Cell fate mediators Notch and Twist in mouse mandibular condylar cartilage.
Cell fate mediators Notch and Twist in mouse mandibular condylar cartilage.
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DOI：
10.1016/j.archoralbio.2010.11.014
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发表时间：
2011-06
影响因子：
3
通讯作者：
Hinton RJ
中科院分区：
文献类型：
--
作者：
Serrano MJ;So S;Svoboda KK;Hinton RJ
本研究的目的是检查 Twist 和 Notch 1 是否存在于下颌髁软骨 (MCC) 中，以及它们的基因表达是否可以被外源性 FGF-2 和 TGF-β2 改变。将胚胎第 17 天 (E17) 收获的 CD-1 小鼠幼崽的半头固定、脱钙并在矢状平面上切片，以便使用共聚焦显微镜对 Notch 和 Twist 进行免疫组织化学检测。 E17 小鼠的其他下颌髁突和邻近支在含有 0、3 或 30 ng/mL FGF-2 的无血清 DMEM 中培养（每个治疗组 10-12 个髁突）。使用含有 0、3 或 30 ng/mL TGF-β2 的培养基重复该实验设计。培养3天后，提取每组的汇集RNA，使用定量实时RT-PCR检查Notch和Twist基因表达。免疫组织化学检查显示，Notch 和 Twist 位于软骨的前软骨细胞层和上软骨细胞层。外源性 FGF-2 上调 E17 小鼠 MCC 外植体中的 Notch1、Twist1 和 Twist2 基因表达，而 TGF-β2 则具有相反的作用。基因表达数据表明，MCC 外植体对已知影响其他组织中 Notch 和 Twist 的生长因子敏感。发现 Twist 和 Notch 免疫反应性的细胞子集提示 FGF-2 和 TGF-β2 作为双能 MCC 细胞群的细胞分化调节剂的作用，这与 Notch 和 Twist 作为这些生长因子在其他组织中的下游介质的作用一致。
The objectives of this study were to examine if Twist and Notch 1 are present in the mandibular condylar cartilage (MCC) and whether their gene expression can be altered by exogenous FGF-2 and TGF-β2. Half-heads from CD-1 mice pups harvested at embryonic day 17 (E17) were fixed, decalcified, and sectioned in the sagittal plane for immunohistochemical detection of Notch and Twist using confocal microscopy. Other mandibular condyles and adjacent ramus from E17 mice were cultured in serum-free DMEM containing 0, 3, or 30 ng/mL of FGF-2 (10–12 condyles per treatment group). This experimental design was repeated with medium containing 0, 3, or 30 ng/mL of TGF-β2. After 3 days of culture, the pooled RNA from each group was extracted for examination of Notch and Twist gene expression using quantitative real-time RT-PCR. Immunohistochemical examination revealed that Notch and Twist were localized to the prechondroblastic and upper chondroblastic layers of the cartilage. Exogenous FGF-2 up-regulated Notch1, Twist1 and Twist2 gene expression in MCC explants from E17 mice, while TGF-β2 had the opposite effect. The gene expression data demonstrate that MCC explants are sensitive to growth factors known to affect Notch and Twist in other tissues. The subset of cells in which Twist and Notch immunoreactivity was found is suggestive of a role for FGF-2 and TGF-β2 as regulators of cell differentiation of the bipotent MCC cell population, consistent with the role of Notch and Twist as downstream mediators of these growth factors in other tissues.
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