Wnt11 expression in rat dental pulp and promotional effects of Wnt signaling on odontoblast differentiation
Wnt11 expression in rat dental pulp and promotional effects of Wnt signaling on odontoblast differentiation
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DOI：
10.1111/cga.12011
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发表时间：
2013-09-01
影响因子：
1.3
通讯作者：
Suda, Hideaki
中科院分区：
文献类型：
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作者：
Koizumi, Yu;Kawashima, Nobuyuki;Suda, Hideaki
无翅型小鼠乳腺肿瘤病毒（MMTV）整合位点家族（Wnt）的先天性异常常伴有牙齿和牙本质异常。本研究旨在探讨Wnt信号对小鼠牙乳头细胞（MDP）成牙本质细胞分化的影响。将小鼠牙乳头细胞培养在含有10%胎牛血清和抗生素的α-改良最低必需培养基中。骨形态发生蛋白2（BMP 2）诱导成牙本质细胞分化，实时荧光定量逆转录聚合酶链反应（RT-PCR）和免疫组化检测成牙本质细胞特异性标志物和Wnt相关信号分子的表达。牙本质涎磷蛋白（Dspp）和牙本质基质蛋白（DSP 1）的表达评价成牙本质细胞的分化。使用抗β-连环蛋白抗体通过免疫细胞化学检测MDP中β-连环蛋白的定位。在BMP 2的存在下，MDPs中的Dspp表达上调。BMP 2处理的MDPs中Wnt 5a、Wnt 11、Lef 1和Tcf 4表达上调。Wnt 11在大鼠牙髓中的表达在体内检测，特别是强的表达Wnt 11检测成牙本质细胞。Wnt拮抗剂IWR-1-endo处理可完全抑制BMP 2诱导的Dspp和DMP 1表达以及碱性磷酸酶活性的增强。在BMP 2处理的MDP中观察到的β-连环蛋白的核转位也被IWR-1-endo处理所否定。这些结果表明，Wnt信号上调成牙本质细胞标记物在MDP中的表达，表明Wnt信号对成牙本质细胞分化的促进作用。
Congenital anomalies of wingless-type mouse mammary tumor virus (MMTV) integration site family (Wnt) are frequently accompanied with tooth and dentin abnormality. The aim of this study was to investigate the effects of Wnt signaling on odontoblast differentiation of mouse dental papilla cells (MDPs). Mouse dental papilla cells were cultured in alpha-modified minimum essential medium containing 10% fetal bovine serum and antibiotics. Odontoblast differentiation was induced by bone morphogenic protein 2 (BMP2), and the expression of odontoblast-specific markers and Wnt-related signaling molecules was analyzed by real-time reverse transcription-polymerase chain reaction and immunohistochemistry. Odontoblast differentiation was evaluated by dentin sialophosphoprotein (Dspp) and dentin matrix protein (DMP) 1 expression. Localization of beta-catenin in MDPs was detected by immunocytochemistry using an anti-beta-catenin antibody. Dspp expression in MDPs was upregulated in the presence of BMP2. Wnt5a, Wnt11, Lef1 and Tcf4 expression was upregulated in BMP2-treated MDPs. Wnt11 expression was detected in rat dental pulp in vivo, and particularly strong expression of Wnt11 was detected in odontoblasts. Enhanced Dspp and DMP1 expression and alkaline phosphatase activity induced by BMP2 were completely negated by the Wnt antagonist: IWR-1-endo treatment. Nuclear translocation of beta-catenin observed in BMP2-treated MDPs was also negated by IWR-1-endo treatment. These results indicate that Wnt signaling upregulates odontoblast marker expression in MDPs, suggesting a promoting effect of Wnt signaling on odontoblast differentiation.