A chromEM-staining protocol optimized for cardiac tissue
Three-dimensional (3D) chromatin organization has a key role in defining the transcription program of cells during development. Its alteration is the cause of gene expression changes responsible for several diseases. Thus, we need new tools to study this aspect of gene expression regulation. To this...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-07-01
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| Series: | Frontiers in Cell and Developmental Biology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcell.2023.1123114/full |
| _version_ | 1797786751867027456 |
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| author | Elettra Musolino Christina Pagiatakis Christina Pagiatakis Federica Pierin Daniele Sabatino Giovanna Finzi Rosalba Gornati Giovanni Bernardini Roberto Papait Roberto Papait |
| author_facet | Elettra Musolino Christina Pagiatakis Christina Pagiatakis Federica Pierin Daniele Sabatino Giovanna Finzi Rosalba Gornati Giovanni Bernardini Roberto Papait Roberto Papait |
| author_sort | Elettra Musolino |
| collection | DOAJ |
| description | Three-dimensional (3D) chromatin organization has a key role in defining the transcription program of cells during development. Its alteration is the cause of gene expression changes responsible for several diseases. Thus, we need new tools to study this aspect of gene expression regulation. To this end, ChromEM was recently developed: this is an electron-microscopy staining technique that selectively marks nuclear DNA without altering its structure and, thus, allows better visualization of 3D chromatin conformation. However, despite increasingly frequent application of this staining technique on cells, it has not yet been applied to visualize chromatin ultrastructure in tissues. Here, we provide a protocol to carry out ChromEM on myocardial tissue harvested from the left ventricles of C57BL/6J mice and use this in combination with transmission electron microscopy (TEM) to measure some morphological parameters of peripheral heterochromatin in cardiomyocytes. This protocol could also be used, in combination with electron tomography, to study 3D chromatin organization in cardiomyocytes in different aspects of heart pathobiology (e.g., heart development, cardiac aging, and heart failure) as well as help to set-up ChromEM in other tissues. |
| first_indexed | 2024-03-13T01:12:20Z |
| format | Article |
| id | doaj.art-12d7cfc66a1b4a858356986a45a53978 |
| institution | Directory Open Access Journal |
| issn | 2296-634X |
| language | English |
| last_indexed | 2024-03-13T01:12:20Z |
| publishDate | 2023-07-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cell and Developmental Biology |
| spelling | doaj.art-12d7cfc66a1b4a858356986a45a539782023-07-05T16:40:08ZengFrontiers Media S.A.Frontiers in Cell and Developmental Biology2296-634X2023-07-011110.3389/fcell.2023.11231141123114A chromEM-staining protocol optimized for cardiac tissueElettra Musolino0Christina Pagiatakis1Christina Pagiatakis2Federica Pierin3Daniele Sabatino4Giovanna Finzi5Rosalba Gornati6Giovanni Bernardini7Roberto Papait8Roberto Papait9Department of Biotechnology and Life Sciences, University of Insubria, Varese, ItalyDepartment of Biotechnology and Life Sciences, University of Insubria, Varese, ItalyDepartment of Cardiovascular Medicine, Humanitas Research Hospital–IRCCS, Rozzano (MI), ItalyDepartment of Biotechnology and Life Sciences, University of Insubria, Varese, ItalyDepartment of Pathology, ASST Sette Laghi, Varese, ItalyDepartment of Pathology, ASST Sette Laghi, Varese, ItalyDepartment of Biotechnology and Life Sciences, University of Insubria, Varese, ItalyDepartment of Biotechnology and Life Sciences, University of Insubria, Varese, ItalyDepartment of Biotechnology and Life Sciences, University of Insubria, Varese, ItalyDepartment of Cardiovascular Medicine, Humanitas Research Hospital–IRCCS, Rozzano (MI), ItalyThree-dimensional (3D) chromatin organization has a key role in defining the transcription program of cells during development. Its alteration is the cause of gene expression changes responsible for several diseases. Thus, we need new tools to study this aspect of gene expression regulation. To this end, ChromEM was recently developed: this is an electron-microscopy staining technique that selectively marks nuclear DNA without altering its structure and, thus, allows better visualization of 3D chromatin conformation. However, despite increasingly frequent application of this staining technique on cells, it has not yet been applied to visualize chromatin ultrastructure in tissues. Here, we provide a protocol to carry out ChromEM on myocardial tissue harvested from the left ventricles of C57BL/6J mice and use this in combination with transmission electron microscopy (TEM) to measure some morphological parameters of peripheral heterochromatin in cardiomyocytes. This protocol could also be used, in combination with electron tomography, to study 3D chromatin organization in cardiomyocytes in different aspects of heart pathobiology (e.g., heart development, cardiac aging, and heart failure) as well as help to set-up ChromEM in other tissues.https://www.frontiersin.org/articles/10.3389/fcell.2023.1123114/fullchromEMchromatin structureheterochromatincardiac tissueaging |
| spellingShingle | Elettra Musolino Christina Pagiatakis Christina Pagiatakis Federica Pierin Daniele Sabatino Giovanna Finzi Rosalba Gornati Giovanni Bernardini Roberto Papait Roberto Papait A chromEM-staining protocol optimized for cardiac tissue Frontiers in Cell and Developmental Biology chromEM chromatin structure heterochromatin cardiac tissue aging |
| title | A chromEM-staining protocol optimized for cardiac tissue |
| title_full | A chromEM-staining protocol optimized for cardiac tissue |
| title_fullStr | A chromEM-staining protocol optimized for cardiac tissue |
| title_full_unstemmed | A chromEM-staining protocol optimized for cardiac tissue |
| title_short | A chromEM-staining protocol optimized for cardiac tissue |
| title_sort | chromem staining protocol optimized for cardiac tissue |
| topic | chromEM chromatin structure heterochromatin cardiac tissue aging |
| url | https://www.frontiersin.org/articles/10.3389/fcell.2023.1123114/full |
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