Inhibition of TRPC5 channels by intracellular ATP.
TRPC5 channels are Ca(2+)-permeable nonselective cation channels activated by G-protein-coupled receptors, although the mechanisms responsible for channel activation and regulation are poorly understood. Carbachol-activated TRPC5 currents were recorded by the whole-cell patch clamp technique from human embryonic kidney 293 cells transiently transfected with TRPC5 and the M1 muscarinic receptor. Some published studies of TRPC5 currents have included ATP and/or GTP in the patch pipette, whereas others used an ATP- and GTP-free pipette solution. We initially included these two nucleotides in the patch pipette but found that TRPC5 currents were absent or were very small. Recordings made with an ATP- and GTP-free pipette solution produced large and robust TRPC5 currents. Under these conditions, treatment of cells with Pasteurella multocida toxin, a selective inhibitor of Galpha(q), almost abolished TRPC5 currents indicating that Galpha(q) is necessary for activation of TRPC5 by the M1 receptor. To study the effect of intracellular ATP on TRPC5 channels, an intracellular perfusion system was used. Perfusion of ADP or control pipette solution had no effect, whereas perfusion of ATP or AMP-PNP, a nonhydrolyzable analog of ATP, significantly inhibited TRPC5 currents. Thus, the effects of ATP have structural specificity and probably involve a direct effect on the channel rather than a phosphorylation-mediated effect. The activity of TRPC5 channels may be linked to cellular metabolism via changes in ATP levels and could be involved in Ca(2+) overload occurring after ischemia when ATP is depleted.
Dattilo
Michael
M
Department of Physiology and Pharmacology, Box 31, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, New York 11203, USA.
Penington
Nicholas J
NJ
Williams
Keith
K
D000255:Adenosine Triphosphate / Q000502:physiology*; D002460:Cell Line; D006801:Humans; D050052:TRPC Cation Channels / Q000037:antagonists & inhibitors*
D050052:TRPC Cation Channels; C491780:TRPC5 protein, human; D000255:Adenosine Triphosphate