Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, Hubei, China.
PLoS One. 2013 Oct 16;8(10):e78203. doi: 10.1371/journal.pone.0078203. eCollection 2013.
The transient receptor potential vanilloid receptor 1 (TRPV1) channel is a nonselective cation channel activated by a variety of exogenous and endogenous physical and chemical stimuli, such as temperature (≥42 °C), capsaicin, a pungent compound in hot chili peppers, and allyl isothiocyanate. Large-conductance calcium- and voltage-activated potassium (BK) channels regulate the electric activities and neurotransmitter releases in excitable cells, responding to changes in membrane potentials and elevation of cytosolic calcium ions (Ca(2+)). However, it is unknown whether the TRPV1 channels are coupled with the BK channels. Using patch-clamp recording combined with an infrared laser device, we found that BK channels could be activated at 0 mV by a Ca(2+) influx through TRPV1 channels not the intracellular calcium stores in submilliseconds. The local calcium concentration around BK is estimated over 10 μM. The crosstalk could be affected by 10 mM BAPTA, whereas 5 mM EGTA was ineffectual. Fluorescence and co-immunoprecipitation experiments also showed that BK and TRPV1 were able to form a TRPV1-BK complex. Furthermore, we demonstrated that the TRPV1-BK coupling also occurs in dosal root ganglion (DRG) cells, which plays a critical physiological role in regulating the "pain" signal transduction pathway in the peripheral nervous system.
瞬时受体电位香草酸受体 1(TRPV1)通道是一种非选择性阳离子通道,可被多种外源性和内源性物理和化学刺激激活,如温度(≥42°C)、辣椒素、热辣椒中的一种刺激性化合物,以及丙烯基异硫氰酸酯。大电导钙和电压激活钾(BK)通道调节兴奋细胞的电活动和神经递质释放,响应膜电位变化和细胞浆钙离子(Ca2+)浓度升高。然而,TRPV1 通道是否与 BK 通道偶联尚不清楚。使用膜片钳记录结合红外激光装置,我们发现 BK 通道可在 0 mV 时通过 TRPV1 通道而不是细胞内钙库在亚毫秒内激活 Ca2+内流。BK 周围的局部钙离子浓度估计超过 10 μM。这种串扰可受 10 mM BAPTA 影响,而 5 mM EGTA 无效。荧光和共免疫沉淀实验也表明 BK 和 TRPV1 能够形成 TRPV1-BK 复合物。此外,我们证明 TRPV1-BK 偶联也发生在背根神经节(DRG)细胞中,在调节外周神经系统中“疼痛”信号转导途径方面发挥着关键的生理作用。