20-hydroxyeicosatetraenoic acid (20-HETE) activates mouse TRPC6 channels expressed in HEK293 cells
20-hydroxyeicosatetraenoic acid (20-HETE) activates mouse TRPC6 channels expressed in HEK293 cells
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DOI：
10.1074/jbc.m304437200
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发表时间：
2003-08-22
影响因子：
4.8
通讯作者：
Payet, MD
中科院分区：
文献类型：
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作者：
Basora, N;Boulay, G;Payet, MD
在本研究中，我们发现，类二十烷酸化合物，20-羟基二十碳四烯酸（20-HETE），一种重要的花生四烯酸代谢物，激活小鼠TRPC 6在一个稳定的，过度表达HEK 293细胞系，Hek-t6.11。应用20-HETE迅速诱导内向，非选择性电流在全细胞记录，这是抑制N-甲基-D-葡糖胺，1.8 mM的Ca 2+，和100 μ M的Gd 3+，但仍不受氟芬那酯和吲哚美辛。在低浓度的20-HETE（1-10 μ M）下获得的电流-电压关系显示轻微的向内整流，而测试的最高浓度的20-HETE（30 μ M）显示向外整流，如先前使用100 μ M 1-油酰基-2-乙酰基-sn-甘油对这些通道所示。剂量-反应曲线表明20-HETE激活TRPC 6通道，EC 50 = 0.8 μ M。使用由内而外的贴片进行的单通道分析显示，20-HETE使小鼠TRPC 6通道的开放概率增加了约3倍，并且这是以膜界定的方式。有趣的是，20-HETE没有引起细胞内Ca 2+浓度的变化。因此，我们已经确定了花生四烯酸代谢物，20-HETE，作为一种新的激活剂的TRP家族成员，TRPC 6。
In the present study, we show that the eicosanoid compound, 20-hydroxyeicosatetraenoic acid (20-HETE), an important arachidonic acid metabolite, activates mouse TRPC6 in a stable, overexpressing HEK293 cell line, Hek-t6.11. Application of 20-HETE rapidly induced an inward, non-selective current in whole-cell recordings, which was inhibited by N-methyl-D-glucamine, 1.8 mM Ca2+, and 100 muM Gd3+ but remained unaffected by flufenamate and indomethacin. The current-voltage relationship obtained at low concentrations of 20-HETE (1-10 muM) demonstrated slight inward rectification, whereas the highest concentration of 20-HETE tested ( 30 muM) showed outward rectification, as shown previously for these channels using 100 muM 1-oleoyl-2-acetyl-sn-glycerol. Dose-response curves indicate that 20-HETE activated TRPC6 channels with an EC50 = 0.8 muM. Single channel analysis using inside-out patches revealed that 20-HETE increased open probability of mouse TRPC6 channels similar to3-fold, and this was in a membrane-delimited fashion. Interestingly, 20-HETE did not provoke changes in intracellular Ca2+ concentrations. Thus, we have identified an arachidonic acid metabolite, 20-HETE, as a novel activator for a TRP family member, TRPC6.