The analysis for role of enamel and enamel related protein in, during body formation

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The expression of amelogenin in rats upper molar were studied by using in situ PCR method that detected amplified gene in histology to examine a role of amelogenin for tissue formation. Results of this study, the expression of amelogenin were confirmed in the ameloblasts and initial undifferentiational odontoblasts. Furthermore, the faintly amelogenin gene expression were observed not only in the undifferentiational but also in some degree of maturational odontoblasts. However, a lot of problems of tissue sections such as removing sections from a slide grasses and a destruction of the tissue were happened in this method because of repeating the temperatures change from high temperatures to a low temperatures quickly, Therefore a new gene amplified method was selected in stead of PCR. Recently, the LAMP method that was used four primers on gene amplification. The character of this methods are used a constant temperature of 75℃, and the amplification efficiency of this method is from ten to hundreds of times higher than a general PCR methods.In this study, in situ LAMP method that was detected amplified gene expression in the tissue section, same as in situ PCR method was tried to detect the gene expression of amelogenin in odontoblasts and pulp cells. Previously, some researchers were reported that using in situ LAMP method to soft tissue. However there were no reports for using hard tissues. Therefore the in situ LAMP system for hard tissues has not been established now, and this study was enforced to establish a system for hard tissues with in situ LAMP method.Moreover, one of main enamel proteases, KLK4 was also examined in this study. From the results of this study, KLK4 gene expression and protein localization were observed not only in ameloblasts, but also in oral cancer cells. This result indicated that amelogenin might has a several role for several areas of cells.