VAMP2-dependent exocytosis regulates plasma membrane insertion of TRPC3 channels and contributes to agonist-stimulated Ca2+ influx

Brij B. Singh, Timothy P. Lockwich, Bidhan C. Bandyopadhyay, Xibao Liu, Sunitha Bollimuntha, So Ching Brazer, Christian Combs, Sunit Das, A. G.Miriam Leenders, Zu Hang Sheng, Mark A. Knepper, Suresh V. Ambudkar, Indu S. Ambudkar

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173 Scopus citations

Abstract

The mechanism(s) involved in agonist-stimulation of TRPC3 channels is not yet known. Here we demonstrate that TRPC3-N terminus interacts with VAMP2 and αSNAP. Further, endogenous and exogenously expressed TRPC3 colocalized and coimmunoprecipitated with SNARE proteins in neuronal and epithelial cells. Imaging of GFP-TRPC3 revealed its localization in the plasma membrane region and in mobile intracellular vesicles. Recovery of TRPC3-GFP fluorescence after photobleaching of the plasma membrane region was decreased by brefeldin-A or BAPTA-AM. Cleavage of VAMP2 with tetanus toxin (TeNT) did not prevent delivery of TRPC3 to the plasma membrane region but reduced its surface expression. TeNT also decreased carbachol and OAG, but not thapsigargin, stimulated Ca 2+ influx. Importantly, carbachol, not thapsigargin, increased surface expression of TRPC3 that was attenuated by TeNT and not by BAPTA. In aggregate, these data suggest that VAMP2-dependent exocytosis regulates plasma membrane insertion of TRPC3 channels and contributes to carbachol-stimulation of Ca2+ influx.

Original languageEnglish (US)
Pages (from-to)635-646
Number of pages12
JournalMolecular Cell
Volume15
Issue number4
DOIs
StatePublished - Aug 27 2004
Externally publishedYes

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ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Singh, B. B., Lockwich, T. P., Bandyopadhyay, B. C., Liu, X., Bollimuntha, S., Brazer, S. C., Combs, C., Das, S., Leenders, A. G. M., Sheng, Z. H., Knepper, M. A., Ambudkar, S. V., & Ambudkar, I. S. (2004). VAMP2-dependent exocytosis regulates plasma membrane insertion of TRPC3 channels and contributes to agonist-stimulated Ca2+ influx. Molecular Cell, 15(4), 635-646. https://doi.org/10.1016/j.molcel.2004.07.010