Intercellular Odontoblast Communication via ATP Mediated by Pannexin-1 Channel and Phospholipase C-coupled Receptor Activation.
Intercellular Odontoblast Communication via ATP Mediated by Pannexin-1 Channel and Phospholipase C-coupled Receptor Activation.
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DOI：
10.3389/fphys.2015.00326
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发表时间：
2015
影响因子：
4
通讯作者：
Shibukawa Y
中科院分区：
文献类型：
--
作者：
Sato M;Furuya T;Kimura M;Kojima Y;Tazaki M;Sato T;Shibukawa Y
在成牙本质细胞机械刺激过程中，通过pannexin-1通道释放的细胞外ATP介导成牙本质细胞间的细胞间通讯。近年来，成牙本质细胞系，如小鼠成牙本质细胞谱系细胞，已被广泛用于研究生理/病理细胞功能。为了阐明成牙本质细胞系之间是否也通过可扩散的化学物质进行通讯，我们研究了机械刺激后小鼠成牙本质细胞系细胞间的化学通讯。使用装有标准细胞外溶液的玻璃移液管刺激单细胞。我们用fura-2测定了刺激细胞以及附近细胞的胞内游离Ca ~（2+）浓度（[Ca ~（2+）]i）。对单个成牙本质细胞的直接机械刺激增加[Ca 2 +]i，这表明对辣椒平敏感。此外，我们观察到增加[Ca 2 +]i不仅在机械刺激的成牙本质细胞，但也在附近的成牙本质细胞。我们可以观察到机械刺激诱导的[Ca 2 +]i增加刺激的人胚肾（HEK）293细胞，但不是在附近的HEK 293细胞。三磷酸腺苷（ATP）释放通道（pannexin-1）抑制剂以浓度和空间依赖性方式抑制附近成牙本质细胞[Ca 2 +]i的增加，但不抑制受刺激的成牙本质细胞[Ca 2 +]i的增加。此外，在磷脂酶C（PLC）抑制剂的存在下，在附近的成牙本质细胞中的[Ca 2 +]i的增加，机械刺激后的单个成牙本质细胞，被废除。在受刺激的成牙本质细胞附近，我们可以记录到一些由成牙本质细胞诱发的内向电流，但只有4.8%的成牙本质细胞可以观察到这种电流。这项研究的结果表明，ATP通过泛连接蛋白-1从机械刺激的成牙本质细胞中释放，该成牙本质细胞通过PLC偶联核苷酸受体的主要激活向附近的成牙本质细胞传递信号。
Extracellular ATP released via pannexin-1 channels, in response to the activation of mechanosensitive-TRP channels during odontoblast mechanical stimulation, mediates intercellular communication among odontoblasts in dental pulp slice preparation dissected from rat incisor. Recently, odontoblast cell lines, such as mouse odontoblast lineage cells, have been widely used to investigate physiological/pathological cellular functions. To clarify whether the odontoblast cell lines also communicate with each other by diffusible chemical substance(s), we investigated the chemical intercellular communication among cells from mouse odontoblast cell lines following mechanical stimulation. A single cell was stimulated using a glass pipette filled with standard extracellular solution. We measured intracellular free Ca2+ concentration ([Ca2+]i) by fura-2 in stimulated cells, as well as in cells located nearby. Direct mechanical stimulation to a single odontoblast increased [Ca2+]i, which showed sensitivity to capsazepine. In addition, we observed increases in [Ca2+]i not only in the mechanically stimulated odontoblast, but also in nearby odontoblasts. We could observe mechanical stimulation-induced increase in [Ca2+]i in a stimulated human embryo kidney (HEK) 293 cell, but not in nearby HEK293 cells. The increase in [Ca2+]i in nearby odontoblasts, but not in the stimulated odontoblast, was inhibited by adenosine triphosphate (ATP) release channel (pannexin-1) inhibitor in a concentration- and spatial-dependent manner. Moreover, in the presence of phospholipase C (PLC) inhibitor, the increase in [Ca2+]i in nearby odontoblasts, following mechanical stimulation of a single odontoblast, was abolished. We could record some inward currents evoked from odontoblasts near the stimulated odontoblast, but the currents were observed in only 4.8% of the recorded odontoblasts. The results of this study showed that ATP is released via pannexin-1, from a mechanically stimulated odontoblast, which transmits a signal to nearby odontoblasts by predominant activation of PLC-coupled nucleotide receptors.