Cloning and pharmacological characterization of mouse TRPV1
Cloning and pharmacological characterization of mouse TRPV1
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DOI：
10.1016/j.neulet.2004.07.058
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发表时间：
2004-11-03
影响因子：
2.5
通讯作者：
Greenfeder, S
中科院分区：
文献类型：
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作者：
Correll, CC;Phelps, PT;Greenfeder, S
瞬时受体电位阳离子通道V1（TRPV 1）在外周伤害感受神经元中表达，并通过各种试剂（包括辣椒素、有害热、低细胞外pH和蛋白激酶C（PKC）的直接磷酸化）进行多模式激活。我们克隆并异源表达了小鼠TRPV 1（mTRPV 1），并利用基于FLIPR的钙成像来表征其功能，以测量对各种小分子激动剂、低pH和通过PKC直接磷酸化的功能反应。各种TRPV 1激动剂以激动剂效力的等级顺序激活mTRPV 1，（树脂毒素（RTX）= arvanil >辣椒素=奥瓦尼> OLDA > PPAHV）（EC 50值分别为0.15 +/- 0.04 nM、0.27 +/- 0.07 nM、9.1 +/- 1.2 nM、3.7 +/- 0.3 nM、258 +/- 105 nM和667 +/- 151 nM）。此外，mTRPV 1被激活的PKC激活剂佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）的低或添加。TRPV 1拮抗剂I-RTX或BCTC均能阻断辣椒素、pH和PKC诱导的mTRPV 1反应（IC 50（I-RTX）分别为0.35 +/- 0.12 nM、1.9 +/- 0.7 nM和0.80 +/- 0.68 nM，IC 50（BCTC）分别为1.3 +/- 0.36 nM、0.59 +/- 0.16 nM和0.37 +/- 0.15 nM）。然而.拮抗剂辣椒平仅能抑制辣椒素诱发的mTRPV 1反应，IC 50为1426 +/- 316 nM。用大鼠TRPV 1获得了类似的结果，而辣椒平阻断了人TRPV 1激活的所有模式。因此，mTRPV 1阳离子通道的分子药理学特征更接近大鼠TRPV 1，而不是人或豚鼠TRPV 1，分子药理学表明辣椒平可能是小鼠体内炎性疼痛模型的无效TRPV 1拮抗剂。(C)2004爱思唯尔爱尔兰有限公司保留所有权利。
The Transient Receptor Potential cation channel V1 (TRPV1) is expressed in peripheral nociceptive neurons and is subject to polymodal activation via various agents including capsaicin, noxious heat, low extracellular pH, and direct phosphorylation by protein kinase C (PKC). We have cloned and heterologously expressed mouse TRPV1 (mTRPV1) and characterized its function utilizing FLIPR-based calcium imaging to measure functional responses to various small molecule agonists, low pH and direct phosphorylation via PKC. The various TRPV1 agonists activated mTRPV1 with a rank order of agonist potency of (resiniferatoxin (RTX) = arvanil > capsaicin =olvanil > OLDA > PPAHV) (EC50 values of 0.15 +/- 0.04 nM, 0.27 +/- 0.07 nM, 9.1 +/- 1.2 nM, 3.7 +/- 0.3 nM, 258 +/- 105 nM, and 667 +/- 151 nM, respectively). Additionally, mTRPV1 was activated by either low or with addition of the PKC activator phorbol 12-myristate 13-acetate (PMA). The TRPV1 antagonists iodinated-resiniferatoxin (I-RTX) or BCTC were both able to block capsaicin, pH and PKC-induced responses of mTRPV1 (IC50 (I-RTX) = 0.35 +/- 0.12 nM, 1.9 +/- 0.7 nM, and 0.80 +/- 0.68 nM, IC50 (BCTC) = 1.3 +/- 0.36 nM, 0.59 +/- 0.16 nM, and 0.37 +/- 0.15 nM, respectively). However. the antagonist capsazepine was only able to inhibit a capsaicin-evoked response of mTRPV1 with an IC50 of 1426 +/- 316 nM. Comparable results were achieved with rat TRPV1, while capsazepine blocked all modes of human TRPV1 activation. Thus, the mTRPV1 cation channel has a molecular pharmacological profile more akin to rat TRPV 1 than either human or guinea pig TRPV1 and the molecular pharmacology suggests that capsazepine may be an ineffective TRPV 1 antagonist for in vivo models of inflammatory pain in the mouse. (C) 2004 Elsevier Ireland Ltd. All rights reserved.