Skip to main content
SpringerLink
Log in

Sp7/osterix positively regulates dlx2b and bglap to affect tooth development and bone mineralization in zebrafish larvae

  • Published:
Journal of Biosciences Aims and scope Submit manuscript

Abstract

Osterix (or Sp7) is an important transcription factor that promotes osteoblast differentiation by modulating the expression of a range of target genes. Although many studies have focused on Osterix/Sp7 regulatory mechanisms, the detailed functions have not been fully elucidated. Toward this end, in this study, we used CRISPR/Cas9 technology to knock out the zebrafish sp7 gene, and then analyzed its phenotype and biological function. Two knockout sp7 mutant lines were successfully obtained. The bone mineralization level was significantly reduced in the zebrafish sp7−/− homozygote, resulting in abnormal tooth development in the larvae. Quantitative real-time polymerase chain reaction showed that loss of sp7 led to down-regulated expression of the dlx2b and bglap genes related to tooth development and bone mineralization, respectively. Moreover, cell transfection experiments demonstrated that Sp7 directly regulates the expression of dlx2b and bglap through Sp7-binding sites on the promoter regions of these two genes. Overall, this study provides new insight into the role of Sp7 in bone mineralization and tooth development.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  • Arnold WH, Naumova KI, Naumova EA and Gaengler P 2008 Comparative qualitative and quantitative assessment of biomineralization of tooth development in man and zebrafish (Danio rerio). Anat. Rec. 291 571–576

    Article  Google Scholar 

  • Baek WY, Lee MA, Jung JW, Kim SY, Akiyama H, de Crombrugghe B and Kim JE 2009 Positive regulation of adult bone formation by osteoblast-specific transcription factor osterix. J. Bone Miner. Res. 24 1055–1065

    Article  CAS  Google Scholar 

  • Gibert Y, Samarut E, Pasco-Viel E, Bernard L, Borday-Birraux V, Sadier A, Labbe C, Viriot L and Laudet V 2015 Altered retinoic acid signalling underpins dentition evolution. Proc. Biol. Sci. 282 https://doi.org/10.1098/rspb.2014.2764.

    Article  PubMed  PubMed Central  Google Scholar 

  • Hosoya A, Yukita A, Ninomiya T, Hiraga T, Yoshiba K, Yoshiba N, Kasahara E and Nakamura H 2013 Localization of SUMOylation factors and Osterix in odontoblast lineage cells during dentin formation and regeneration. Histochem. Cell Biol. 140 201–211

    Article  CAS  Google Scholar 

  • Hsu PD, Scott DA, Weinstein JA, Ran FA, Konermann S, Agarwala V, Li Y, Fine EJ, Wu X, Shalem O, Cradick TJ, Marraffini LA, Bao G and Zhang F 2013 DNA targeting specificity of RNA-guided Cas9 nucleases. Nat. Biotechnol. 31 827–832

    Article  CAS  Google Scholar 

  • Jackman WR, Draper BW and Stock DW 2004 Fgf signaling is required for zebrafish tooth development. Dev. Biol. 274 139–157

    Article  CAS  Google Scholar 

  • Kague E, Witten PE, Soenens M, Campos CL, Lubiana T, Fisher S, Hammond C, Brown KR, Passos-Bueno MR and Huysseune A 2018 Zebrafish sp7 mutants show tooth cycling independent of attachment, eruption and poor differentiation of teeth. Dev. Biol. 435 176–184

    Article  CAS  Google Scholar 

  • Kawai S, Yamauchi M and Amano A 2017 Zinc-finger transcription factor Odd-skipped related 1 regulates cranial bone formation. J. Bone Miner. Metab. 36 640–647

    Article  Google Scholar 

  • Koga T, Matsui Y, Asagiri M, Kodama T, de Crombrugghe B, Nakashima K and Takayanagi H 2005 NFAT and Osterix cooperatively regulate bone formation. Nat. Med. 11 880–885

    Article  CAS  Google Scholar 

  • Lee MH, Kwon TG, Park HS, Wozney JM and Ryoo HM 2003 BMP-2-induced Osterix expression is mediated by Dlx5 but is independent of Runx2. Biochem. Biophys. Res. Commun. 309 689–694

    Article  CAS  Google Scholar 

  • Merlo GR, Zerega B, Paleari L, Trombino S, Mantero S and Levi G 2000 Multiple functions of Dlx genes. Int. J. Dev. Biol. 44 619–626

    CAS  PubMed  Google Scholar 

  • Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR and de Crombrugghe B 2002 The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell 108 17–29

    Article  CAS  Google Scholar 

  • Nishimura R, Wakabayashi M, Hata K, Matsubara T, Honma S, Wakisaka S, Kiyonari H, Shioi G, Yamaguchi A, Tsumaki N, Akiyama H and Yoneda T 2012 Osterix regulates calcification and degradation of chondrogenic matrices through matrix metalloproteinase 13 (MMP13) expression in association with transcription factor Runx2 during endochondral ossification. J. Biol. Chem. 287 33179–33190

    Article  CAS  Google Scholar 

  • Ortuno MJ, Susperregui AR, Artigas N, Rosa JL and Ventura F 2010 Osterix induces Col1a1 gene expression through binding to Sp1 sites in the bone enhancer and proximal promoter regions. Bone 52 2013 548–556

    Google Scholar 

  • Perez-Campo FM, Santurtun A, Garcia-Ibarbia C, Pascual MA, Valero C, Garces C, Sanudo C, Zarrabeitia MT and Riancho JA 2016 Osterix and RUNX2 are transcriptional regulators of sclerostin in human bone. Calcif .Tissue Int. 99 302–309

    Article  CAS  Google Scholar 

  • Ran FA, Hsu PD, Lin CY, Gootenberg JS, Konermann S, Trevino AE, Scott DA, Inoue A, Matoba S, Zhang Y and Zhang F 2013 Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity. Cell 154 1380–1389

    Article  CAS  Google Scholar 

  • Reinholt FP, Hultenby K, Oldberg A and Heinegard D 1990 Osteopontin–a possible anchor of osteoclasts to bone. Proc. Natl. Acad. Sci. USA 87 4473–4475

    Article  CAS  Google Scholar 

  • Renn J and Winkler C 2014 Osterix/Sp7 regulates biomineralization of otoliths and bone in medaka (Oryzias latipes). Matrix Biol. 34 193–204

    Article  CAS  Google Scholar 

  • Slavkin HC, Hu CC, Sakakura Y, Diekwisch T, Chai Y, Mayo M, Bringas P Jr, Simmer J, Mak G, Sasano Y, et al. 1992 Gene expression, signal transduction and tissue-specific biomineralization during mammalian tooth development. Crit. Rev. Eukaryot. Gene Expr. 2 315–329

    CAS  PubMed  Google Scholar 

  • Tang W, Li Y, Osimiri L and Zhang C 2011 Osteoblast-specific transcription factor Osterix (Osx) is an upstream regulator of Satb2 during bone formation. J. Biol. Chem. 286 32995–33002

    Article  CAS  Google Scholar 

  • Termine JD, Kleinman HK, Whitson SW, Conn KM, McGarvey ML and Martin GR 1981 Osteonectin, a bone-specific protein linking mineral to collagen. Cell 26 99–105

    Article  CAS  Google Scholar 

  • Thomas BL, Porteus MH, Rubenstein JL and Sharpe PT 1995 The spatial localization of Dlx-2 during tooth development. Connect. Tissue Res. 32 27–34

    Article  CAS  Google Scholar 

  • Verreijdt L, Debiais-Thibaud M, Borday-Birraux V, Van der Heyden C, Sire JY and Huysseune A 2006 Expression of the dlx gene family during formation of the cranial bones in the zebrafish (Danio rerio): differential involvement in the visceral skeleton and braincase. Dev. Dyn. 235 1371–1389

    Article  CAS  Google Scholar 

  • Walker MB and Kimmel CB 2007 A two-color acid-free cartilage and bone stain for zebrafish larvae. Biotech. Histochem. 82 23–28

    Article  CAS  Google Scholar 

  • Weinreb M, Shinar D and Rodan GA 1990 Different pattern of alkaline phosphatase, osteopontin, and osteocalcin expression in developing rat bone visualized by in situ hybridization. J. Bone Miner. Res. 5 831–842

    Article  CAS  Google Scholar 

  • Wise SB and Stock DW 2010 bmp2b and bmp4 are dispensable for zebrafish tooth development. Dev. Dyn. 239 2534–2546

    Article  CAS  Google Scholar 

  • Yu T, Graf M, Renn J, Schartl M, Larionova D, Huysseune A, Witten PE and Winkler C 2017 A vertebrate-specific and essential role for osterix in osteogenesis revealed by gene knockout in the teleost medaka. Development 144 265–271

    Article  CAS  Google Scholar 

  • Yuan Q, Zhao M, Tandon B, Maili L, Liu X, Zhang A, Baugh EH, Tran T, Silva RM, Hecht JT, Swindell EC, Wagner DS and Letra A 2017 Role of WNT10A in failure of tooth development in humans and zebrafish. Mol. Genet. Genomic Med. 5 730–741

    Article  CAS  Google Scholar 

  • Yun-Feng W, Matsuo N, Sumiyoshi H and Yoshioka H 2010 Sp7/Osterix up-regulates the mouse pro-alpha3(V) collagen gene (Col5a3) during the osteoblast differentiation. Biochem. Biophys. Res. Commun. 394 503–508

    Article  Google Scholar 

  • Zhang C, Cho K, Huang Y, Lyons JP, Zhou X, Sinha K, McCrea PD and de Crombrugghe B 2008 Inhibition of Wnt signaling by the osteoblast-specific transcription factor Osterix. Proc. Natl. Acad. Sci. USA 105 6936–6941

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by Grants from Suzhou people’s Welfare and Science Items (SYS201659).

Author information

Author notes

  1. These authors contributed equally to this work.

Authors and Affiliations

  1. Jiangsu Key Laboratory of Oral Diseases, Nanjing Medical University, Nanjing, China

    Zhongjian Chen & Hongbing Jiang

  2. Department of Oral and Maxillofacial Surgery, The Affiliated Hospital of Stomatology, Nanjing Medical University, Nanjing, China

    Zhongjian Chen & Hongbing Jiang

  3. The Affiliated Stemmatological Hospital of Soochow University, Suzhou, China

    Zhongjian Chen & Zhiyun Song

  4. School of Biology and Basic Medical Sciences, Soochow University, Suzhou, China

    Jinjing Yang & Jian Huang

Authors
  1. Zhongjian Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Zhiyun Song
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jinjing Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jian Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Hongbing Jiang
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hongbing Jiang.

Additional information

Editorial Responsibiility: BJ Rao.

Corresponding editor: BJ Rao

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, Z., Song, Z., Yang, J. et al. Sp7/osterix positively regulates dlx2b and bglap to affect tooth development and bone mineralization in zebrafish larvae. J Biosci 44, 127 (2019). https://doi.org/10.1007/s12038-019-9948-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s12038-019-9948-5

Keywords

Search

Navigation