A ChromEM-staining Protocol Optimized for Cardiac Tissue

Overview

Journal Front Cell Dev Biol

Specialty Cell Biology

Date 2023 Jul 21

PMID 37476155

Authors

Elettra Musolino

Christina Pagiatakis

Federica Pierin

Daniele Sabatino

Giovanna Finzi

Rosalba Gornati

Giovanni Bernardini

Roberto Papait

Affiliations

Soon will be listed here.

Abstract

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.

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