TY - JOUR
T1 - VAMP2-dependent exocytosis regulates plasma membrane insertion of TRPC3 channels and contributes to agonist-stimulated Ca2+ influx
AU - Singh, Brij B.
AU - Lockwich, Timothy P.
AU - Bandyopadhyay, Bidhan C.
AU - Liu, Xibao
AU - Bollimuntha, Sunitha
AU - Brazer, So Ching
AU - Combs, Christian
AU - Das, Sunit
AU - Leenders, A. G.Miriam
AU - Sheng, Zu Hang
AU - Knepper, Mark A.
AU - Ambudkar, Suresh V.
AU - Ambudkar, Indu S.
N1 - Funding Information:
We thank Dr. Martha Nowycky for her helpful comments. We are extremely grateful to Dr. Craig Montell for generously providing us with the anti-TRPC3 antibody and Dr. Michael Zhu for HA-TRPC3 and exo-HA-TRPC3 plasmids. Financial support for Brij Singh was provided in part by ND-BRIN(NIH).
PY - 2004/8/27
Y1 - 2004/8/27
N2 - 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.
AB - 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.
UR - https://www.scopus.com/pages/publications/4344685732
UR - https://www.scopus.com/inward/citedby.url?scp=4344685732&partnerID=8YFLogxK
U2 - 10.1016/j.molcel.2004.07.010
DO - 10.1016/j.molcel.2004.07.010
M3 - Article
C2 - 15327778
AN - SCOPUS:4344685732
SN - 1097-2765
VL - 15
SP - 635
EP - 646
JO - Molecular Cell
JF - Molecular Cell
IS - 4
ER -