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Simultaneous Fluorescent Identification of Odontoblasts and Ameloblasts.
K Isono1, E Takahashi2, I Miyoshi3
1Department of Stem Cells and Developmental Biology, Division of Fundamental Medicine, Mie University Graduate School of Medicine, Tsu, Mie, Japan.
Journal of Dental Research
|December 8, 2020
Summary
New mouse models accurately label odontoblasts and ameloblasts using fluorescent reporters, aiding tooth development and regeneration research. These models allow precise tracking of cell differentiation and isolation for further study.
Area of Science:
- Developmental Biology
- Regenerative Medicine
- Genetics
Background:
- Accurate identification of odontoblasts and ameloblasts is crucial for studying tooth development and regeneration.
- Existing transgenic models for labeling these cells may not perfectly reflect endogenous gene expression patterns.
Purpose of the Study:
- To establish novel mouse models for precise labeling and tracking of odontoblasts and ameloblasts.
- To validate the accuracy of reporter gene expression against endogenous markers.
- To investigate the roles of specific genes in tooth development and mineralization.
Main Methods:
- Generated two mouse lines: one with tdTomato knocked into the Amelogenin (Amelx) gene and another with green fluorescent protein (GFP) knocked into the Dentin sialophosphoprotein (Dspp) gene.
- Utilized micro-computed tomography (micro-CT) to analyze enamel volume and dentin mineral density.
- Employed fluorescence-activated cell sorting (FACS) with antibodies against cell surface markers (CD90, CD166, CD49f, Epcam1) to isolate and identify cell populations.
Main Results:
- tdTomato and GFP were highly expressed in secretory ameloblasts and odontoblasts, respectively, closely mirroring endogenous expression.
- DSPP and AMELX were correctly localized within dentin and enamel matrices, respectively.
- Analysis revealed significant alterations in enamel volume and dentin mineral density in the reporter mouse lines, indicating their utility in studying tooth formation.
- Specific cell surface marker combinations successfully distinguished and enriched GFP+ odontoblasts and tdTomato+ ameloblasts from developing tooth germ.
Conclusions:
- The established reporter mouse models provide accurate and reliable tools for studying odontoblast and ameloblast differentiation.
- These models facilitate in vivo and in vitro investigations of tooth development and regeneration.
- The ability to easily distinguish and isolate these cell types opens new avenues for regenerative dentistry research.
