Rapid vesicular translocation and insertion of TRP channels

Authors:
Bezzerides VJ, Ramsey IS, Kotecha S, Greka A and Clapham DE
In:
Source: Nat Cell Biol
Publication Date: (2004)
Issue: 6(8): 709-720
Research Area:
Neurobiology
Cells used in publication:
Neuron, hippo/cortical, rat
Species: rat
Tissue Origin: brain
Platform:
Nucleofector® I/II/2b
Experiment
Primary rat hippocampal neurons were transfected by nucleofection with either one or combinations of expression plasmids for EGFP, a dominant negative mutant of TRPC5 fused to EGFP, and phosphatidylinositol 4-phosphate 5-kinase (PIP(5)K alpha). The effects on the neurite outgrowth rate were measured.
Abstract
The broadly expressed transient receptor potential (TRP) family of ion channels are permeant to cations, most resulting in increased intracellular calcium. However, their regulation and gating is not well understood. Here, we report that growth factor stimulation initiates the rapid translocation of the transient receptor potential ion channel, TRPC5, from vesicles held in reserve just under the plasma membrane. This process, which we term 'rapid vesicular insertion of TRP' (RiVIT), dramatically increases membrane-associated TRPC5 channels and functional TRPC5 current, resulting in tight spatial-temporal control of these Ca(2+)-permeant nonselective channels. Epidermal growth factor (EGF)-induced incorporation of functional TRP channels requires phosphatidylinositide 3-kinase (PI(3)K), the Rho GTPase Rac1 and phosphatidylinositol 4-phosphate 5-kinase (PIP(5)K alpha). The increase in TRPC5 availability affects neurite extension rates in cultured hippocampal neurons, and may be a general mechanism for initiating Ca(2+) influx and cell morphological changes in response to stimuli.