TODENTJ-25-03-2019_TODENTJ-4-223 1553505216 Bentham Open doi@benthamopen.net Bentham Open The Open Dentistry Journal TODENTJ 1874-2106 12 15 2010 4 1 Andrew Cho Shigeki Suzuki Junko Hatakeyama Naoto Haruyama Ashok B Kulkarni Tooth and bone specimen require extensive demineralization for careful analysis of cell morphology, as well as gene and protein expression levels. The LacZ gene, which encodes the ß-galactosidase enzyme, is often used as a reporter gene to study gene-structure function, tissue-specific expression by a promoter, cell lineage and fate. This reporter gene is particularly useful for analyzing the spatial and temporal gene expression pattern, by expressing the LacZ gene under the control of a promoter of interest. To analyze LacZ activity, and the expression of other genes and their protein products in teeth and bones, it is necessary to carry out a complete demineralization of the specimen before cutting sections. However, strong acids, such as formic acid used for tooth demineralization, destroy the activities of enzymes including those of ß-galactosidase. Therefore, most protocols currently use mild acids such as 0.1 M ethylene diamine tetra-acetic acid (EDTA) for demineralization of tooth and bone specimen, which require a longer period of treatment for complete demineralization. A method by which hard tissue specimens such as teeth and bones can be rapidly, but gently, decalcified is necessary to save time and effort. Here, we report a suitable method for rapid demineralization of mouse teeth in 0.1M EDTA at 42˚C without any loss of ß-galactosidase activity. 12 15 2010 223 229 TODENTJ-4-223 1 10.2174/crossmark_policy opendentistryjournal.com benthamopen.com true Peer Reviewed Single blind Checked with iThenticate 2010-7-21 2010-9-8 2010-9-9 2010-12-15 http://creativecommons.org/licenses/by-nc/3.0/ 10.2174/1874210601004010223 1553505216 https://opendentistryjournal.com/VOLUME/4/PAGE/223/ https://opendentistryjournal.com/contents/volumes/V4/TODENTJ-4-223/TODENTJ-4-223.pdf https://opendentistryjournal.com/contents/volumes/V4/TODENTJ-4-223/TODENTJ-4-223.pdf https://opendentistryjournal.com/contents/volumes/V4/TODENTJ-4-223/TODENTJ-4-223.pdf https://opendentistryjournal.com/contents/volumes/V4/TODENTJ-4-223/TODENTJ-4-223.pdf https://opendentistryjournal.com/contents/volumes/V4/TODENTJ-4-223/TODENTJ-4-223.pdf https://opendentistryjournal.com/contents/volumes/V4/TODENTJ-4-223/TODENTJ-4-223.pdf https://opendentistryjournal.com/contents/volumes/V4/TODENTJ-4-223/TODENTJ-4-223.pdf https://opendentistryjournal.com/contents/volumes/V4/TODENTJ-4-223/TODENTJ-4-223.xml Histochem Cell Biol Shibata Y 113 153 2000 Shibata Y, Fujita S, Takahashi H, Yamaguchi A, Koji T. 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