Establishment and characterization of rat dental epithelial derived ameloblast-lineage clones.
Teeth are the hardest tissues covered with enamel produced by ameloblasts. The ameloblast differentiation is controlled by sequential epithelial-mesenchymal interactions during tooth morphogenesis. However, the molecular mechanism of ameloblast differentiation remains unclear. To address this question, we developed an in vitro assay system to evaluate the molecular mechanism of amelogenesis. First, we established dental epithelium-derived clones from 6-day-old rat incisors and established that cells of the clone SRE-G5 were the largest producers of amelogenin mRNA. Next, we analyzed the effects of several chemicals on the amelogenin expression in SRE-G5 cells. Only mitogen-activated protein kinase (MAPK) activators enhanced amelogenin mRNA expression. This finding corresponded to the immunohistochemical data showing the presence of phosphorylated forms of p38, c-Jun N-terminal kinase (JNK), and extracellular signal-regulated kinase (ERK) during ameloblast differentiation. To examine the roles of MAPK signals, we compared the effects of anisomycin and sodium salicylate on the expression of tooth-related differentiation markers. Both anisomycin and sodium salicylate induced amelogenin, Abcg2, and Bmp4 mRNA and down-regulated p75NGFR mRNA. On the other hand, ALP, ectodin, Bmp2 and Fgf8 mRNA were up-regulated only by anisomycin. These results indicate that MAPK signaling functions, at least in part, as the inducer of ameloblast differentiation.
Abe
Kaori
K
Department of Molecular Biology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, Japan.
Miyoshi
Keiko
K
Muto
Taro
T
Ruspita
Intan
I
Horiguchi
Taigo
T
Nagata
Toshihiko
T
Noma
Takafumi
T
D000565:Ameloblasts / Q000166:cytology* / Q000378:metabolism*; D000566:Amelogenesis / Q000502:physiology*; D053523:Amelogenin / Q000378:metabolism*; D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D004847:Epithelial Cells / Q000166:cytology* / Q000378:metabolism*; D007180:Incisor / Q000166:cytology / Q000378:metabolism; D051381:Rats
D053523:Amelogenin