MicroRNA（miRNA）在嚙齒類動物牙齒發(fā)育過程中扮演著重要的調(diào)控角色,，但我們對其在大型哺乳類動物的牙齒發(fā)育中的作用知之甚少。現(xiàn)在,，首都醫(yī)科大學副校長王松靈教授領(lǐng)銜的團隊利用Seq-Array?（芯片與二代測序相結(jié)合策略）技術(shù)方法鑒定出小型豬在下乳牙發(fā)育過程中的特定miRNA組和表達譜，此項研究也為探索小型豬牙齒發(fā)育的分子機制提供了重要信息,。該研究成果發(fā)表在2012年12月刊PLoS ONE上,。
王松靈教授團隊首先使用二代深度測序技術(shù)大規(guī)模篩查小型豬（Sus scrofa）在下乳牙發(fā)育過程中的miRNA表達譜，鑒別出637條miRNA序列,。深度測序獲取的結(jié)果,可以廣泛地覆蓋不同發(fā)育時期和不同來源樣品中的全部miRNA,，而不需要任何已知的序列信息,。然而，由于樣品處理相對的高成本與低通量特性,，如果用測序進行重復性的驗證和（或是）表達譜分析將會發(fā)現(xiàn)花費很長時間,，而且價格昂貴。
代替重復測序,，使用芯片可以進行高通量的表達譜分析,。芯片上可以涵蓋完整的測序發(fā)現(xiàn)的序列信息，系統(tǒng)的分析表達譜,，并進行差異表達分析,。芯片具有快速、可重復,，并且性價比高的特點。
研究人員將測序發(fā)現(xiàn)的候選miRNA,，和105條已知的Sus scrofamiRNA定制合成在miRNA芯片上用于分析牙齒發(fā)育的四個時期：牙芽期,，帽形期，鐘形早期,，鐘形晚期的miRNA表達譜,。芯片分析結(jié)果顯示在四個發(fā)育時期有166個miRNA存在差異表達。生物信息學分析鑒別出18個關(guān)鍵miRNA,，包括 let-7f,，miR-128, miR-200b, 和miR-200c，在牙齒發(fā)育過程中也許扮演著關(guān)鍵角色,。(生物谷Bioon.com)
doi: 10.1371/journal.pone.0052256.
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MicroRNAome and Expression Profile of Developing Tooth Germ in Miniature Pigs.
Li A, Song T, Wang F, Liu D, Fan Z, Zhang C, He J, Wang S.
MicroRNAs (miRNAs) play important roles in the regulation of rodent tooth development, but little is known about their role in tooth development in large mammals. We identified 637 unique miRNA sequences in a large-scale screen for miRNA expression profiles in the developing lower deciduous molars of miniature pigs (Sus scrofa) using Illumina Solexa deep sequencing. These candidate miRNAs and another 105 known Sus scrofa miRNAs were included in the custom-designed microarray and used to analyze the miRNA expression profile in the bud, cap, early bell, and late bell stages of tooth development. Microarray analysis revealed 166 transcripts that were differentially expressed in the four stages. Bioinformatic analysis identified 18 key miRNAs, including let-7f, miR-128, miR-200b, and miR-200c, that might play key roles in tooth development. Taken together, our results not only identified the specific microRNAome and expression profile in developing lower deciduous molars of the miniature pig, but they also provided useful information for investigating the molecular mechanism of tooth development in the miniature pig.
