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ナカシマ カズヒサ
Nakashima Kazuhisa
中島 和久 所属 鶴見大学 歯学部 歯学科 薬理学 職種 准教授 |
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| 論文種別 | 【査読あり】 研究論文（学術雑誌） |
| 言語種別 | 英語 |
| 査読の有無 | 査読あり |
| 表題 | Improved methods for detection of beta-galactosidase (lacZ) activity in hard tissue |
| 掲載誌名 | 正式名：HISTOCHEMISTRY AND CELL BIOLOGY ISSNコード：09486143 |
| 出版社 | SPRINGER |
| 巻・号・頁 | 137(6),pp.841-847 |
| 著者・共著者 | Akemi Shimada,Koichiro Komatsu,Kazuhisa Nakashima,Ernst Poeschl,Akira Nifuji |
| 発行年月 | 2012/06 |
| 概要 | The beta-galactosidase gene () of is widely used as a reporter gene. The expression of can be detected by enzyme-based histochemical staining using chromogenic substrates such as 5-bromo-4-chloro-3-indolyl-beta--galactoside (X-gal). Because the enzymatic activity of is vulnerable to high temperatures and acid treatment for demineralization, detection of on paraffinized sections is difficult, especially for hard tissues, which require demineralization before sectioning in paraffin. To circumvent this problem, whole-mount X-gal staining before sectioning is performed. However, detection of activity in the center of larger portions of hard whole adult tissues is challenging. In this study, focusing on fixation procedures, we determined the conditions conducive to improved detection of activity in deeper areas of whole tissues. We used an annexin a5 ()- reporter mouse model in which the expression in hard tissue is indicated by activity. We found that activity could be detected throughout the periodontal ligament of adult mice when fixed in 100% acetone, whereas it was not detected in the periodontal ligament around the root apex fixed in glutaraldehyde and paraformaldehyde. This stain |
| DOI | 10.1007/s00418-012-0936-1 |