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Fam20C overexpression in odontoblasts regulates dentin formation and odontoblast differentiation

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Abstract

FAM20C phosphorylates secretory proteins at S-x-E/pS motifs, and previous studies of Fam20C-dificient mice revealed that FAM20C played essential roles in bone and tooth formation. Inactivation of FAM20C in mice led to hypophosphatemia that masks direct effect of FAM20C in these tissues, and consequently the direct role of FAM20C remains unknown. Our previous study reported that osteoblast/odontoblast-specific Fam20C transgenic (Fam20C-Tg) mice had normal serum phosphate levels and that osteoblastic FAM20C-mediated phosphorylation regulated bone formation and resorption. Here, we investigated the direct role of FAM20C in dentin using Fam20C-Tg mice. The tooth of Fam20C-Tg mice contained numerous highly phosphorylated proteins, including SIBLINGs, compared to that of wild-type mice. In Fam20C-Tg mice, coronal dentin volume decreased and mineral density unchanged at early age, while the volume unchanged and the mineral density elevated at maturity. In these mice, radicular dentin volume and mineral density decreased at all ages, and histologically, the radicular dentin had wider predentin and abnormal apical-side dentin with embedded cells and argyrophilic canaliculi. Immunohistochemical analyses revealed that abnormal apical-side dentin had bone and dentin matrix properties accompanied with osteoblast-lineage cells. Further, in Fam20C-Tg mice, DSPP content which is important for dentin formation, was reduced in dentin, especially radicular dentin, which might lead to defects mainly in radicular dentin. Renal subcapsular transplantations of tooth germ revealed that newly formed radicular dentin replicated apical abnormal dentin of Fam20C-Tg mice, corroborating that FAM20C overexpression indeed caused the abnormal dentin. Our findings indicate that odontoblastic FAM20C-mediated phosphorylation in the tooth regulates dentin formation and odontoblast differentiation.

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Data availability

The data used to support the findings of this study are available from the corresponding author upon request.

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Acknowledgements

The phosphoproteomic analysis was performed by Medical ProteoScope (Yokohama, Japan), and the authors would like to acknowledge the support of the staff there.

Funding

This work was supported by Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Numbers JP17H04368, JP26670801, JP19K18944, and JP21K16930.

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Author notes

  1. Kohei Naniwa and Katsutoshi Hirose are contributed equally.

Authors and Affiliations

  1. Department of Oral and Maxillofacial Pathology, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Kohei Naniwa, Katsutoshi Hirose, Yu Usami & Satoru Toyosawa

  2. Department of Oral & Maxillofacial Oncology and Surgery, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Kohei Naniwa & Narikazu Uzawa

  3. Department of Molecular and Cellular Biochemistry, Osaka University Graduate School of Dentistry, 1-8 Yamadaoka, Suita, Osaka, 565-0871, Japan

    Kenji Hata

  4. Bruker Japan K.K. BioSpin Division, Application Department, 3-9 Kanagawaku Moriyacho, Yokohama, Kanagawa, 221-0022, Japan

    Rikita Araki

  5. Department of Molecular Bone Biology, Nagasaki University Graduate School of Biomedical Sciences, 1-7-1 Sakamoto, Nagasaki, 852–8588, Japan

    Toshihisa Komori

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Contributions

All authors contributed to this work, and reviewed and approved the manuscript for submission. KN and KH contributed equally to this work. KN: Investigation, Data Curation, Visualization. KH: Methodology, Validation, Investigation, Resources, Data Curation, Writing—Original Draft, Writing—Review and Editing, Visualization, Supervision, Project Administration, Funding Acquisition. YU: Resources. KH: Investigation, Resources, Supervision. RA: Investigation, Visualization. NU: Supervision. TK: Methodology, Supervision. ST: Conceptualization, Methodology, Validation, Resources, Data Curation, Writing—Original Draft, Writing—Review and Editing, Visualization, Supervision, Project Administration, Funding Acquisition.

Corresponding author

Correspondence to Satoru Toyosawa.

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All the authors have no relevant financial or non-financial interests to disclose.

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All animal experiments were reviewed and approved by the Intramural Animal Use and Care Committee of Osaka University Graduate School of Dentistry (Permit Number R-01-006-0).

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Naniwa, K., Hirose, K., Usami, Y. et al. Fam20C overexpression in odontoblasts regulates dentin formation and odontoblast differentiation. J Mol Histol 54, 329–347 (2023). https://doi.org/10.1007/s10735-023-10123-y

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