New population of odontoblasts responsible for tooth root formation

• We identified and characterized apical odontoblasts for the formation of tooth roots.
• Apical odontoblasts are differentiating odontoblasts, located in the apical side of developing roots.
• Several transcription factors are temporospatially expressed in apical odontoblasts.
• Apical odontoblasts may be easily damaged to genetic alteration as well as irradiation.

Root formation is initiated with the extension of Hertwig’s epithelial root sheath (HERS) after crown morphogenesis. To date, little is known about the molecular and cellular mechanisms controlling root formation. Recently we found rootless molars are formed in the dental mesenchyme-specific β-catenin conditional knockout mice. The striking root phenotypes of these mutant mice result from the disrupted differentiation of differentiating odontoblasts, caused by ablation of β-catenin during initiation of root formation. Here we show the cellular and molecular characteristics of differentiating odontoblasts using histochemistry and immunohistochemistry. These cells were not found in crown formation, but appeared only in the apical end of developing tooth, thus we have named these cells “apical odontoblasts” (AOds). AOds appeared immediately after HERS formation and were always present on the apical side of developing roots until root formation was complete. These findings indicate that AOds are closely associated with the transition from crown to root and with root elongation. In AOds, several transcription factors, including Nfic, Creb3l1, and Osx, as well as β-catenin and alkaline phosphatase were expressed but Phex and Dspp were not expressed. Taken together, our results indicate that AOds are the principal cells responsible for tooth root formation. These findings may contribute to the further understanding of the mechanisms underlying tooth root formation and root regeneration.