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UC Davis researchers discover key to body's ability to detect subtle temperature changes

In the current study, Zheng and his colleagues focused on a group of ion channels called transient receptor potential (TRP) channels. In all, there are more than 20 TRP channels. Zhengs group studied four of the six channels that have been shown to be involved in sensing temperature.

Previous studies concluded that different thermosensitive TRP channel subunits did not coassemble, Zheng said. He realized, however, that there were some technical limitations to the previous work. So, he and his colleagues decided to use a technique they developed last year, called spectra FRET. Spectra FRET, or spectroscopy-based fluorescence resonance energy transfer, allows the researcher to observe interactions between different channel subunits under a microscope.

"This technique allows us to look at the channel subunit composition in real-time in live cells," Zheng said.

In the current experiments, cDNA coding for particular subunits is linked to cDNA coding for fluorescent proteins and then added to a culture of human embryonic kidney cells. The cells take up the DNA and then express the channel proteins, each now having a fluorescent protein tag. The researchers then observed the cells in the spectra FRET apparatus.

"Using spectra FRET, we were able to focus on just the signal from the plasma membrane," Zheng explained. "What we found was that the subunits of one kind of heat-sensitive channel coassembled with subunits of other heat-sensitive channels to form new channels. This means that instead of four heat-sensitive channels we have a potential of 256 heat-sensitive channels with potentially different temperature sensitivity ranges."

Zheng and his colleagues then confirmed their results using a technique called patch clamping that allowed them to record the electrical current flowing through individual open channels.

"Using these single-molecule recordings, we see many different channel types," Zheng
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Contact: Carole Gan
carole.gan@ucdmc.ucdavis.edu
916-734-9047
University of California, Davis - Health System
26-Feb-2007


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