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The Potential Role Of TRPC3‐VAMP2 Interaction In Neurosecretion
Author(s) -
Bollimuntha Sunitha,
Watt John A,
Singh Brij B
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a117
Subject(s) - trpc3 , biology , microbiology and biotechnology , neurosecretion , calcium binding protein , neuroscience , medicine , calcium , biochemistry , receptor , trpc , transient receptor potential channel
AIMS To identify the interaction of TRPC3 with vesicle associated membrane protein 2 (VAMP2) and SNARE proteins and to understand its role in neurosecretion METHODS Yeast Two hybrid of TRPC3 with rat brain cDNA library showed VAMP2 interaction with its N‐terminus. Following this, immunoprecipitation analysis was done on PC12 cells expressing TRPC3 or N‐terminal deletion mutants to study the interactions with VAMP2 and other SNARE complex proteins. Function of TRPC3 and N‐terminal deletion mutants was done by calcium influx measurements on Fura‐2 loaded cells. Confocal and electron microscopy were done on rat brain sections to study localization of TRPC3 with VAMP2, Vasopressin (VP) and Oxytocin (OT) in neurosecretory vesicles of the supra optic nucleus (SON). Competitive ELISA was done to study the 1‐oleoyl‐2‐acetyl‐ sn ‐glycerol (OAG) evoked VP and OT release from neural lobe explants . RESULTS N‐terminal coiled coil domain of TRPC3 interacted with VAMP2. TRPC3 also interacted with α‐SNAP, synaptophysin and PSD95. N‐terminal deletion mutants had reduced calcium influx. TRPC3 colocalized with VP, OT and VAMP2 in the SON. A potentiation of VP and OT release upon stimulation of TRPC3 with OAG was seen. CONCLUSION The interaction of TRPC3 with VAMP2 is essential for critical physiological events like exocytosis and neurosecretion apart from regular calcium signaling. Supported by ‐ NCRR (NIH) & NDEPSCoR (NSF)