Expression of fibroblast growth factor receptor1, -2c, and -3c transcripts in mouse molars after tooth eruption

A previous study suggested that fibroblast growth factor (FGF) signaling plays an important role in dentin formation during tooth development. In this study, to examine dentin formation after tooth eruption involving secondary and tertiary dentin, we analyzed the expression patterns and expressing cells of Fgfr1, -2c, and -3c in mouse maxillary first molars (M1). Since it is difficult to recover the mRNAs from mineralized tissues, we tested methods for extraction after fixation and decalcification of teeth. We successfully obtained consistent results with quantitative real-time PCR (qPCR) using β-actin transcripts for validation. qPCR for Dentin sialo phosphoprotein (Dspp), Fgfr1, -2c, and -3c transcripts was performed on mice at ages of 2-20 weeks. The results showed that the highest expression levels of Dspp and Fgfr2c occurred at 2 weeks old followed by lower expression levels after 4 weeks old. However, the expression levels of Fgfr1 and Fgfr3c were constant throughout the experimental period. By in situ hybridization, Dspp, Fgfr1, and Fgfr3c transcripts were detected in odontoblasts at ages of 2 and 4 weeks. Also, Dspp and Fgfr1 transcripts were detected in odontoblasts facing reactionary dentin at 8 weeks old. These results suggest that FGF-FGFR signaling might be involved in the regulation of odontoblasts even after tooth eruption, including secondary and tertiary dentin formation. Moreover, our modified method for extracting mRNA from mineralized tissues after fixation and decalcification successfully produced consistent results.

By utilizing our modified RNA extraction method, we determined the expression of FGFRs after tooth eruption, which suggested the commitment of FGF signaling to the homeostasis of odontoblasts.

内容要旨・審査要旨・論文本文の公開
本論文は，著者Tsuyoshi Kanoの学位論文として提出され，学位審査・授与の対象となっている。
This is a post-peer-review, pre-copyedit version of an article published in Anatomical Science International. The final authenticated version is available online at: https://doi.org/10.1007/s12565-020-00594-4