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Silencing markers are retained on pericentric heterochromatin during murine primordial germ cell development
BACKGROUND: In the nuclei of most mammalian cells, pericentric heterochromatin is characterized by DNA methylation, histone modifications such as H3K9me3 and H4K20me3, and specific binding proteins like heterochromatin-binding protein 1 isoforms (HP1 isoforms). Maintenance of this specialized chroma...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5346203/ https://www.ncbi.nlm.nih.gov/pubmed/28293300 http://dx.doi.org/10.1186/s13072-017-0119-3 |
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author | Magaraki, Aristea van der Heijden, Godfried Sleddens-Linkels, Esther Magarakis, Leonidas van Cappellen, Wiggert A. Peters, Antoine H. F. M. Gribnau, Joost Baarends, Willy M. Eijpe, Maureen |
author_facet | Magaraki, Aristea van der Heijden, Godfried Sleddens-Linkels, Esther Magarakis, Leonidas van Cappellen, Wiggert A. Peters, Antoine H. F. M. Gribnau, Joost Baarends, Willy M. Eijpe, Maureen |
author_sort | Magaraki, Aristea |
collection | PubMed |
description | BACKGROUND: In the nuclei of most mammalian cells, pericentric heterochromatin is characterized by DNA methylation, histone modifications such as H3K9me3 and H4K20me3, and specific binding proteins like heterochromatin-binding protein 1 isoforms (HP1 isoforms). Maintenance of this specialized chromatin structure is of great importance for genome integrity and for the controlled repression of the repetitive elements within the pericentric DNA sequence. Here we have studied histone modifications at pericentric heterochromatin during primordial germ cell (PGC) development using different fixation conditions and fluorescent immunohistochemical and immunocytochemical protocols. RESULTS: We observed that pericentric heterochromatin marks, such as H3K9me3, H4K20me3, and HP1 isoforms, were retained on pericentric heterochromatin throughout PGC development. However, the observed immunostaining patterns varied, depending on the fixation method, explaining previous findings of a general loss of pericentric heterochromatic features in PGCs. Also, in contrast to the general clustering of multiple pericentric regions and associated centromeres in DAPI-dense regions in somatic cells, the pericentric regions of PGCs were more frequently organized as individual entities. We also observed a transient enrichment of the chromatin remodeler ATRX in pericentric regions in embryonic day 11.5 (E11.5) PGCs. At this stage, a similar and low level of major satellite repeat RNA transcription was detected in both PGCs and somatic cells. CONCLUSIONS: These results indicate that in pericentric heterochromatin of mouse PGCs, only minor reductions in levels of some chromatin-associated proteins occur, in association with a transient increase in ATRX, between E11.5 and E13.5. These pericentric heterochromatin regions more frequently contain only a single centromere in PGCs compared to the surrounding soma, indicating a difference in overall organization, but there is no de-repression of major satellite transcription. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13072-017-0119-3) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5346203 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-53462032017-03-14 Silencing markers are retained on pericentric heterochromatin during murine primordial germ cell development Magaraki, Aristea van der Heijden, Godfried Sleddens-Linkels, Esther Magarakis, Leonidas van Cappellen, Wiggert A. Peters, Antoine H. F. M. Gribnau, Joost Baarends, Willy M. Eijpe, Maureen Epigenetics Chromatin Research BACKGROUND: In the nuclei of most mammalian cells, pericentric heterochromatin is characterized by DNA methylation, histone modifications such as H3K9me3 and H4K20me3, and specific binding proteins like heterochromatin-binding protein 1 isoforms (HP1 isoforms). Maintenance of this specialized chromatin structure is of great importance for genome integrity and for the controlled repression of the repetitive elements within the pericentric DNA sequence. Here we have studied histone modifications at pericentric heterochromatin during primordial germ cell (PGC) development using different fixation conditions and fluorescent immunohistochemical and immunocytochemical protocols. RESULTS: We observed that pericentric heterochromatin marks, such as H3K9me3, H4K20me3, and HP1 isoforms, were retained on pericentric heterochromatin throughout PGC development. However, the observed immunostaining patterns varied, depending on the fixation method, explaining previous findings of a general loss of pericentric heterochromatic features in PGCs. Also, in contrast to the general clustering of multiple pericentric regions and associated centromeres in DAPI-dense regions in somatic cells, the pericentric regions of PGCs were more frequently organized as individual entities. We also observed a transient enrichment of the chromatin remodeler ATRX in pericentric regions in embryonic day 11.5 (E11.5) PGCs. At this stage, a similar and low level of major satellite repeat RNA transcription was detected in both PGCs and somatic cells. CONCLUSIONS: These results indicate that in pericentric heterochromatin of mouse PGCs, only minor reductions in levels of some chromatin-associated proteins occur, in association with a transient increase in ATRX, between E11.5 and E13.5. These pericentric heterochromatin regions more frequently contain only a single centromere in PGCs compared to the surrounding soma, indicating a difference in overall organization, but there is no de-repression of major satellite transcription. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13072-017-0119-3) contains supplementary material, which is available to authorized users. BioMed Central 2017-03-11 /pmc/articles/PMC5346203/ /pubmed/28293300 http://dx.doi.org/10.1186/s13072-017-0119-3 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Magaraki, Aristea van der Heijden, Godfried Sleddens-Linkels, Esther Magarakis, Leonidas van Cappellen, Wiggert A. Peters, Antoine H. F. M. Gribnau, Joost Baarends, Willy M. Eijpe, Maureen Silencing markers are retained on pericentric heterochromatin during murine primordial germ cell development |
title | Silencing markers are retained on pericentric heterochromatin during murine primordial germ cell development |
title_full | Silencing markers are retained on pericentric heterochromatin during murine primordial germ cell development |
title_fullStr | Silencing markers are retained on pericentric heterochromatin during murine primordial germ cell development |
title_full_unstemmed | Silencing markers are retained on pericentric heterochromatin during murine primordial germ cell development |
title_short | Silencing markers are retained on pericentric heterochromatin during murine primordial germ cell development |
title_sort | silencing markers are retained on pericentric heterochromatin during murine primordial germ cell development |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5346203/ https://www.ncbi.nlm.nih.gov/pubmed/28293300 http://dx.doi.org/10.1186/s13072-017-0119-3 |
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