Abstract
Cytodifferentiation of odontogenic cells, a late stage event in odontogenesis is based on gene regulation. However, studies on the identification of the involved genes are scarce. The present study aimed to search for molecules for the cytodifferentiation of ameloblastic cells in rats. Differential display-PCR revealed a differentially expressed gene between cap/early bell stage and hard tissue formation stage in molars. This gene was identified as N-myc Downregulated Gene 1 (Ndrg1), which is the first report in tooth development. Real time PCR and western blotting confirmed that the mRNA level of Ndrg1 was higher during enamel formation than the cap stage. Ndrg1 expression was upregulated in the early bell, crown, and root stages in a time-dependent manner. These patterns of expression were similar in Ndrg2, but Ndrg3 and Ndrg4 levels did not change during the developmental stages. Immunofluorescence revealed that strong immunoreactivity against Ndrg1 were detected in differentiated ameloblasts only, not inner enamel epithelium, odontoblasts and ameloblastic cells in defected enamel regions. Alkaline phosphatase and alizarin red s stains along with real time PCR, revealed that Ndrg1 and Ndrg2 were involved in cytodifferentiation and enamel matrix mineralization by selectively regulating amelogenin and ameloblastin genes in SF2 ameloblastic cells. These results suggest that Ndrg may play a crucial functional role in the cytodifferentiation of ameloblasts for amelogenesis.
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Acknowledgements
This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MSIT) (No. 2019R1A5A2027521), (No. 2019R1A2C1003520), and Basic Science Research Program funded by the Ministry of Education (RS-2023-00244054). The authors report no conflicts of interest related to this study.
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All authors have read and approved the final version of the manuscript. DW.Y and JH.K contributed to conception, data acquisition, and analysis; MS.K and SH .K contributed to design and data interpretation and drafted and critically revised the manuscript.
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Yang, DW., Kang, JH., Kim, MS. et al. Regulatory role of N-myc downregulated genes in amelogenesis in rats. J Mol Histol 55, 149–157 (2024). https://doi.org/10.1007/s10735-024-10182-9
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DOI: https://doi.org/10.1007/s10735-024-10182-9