GSK3β regulates ameloblast differentiation via Wnt and TGF-β pathways

Wnt and TGF-β signaling pathways participate in regulating a variety of cell fates during organogenesis, including tooth development. Despite well-documented, the specific mechanisms, especially how these two pathways act coordinately in regulating enamel development, remain unknown. In this study, we identified Glycogen Synthase Kinase 3 beta (GSK3β), a negative regulator of Wnt signal pathway, participated in ameloblast differentiation via Wnt and TGF-β pathways during enamel development. In vitro rat mandible culture treated with specific GSK3β inhibitor SB415286 displayed enamel defects, accompanied by disrupted ameloblasts polarization, while odontoblasts and dentin appeared to be unaffected. Moreover, after GSK3β knockdown by lentivirus-mediated RNA silencing, HAT-7 cells displayed abnormal cell polarity and cell adhesion, and failed to synthesize appreciable amounts of ameloblast-specific proteins. More importantly, inactivation of GSK3β caused upregulated Wnt and downregulated TGF-β pathway, while reactivation of TGF-β signaling or suppression of Wnt signaling partially rescued the differentiation defects of ameloblasts caused by the GSK3β knock-down. Taken together, these results suggested that GSK3β was essential for ameloblasts differentiation, which might be indirectly mediated through Wnt and TGF-β signaling pathways.