Myosin X regulates netrin receptors and functions in axonal path-finding

Article metrics


Netrins regulate axon path-finding during development, but the underlying mechanisms are not well understood. Here, we provide evidence for the involvement of the unconventional myosin X (Myo X) in netrin-1 function. We find that Myo X interacts with the netrin receptor deleted in colorectal cancer (DCC) and neogenin, a DCC-related protein. Expression of Myo X redistributes DCC to the cell periphery or to the tips of neurites, whereas its silencing prevents DCC distribution in neurites. Moreover, expression of DCC, but not neogenin, stimulates Myo X-mediated formation and elongation of filopodia, suggesting that Myo X function may be differentially regulated by DCC and neogenin. The involvement of Myo X in netrin-1 function was further supported by the effects of inhibiting Myo X function in neurons. Cortical explants derived from mouse embryos expressing a motor-less Myo X exhibit reduced neurite outgrowth in response to netrin-1 and chick commissural neurons expressing the motor-less Myo X, or in which Myo X is silenced using microRNA (miRNA), show impaired axon projection in vivo. Taken together, these results identify a novel role for Myo X in regulating netrin-1 function.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1: Myo X interaction with DCC and neogenin.
Figure 2: Regulation of DCC distribution in neurons by Myo X.
Figure 3: Distinct regulation by DCC and neogenin of Myo X-mediated filopodium elongation in NLT cells.
Figure 4: Involvement of Myo X in netrin-1-induced cortical neurite outgrowth.
Figure 5: Impaired commissural neuronal axon projections in chicken embryos expressing motor-less Myo X or miRNA.


  1. 1

    Keino-Masu, K. et al. Deleted in Colorectal Cancer (DCC) encodes a netrin receptor. Cell 87, 175–185 (1996).

  2. 2

    Kolodziej, P. A. et al. frazzled encodes a Drosophila member of the DCC immunoglobulin subfamily and is required for CNS and motor axon guidance. Cell 87, 197–204 (1996).

  3. 3

    Chan, S. S. et al. UNC-40, a C. elegans homolog of DCC (Deleted in Colorectal Cancer), is required in motile cells responding to UNC-6 netrin cues. Cell 87, 187–195 (1996).

  4. 4

    Rajagopalan, S. et al. Neogenin mediates the action of repulsive guidance molecule. Nature Cell Biol. 6, 756–762 (2004).

  5. 5

    Matsunaga, E. et al. RGM and its receptor neogenin regulate neuronal survival. Nature Cell Biol. 6, 749–755 (2004).

  6. 6

    Ming, G. L. et al. cAMP-dependent growth cone guidance by netrin-1. Neuron 19, 1225–1235 (1997).

  7. 7

    Ming, G. et al. Phospholipase C-γ and phosphoinositide 3-kinase mediate cytoplasmic signaling in nerve growth cone guidance. Neuron 23, 139–148 (1999).

  8. 8

    Forcet, C. et al. Netrin-1-mediated axon outgrowth requires deleted in colorectal cancer-dependent MAPK activation. Nature 417, 443–447 (2002).

  9. 9

    Li, X., Saint-Cyr-Proulx, E., Aktories, K. & Lamarche-Vane, N. Rac1 and Cdc42 but not RhoA or Rho kinase activities are required for neurite outgrowth induced by the netrin-1 receptor DCC (Deleted in Colorectal Cancer) in N1E-115 reuroblastoma cells. J. Biol. Chem. 277, 15207–15214 (2002).

  10. 10

    Xie, Y. et al. Phosphatidylinositol transfer protein-α in netrin-1-induced PLC signalling and neurite outgrowth. Nature Cell Biol. 7, 1124–1132 (2005).

  11. 11

    Xie, Y. et al. DCC-dependent phospholipase C signaling in netrin-1-induced neurite elongation. J. Biol. Chem. 281, 2605–2611 (2006).

  12. 12

    Chang, C. et al. MIG-10/lamellipodin and AGE-1/PI3K promote axon guidance and outgrowth in response to slit and netrin. Curr. Biol. 16, 854–862 (2006).

  13. 13

    Ren, X. R. et al. Focal adhesion kinase in netrin-1 signaling. Nature Neurosci. 7, 1204–1212 (2004).

  14. 14

    Li, W. et al. Activation of FAK and Src are receptor-proximal events required for netrin signaling. Nature Neurosci. 7, 1213–1221 (2004).

  15. 15

    Liu, G. et al. Netrin requires focal adhesion kinase and Src family kinases for axon outgrowth and attraction. Nature Neurosci. 7, 1222–1232 (2004).

  16. 16

    Hu, G. et al. Mammalian homologs of seven in absentia regulate DCC via the ubiquitin-proteasome pathway. Genes Dev. 11, 2701–2714 (1997).

  17. 17

    Sousa, A. D. & Cheney, R. E. Myosin-X: a molecular motor at the cell's fingertips. Trends Cell Biol. 15, 533–539 (2005).

  18. 18

    Berg, J. S., Derfler, B. H., Pennisi, C. M., Corey, D. P. & Cheney, R. E. Myosin-X, a novel myosin with pleckstrin homology domains, associates with regions of dynamic actin. J. Cell Sci. 113, 3439–3451 (2000).

  19. 19

    Berg, J. S. & Cheney, R. E. Myosin-X is an unconventional myosin that undergoes intrafilopodial motility. Nature Cell Biol. 4, 246–250 (2002).

  20. 20

    Zhang, H. et al. Myosin-X provides a motor-based link between integrins and the cytoskeleton. Nature Cell Biol. 6, 523–531 (2004).

  21. 21

    Sousa, A. D., Berg, J. S., Robertson, B. W., Meeker, R. B. & Cheney, R. E. Myo10 in brain: developmental regulation, identification of a headless isoform and dynamics in neurons. J. Cell Sci. 119, 184–194 (2006).

  22. 22

    Metin, C., Deleglise, D., Serafini, T., Kennedy, T. E. & Tessier-Lavigne, M. A role for netrin-1 in the guidance of cortical efferents. Development 124, 5063–5074 (1997).

  23. 23

    Braisted, J. E. et al. Netrin-1 promotes thalamic axon growth and is required for proper development of the thalamocortical projection. J. Neurosci. 20, 5792–5801 (2000).

  24. 24

    Culotti, J. G. & Merz, D. C. DCC and netrins. Curr. Opin. Cell Biol. 10, 609–613 (1998).

  25. 25

    Huang, X., Cheng, H. J., Tessier-Lavigne, M. & Jin, Y. MAX-1, a novel PH/MyTH4/FERM domain cytoplasmic protein implicated in netrin-mediated axon repulsion. Neuron 34, 563–576 (2002).

  26. 26

    Li, H. S. et al. Vertebrate slit, a secreted ligand for the transmembrane protein roundabout, is a repellent for olfactory bulb axons. Cell 96, 807–818 (1999).

  27. 27

    Ren, X. R. et al. Regulation of CDC42 GTPase by proline-rich tyrosine kinase 2 interacting with PSGAP, a novel pleckstrin homology and Src homology 3 domain containing rhoGAP protein. J. Cell Biol. 152, 971–984 (2001).

  28. 28

    Wang, Q. et al. Regulation of the formation of osteoclastic actin rings by proline-rich tyrosine kinase 2 interacting with gelsolin. J. Cell Biol. 160, 565–575 (2003).

  29. 29

    Niwa, H., Yamamura, K. & Miyazaki, J. Efficient selection for high-expression transfectants with a novel eukaryotic vector. Gene 108, 193–199 (1991).

  30. 30

    Saito, T. & Nakatsuji, N. Efficient gene transfer into the embryonic mouse brain using in vivo electroporation. Dev. Biol. 240, 237–246 (2001).

  31. 31

    Richards, L. J., Koester, S. E., Tuttle, R. & O'Leary, D. D. Directed growth of early cortical axons is influenced by a chemoattractant released from an intermediate target. J. Neurosci. 17, 2445–2458 (1997).

  32. 32

    Finger, J. H. et al. The netrin 1 receptors Unc5h3 and Dcc are necessary at multiple choice points for the guidance of corticospinal tract axons. J. Neurosci. 22, 10346–10356 (2002).

Download references


This study was supported in part by grants from the National Institutes of Health (L.M. and W.C.X.), Muscular Dystrophy Association (L.M.), Philips Morris Research Program (L.M.), and National Nature Science Foundation of China (Y.Q.D. and W.C. X.).

Author information

All authors contributed to the experimental work and/or data analysis. Y.-Q.D. and W.-C.X. contributed to project planning.

Correspondence to Yu-Qiang Ding or Wen-Cheng Xiong.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Information

Supplementary Figures S1, S2, S3 and S4 (PDF 1036 kb)

Rights and permissions

Reprints and Permissions

About this article

Further reading