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Requirement of TRPC channels in netrin-1-induced chemotropic turning of nerve growth cones

Nature volume 434, pages 898–904 (2005)Cite this article

Abstract

Ion channels formed by the TRP (transient receptor potential) superfamily of proteins act as sensors for temperature, osmolarity, mechanical stress and taste1,2. The growth cones of developing axons are responsible for sensing extracellular guidance factors, many of which trigger Ca2+ influx at the growth cone3,4; however, the identity of the ion channels involved remains to be clarified. Here, we report that TRP-like channel activity exists in the growth cones of cultured Xenopus neurons and can be modulated by exposure to netrin-1 and brain-derived neurotrophic factor, two chemoattractants for axon guidance. Whole-cell recording from growth cones showed that netrin-1 induced a membrane depolarization, part of which remained after all major voltage-dependent channels were blocked. Furthermore, the membrane depolarization was sensitive to blockers of TRP channels. Pharmacological blockade of putative TRP currents or downregulation of Xenopus TRP-1 (xTRPC1) expression with a specific morpholino oligonucleotide abolished the growth-cone turning and Ca2+ elevation induced by a netrin-1 gradient. Thus, TRPC currents reflect early events in the growth cone's detection of some extracellular guidance signals, resulting in membrane depolarization and cytoplasmic Ca2+ elevation that mediates the turning of growth cones.

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Figure 1: Membrane depolarization and putative TRPC currents induced by netrin-1 and BDNF.
Figure 2: Growth-cone turning induced by netrin-1 or BDNF, and the effect of SKF-96365.
Figure 3: Effects of xTRPC1 MO on TRPC currents and growth-cone turning induced by netrin-1.
Figure 4: xTRPC1 mediates netrin-1-induced Ca2+ elevation in the growth cone.
Figure 5: xTRPC1 mediates netrin-1-induced membrane depolarization.

References

  1. Clapham, D. E. TRP channels as cellular sensors. Nature 426, 517–524 (2003)

    Article  ADS  CAS  Google Scholar 

  2. Montell, C., Birnbaumer, L. & Flockerzi, V. The TRP channels, a remarkably functional family. Cell 108, 595–598 (2002)

    Article  CAS  Google Scholar 

  3. Hong, K., Nishiyama, M., Henley, J., Tessier-Lavigne, M. & Poo, M. Calcium signalling in the guidance of nerve growth by netrin-1. Nature 403, 93–98 (2000)

    Article  ADS  CAS  Google Scholar 

  4. Henley, J. & Poo, M. M. Calcium mediates bidirectional growth cone turning induced by myelin-associated glycoprotein. Neuron 44, 909–916 (2004)

    Article  CAS  Google Scholar 

  5. Henley, J. & Poo, M. M. Guiding neuronal growth cones using Ca2+ signals. Trends Cell Biol. 14, 320–330 (2004)

    Article  CAS  Google Scholar 

  6. Nishiyama, M. et al. Cyclic AMP/GMP-dependent modulation of Ca2+ channels sets the polarity of nerve growth-cone turning. Nature 423, 990–995 (2003)

    Article  ADS  CAS  Google Scholar 

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

    Article  MathSciNet  CAS  Google Scholar 

  8. Kim, S. J. et al. Activation of the TRPC1 cation channel by metabotropic glutamate receptor mGluR1. Nature 426, 285–291 (2003)

    Article  ADS  CAS  Google Scholar 

  9. Li, H. S., Xu, X. Z. & Montell, C. Activation of a TRPC3-dependent cation current through the neurotrophin BDNF. Neuron 24, 261–273 (1999)

    Article  CAS  Google Scholar 

  10. Zhu, X., Jiang, M. & Birnbaumer, L. Receptor-activated Ca2+ influx via human Trp3 stably expressed in human embryonic kidney (HEK)293 cells. Evidence for a non-capacitative Ca2+ entry. J. Biol. Chem. 273, 133–142 (1998)

    Article  CAS  Google Scholar 

  11. Schwarz, G., Droogmans, G. & Nilius, B. Multiple effects of SKF 96365 on ionic currents and intracellular calcium in human endothelial cells. Cell Calcium 15, 45–54 (1994)

    Article  CAS  Google Scholar 

  12. Greka, A., Navarro, B., Oancea, E., Duggan, A. & Clapham, D. E. TRPC5 is a regulator of hippocampal neurite length and growth cone morphology. Nature Neurosci. 6, 837–845 (2003)

    Article  CAS  Google Scholar 

  13. Song, H. J., Ming, G. L. & Poo, M. M. cAMP-induced switching in turning direction of nerve growth cones. Nature 388, 275–279 (1997)

    Article  ADS  CAS  Google Scholar 

  14. Jung, S. et al. Lanthanides potentiate TRPC5 currents by an action at extracellular sites close to the pore mouth. J. Biol. Chem. 278, 3562–3571 (2003)

    Article  CAS  Google Scholar 

  15. Inoue, R. et al. The transient receptor potential protein homologue TRP6 is the essential component of vascular alpha(1)-adrenoceptor-activated Ca(2 + )-permeable cation channel. Circ. Res. 88, 325–332 (2001)

    Article  CAS  Google Scholar 

  16. Gomez, T. M., Robles, E., Poo, M. & Spitzer, N. C. Filopodial calcium transients promote substrate-dependent growth cone turning. Science 291, 1983–1987 (2001)

    Article  ADS  CAS  Google Scholar 

  17. Li, Y. et al. Essential role of TRPC channels in the guidance of nerve growth cones by brain-derived neurotrophic factor. Nature doi:10.1038/nature03477 (this issue)

  18. Bobanovic, L. K. et al. Molecular cloning and immunolocalization of a novel vertebrate trp homologue from Xenopus . Biochem. J. 340, 593–599 (1999)

    Article  CAS  Google Scholar 

  19. Brereton, H. M., Harland, M. L., Auld, A. M. & Barritt, G. J. Evidence that the TRP-1 protein is unlikely to account for store-operated Ca2+ inflow in Xenopus laevis oocytes. Mol. Cell. Biochem. 214, 63–74 (2000)

    Article  CAS  Google Scholar 

  20. Lintschinger, B. et al. Coassembly of Trp1 and Trp3 proteins generates diacylglycerol- and Ca2+-sensitive cation channels. J. Biol. Chem. 275, 27799–27805 (2000)

    CAS  PubMed  Google Scholar 

  21. Strubing, C., Krapivinsky, G., Krapivinsky, L. & Clapham, D. E. TRPC1 and TRPC5 form a novel cation channel in mammalian brain. Neuron 29, 645–655 (2001)

    Article  CAS  Google Scholar 

  22. Strubing, C., Krapivinsky, G., Krapivinsky, L. & Clapham, D. E. Formation of novel TRPC channels by complex subunit interactions in embryonic brain. J. Biol. Chem. 278, 39014–39019 (2003)

    Article  Google Scholar 

  23. Xu, X. Z., Li, H. S., Guggino, W. B. & Montell, C. Coassembly of TRP and TRPL produces a distinct store-operated conductance. Cell 89, 1155–1164 (1997)

    Article  CAS  Google Scholar 

  24. Amiri, H., Schultz, G. & Schaefer, M. FRET-based analysis of TRPC subunit stoichiometry. Cell Calcium 33, 463–470 (2003)

    Article  CAS  Google Scholar 

  25. Schaefer, M., Plant, T. D., Stresow, N., Albrecht, N. & Schultz, G. Functional differences between TRPC4 splice variants. J. Biol. Chem. 277, 3752–3759 (2002)

    Article  CAS  Google Scholar 

  26. Goel, M., Sinkins, W. G. & Schilling, W. P. Selective association of TRPC channel subunits in rat brain synaptosomes. J. Biol. Chem. 277, 48303–48310 (2002)

    Article  CAS  Google Scholar 

  27. Hofmann, T., Schaefer, M., Schultz, G. & Gudermann, T. Subunit composition of mammalian transient receptor potential channels in living cells. Proc. Natl Acad. Sci. USA 99, 7461–7466 (2002)

    Article  ADS  CAS  Google Scholar 

  28. Lohof, A. M. et al. Asymmetric modulation of cytosolic cAMP activity induces growth cone turning. J. Neurosci. 12, 1253–1261 (1992)

    Article  CAS  Google Scholar 

  29. Gu, X. & Spitzer, N. C. Low-threshold Ca2+ current and its role in spontaneous elevations of intracellular Ca2+ in developing Xenopus neurons. J. Neurosci. 13, 4936–4948 (1993)

    Article  CAS  Google Scholar 

  30. Schweitz, H. et al. Calcicludine, a venom peptide of the Kunitz-type protease inhibitor family, is a potent blocker of high-threshold Ca2+ channels with a high affinity for L-type channels in cerebellar granule neurons. Proc. Natl Acad. Sci. USA 91, 878–882 (1994)

    Article  ADS  CAS  Google Scholar 

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Acknowledgements

We thank G. J. Barritt for the gift of xTRP-1 antibody. This work was supported by a grant from NIH and a NSF predoctoral fellowship (to G.X.W.)

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Authors and Affiliations

  1. Division of Neurobiology, Department of Molecular and Cell Biology, Helen Wills Neuroscience Institute, University of California, Berkeley, California, 94720, USA

    Gordon X. Wang & Mu-ming Poo

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  1. Gordon X. Wang
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  2. Mu-ming Poo
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Correspondence to Mu-ming Poo.

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The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Figure S1

Comparison of neurite growth in Xenopus spinal neurons with or without loading of xTRPC1 morpholino oligo (MO). (JPG 24 kb)

Supplementary Figure S2

Western blot of xTRPC1 on Xenopus spinal cord extracts. (JPG 24 kb)

Supplementary Figure S3

Voltage-dependent channels are not affected by the blockade of TRPC1. (JPG 37 kb)

Supplementary Figure Legends

Legends to accompany the above Supplementary Figures. (DOC 23 kb)

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Wang, G., Poo, Mm. Requirement of TRPC channels in netrin-1-induced chemotropic turning of nerve growth cones. Nature 434, 898–904 (2005). https://doi.org/10.1038/nature03478

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