Ligand-induced dynamic membrane changes and cell deletion conferred by vanilloid receptor 1
Ligand-induced dynamic membrane changes and cell deletion conferred by vanilloid receptor 1
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DOI：
10.1074/jbc.m008392200
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发表时间：
2001-04-06
影响因子：
4.8
通讯作者：
Iadarola, MJ
中科院分区：
文献类型：
--
作者：
Olah, Z;Szabo, T;Iadarola, MJ
研究了香草酸诱导的细胞毒性的真实的时间动力学和表达野生型香草酸受体（VR 1）的伤害性神经元的特异性缺失。VR 1的C-末端用27-kDa增强型绿色荧光蛋白（eGFP）或12-氨基酸的β-表位标记。暴露于树脂毒素后，VR 1 eGFP或VR 1 β表达细胞表现出与表达未标记VR 1的细胞相似的药理学反应。在香草素暴露的几秒钟内，细胞内游离钙（[Ca 2 +]（i））在表达VR 1的细胞中升高。VR 1的功能库也定位于内质网，其在不存在细胞外钙的情况下，也能够在激动剂处理后释放钙。共聚焦成像揭示了树脂毒素治疗：诱导线粒体和内质网囊泡化（类似于1 min），核膜破裂（5-10 min）和细胞裂解（1-2 h），伤害感受性初级感觉神经元内源性表达VR 1，树脂毒素处理诱导[Ca 2 +]（i）的突然增加和细胞选择性的线粒体破坏，因为神经胶质和非VR 1表达神经元不受影响。细胞毒性的早期标志之后是VR 1表达细胞的特异性缺失。这些数据表明，香草素破坏细胞体内的重要细胞器，如果给予感觉神经节，可以用来快速和选择性地删除伤害性神经元。
The real time dynamics of vanilloid-induced cytotoxicity and the specific deletion of nociceptive neurons expressing the wild-type vanilloid receptor (VR1) were investigated. VR1 was C-terminally tagged with either the 27-kDa enhanced green fluorescent protein (eGFP) or a 12-amino acid epsilon -epitope. Upon exposure to resiniferatoxin, VR1eGFP- or VR1 epsilon -expressing cells exhibited pharmacological responses similar to those of cells expressing the untagged VR1. Within seconds of vanilloid exposure, the intracellular free calcium ([Ca2+](i)) was elevated in cells expressing VR1, A functional pool of VR1 also was localized to the endoplasmic reticulum that, in the absence of extracellular calcium, also was capable of releasing calcium upon agonist treatment. Confocal imaging disclosed that resiniferatoxin treatment: induced vesiculation of the mitochondria and the endoplasmic reticulum (similar to1 min), nuclear membrane disruption (5-10 min), and cell lysis (1-2 h), Nociceptive primary sensory neurons endogenously express VR1, and resiniferatoxin treatment induced a sudden increase in [Ca2+](i) and mitochondrial disruption which was cell-selective, as glia and non-VR1-expressing neurons were unaffected. Early hallmarks of cytotoxicity were followed by specific deletion of VR1-expressing cells. These data demonstrate that vanilloids disrupt vital organelles within the cell body and, if administered to sensory ganglia, may be employed to rapidly and selectively delete nociceptive neurons.