin. Mol. Biol. Cell 13, 96–109 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Nakata, T. & Hirokawa, N. Microtubules provide directional cues for polarized axonal transport through interaction with kinesin motor head. J. Cell Biol. 162, 1045–1055 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Wilkinson, D.G. Multiple roles of EPH receptors and ephrins in neural development. Nat. Rev. Neurosci. 2, 155–164 (2001).

    Article  CAS  PubMed  Google Scholar 

  46. Kullander, K. & Klein, R. Mechanisms and functions of Eph and ephrin signalling. Nat. Rev. Mol. Cell Biol. 3, 475–486 (2002).

    Article  CAS  PubMed  Google Scholar 

  47. Yamaguchi, Y. & Pasquale, E.B. Eph receptors in the adult brain. Curr. Opin. Neurobiol. 14, 288–296 (2004).

    Article  CAS  PubMed  Google Scholar 

  48. Birgbauer, E., Oster, S.F., Severin, C.G. & Sretavan, D.W. Retinal axon growth cones respond to EphB extracellular domains as inhibitory axon guidance cues. Development 128, 3041–3048 (2001).

    CAS  PubMed  Google Scholar 

  49. Zhou, X., Suh, J., Cerretti, D.P., Zhou, R. & DiCicco-Bloom, E. Ephrins stimulate neurite outgrowth during early cortical neurogenesis. J. Neurosci. Res. 66, 1054–1063 (2001).

    Article  CAS  PubMed  Google Scholar 

  50. Cowan, C.A. & Henkemeyer, M. Ephrins in reverse, park and drive. Trends Cell Biol. 12, 339–346 (2002).

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank M. Greenberg for Eph receptor reagents, G. Banker for CD8, A. Hung for GRIP1(L732Y) and A. Akhmanova for pSuper-TRE. C.C.H. is a recipient of a long-term fellowship from Human Frontier Science Program Organization. I.M.E. and M.H. are supported by a National Institute of Mental Health grant and MH66332, respectively. M.S. is an Investigator of the Howard Hughes Medical Institute.

Author information

Authors and Affiliations

  1. The Picower Center for Learning and Memory, RIKEN-MIT Neuroscience Research Center, Howard Hughes Medical Institute, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA

    Casper C Hoogenraad, Aaron D Milstein & Morgan Sheng

  2. Division of Biomedical Sciences, University of California Riverside, Riverside, 92521, California, USA

    Iryna M Ethell

  3. Center for Developmental Biology and Kent Waldrep Center for Basic Research on Nerve Growth and Regeneration, University of Texas Southwestern Medical Center, Dallas, 75390, Texas, USA

    Mark Henkemeyer

Authors
  1. Casper C Hoogenraad
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Aaron D Milstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Iryna M Ethell
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mark Henkemeyer
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Morgan Sheng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Morgan Sheng.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

MAP2 staining in GRIP1-siRNA expressing hippocampal neurons. (PDF 1440 kb)

Supplementary Fig. 2

RNAi knock-down of GRIP1 impairs dendrite morphogenesis of pyramidal neurons in hippocampal slice cultures. (PDF 667 kb)

Supplementary Fig. 3

GRIP1 is essential for the formation and growth of dendrites but not axons in developing hippocampal neurons. (PDF 590 kb)

Supplementary Fig. 4

Reduced NMDA receptor clusters in GRIP1-knockdown neurons. (PDF 757 kb)

Supplementary Fig. 5

Brefeldin A treatment: EphB2 is trapped in the Golgi apparatus in GRIP1 siRNA expressing hippocampal neurons. (PDF 978 kb)

Supplementary Methods (PDF 47 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hoogenraad, C., Milstein, A., Ethell, I. et al. GRIP1 controls dendrite morphogenesis by regulating EphB receptor trafficking. Nat Neurosci 8, 906–915 (2005). https://doi.org/10.1038/nn1487

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nn1487

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