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Abstract
While major changes in cellular morphology during apoptosis have been well described, the subcellular changes in nuclear architecture involved in this process remain poorly understood. Imaging of nucleosomes in cortical neurons in vitro before and during apoptosis revealed that chromatin compaction precedes the activation of caspase-3 and nucleus shrinkage. While this early chromatin compaction remained unaffected by pharmacological blockade of the final execution of apoptosis through caspase-3 inhibition, interfering with the chromatin dynamics by modulation of actomyosin activity prevented apoptosis, but resulted in necrotic-like cell death instead. With super-resolution imaging at different phases of apoptosis in vitro and in vivo, we demonstrate that chromatin compaction occurs progressively and can be classified into five stages. In conclusion, we show that compaction of chromatin in the neuronal nucleus precedes apoptosis execution. These early changes in chromatin structure critically affect apoptotic cell death and are not part of the final execution of the apoptotic process in developing cortical neurons.
Single-molecule imaging in developing cortical neurons shows that chromatin compaction precedes apoptosis and is an essential part of it, but can be uncoupled from the following apoptotic process.
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1 University Medical Center of the Johannes Gutenberg University, Institute of Physiology, Mainz, Germany (GRID:grid.410607.4)
2 Institute of Molecular Biology gGmbH (IMB), Mainz, Germany (GRID:grid.424631.6) (ISNI:0000 0004 1794 1771)
3 Institute of Molecular Biology gGmbH (IMB), Mainz, Germany (GRID:grid.424631.6) (ISNI:0000 0004 1794 1771); University of Heidelberg, Kirchhoff Institute for Physics (KIP), Heidelberg, Germany (GRID:grid.7700.0) (ISNI:0000 0001 2190 4373); University of Heidelberg, Interdisciplinary Center for Scientific Computing (IWR), Heidelberg, Germany (GRID:grid.7700.0) (ISNI:0000 0001 2190 4373)