Developmental studies on the acquisition of perception conducting pathways via TRP channels in rat molar odontoblasts using immunohistochemistry and RT-qPCR
Developmental studies on the acquisition of perception conducting pathways via TRP channels in rat molar odontoblasts using immunohistochemistry and RT-qPCR
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DOI：
10.1007/s12565-019-00517-y
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发表时间：
2019-12
影响因子：
1.2
通讯作者：
Aoi Tanaka;Y. Shibukawa;Masahito Yamamoto;S. Abe;Hitoshi Yamamoto;S. Shintani
中科院分区：
文献类型：
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作者：
Aoi Tanaka;Y. Shibukawa;Masahito Yamamoto;S. Abe;Hitoshi Yamamoto;S. Shintani
成牙本质细胞作为牙本质形成和感觉受体。近年来研究发现，瞬时受体电位锚蛋白（TRPA）1、TRP香草酸（TRPV）4和泛连接蛋白1（PANX-1）在成牙本质细胞感觉感受中起重要作用。然而，成牙本质细胞何时开始具有感觉接收功能尚不清楚。本研究的目的是阐明成牙本质细胞作为感觉受体的发育。取出生后0 ~ 12天Wistar大鼠下颌第一磨牙切片，行苏木精-伊红染色。免疫组化，我们使用抗牙本质唾液蛋白（DSP），抗TRPA 1，抗TRPV 4，抗PANX-1，和抗神经丝（NF）抗体。此外，我们还通过逆转录定量聚合酶链反应（RT-qPCR）研究了TRPA 1和TRPV 4的表达。PN 0时，未分化的成牙本质细胞对抗DSP、抗TRPA 1、抗TRPV 4或抗PANX-1抗体无免疫反应。然而，这些抗体的免疫阳性反应增加成牙本质细胞分化在PN 3和PN 6。在成牙本质细胞开始形成根面牙本质的PN 12时，抗NF抗体在成牙本质细胞附近出现免疫阳性反应，且出现时间晚于其他抗体。RT-qPCR结果显示，PN 6组TRPA 1的表达显著低于PN 0组（p< 0.05）和PN 3组（p< 0.01）。PN 6时TRPV 4的表达显著低于PN 0（p< 0.01）和PN 3（p< 0.01）。本研究的结果表明，成牙本质细胞可能获得感觉受体功能后，开始形成根牙本质，当TRPA 1，TRPV 4，PANX-1通道，和神经纤维完全形成。
Odontoblasts act as dentin formation and sensory receptors. Recently, it was reported that transient receptor potential ankyrin (TRPA) 1, TRP vanilloid (TRPV) 4 and pannexin 1 (PANX-1) play important roles in odontoblast sensory reception. However, it is not known when odontoblasts begin to possess a sense reception function. The aim of this study was to clarify the development of odontoblasts as sense receptors. Sections of mandibular first molars from postnatal day (PN) 0 to PN12 Wistar rats were prepared for hematoxylin–eosin staining. Immunohistochemically, we used anti-dentin sialoprotein (DSP), anti-TRPA1, anti-TRPV4, anti-PANX-1, and anti-neurofilament (NF) antibodies. In addition, we investigatedTRPA1andTRPV4expression by reverse transcriptional quantitative polymerase chain reaction (RT-qPCR). At PN0, undifferentiated odontoblasts showed no immunoreaction to anti-DSP, anti-TRPA1, anti-TRPV4, or anti-PANX-1 antibodies. However, immunopositive reactions of these antibodies increased during odontoblast differentiation at PN3 and PN6. An immunopositive reaction of the anti-NF antibody appeared in the odontoblast neighborhood at PN12, when the odontoblasts began to form root dentin, and this appeared later than that of the other antibodies. By RT-qPCR, expression ofTRPA1at PN6 was significantly lower than that at PN0 (p< 0.05) and PN3 (p< 0.01). Expression ofTRPV4at PN6 was significantly lower than that at PN0 (p< 0.01) and PN3 (p< 0.01). The results of this study suggest that odontoblasts may acquire sensory receptor function after beginning to form root dentin, when TRPA1, TRPV4, PANX-1 channels, and nerve fibers are completely formed.