Expression of clusterin (Clu) in the murine first molar tooth germ was markedly increased at postnatal developmental stages. The time-course of expression of this gene paralleled those of other genes encoding proteins involved during the secretory phase of odontogenesis, as described previously. Immunohistochemical studies of clusterin in murine molar tooth germs suggested this protein to be located in outer enamel epithelium, regressing enamel organ, secretory ameloblasts, and the dental epithelium connecting the tooth to the oral epithelium at an early eruptive stage. Immunolabelling of transforming growth factor beta-1 (TGF-β1) revealed it to be located close to clusterin. The levels of expression of Clu and Tgfb1 were markedly decreased following in-vivo transfection with anti-miR-214. In contrast, the expression of several genes associated with regulation of growth and development were increased by this treatment. We suggest that clusterin has functions during secretory odontogenesis and the early eruptive phase. Bioinformatic analysis after treatment with anti-miR-214 suggested that, whilst cellular activities associated with tooth mineralization and eruption were inhibited, activities associated with an alternative developmental activity (i.e. biosynthesis of contractile proteins) appeared to be stimulated. These changes probably occur through regulation mediated by a common cluster of transcription factors and support suggestions that microRNAs (miRNAs) are highly significant as regulators of differentiation during odontogenesis.

译文

在出生后发育阶段,鼠第一磨牙牙胚中簇蛋白 (Clu) 的表达显着增加。如前所述,该基因的表达时程与其他编码齿状细胞分泌阶段涉及的蛋白质的基因的表达时程平行。鼠磨牙牙胚中clusterin的免疫组织化学研究表明,该蛋白位于釉质外上皮,退化的釉质器官,分泌成釉细胞以及在萌发初期将牙齿连接到口腔上皮的牙齿上皮中。转化生长因子 β1 (TGF-β1) 的免疫标记显示其位于聚集蛋白附近。用anti-miR-214体内转染后,Clu和Tgfb1的表达水平显着降低。相反,通过这种处理增加了与生长和发育调节相关的几个基因的表达。我们建议簇蛋白在分泌型牙本质发生和早期爆发阶段具有功能。用anti-miR-214治疗后的生物信息学分析表明,尽管与牙齿矿化和萌发相关的细胞活动受到抑制,但与替代发育活动 (即收缩蛋白的生物合成) 相关的活动似乎受到刺激。这些变化可能是通过共同的转录因子簇介导的调节而发生的,并支持microRNAs (miRNAs) 作为牙形成过程中分化的调节因子非常重要的建议。

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