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A second pathway for antidepressants
Date:2/7/2011

itous of these transmembrane proteins, gating the passage of potassium ions through neural membranes, which sets the excitability of the neuron. Earlier studies had shown that when the TREK1 gene is "knocked out" of mice, the mice display a depression-resistant phenotype that mimics the behavior of mice treated with fluoxetine and that the antidepressant inhibits the activity of the TREK1 channel. While these results pointed to a possible role for the TREK1 ion channel in the beneficial response to fluoxetine, the mechanism behind this activity was unclear.

"Studying what the different protein parts of an ion channel do is a huge challenge," Isacoff says. "Over the years, my group has developed techniques by which the domains of channel proteins can be labeled with site-specific fluorescent dyes. Structural rearrangements of the labeled sites in the channel can then be detected through changes in the fluorescence."

Isacoff and his group separated the C-terminal domain from the rest of the protein and tagged it with a green fluorescent protein (GFP) - a fluorescent protein from jellyfish commonly used to paint cells green for biological studies. Whereas the pore of the TREK1 ion channel is embedded in the plasma membrane of a neuron, the C-terminal is a short tail that protrudes out into the surrounding cytoplasm.

Using voltage clamps to measure electrical currents through the channel and fluorescence to monitor the disposition of the C-terminal domain, Isacoff and his group found that when the C-terminal tail is fully bound to the plasma membrane, the TREK1 potassium channel opens more; when the tail is unbound from the plasma membrane, the ion channel tends to close.

"We found that fluoxetine causes the isolated C-terminal domain to unbind from the membrane and also causes an inhibition of current from the full TREK1 channel," Isacoff says.

The next step will be to see how the C-terminal tail is affected by the pre
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Contact: Lynn Yarris
lcyarris@lbl.gov
510-486-5375
DOE/Lawrence Berkeley National Laboratory
Source:Eurekalert  

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