Localization of Putative Stem Cells in Dental Epithelium and Their Association with Notch and Fgf Signaling

TLDR

The continuously growing mouse incisor, whose epithelium growth is regulated by mesenchyme, serves as an excellent model for studying stem cell lineage mechanisms. The study proposes that mesenchymal FGF signaling regulates the Notch pathway in dental epithelial stem cells by stimulating lunatic fringe, thereby coupling stem cell proliferation and fate decisions. The authors cultured the incisor apex, labeled epithelial lineages with BrdU and DiI, and applied FGF‑10 beads to the cervical loop epithelium to stimulate proliferation and lunatic fringe expression. Stem cells are located in the cervical loop epithelium, with slowly dividing Notch1‑expressing stellate reticulum cells adjacent to lunatic fringe‑expressing basal cells; FGF‑3/10 from the mesenchyme promote proliferation and lunatic fringe, supporting the proposed FGF‑Notch regulatory model.

Abstract

The continuously growing mouse incisor is an excellent model to analyze the mechanisms for stem cell lineage. We designed an organ culture method for the apical end of the incisor and analyzed the epithelial cell lineage by 5-bromo-2′-deoxyuridine and DiI labeling. Our results indicate that stem cells reside in the cervical loop epithelium consisting of a central core of stellate reticulum cells surrounded by a layer of basal epithelial cells, and that they give rise to transit-amplifying progeny differentiating into enamel forming ameloblasts. We identified slowly dividing cells among the Notch1-expressing stellate reticulum cells in specific locations near the basal epithelial cells expressing lunatic fringe, a secretory molecule modulating Notch signaling. It is known from tissue recombination studies that in the mouse incisor the mesenchyme regulates the continuous growth of epithelium. Expression of Fgf-3 and Fgf-10 were restricted to the mesenchyme underlying the basal epithelial cells and the transit-amplifying cells expressing their receptors Fgfr1b and Fgfr2b. When FGF-10 protein was applied with beads on the cultured cervical loop epithelium it stimulated cell proliferation as well as expression of lunatic fringe. We present a model in which FGF signaling from the mesenchyme regulates the Notch pathway in dental epithelial stem cells via stimulation of lunatic fringe expression and, thereby, has a central role in coupling the mitogenesis and fate decision of stem cells.