Identification of differential microRNA expression during tooth morphogenesis in the heterodont dentition of miniature pigs, SusScrofa
Identification of differential microRNA expression during tooth morphogenesis in the heterodont dentition of miniature pigs, SusScrofa
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小型猪异齿牙列牙齿形态发生过程中差异 microRNA 表达的鉴定
DOI：
10.1186/s12861-015-0099-0
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发表时间：
2015-12-29
影响因子：
--
通讯作者：
Wang, Songlin
中科院分区：
文献类型：
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作者：
Li, Ang;Li, Ye;Wang, Songlin
背景研究发现microRNAs（miRNAs）在牙齿发育的调控中起着重要作用，并且很可能增加了遗传网络的复杂性，从而导致牙齿的复杂性增加。但目前还没有基于miRNAs表达谱对牙齿形态发生相关的关键microRNAs进行研究。与小鼠相比，猪模型具有丰富的牙齿类型，这与人类的牙齿模式相似。因此，我们使用小型猪作为大动物模型，探讨差异表达的miRNA表达在牙齿形态发生在早期发育阶段的牙胚。ResultsA定制设计的miRNA芯片与742 miRNA基因探针被用来分析表达谱的四种类型的牙齿在三个阶段的牙齿发育。在591个可检测的miRNA转录本中，有212个miRNA在所有类型的牙胚中持续表达，但在4种不同类型的牙胚中各胚胎期的miRNA转录本数量差异有统计学意义（p< 0.01）。系统聚类和主成分分析结果表明，miRNA的表达随类型和时间的变化而发生全局性变化。通过聚类分析，我们预测了11个独特的miRNA序列，分别属于mir-103和mir-107，mir-133 a和mir-133 b，以及mir-127 isomiR家族。实时荧光定量RT-PCR和原位杂交结果显示，5种具有代表性的miRNAs可能分别在切牙、犬齿、双尖牙和磨牙的不同发育阶段发挥重要作用。结论本研究表明，ssc-miR-103和ssc-miR-107，ssc-miR-133 a和ssc-miR-133 b，ssc-miR-127，可能在小型猪牙齿发育的不同阶段对不同类型的牙齿起着关键的调节作用，因此可能在小型猪牙齿发育的早期形态发生中起着关键的作用。
BackgroundIt has been found that microRNAs (miRNAs) play important roles in the regulation of tooth development, and most likely increase the complexity of the genetic network, thus lead to greater complexity of teeth. But there has been no research about the key microRNAs associated with tooth morphogenesis based on miRNAs expression profiles. Compared to mice, the pig model has plentiful types of teeth, which is similar with the human dental pattern. Therefore, we used miniature pigs as large-animal models to investigate differentially expressed miRNAs expression during tooth morphogenesis in the early developmental stages of tooth germ.ResultsA custom-designed miRNA microarray with 742 miRNA gene probes was used to analyze the expression profiles of four types of teeth at three stages of tooth development. Of the 591 detectable miRNA transcripts, 212 miRNAs were continuously expressed in all types of tooth germ, but the numbers of miRNA transcript among the four different types of teeth at each embryonic stage were statistically significant differences (p< 0.01). The hierarchical clustering and principal component analysis results suggest that the miRNA expression was globally altered by types and temporal changes. By clustering analysis, we predicted 11 unique miRNA sequences that belong to mir-103 and mir-107, mir-133a and mir-133b, and mir-127 isomiR families. The results of real-time reverse-transcriptase PCR andin situhybridization experiments revealed that five representative miRNAs may play important roles during different developmental stages of the incisor, canine, biscuspid, and molar, respectively.ConclusionsThe present study indicated that these five miRNAs, including ssc-miR-103 and ssc-miR-107, ssc-miR-133a and ssc-miR-133b, and ssc-miR-127, may play key regulatory roles in different types of teeth during different stages and thus may play critical roles in tooth morphogenesis during early development in miniature pigs.