Potential function of TGF-β isoforms in maturation-stage ameloblasts

Publication date: Available online 3 January 2019

Source: Journal of Oral Biosciences

Author(s): Miu Okubo, Risako Chiba, Takeo Karakida, Hajime Yamazaki, Ryuji Yamamoto, Saeko Kobayashi, Takahiko Niwa, Henry C. Margolis, Takatoshi Nagano, Yasuo Yamakoshi, Kazuhiro Gomi

Abstract

Objectives

To investigate potential functions of transforming growth factor-beta (TGF-β) isoforms in maturation-stage ameloblasts during amelogenesis.

Methods

In vivo activation of TGF-β was characterized by using matrix metalloproteinase 20 null (Mmp20^-/-) and wild-type (Mmp20^+/+) mice. Using mHAT9d cells cultured in the presence of each TGF-β isoform, (1) cell proliferation was determined by MTS assay, (2) immunostaining with anti-cleaved caspase-3 monoclonal antibody was performed and apoptotic indices were measured, (3) gene expression was analyzed by RT-qPCR, and (4) the uptake of amelogenin into mHAT9d cells was directly observed using a fluorescence microscope.

Results

TGF-β1 and TGF-β3 were present in the enamel matrix of developing teeth which were activated by MMP20 in vivo. A genetic study revealed that the three TGF-β isoforms upregulate kallikrein 4 (KLK4) mRNA levels but downregulate carbonic anhydrase II. Moreover, TGF-β1 and TGF-β2 significantly upregulated the mRNA level of amelotin, whereas TGF-β3 dramatically downregulated the mRNA levels of odontogenic ameloblast-associated protein (ODAM), family with sequence similarity 83 member H (FAM83H), and alkaline phosphatase (ALP). Immunostaining analysis showed that the apoptosis of mHAT9d cells is induced by three TGF-β isoforms, with TGF-β3 being most effective. Both TGF-β1 and TGF-β3 induced endocytosis of amelogenin.

Conclusions

We propose that TGF-β is regulated in an isoform-specific manner to perform multiple biological functions such as gene expression related to the structure of basal lamina/ameloblasts, mineral ion transport, apoptosis, and endocytosis in maturation-stage ameloblasts.

Graphical abstract

http://bit.ly/2R93Udd