Global and gene-specific histone modification profiles of mouse multipotent adult germline stem cells
| Autor: | Marieke Wolf, Ralf Dressel, D. V. Krishna Pantakani, Wolfgang Engel, Ulrich Zechner, Jessica Nolte, Tatjana Khromov |
|---|---|
| Rok vydání: | 2010 |
| Předmět: |
Male
Homeobox protein NANOG Chromatin Immunoprecipitation Embryology Adult Germline Stem Cells Blotting Western Fluorescent Antibody Technique Biology Methylation Polymerase Chain Reaction Cell Line Epigenesis Genetic Histones Mice 03 medical and health sciences 0302 clinical medicine SOX2 Genetics Animals Epigenetics 10. No inequality Molecular Biology 030304 developmental biology Homeodomain Proteins 0303 health sciences Genome Multipotent Stem Cells SOXB1 Transcription Factors Obstetrics and Gynecology Acetylation Nanog Homeobox Protein Cell Biology Flow Cytometry Molecular biology Spermatogonia Chromatin Reproductive Medicine embryonic structures H3K4me3 Octamer Transcription Factor-3 Chromatin immunoprecipitation 030217 neurology & neurosurgery Developmental Biology Bivalent chromatin |
| Zdroj: | Molecular Human Reproduction. 17:166-174 |
| ISSN: | 1460-2407 1360-9947 |
| DOI: | 10.1093/molehr/gaq085 |
| Popis: | We previously reported the generation of multipotent adult germline stem cells (maGSCs) from spermatogonial stem cells (SSCs) isolated from adult mouse testis. In a later study, we substantiated the pluripotency of maGSCs by demonstrating their close similarity to pluripotent male embryonic stem cells (ESCs) at the epigenetic level of global and gene-specific DNA methylation. Here, we extended the comparative epigenetic analysis of maGSCs and male ESCs by investigating the second main epigenetic modification in mammals, i.e. global and gene-specific modifications of histones (H3K4 trimethylation, H3K9 acetylation, H3K9 trimethylation and H3K27 trimethylation). Using immunofluorescence staining, flow cytometry and western blot analysis, we show that maGSCs are very similar to male ESCs with regard to global levels and nuclear distribution patterns of these modifications. Chromatin immunoprecipitation real-time PCR analysis of these modifications at the gene-specific level further revealed modification patterns of the pluripotency marker genes Oct4, Sox2 and Nanog in maGSCs that are nearly identical to those of male ESCs. These genes were enriched for activating histone modifications including H3K4me3 and H3K9ac and depleted of repressive histone modifications including H3K27me3 and H3K9me3. In addition, Hoxa11, a key regulator of early embryonic development showed the ESC-typical bivalent chromatin conformation with enrichment of both the activating H3K4me3 and the repressive H3K27me3 modification also in maGSCs. Collectively, our results demonstrate that maGSCs also closely resemble ESCs with regard to their chromatin state and further evidence their pluripotent nature. |
| Databáze: | OpenAIRE |
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