Translocation of TRPV2 channel induced by focal administration of mechanical stress.
Translocation of TRPV2 channel induced by focal administration of mechanical stress.
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DOI：
10.14814/phy2.12296
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发表时间：
2015-02-01
影响因子：
2.5
通讯作者：
Kojima I
中科院分区：
文献类型：
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作者：
Nagasawa M;Kojima I
在HT 1080细胞中研究了局部机械应力对TRPV 2定位的影响，其中在TRPV通道家族的成员中仅检测到TRPV 2的mRNA。通过使用玻璃移液管添加负压来施加机械应力。当施加局部机械应力时，移液管下方的亚质膜Ca 2+浓度（[Ca 2 +]s）增加，其在整个细胞中传播。[Ca 2 +]s的增加被钌红或敲低TRPV 2阻断。通过去除细胞外Ca 2+、加入磷脂酰肌醇3-激酶抑制剂LY 29034和转染显性阴性Rac，未观察到[Ca 2 +]s升高。在表达GFP-TRPV 2和RFP-Akt的细胞中，局部机械应力的施用诱导GFP-TRPV 2在移液管下方的积累。RFP-Akt也积累到同一位点。钆阻断了由局灶性机械应力引起的[Ca 2 +]s的升高，也减弱了TRPV 2的积累。当GFP-TRPV 1、GFP-TRPV 3、GFP-TRPV 4、GFP-TRPV 5或GFP-TRPV 6异位转染到HT 1080细胞中时，只有GFP-TRPV 4响应于局部机械应力而积聚在移液管下方。这些结果表明TRPV 2易位到接受局部机械应力的部位并增加[Ca 2 +]s。
The effect of focal mechanical stress on the localization of TRPV2 was investigated in HT1080 cells, where only mRNA for TRPV2 was detected among members of the TRPV channel family. Mechanical stress was applied by adding negative pressure using a glass pipette. When focal mechanical stress was applied, subplasma membrane Ca2+ concentration ([Ca2+]s) was increased beneath the pipette, which propagated throughout the cell. The increase in [Ca2+]s was blocked by ruthenium red or by knocking down TRPV2. Elevation of [Ca2+]s was not observed by removal of extracellular Ca2+, by an addition of a phosphatidylinositol 3-kinase inhibitor LY29034, and by transfection of dominant-negative Rac. In cells expressing GFP-TRPV2 and RFP-Akt, administration of focal mechanical stress induced accumulation of GFP-TRPV2 beneath the pipette. RFP-Akt was also accumulated to the same site. Gadolinium blocked the elevation of [Ca2+]s induced by focal mechanical stress and also attenuated accumulation of TRPV2. When GFP-TRPV1, GFP-TRPV3, GFP-TRPV4, GFP-TRPV5, or GFP-TRPV6 was transfected ectopically in HT1080 cells, only GFP-TRPV4 was accumulated beneath the pipette in response to the focal mechanical stress. These results indicate that TRPV2 translocates to the site receiving a focal mechanical stress and increases [Ca2+]s.