Molecular Laboratory for Gene Therapy and Tooth Regeneration, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, School of Stomatology, Capital Medical University, Tian Tan Xi Li No.4, Beijing 100050, China.
MOE Key Laboratory of Bioinformatics, School of Life Sciences, Tsinghua University, Beijing 100084, China.
J Proteomics. 2018 Feb 10;172:57-67. doi: 10.1016/j.jprot.2017.10.013. Epub 2017 Nov 10.
Taking advantage of genetic manipulation tools and accessibility, almost all molecular knowledge on vertebrate tooth development was obtained from rodent models that only have one dentition in their entire lives. Whether the tooth development in other vertebrates such as swine or human follows the same rules remains elusive. Rodent dentitions differ considerably from human dentitions, therefore limiting the application of knowledge from rodent tooth to human tooth. Signal-mediated communication between cells and complex gene and protein regulatory networks are key components of tooth development. By combining isobaric tandem mass tag (TMT) labeling with liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) technology, we constructed the proteomic profile of deciduous molars at embryonic days 40 and 50 in miniature pig (Sus scrofa). During the ten days of prenatal development of the miniature pig, the morphology of the lower deciduous molar moves from the early cap to the bell stage. Thus, we identified proteins that are associated with these developing stages and identified differentially regulated proteins (DRPs) that are potential or novel drivers of tooth morphogenesis. Three candidate proteins were validated via qRT-PCR, western blotting analysis, and the location of those proteins in tooth germ were observed by immunohistochemical staining. Multiple signaling pathways and protein interaction network revealed potential mechanisms of early tooth programming in a large mammal. Bioinformatic analysis also showed that cross interaction of Wnt and Sonic hedgehog pathways may play a key role in deciduous development during cap to bell transition in miniature pig.
We performed the most comprehensive study of the whole tooth germ proteome in mammals to date. The high-throughput proteomic analysis identifies differentially regulated proteins and pathways that will help elucidate the mechanisms of tooth development.
利用遗传操作工具和可及性，脊椎动物牙齿发育的几乎所有分子知识都来自于啮齿动物模型，这些模型在其一生中只有一次牙齿发育。其他脊椎动物（如猪或人类）的牙齿发育是否遵循相同的规则仍不清楚。啮齿动物的牙齿与人类的牙齿有很大的不同，因此限制了从啮齿动物牙齿到人类牙齿的知识的应用。细胞之间的信号介导的通讯和复杂的基因和蛋白质调控网络是牙齿发育的关键组成部分。通过结合等压串联质量标签（TMT）标记与液相色谱-质谱/质谱（LC-MS/MS）技术，我们构建了小型猪（Sus scrofa）胚胎第 40 天和第 50 天乳磨牙的蛋白质组图谱。在小型猪产前发育的十天中，下颌乳磨牙的形态从早期帽状期到钟状期。因此，我们鉴定了与这些发育阶段相关的蛋白质，并鉴定了差异调节蛋白（DRPs），这些蛋白可能是牙齿形态发生的潜在或新的驱动因素。通过 qRT-PCR、western blot 分析验证了 3 个候选蛋白，并通过免疫组织化学染色观察这些蛋白在牙胚中的位置。多个信号通路和蛋白质相互作用网络揭示了大型哺乳动物早期牙齿编程的潜在机制。生物信息学分析还表明，Wnt 和 Sonic hedgehog 通路的交叉相互作用可能在小型猪帽状期到钟状期过渡期间的乳齿发育中发挥关键作用。
我们进行了迄今为止哺乳动物全牙胚蛋白质组的最全面研究。高通量蛋白质组学分析鉴定了差异调节蛋白和通路，这将有助于阐明牙齿发育的机制。
