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


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.


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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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Correspondence to Mu-ming Poo.

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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).

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