Construction of a cDNA library for miniature pig mandibular deciduous molars

BMC Developmental Biology

Songlin Wang¹  Junqi He¹  Chunmei Zhang²  Shan Cheng¹  Xuejiu Wang²  Zhipeng Fan²  Dayong Liu²  Yilin Xie²  Fu Wang²  Ang Li²  Fulan Wei²  Tingting Wu²  Tieli Song³ 

[1] Department of Biochemistry and Molecular Biology, Capital Medical University School of Basic Medical Sciences, Beijing 100069, China;Molecular Laboratory for Gene Therapy and Tooth Regeneration, Beijing Key Laboratory of Tooth Regeneration and Function Reconstruction, Capital Medical University School of Stomatology, Tian Tan Xi Li No.4, Beijing 100050, China;Department of Stomatology, Beijing Tongren Hospital Affiliated to Capital Medical University, Dong Jiao Min Xiang No.1, Beijing 100730, China

关键词: Miniature pig;    Sequence;    Unigene;    Histology;    Development;    Tooth;   

Others  :  820856
DOI  :  10.1186/1471-213X-14-16

 received in 2013-08-20, accepted in 2014-04-09,  发布年份 2014

【 摘 要 】

Background

The miniature pig provides an excellent experimental model for tooth morphogenesis because its diphyodont and heterodont dentition resembles that of humans. However, little information is available on the process of tooth development or the exact molecular mechanisms controlling tooth development in miniature pigs or humans. Thus, the analysis of gene expression related to each stage of tooth development is very important.

Results

In our study, after serial sections were made, the development of the crown of the miniature pigs’ mandibular deciduous molar could be divided into five main phases: dental lamina stage (E33-E35), bud stage (E35-E40), cap stage (E40-E50), early bell stage (E50-E60), and late bell stage (E60-E65). Total RNA was isolated from the tooth germ of miniature pig embryos at E35, E45, E50, and E60, and a cDNA library was constructed. Then, we identified cDNA sequences on a large scale screen for cDNA profiles in the developing mandibular deciduous molars (E35, E45, E50, and E60) of miniature pigs using Illumina Solexa deep sequencing. Microarray assay was used to detect the expression of genes. Lastly, through Unigene sequence analysis and cDNA expression pattern analysis at E45 and E60, we found that 12 up-regulated and 15 down-regulated genes during the four periods are highly conserved genes homologous with known Homo sapiens genes. Furthermore, there were 6 down-regulated and 2 up-regulated genes in the miniature pig that were highly homologous to Homo sapiens genes compared with those in the mouse.

Conclusion

Our results not only identify the specific transcriptome and cDNA profile in developing mandibular deciduous molars of the miniature pig, but also provide useful information for investigating the molecular mechanism of tooth development in the miniature pig.

【 授权许可】

   
2014 Song et al.; licensee BioMed Central Ltd.

【 图 表 】

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