A role for hTRPC1 and lipid raft domains in store-mediated calcium entry in human platelets.
We have previously suggested that store-mediated Ca2+ entry (SMCE) in human platelets may be activated by a secretion-like coupling model, involving de novo coupling of the type II inositol 1,4,5-trisphosphate receptor (IP(3)RII) to the putative Ca2+ entry channel, hTRPC1. In other cells, hTRPC1 has been reported to be associated with cholesterol-rich lipid raft domains (LRDs) in the plasma membrane. Here we have shown that hTRPC1 is largely associated with detergent-resistant platelet membranes, from which it is partially released when the cells are depleted of cholesterol by treatment with methyl-beta-cyclodextrin (MBCD). MBCD treatment inhibited thapsigargin (TG)-evoked SMCE in a concentration-dependent manner, reducing it to 38.1+/-4.1% at a concentration of 10mM. Similarly, the Ca2+ entry evoked by thrombin (1unit/ml) was reduced to 48.2+/-4.5% of control following MBCD (10mM) treatment. Thrombin- and TG-evoked coupling between IP(3)RII and hTRPC1 was also reduced following cholesterol depletion. These results suggest that hTRPC1 is associated with LRDs in human platelets and that these domains are important for its participation in SMCE.
Brownlow
Sharon L
SL
Department of Physiology, University of Cambridge, Downing Street, Cambridge CB2 3EG, UK.
Harper
Alan G S
AG
Harper
Matthew T
MT
Sage
Stewart O
SO
D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000097:blood / Q000502:physiology*; D003505:Cyclodextrins / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D006801:Humans; D021962:Membrane Microdomains / Q000187:drug effects / Q000502:physiology*; D050052:TRPC Cation Channels; D047392:beta-Cyclodextrins
D015220:Calcium Channels; D003505:Cyclodextrins; D050052:TRPC Cation Channels; D047392:beta-Cyclodextrins; C108732:methyl-beta-cyclodextrin; C096351:transient receptor potential cation channel, subfamily C, member 1; D002118:Calcium