Purinoceptor-mediated calcium signaling in primary neuron-glia trigeminal cultures
Purinoceptor-mediated calcium signaling in primary neuron-glia trigeminal cultures
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DOI：
10.1016/j.ceca.2007.10.003
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发表时间：
2008-06-01
期刊：
影响因子：
4
通讯作者：
Abbracchio, Maria P.
中科院分区：
文献类型：
--
作者：
Ceruti, Stefania;Furnagalli, Marta;Abbracchio, Maria P.
细胞外核苷酸受体（P2X 钙通道和磷脂酶 C 偶联 P2Y 受体）在疼痛信号传导中发挥关键作用，但对其在三叉神经节中的功能知之甚少，三叉神经节的过度激活会导致偏头痛的发生。在这里，我们表征了原代小鼠神经元-胶质细胞三叉神经元培养物中通过 P2X(3) 和 P2Y 受体的钙信号传导。与完整神经节的比较表明，在分离的培养物中，感觉神经元至少部分保留了它们与卫星神经胶质细胞的物理关系。 RT-PCR 表明 P2X(2)/P2X(3) 的表达（通过免疫细胞化学证实）和所有克隆的 P2Y 受体的表达。使用亚型选择性 P2 激动剂/拮抗剂进行的单细胞钙成像显示，神经元和神经胶质细胞上存在功能性神经元 P2X(3)，以及 ADP 敏感的 P2Y(1,12,13) 和 UTP 激活的 P2Y(2)/P2Y(4) 受体。神经胶质细胞的钙反应要高得多，神经胶质细胞也对 UDP 做出反应，表明 P2Y(6) 受体具有功能。为了研究三叉神经节 P2 受体是否在促炎剂治疗后受到调节，将培养物急性（最多 3 分钟）或长期（24 小时）暴露于缓激肽。这导致致痛 P2X(3) 受体介导的钙反应增强，随后在 24 小时下调。在该暴露时间，卫星胶质细胞中的 P2Y 受体反应反而上调，表明 P2 受体在疼痛信号传导中存在复杂的调节。 (C) 2007 Elsevier Ltd. 保留所有权利。
Receptors for extracellular nucleotides (the P2X-calcium channels and the phospholipase C-coupled P2Y receptors) play key roles in pain signaling, but little is known on their function in trigerninal ganglia, whose hyperactivation leads to the development of migraine pain. Here we characterize calcium signaling via P2X(3) and P2Y receptors in primary mouse neuron-glia trigerninal cultures. Comparison with intact ganglion showed that, in dissociated cultures, sensory neurons retain, at least in part, their physical relationships with satellite glia. RT-PCR indicated expression of P2X(2)/P2X(3) (confirmed by immunocytochemistry) and of all cloned P2Y receptors. Single-cell calcium imaging with subtype-selective P2-agonists/antagonists revealed presence of functional neuronal P2X(3), as well as of ADP-sensitive P2Y(1,12,13) and UTP-activated P2Y(2)/P2Y(4) receptors on both neurons and glia. Calcium responses were much higher in glia, that also responded to UDP, suggesting functional P2Y(6) receptors. To study whether trigeminal ganglia P2 receptors are modulated upon treatment with pro-inflammatory agents, cultures were acutely (up to 3 min) or chronically (24 h) exposed to bradykinin. This resulted in potentiation of algogenic P2X(3) receptor-mediated calcium responses followed by their down-regulation at 24 h. At this exposure time, P2Y receptors responses in satellite glia were instead upregulated, suggesting a complex modulation of P2 receptors in pain signaling. (C) 2007 Elsevier Ltd. All rights reserved.