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J. Biol. Chem., Vol. 283, Issue 1, 416-426, January 4, 2008

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TRPC4 in Rat Dorsal Root Ganglion Neurons Is Increased after Nerve Injury and Is Necessary for Neurite Outgrowth^*

From the Neuroscience Centre, Institute of Cell and Molecular Sciences, Queen Mary University of London, London E1 2AT, United Kingdom

Canonical transient receptor potential (TRPC) receptors are Ca²⁺-permeable cation channels that have a variety of physiological functions and may be involved in neuronal development and plasticity. We investigated the expression profile of TRPC channels in adult rat dorsal root ganglia (DRG) after nerve injury and examined the role of TRPC4 in neurite outgrowth in cultured DRG neurons. Sciatic nerve transection and microinjection of dibutyryl cAMP were employed to induce axonal regeneration in vivo. TRPC4 mRNA was significantly increased whereas TRPC1, TRPC3, TRPC6, and TRPC7 remained unaltered after nerve injury or dibutyryl-cAMP microinjection. The increases in TRPC4 transcript and protein were transient with maximal levels reached at 2 or 7 days, respectively. In addition, TRPC4 transcript in ND7/23 and NDC cells, hybrid cell lines derived from neonatal DRG and neuroblastoma, was substantially increased on differentiation, characterized by neurite outgrowth. In adult DRG, TRPC4 immunoreactivity was found in small and large neurons, and nerve injury increased the number of TRPC4-immunoreactive cells, particularly in large neurons. TRPC4 immunoreactivity was present in growth cones at various stages of DRG neurite outgrowth in vitro. Suppression of TRPC4 by a specific small interfering RNA or antisense significantly reduced the length of neurites in cultured DRG neurons. Expression of short hairpin RNA significantly down-regulated TRPC4 protein level and shortened neurite lengths in differentiated ND7/23 cells. The reduction in neurite lengths in ND7/23 cells was rescued by overexpression of human TRPC4. Our results suggest that TRPC4 contributes to axonal regeneration after nerve injury.

Received for publication, April 16, 2007 , and in revised form, September 4, 2007.

^* This work was supported by grants from St. Bartholomew's and the Royal London Charitable Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

¹ To whom correspondence should be addressed: 4 Newark St., London E1 2AT, UK. Fax: 44-207-882-2180; E-mail: m.liu{at}qmul.ac.uk.

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