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In situ detection of methylated DNA by histo endonuclease-linked detection of methylated DNA sites: a new principle of analysis of DNA methylation.
http://hdl.handle.net/10069/22183
http://hdl.handle.net/10069/2218394728269-b8b5-49ff-8be8-f4cf0040aabe
名前 / ファイル | ライセンス | アクション |
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HCB130_917.pdf (350.5 kB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2009-09-18 | |||||
タイトル | ||||||
タイトル | In situ detection of methylated DNA by histo endonuclease-linked detection of methylated DNA sites: a new principle of analysis of DNA methylation. | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | DNA methylation | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Restriction enzyme | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Apoptosis | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Epigenetics | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Spermatogenesis | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||
資源タイプ | journal article | |||||
著者 |
Koji, Takehiko
× Koji, Takehiko× Kondo, Shiho× Hishikawa, Yoshitaka× An, Shucai× Sato, Yoko |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | For a better understanding of epigenetic regulation of cell differentiation, it is important to analyze DNA methylation at a specific site. Although previous studies described methylation of isolated DNA extracted from cells and tissues using a combination of appropriate restriction endonucleases, no application to tissue cell level has been reported. Here, we report a new method, named histo endonuclease-linked detection of methylation sites of DNA (HELMET), designed to detect methylation sites of DNA with a specific sequences in a tissue section. In this study, we examined changes in the methylation level of CCGG sites during spermatogenesis in paraffin-embedded sections of mouse testis. In principle, the 3'-OH ends of DNA strand breaks in a section were firstly labeled with a mixture of dideoxynucleotides by terminal deoxynucleotidyl transferase (TdT), not to be further elongated by TdT. Then the section was digested with Hpa II, resulting in cutting the center portion of non-methylated CCGG. The cutting sites were labeled with biotin-16-dUTP by TdT. Next, the section was treated with Msp I, which can cut the CCGG sequence irrespective of the presence or absence of methylation of the second cytosine, and the cutting sites were labeled with digoxigenin-11-dUTP by TdT. Finally, both biotin and digoxigenin were visualized by enzyme- or fluorescence-immunohistochemistry. Using this method, we found hypermethylation of CCGG sites in most of the germ cells although non-methylated CCGG were colocalized in elongated spermatids. Interestingly, some TUNEL-positive germ cells, which are frequent in mammalian spermatogenesis, became markedly Hpa II-reactive, indicating that the CCGG sites may be demethylated during apoptosis. | |||||
書誌情報 |
Histochemistry and cell biology 巻 130, 号 5, p. 917-925, 発行日 2008-11 |
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ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 09486143 | |||||
EISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 1432-119X | |||||
書誌レコードID | ||||||
収録物識別子タイプ | NCID | |||||
収録物識別子 | AA11033302 | |||||
PubMed番号 | ||||||
関連タイプ | isVersionOf | |||||
識別子タイプ | PMID | |||||
関連識別子 | 18797917 | |||||
DOI | ||||||
関連タイプ | isVersionOf | |||||
識別子タイプ | DOI | |||||
関連識別子 | 10.1007/s00418-008-0487-7 | |||||
権利 | ||||||
権利情報 | c Springer-Verlag 2008 | |||||
権利 | ||||||
権利情報 | The original publication is available at www.springerlink.com | |||||
著者版フラグ | ||||||
出版タイプ | AM | |||||
出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||
引用 | ||||||
内容記述タイプ | Other | |||||
内容記述 | Histochemistry and Cell Biology, 130(5), pp.917-925; 2008 |