Journal of Molecular Histology

, Volume 49, Issue 5, pp 545–553 | Cite as

Expression of BMP2/4/7 during the odontogenesis of deciduous molars in miniature pig embryos

  • Zhenhua Gao
  • Lingxiao Wang
  • Fu Wang
  • Chunmei Zhang
  • Jinsong Wang
  • Junqi He
  • Songlin WangEmail author
Original Paper


Bone morphogenetic proteins (BMPs) play important roles in tooth development. However, their expression has not been studied in miniature pigs, which have many anatomical similarities in oral and maxillofacial region compared to human. This study investigated BMP2/4/7 expression patterns during deciduous molar development in miniature pigs on embryonic days (E) 40, 50, and 60. The mandibles were fixed, decalcified, and embedded before sectioning. H&E staining, immunohistochemistry, in situ hybridization using specific radionuclide-labeled cRNA probes, and real-time PCR were used to detect the BMP expression patterns during morphogenesis of the third deciduous molar. H&E staining showed that for the deciduous third molar, E40 represented the cap stage, E50 represented the early bell stage, and E60 represented the late bell stage or secretory stage. BMP2 was expressed in both the enamel organ and in the dental mesenchyme on E40 and E50 and was expressed mainly in pre-odontoblasts on E60. BMP7 expression was similar to BMP2 expression, but BMP7 was also expressed in the inner enamel epithelium on E60. On E40, BMP4 was expressed mainly in the epithelium, with some weak expression in the mesenchyme. On E50, BMP4 expression was stronger in the mesenchyme but weaker in the epithelium. On E60, BMP4 was expressed mainly in the mesenchyme. These data indicated that BMP2/4/7 showed differential spatial and temporal expression during the morphogenesis and odontogenesis of deciduous molars, suggesting that these molecules were associated with tooth morphogenesis and cell differentiation.


BMP2/4/7 Tooth Development Miniature pig 





Deciduous molar


Dental papilla


Hematoxylin & eosin


Inner enamel epithelium




Outer enamel epithelium




Stellate reticulum



This work was supported by grants from National Natural Science Foundation of China (Grant No. 81600884), Beijing Municipal Administration of Hospitals ‘Youth Programme, code: QML20171502’, Beijing Municipal Organization Department of outstanding talent project (Grant No. 2016000021 469G205). Beijing Municipality Government grants (Beijing Scholar Program- PXM2018_014226_000021, PXM2017_014226_000023, PXM2018_193312_000006_0028S643_FCG PXM2016_014226_000034, PXM2016_014226_000006).

Compliance with ethical standards

Conflict of interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Supplementary material

10735_2018_9792_MOESM1_ESM.docx (2.6 mb)
Supplementary Figures (DOCX 2629 KB)
10735_2018_9792_MOESM2_ESM.tif (85 kb)
Supplementary Table 1 (TIF 84 KB)
10735_2018_9792_MOESM3_ESM.tif (79 kb)
Supplementary Table 2 (TIF 78 KB)


  1. Aberg T, Wozney J, Thesleff I (1997) Expression patterns of bone morphogenetic proteins (Bmps) in the developing mouse tooth suggest roles in morphogenesis and cell differentiation. Dev Dyn 210:383–396CrossRefPubMedGoogle Scholar
  2. Abu-Serriah MM, Odell E, Lock C, Gillar A, Ayoub AF, Fleming RH (2004) Histological assessment of bioengineered new bone in repairing osteoperiosteal mandibular defects in sheep using recombinant human bone morphogenetic protein-7. Br J Oral Maxillofac Surg 42:410–418. CrossRefPubMedGoogle Scholar
  3. Balic A, Thesleff I (2015) Tissue interactions regulating tooth development and renewal. Curr Top Dev Biol 115:157–186. CrossRefPubMedGoogle Scholar
  4. Deng W, Yang D, Zhao B, Ouyang Z, Song J, Fan N, Liu Z, Zhao Y, Wu Q, Nashun B, Tang J, Wu Z, Gu W, Lai L (2011) Use of the 2A peptide for generation of multi-transgenic pigs through a single round of nuclear transfer. PLoS ONE 6:e19986. CrossRefPubMedPubMedCentralGoogle Scholar
  5. Dong X, Shen B, Ruan N, Guan Z, Zhang Y, Chen Y, Hu X (2014) Expression patterns of genes critical for BMP signaling pathway in developing human primary tooth germs. Histochem Cell Biol 142:657–665. CrossRefPubMedGoogle Scholar
  6. Ekser B, Ezzelarab M, Hara H, Van DJ, Wijkstrom M, Bottino R, Trucco M, Cooper DK (2012) Clinical xenotransplantation: the next medical revolution? Lancet 379:672–683. CrossRefPubMedGoogle Scholar
  7. Feng JQ, Zhang J, Tan X, Lu Y, Guo D, Harris SE (2002) Identification of cis-DNA regions controlling Bmp4 expression during tooth morphogenesis in vivo. J Dent Res 81:6–10CrossRefPubMedGoogle Scholar
  8. Feng J, Jing J, Li J, Zhao H, Punj V, Zhang T, Xu J, Chai Y (2017) BMP signaling orchestrates a transcriptional network to control the fate of mesenchymal stem cells in mice. Development 144:2560–2569. CrossRefPubMedPubMedCentralGoogle Scholar
  9. Hasegawa K, Wada H, Nagata K, Fujiwara H, Wada N, Someya H, Mikami Y, Sakai H, Kiyoshima T (2016) Facioscapulohumeral muscular dystrophy (FSHD) region gene 1 (FRG1) expression and possible function in mouse tooth germ development. J Mol Histol 47:375–387. CrossRefPubMedGoogle Scholar
  10. Kriangkrai R, Iseki S, Eto K, Chareonvit S (2006) Dual odontogenic origins develop at the early stage of rat maxillary incisor development. Anat Embryol (Berlin) 211:101–108. CrossRefGoogle Scholar
  11. Li S, Pan Y (2017) Differential expression of transforming growth factor-beta1 connective tissue growth factor, phosphorylated-SMAD2/3 and phosphorylated-ERK1/2 during mouse tooth development. J Mol Histol 48:347–355. CrossRefPubMedGoogle Scholar
  12. Li A, Song T, Wang F, Liu D, Fan Z, Zhang C, He J, Wang S (2012) Micrornaome and expression profile of developing tooth germ in miniature pigs. PLoS ONE 7:e52256. CrossRefPubMedPubMedCentralGoogle Scholar
  13. Li A, Li Y, Song T, Fu W, Liu D, Fan Z, Zhang C, Wang J, He J, Wang S (2015a) Identification of differential microrna expression during tooth morphogenesis in the heterodont dentition of miniature pigs, susscrofa. BMC Dev Biol 15:1–12. CrossRefGoogle Scholar
  14. Li J, Feng J, Liu Y, Ho TV, Grimes W, Ho HA, Park S, Wang S, Chai Y (2015b) BMP-SHH signaling network controls epithelial stem cell fate via regulation of its niche in the developing tooth. Dev Cell 33:125–135. CrossRefPubMedPubMedCentralGoogle Scholar
  15. Li L, Tang Q, Nakamura T, Suh JG, Ohshima H, Jung HS (2016) Fine tuning of Rac1 and RhoA alters cuspal shapes by remolding the cellular geometry. Sci Rep 6:37828. CrossRefPubMedPubMedCentralGoogle Scholar
  16. Li Y, Gong Y, Wu X, Wang F, Xie Y, Zhu Z, Su Y, Wang J, Zhang C, He J, Deng H, Wang S (2017) Quantitative proteomic analysis of deciduous molars during cap to bell transition in miniature pig. J Proteom 172:57–67. CrossRefGoogle Scholar
  17. Li Q, Guo Y, Yao M, Li J, Chen Y, Liu Q, Chen Y, Zeng Y, Ji B, Feng Y (2018) Methylation of Cdkn1c may involve in the regulation of tooth development through cell cycle inhibition. J Mol Histol. (Epub ahead of print)CrossRefPubMedPubMedCentralGoogle Scholar
  18. Lian M, Zhang Y, Shen Q, Xing J, Lu X, Huang D, Cao P, Shen S, Zheng K, Zhang J, Chen J, Wang Y, Feng G, Feng X (2016) JAB1 accelerates odontogenic differentiation of dental pulp stem cells. J Mol Histol 47:317–324. CrossRefPubMedGoogle Scholar
  19. Lin D, Huang Y, He F, Gu S, Zhang G, Chen Y, Zhang Y (2007) Expression survey of genes critical for tooth development in the human embryonic tooth germ. Dev Dyn 236:1307–1312. CrossRefPubMedGoogle Scholar
  20. Litsiou A, Hanson S, Streit A (2005) A balance of FGF, BMP and WNT signalling positions the future placode territory in the head. Development 132:4051–4062. CrossRefPubMedGoogle Scholar
  21. Liu Y, Zheng Y, Ding G, Fang D, Zhang C, Bartold PM, Gronthos S, Shi S, Wang S (2008) Periodontal ligament stem cell-mediated treatment for periodontitis in miniature swine. Stem Cell 26:1065–1073. CrossRefGoogle Scholar
  22. Shi L, Li L, Wang D, Li S, Chen Z, An Z (2016) Spatiotemporal expression of caveolin-1 and EMMPRIN during mouse tooth development. J Mol Histol 47:337–344. CrossRefPubMedGoogle Scholar
  23. Song T, Wu T, Wei F, Li A, Wang F, Xie Y, Liu D, Fan Z, Wang X, Cheng S, Zhang C, He J, Wang S (2014) Construction of a cdna library for miniature pig mandibular deciduous molars. BMC Dev Biol 14:16. CrossRefPubMedPubMedCentralGoogle Scholar
  24. Vainio S, Karavanova I, Jowett A, Thesleff I (1993) Identification of BMP-4 as a signal mediating secondary induction between epithelial and mesenchymal tissues during early tooth development. Cell 75(1):45–58CrossRefPubMedGoogle Scholar
  25. Wang Y, Li L, Zheng Y, Yuan G, Yang G, He F, Chen Y (2012) Bmp activity is required for tooth development from the lamina to bud stage. J Dent Res 91:690–695. CrossRefPubMedPubMedCentralGoogle Scholar
  26. Wang F, Xiao J, Cong W, Li A, Song T, Wei F, Xu J, Zhang C, Fan Z, Wang S (2014a) Morphology and chronology of diphyodont dentition in miniature pigs, Sus Scrofa. Oral Dis 20:367–379. CrossRefPubMedGoogle Scholar
  27. Wang F, Xiao J, Cong W, Li A, Wei F, Xu J, Zhang C, Fan Z, He J, Wang S (2014b) Stage-specific differential gene expression profiling and functional network analysis during morphogenesis of diphyodont dentition in miniature pigs, Sus Scrofa. BMC Genom 15:103. CrossRefGoogle Scholar
  28. Wu X, Li Y, Wang F, Hu L, Li Y, Wang J, Zhang C, Wang S (2017) Spatiotemporal expression of wnt/β-catenin signaling during morphogenesis and odontogenesis of deciduous molar in miniature pig. Int J Biol Sci 13:1082. CrossRefPubMedPubMedCentralGoogle Scholar
  29. Yang G, Jiang B, Cai W, Liu S, Zhao S (2016) Hyaluronan and hyaluronan synthases expression and localization in embryonic mouse molars. J Mol Histol 47:413–420. CrossRefPubMedGoogle Scholar
  30. Yuan G, Yang G, Zheng Y, Zhu X, Chen Z, Zhang Z, Chen Y (2015) The non-canonical BMP and Wnt/β-catenin signaling pathways orchestrate early tooth development. Development 142:128–139. CrossRefPubMedPubMedCentralGoogle Scholar
  31. Yun HM, Park KR, Quang TH, Oh H, Hong JT, Kim YC, Kim EC (2015) 2,4,5-Trimethoxyldalbergiquinol promotes osteoblastic differentiation and mineralization via the BMP and Wnt/β-catenin pathway. Cell Death Dis 6:e1819. CrossRefPubMedPubMedCentralGoogle Scholar
  32. Zhang W, Zhang X, Ling J, Wei X, Jian Y (2016) Osteo-/odontogenic differentiation of BMP2 and VEGF gene-co-transfected human stem cells from apical papilla. Mol Med Rep 13:3747–3754. CrossRefPubMedPubMedCentralGoogle Scholar
  33. Zhou Z, Yin Y, Jiang F, Niu Y, Wan S, Chen N, Shen M (2016) CBX7 deficiency plays a positive role in dentin and alveolar bone development. J Mol Histol 47:401–411. CrossRefPubMedGoogle Scholar
  34. Zhou M, Guo S, Yuan L, Zhang Y, Zhang M, Chen H, Lu M, Yang J, Ma J (2017) Blockade of LGR4 inhibits proliferation and odonto/osteogenic differentiation of stem cells from apical papillae. J Mol Histol 48:389–401. CrossRefPubMedPubMedCentralGoogle Scholar
  35. Zouvelou V, Luder HU, Mitsiadis TA, Graf D (2009) Deletion of BMP7 affects the development of bones, teeth, and other ectodermal appendages of the orofacial complex. J Exp Zool B 312B:361–374. CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Zhenhua Gao
    • 1
    • 2
  • Lingxiao Wang
    • 2
  • Fu Wang
    • 1
  • Chunmei Zhang
    • 1
  • Jinsong Wang
    • 1
  • Junqi He
    • 3
  • Songlin Wang
    • 1
    • 3
    Email author
  1. 1.Molecular Laboratory for Gene Therapy and Tooth Regeneration, Beijing Key Laboratory of Tooth Regeneration and Function ReconstructionCapital Medical University School of StomatologyBeijingChina
  2. 2.Outpatient Department of Oral and Maxillofacial SurgeryCapital Medical University School of StomatologyBeijingChina
  3. 3.Department of Biochemistry and Molecular BiologyCapital Medical University School of Basic Medical SciencesBeijingChina

Personalised recommendations