TRPV1 channels are gated by a variety of thermal, chemical, and mechanical stimuli. We used optical recording of Ca2+ influx through TRPV1 to measure activity and mobility of single TRPV1 molecules in isolated dorsal root ganglion neurons and cell lines. The opening of single TRPV1 channels produced sparklets, representing localized regions of elevated Ca2+. Unlike sparklets reported for L-type Ca2+ channels, TRPV4 channels, and AchR channels, TRPV1 channels diffused laterally in the plasma membrane as they gated. Mobility was highly variable from channel-to-channel and, to a smaller extent, from cell to cell. Most surprisingly, we found that mobility decreased upon channel activation by capsaicin, but only in the presence of extracellular Ca2+. We propose that decreased mobility of open TRPV1 could act as a diffusion trap to concentrate channels in cell regions with high activity.
Animal experimentation: Many local labs generating transgenic mice have a surplus of wild-type littermates that are euthanized after phenotyping. We have arranged to be present during the euthanasia process and immediately take the carcass for harvesting of the tissue we require. These arrangements are consistent with the principle of reducing the total number of animals used in biomedical experimentation and are approved by AALAC and the University of Washington IACUC.
- Richard Aldrich, The University of Texas at Austin, United States
© 2015, Senning & Gordon
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