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Cell and Tissue Research

, Volume 361, Issue 2, pp 457–466 | Cite as

Microtubule-associated protein tau (Mapt) is expressed in terminally differentiated odontoblasts and severely down-regulated in morphologically disturbed odontoblasts of Runx2 transgenic mice

  • Toshihiro Miyazaki
  • Tomomi T. Baba
  • Masako Mori
  • Takeshi Moriishi
  • Toshihisa Komori
Short Communication
  • 464 Downloads

Abstract

Runx2 is an essential transcription factor for osteoblast and odontoblast differentiation and the terminal differentiation of chondrocytes. We have previously shown that the terminal differentiation of odontoblasts is inhibited in Runx2 transgenic {Tg(Col1a1-Runx2)} mice under the control of the 2.3-kb Col1a1 promoter, which directs the transgene expression to osteoblasts and odontoblasts. Odontoblasts show severe reductions in Dspp and nestin expression and lose their characteristic polarized morphology, including a long process extending to dentin, in Tg(Col1a1-Runx2) mice. We study the molecular mechanism of odontoblast morphogenesis by comparing gene expression in the molars of wild-type and Tg(Col1a1-Runx2) mice, focusing on cytoskeleton-related genes. Using microarray, we found that the gene expression of microtubule-associated protein tau (Mapt), a neuronal phosphoprotein with important roles in neuronal biology and microtubule dynamics and assembly, was high in wild-type molars but severely reduced in Tg(Col1a1-Runx2) molars. Immunohistochemical analysis revealed that Mapt was specifically expressed in terminally differentiated odontoblasts including their processes in wild-type molars but its expression was barely detectable in Tg(Col1a1-Runx2) molars. Double-staining of Mapt and Runx2 showed their reciprocal expression in odontoblasts. Mapt and tubulin co-localized in odontoblasts in wild-type molars. Immunoelectron microscopic analysis demonstrated Mapt lying around α-tubulin-positive filamentous structures in odontoblast processes. Thus, Mapt is a useful marker for terminally differentiated odontoblasts and might play an important role in odontoblast morphogenesis.

Keywords

Microtubule-associated protein tau (Mapt) Odontoblast Cytoskeleton α-Tubulin Runx2 

Notes

Acknowledgments

We thank Y. Matsuo for technical assistance.

Supplementary material

441_2015_2135_MOESM1_ESM.pdf (72 kb)
Supplemental Table 1 (PDF 71 kb)
441_2015_2135_MOESM2_ESM.pdf (93 kb)
Supplemental Table 2 (PDF 92 kb)
441_2015_2135_MOESM3_ESM.pdf (86 kb)
Supplemental Table 3 (PDF 85 kb)
441_2015_2135_MOESM4_ESM.pdf (718 kb)
Supplementary Figure 1 Hematoxylin and eosin (H-E) staining of a section of an extracted lower first molar at 2 weeks of age. Intercuspal and root regions indicated by boxes in a are magnified in b, c, respectively. No cellular degradation of the extracted teeth including periodontal tissues and reduced enamel epithelium (Re) is seen (D dentin, Dp dental pulp, Ob odontoblasts, Pl periodontal ligament). Bars 500 μm (a), 100 μm (b, c). (PDF 718 kb)
441_2015_2135_MOESM5_ESM.pdf (754 kb)
Supplementary Figure 2 Immunohistochemical analysis of Mapt. Localization of Mapt in frontal sections of the craniofacial region of wild-type (wt; a, c, d) and Tg(Col1a1-Runx2) (tg; b, e, f) mice at 1 day of age. Boxed regions in a, b are shown at higher magnification in c–f, respectively. Mapt expression (brown) is found in nervous tissues in both mice types, whereas it is located in odontoblasts in tooth germs of wild-type mice but not of Tg(Col1a1-Runx2) mice (arrows in c, e). Bars 1 mm (a, b), 100 μm (c–f). (PDF 754 kb)
441_2015_2135_MOESM6_ESM.pdf (734 kb)
Supplementary Figure 3 Immunohistochemical localization of DSP. Localization of DSP protein in the first molars in wild-type (wt, a, b, e, f) and Tg(Col1a1-Runx2) (tg) (c, d, g, h) mice at 1 day (a–d) and 9 days (e–h) of age. Boxed regions in a, c, e, g are shown at higher magnification in b, d, f, h, respectively. DSP expression is exclusively and strongly detected in differentiated odontoblasts, dentinal tubules and dentin matrices in wild-type mice, whereas it is faintly detected in odontoblasts in Tg(Col1a1-Runx2) mice (Ob odontoblasts). Bars 200 μm (a, c), 500 μm (e, g), 50 μm (b, d), 20 μm (f, h). (PDF 733 kb)
441_2015_2135_MOESM7_ESM.pdf (750 kb)
Supplementary Figure 4 Immunohistochemical localization of tubulin α1A. Localization of tubulin α1A protein in the first molars in wild-type (wt, a, b, e, f) and Tg(Col1a1-Runx2) (tg, c, d, g, h) mice at 1 day (a–d) and 9 days (e–h) of age. Boxed regions in a, c, e, g are shown at higher magnification in b, d, f, h, respectively. Tubulin α1A protein is detected in odontoblasts and in various other cells in both wild-type and Tg(Col1a1-Runx2) mice. The intensity of tubulin α1A staining was similar in odontoblasts of wild-type and Tg(Col1a1-Runx2) molars at 1 day of age, whereas it was reduced in odontoblasts of Tg(Col1a1-Runx2) molars at 9 days of age mainly because of the lack of the tall columnar cell body and polarization. Bars 200 μm (a, c), 500 μm (e, g), 50 μm (b, d), 20 μm (f, h). (PDF 750 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  • Toshihiro Miyazaki
    • 1
  • Tomomi T. Baba
    • 2
  • Masako Mori
    • 1
  • Takeshi Moriishi
    • 1
  • Toshihisa Komori
    • 1
  1. 1.Department of Cell Biology, Unit of Basic Medical SciencesNagasaki University Graduate School of Biomedical SciencesNagasakiJapan
  2. 2.Department of Oral Molecular Biology, Unit of Basic Medical SciencesNagasaki University Graduate School of Biomedical SciencesNagasakiJapan

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