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Tuftelin and HIFs expression in osteogenesis

  • Jan Bobek
  • Veronika Oralova
  • Adela Kratochvilova
  • Ivana Zvackova
  • Herve Lesot
  • Eva MatalovaEmail author
Original Paper

Abstract

Tuftelin was originally discovered and mostly studied in the tooth, but later found also in other organs. Despite its wide distribution among tissues, tuftelin’s function has so far been specified only in the formation of enamel crystals. Nevertheless, in many cases, tuftelin was suggested to be associated with cellular adaptation to hypoxia and recently even with cell differentiation. Therefore, we aimed to investigate tuftelin expression along with hypoxia-inducible factors (HIFs) during the early development of the mandibular/alveolar (m/a) bone, when osteoblasts started to differentiate in vivo and to compare their expression levels in undifferentiated versus differentiated osteoblastic cells in vitro. Immunohistochemistry demonstrated the presence of tuftelin already in osteoblastic precursors which were also HIF1-positive, but HIF2-negative. Nevertheless, HIF2 protein appeared when osteoblasts differentiated, one day later. This is in agreement with observations made with MC3T3-E1 cells, where there was no significant difference in tuftelin and Hif1 expression in undifferentiated vs. differentiated cells, although Hif2 increased upon differentiation induction. In differentiated osteoblasts of the m/a bone, all three proteins accumulated, first, prenatally, in the cytoplasm and later, particularly at postnatal stages, they displayed also peri/nuclear localization. Such a dynamic time–space pattern of tuftelin expression has recently been reported in neurons, which, as the m/a bone, differentiate under less hypoxic conditions as indicated also by a prevalent cytoplasmic expression of HIF1 in osteoblasts. However, unlike what was shown in cultured neurons, tuftelin does not seem to participate in final osteoblastic differentiation and its functions, thus, appears to be tissue specific.

Keywords

Intramembranous Ossification Bone Tuftelin HIF1 HIF2 

Notes

Acknowledgement

This work was supported by the Grant Agency of the Czech Republic (17-14886S).

Author contributions

J. B. samples, analysis (immunohistochemistry), evaluation of results, contribution to manuscript preparation; V. O. samples, analysis (qPCR/bones, immunofluorescence), evaluation of results, contribution to manuscript preparation; A. K. analysis (qPCR/MC3T3-E1 cells); I. Z. preliminary data; H. L. critical review and contribution to manuscript preparation; E. M. head of the project.

Compliance with ethical standards

Conflict of interest

The authors declare they have no competing interests.

Supplementary material

418_2019_1813_MOESM1_ESM.tif (72 kb)
Supplementary material 1 (TIFF 71 kb). qPCR of Glut1 and Pdk1 in the intramembranous/alveolar bone at E13, E14, E15. Glut1 expression in the forming intramembranous alveolar/mandibular bone at E13, E14, E15, and in the undifferentiated/differentiated MC3T3 cells (a). Pdk1 expression at E13, E14, E15 in the intramembranous alveolar/mandibular bone (b). The results are representative of three independent experiments
418_2019_1813_MOESM2_ESM.tif (110 kb)
Supplementary material 2 (TIFF 109 kb). qPCR of Hif1, Hif2, Glut1 and Pdk1 during differentiation of MC3T3-E1 cells. Intervals of 3, 10 and 21 days of cultivation, expression of Hif1 (a), Hif2 (b), Glut1 (c), Pdk1 (d). Statistically significant differences are highlighted (ANOVA, *p < 0.01; **p < 0.0001). The results are representative of three independent experiments
418_2019_1813_MOESM3_ESM.tif (25 kb)
Supplementary material 3 (TIFF 24 kb). qPCR of tuftelin during differentiation of MC3T3-E1 cells. Intervals of 3, 10 and 21 days of cultivation. The results are representative of three independent experiments

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Jan Bobek
    • 1
  • Veronika Oralova
    • 1
  • Adela Kratochvilova
    • 1
  • Ivana Zvackova
    • 1
  • Herve Lesot
    • 1
  • Eva Matalova
    • 1
    • 2
    Email author
  1. 1.Laboratory of Odontogenesis and OsteogenesisInstitute of Animal Physiology and Genetics, v.v.i, Academy of Sciences of the Czech RepublicBrnoCzech Republic
  2. 2.Department of PhysiologyUniversity of Veterinary and Pharmaceutical SciencesBrnoCzech Republic

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