Functional expression of TrpC1: a human homologue of the Drosophila Trp channel.
TrpC1 appears to be a store-operated channel (SOC) when expressed in mammalian cells. In the present study, TrpC1 was expressed in Sf9 insect cells using the baculovirus expression system. Expression of TrpC1 caused an increase in basal cytosolic free Ca2+ concentration ([Ca2+]i) as a function of post-infection time. Basal Ba2+ influx, an index of plasmalemmal Ca2+ permeability, was also increased and was blocked by La3+. Although the thapsigargin-induced change in [Ca2+]i was greater in TrpC1-expressing cells than controls, Ba2+ influx was unaffected by thapsigargin. Whole-cell membrane currents recorded in TrpC1-expressing cells increased as a function of post-infection time and were (1) inwardly rectifying in symmetrical sodium gluconate solutions, (2) non-selective with respect to Na+, Ca2+ and Ba2+, and (3) blocked by La3+. Furthermore TrpC1 currents were unaffected by (1) thapsigargin, (2) dialysis of the cell with Ins(1,4,5)P3 or (3) dialysis of the cell with solutions containing high concentrations of the Ca2+ chelator, EGTA. These results suggest that TrpC1 forms non-selective cation channels that are constitutively active when expressed in Sf9 cells, but insensitive to depletion of the internal Ca2+ stores. Thus TrpC1 may be a subunit of a SOC which alone can form functional channels in Sf9 cells, but which requires additional subunits or cytoplasmic factors present in mammalian cells for expression of SOC activity.
Sinkins
W G
WG
Rammelkamp Center for Education and Research, MetroHealth Campus, Case Western Reserve University, Cleveland, OH, 44109-1998, USA.
Estacion
M
M
Schilling
W P
WP
D000818:Animals; D002118:Calcium / Q000378:metabolism; D015220:Calcium Channels; D002460:Cell Line; D004330:Drosophila; D004594:Electrophysiology; D006801:Humans; D007473:Ion Channels / Q000235:genetics / Q000378:metabolism*; D017136:Ion Transport; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D050052:TRPC Cation Channels
D015220:Calcium Channels; D007473:Ion Channels; D011994:Recombinant Proteins; D050052:TRPC Cation Channels; C096351:transient receptor potential cation channel, subfamily C, member 1; D002118:Calcium