ArticleHistochemistry and cell biology2008
In situ detection of methylated DNA by histo endonuclease-linked detection of methylated DNA sites: a new principle of analysis of DNA methylation.
Article in Histochemistry and cell biology, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. An erratum has been issued. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 34 citations in OpenAlex.
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- In focus in HCB.Histochemistry and cell biology · 2023Article
- In situ sequence-specific visualization of single methylated cytosine on tissue sections using ICON probe and rolling-circle amplification.Histochemistry and cell biology · 2023Article
- Histochemistry, Cytochemistry and Epigenetics.Acta histochemica et cytochemica · 2022Review
- Vitamin K-Dependent γ-Glutamyl Carboxylase in Sertoli Cells Is Essential for Male Fertility in Mice.Molecular and cellular biology · 2021Article
- Global changes in epigenomes during mouse spermatogenesis: possible relation to germ cell apoptosis.Histochemistry and cell biology · 2020Review
- In focus in HCB.Histochemistry and cell biology · 2020Article
- Morphology-oriented epigenetic research.Histochemistry and cell biology · 2018Review
- Analysis of H3K27me3 expression and DNA methylation at CCGG sites in smoking and non-smoking patients with non-small cell lung cancer and their clinical significance.Oncology letters · 2018Article
- Changes in DNA Methylation of Oocytes and Granulosa Cells Assessed by HELMET during Folliculogenesis in Mouse Ovary.Acta histochemica et cytochemica · 2018Article
- Global 5-methylcytosine alterations in DNA during ageing of Quercus robur seeds.Annals of botany · 2015Article
- Keratinocyte Growth Factor Gene Electroporation into Skeletal Muscle as a Novel Gene Therapeutic Approach for Elastase-Induced Pulmonary Emphysema in Mice.Acta histochemica et cytochemica · 2015Article
- Aberrant levels of histone H3 acetylation induce spermatid anomaly in mouse testis.Histochemistry and cell biology · 2015Article
- Roles of epigenome in mammalian spermatogenesis.Reproductive medicine and biology · 2014Review
- Decrease in cytosine methylation at CpG island shores and increase in DNA fragmentation during zebrafish aging.Age (Dordrecht, Netherlands) · 2014Article
- Accelerated proliferation of hepatocytes in rats with iron overload after partial hepatectomy.Histochemistry and cell biology · 2012Article
- Estrogen-dependent regulation of sodium/hydrogen exchanger-3 (NHE3) expression via estrogen receptor β in proximal colon of pregnant mice.Histochemistry and cell biology · 2012Article
- Immunohistochemical Analysis of Histone H3 Modifications in Germ Cells during Mouse Spermatogenesis.Acta histochemica et cytochemica · 2011Article
- 5-Hydroxymethylcytosine in the mammalian zygote is linked with epigenetic reprogramming.Nature communications · 2011Article
- A method to quantify mouse coat-color proportions.PloS one · 2009Article
Corrections and comments
- Erratum issued
Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
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.
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