Modulation of Transient Receptor Vanilloid 1 Activity by Transient Receptor Potential Ankyrin 1
Modulation of Transient Receptor Vanilloid 1 Activity by Transient Receptor Potential Ankyrin 1
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DOI：
10.1124/mol.113.088997
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发表时间：
2014-02-01
影响因子：
3.6
通讯作者：
Zoellner, Christian
中科院分区：
文献类型：
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作者：
Spahn, Viola;Stein, Christoph;Zoellner, Christian
瞬时受体电位香草酸1（TRPV 1）是一种非选择性配体门控阳离子通道，对有害热、质子和化学物质（如辣椒素）产生反应。TRPV 1在感觉神经元中表达，并在与组织损伤、炎症和神经病变相关的疼痛中起关键作用。瞬时受体电位锚蛋白1（TRPA 1）与TRPV 1共表达。它是由引起灼烧感的化合物激活的（例如。例如，在一个实施例中，芥子油）和间接地通过引起伤害感受器兴奋和疼痛超敏性的炎性环境的组分。先前的研究表明TRPV 1和TRPA 1信号通路相互作用。在这里，我们试图研究伤害感受神经元中这种相互作用的分子机制。我们首先使用放射性配体结合研究排除了两个通道的物理相互作用。通过显微荧光法、电生理实验、cAMP测量和定点突变，我们发现芥子油以钙和cAMP/蛋白激酶A（PKA）依赖的方式刺激TRPA 1后，TRPV 1增敏。TRPA 1刺激通过推定的PKA磷酸化位点丝氨酸116增强TRPV 1磷酸化。我们还检测到钙敏感性增加TRPV 1活性后TRPA 1激活背根神经节神经元。HC-030031对TRPA 1的抑制作用（1，2，3，6-四氢-1，3-二甲基-N-[4-（1-甲基乙基）苯基]-2，6-二氧代-7H-嘌呤-7乙酰胺，2-（1，3-二甲基-2，6-二氧代-1，2，3，6-四氢-7H-嘌呤-7-基）-N-（4-异丙基苯基）乙酰胺）（和钙不敏感的TRPA 1突变体D477 A）在其初始刺激后仍然显示出辣椒素诱导的TRPV 1活性增加。这排除了TRPV 1刺激后钙诱导的附加TRPA 1电流。我们的研究表明TRPV 1通过激活TRPA 1而致敏，这涉及腺苷酸环化酶，cAMP增加，随后的PKA易位和激活，以及TRPV 1在PKA磷酸化残基处的磷酸化。这表明TRP通道的交叉致敏有助于增强发炎组织中的疼痛敏感性。
Transient receptor potential vanilloid 1 (TRPV1) is a nonselective ligand-gated cation channel responding to noxious heat, protons, and chemicals such as capsaicin. TRPV1 is expressed in sensory neurons and plays a critical role in pain associated with tissue injury, inflammation, and nerve lesions. Transient receptor potential ankyrin 1 (TRPA1) is coexpressed with TRPV1. It is activated by compounds that cause a burning sensation (e. g., mustard oil) and, indirectly, by components of the inflammatory milieu eliciting nociceptor excitation and pain hypersensitivity. Previous studies indicate an interaction of TRPV1 and TRPA1 signaling pathways. Here we sought to examine the molecular mechanisms underlying such interactions in nociceptive neurons. We first excluded physical interactions of both channels using radioligand binding studies. By microfluorimetry, electrophysiological experiments, cAMP measurements, and site-directed mutagenesis we found a sensitization of TRPV1 after TRPA1 stimulation with mustard oil in a calciumand cAMP/protein kinase A (PKA)-dependent manner. TRPA1 stimulation enhanced TRPV1 phosphorylation via the putative PKA phosphorylation site serine 116. We also detected calcium-sensitive increased TRPV1 activity after TRPA1 activation in dorsal root ganglion neurons. The inhibition of TRPA1 by HC-030031 (1,2,3,6-tetrahydro-1,3-dimethyl-N-[4-(1-methylethyl)phenyl]-2,6-dioxo-7H-purine-7acetamide, 2-(1,3-dimethyl-2,6-dioxo-1,2,3,6-tetrahydro-7H-purin-7-yl)-N-(4-isopropylphenyl) acetamide) after its initial stimulation (and the calcium-insensitive TRPA1 mutant D477A) still showed increased capsaicin-induced TRPV1 activity. This excludes a calcium-induced additive TRPA1 current after TRPV1 stimulation. Our study shows sensitization of TRPV1 via activation of TRPA1, which involves adenylyl cyclase, increased cAMP, subsequent translocation and activation of PKA, and phosphorylation of TRPV1 at PKA phosphorylation residues. This suggests that cross-sensitization of TRP channels contributes to enhanced pain sensitivity in inflamed tissues.