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

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.