Expression of Functional Receptor-coupled TRPC3 Channels in DT40 Triple Receptor InsP3 knockout Cells

Kartik Venkatachalam, Hong Tao Ma, Diana L. Ford, Donald L. Gill

Research output: Contribution to journalArticlepeer-review

137 Scopus citations

Abstract

The TRPC3 channel, an intensively studied member of the widely expressed transient receptor potential (TRP) family, is a Ca2+-conducting channel activated in response to phospholipase C-coupled receptors. Despite scrutiny, the receptor-induced mechanism to activate TRPC3 channels remains unclear. Evidence indicates TRPC3 channels interact directly with intracellular inositol 1,4,5-trisphosphate receptors (InsP3Rs) and that channel activation is mediated through coupling to InsP3Rs. TRPC3 channels were expressed in DT40 chicken B lymphocytes in which all three InsP 3R genes were deleted (DT40InsP3R-k/o). Endogenous B-cell receptors (BCR) coupled through Syk kinase to phospholipase C-γ (PLC-γ) activated the expressed TRPC3 channels in both DT40w/t and DT40InsP3R-k/o cells. The diacylglycerol (DAG) analogue 1-oleoyl-2-acetyl-sn-glycerol (OAG) also activated TRPC3 channels independently of InsP3Rs. BCR-induced TRPC3 activation was blocked by the PLC enzymic inhibitor, U-73122, and also blocked by wortmannin-induced PLC substrate depletion. Neither U-73122 nor wortmannin modified either OAG-induced TRPC3 activation or store-operated channel activation in DT40 cells. Cotransfection of cells with both G protein-coupled M5 muscarinic receptors and TRPC3 channels resulted in successful M5 coupling to open TRPC3 channels mediated by PLC-β. We conclude that TRPC3 channels are activated independently of InsP3Rs through DAG production resulting from receptor-mediated activation of either PLC-γ or PLC-β.

Original languageEnglish (US)
Pages (from-to)33980-33985
Number of pages6
JournalJournal of Biological Chemistry
Volume276
Issue number36
DOIs
StatePublished - Sep 7 2001

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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