Selective interference with TRPC3/6 channels disrupts OX1 receptor signalling via NCX and reveals a distinct calcium influx pathway

TRPC channels play significant roles in the regulation of neuronal plasticity and development. The mechanism by which these nonselective cation channels exert their trophic actions appears to involve entry of Ca2+ into the cells. Using a neuronal cell model (differentiated human IMR32 neuroblastoma cells), we demonstrate a central role for sodium entry via TRPC3/6 channels in receptor-mediated increases in intracellular calcium. These Ne+-dependent Ca2+ influxes, which were observed in a subpopulation of cells, were efficiently blocked by protein kinase C activation, by the Ne+/Ca2+ exchanger inhibitors, and by molecular disruption of TRPC3/6 channel function. On the other hand, another subpopulation of cells showed a Ne+-independent Ca2+ entry upon stimulation of the same receptors, orexin/hypocretin and bradykinin receptors. This second type of response was not affected by the above mentioned treatments, but it was sensitive to polyvalent cations, such as ruthenium red, spermine and Gd3+. The data suggest that a NCX-TRPC channel interaction constitutes an important functional unit in receptor-mediated Ca2+. influx in neuronal cells. (C) 2010 Elsevier Ltd. All rights reserved.