New Modes of GPCR Signalling
New Modes of GPCR Signalling
New Modes of GPCR Signalling
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Role <strong>of</strong> Heat on the Expression and Function <strong>of</strong> KCNQ2<br />
Fang Yu#, Yuncui Wang#, Yanlan Liu, Wanhong Liu, Biwen Peng* and Xiaohua,He*<br />
School <strong>of</strong> Medicine, Wuhan University, Wuhan 430071, China<br />
# Equal contribution to this work<br />
Objective: KCNQ2 (Kv7.2), Voltage-dependent potassium channels, plays a key role in<br />
regulating neuronal excitability. Mutations in neuronal KCNQ2/3 subunits, the<br />
molecular correlates <strong>of</strong> M current, have previously been linked with benign familial<br />
neonatal epilepsy (BFNC). In this study, we measured the temperature sensitivity <strong>of</strong><br />
KCNQ2 channel and determined whether temperature affected the excitability <strong>of</strong> the<br />
nervous system by the regulation <strong>of</strong> KCNQ2 channels.Methods: The wild type and<br />
mutant cDNAs <strong>of</strong> human KCNQ2, R214W and Y284C genes were subcloned into<br />
pEGFP-C1 plasmid, and then were transfected into 293T cells respectively. The<br />
KCNQ2 protein expression at different temperature was detected by fluorescence<br />
confocal microscopy and Western-blotting. While whole-cell patch clamp was<br />
performed to record the effect <strong>of</strong> temperature electrophysiological activity <strong>of</strong> KCNQ2<br />
potassium channel. Results: Total fluorescence intensity values <strong>of</strong> wtKCNQ2,R214W<br />
and Y284C were enhanced with the temperature increasing from 37℃ to 40 ℃<br />
(P
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