Abstract
The transient receptor potential vanilloid (TRPV) family of ion channels represents a group of non-selective cation channels remarkable for their diversity of function in nociception, thermosensation, mechanosensation and osmosensation. At present the bulk of our understanding of these channels comprises how their function is directly regulated by agonists and local environment. We and others have previously shown that TRPV1 channels can be regulated by trafficking. A number of important questions remain. How do the insertion and removal rates of TRPV1 as well as the distribution of the channel over the plasma membrane allow the cell to regulate the action of this channel? Is active transport of TRPV1 an aspect of this spatial regulation? We have addressed these questions by applying total internal reflection fluorescence (TIRF) imaging to TRPV1 channels tagged with a fluorescent protein. With TIRF imaging it becomes possible to discern small clusters of TRPV1 when the channel is expressed at low levels in cultured F-11 cells, and we implement this approach to address mechanisms of TRPV1 control in cells. Funding provided through the University of Washington training grant in Cardiovascular Pathology (NIH).
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