Abstract
The interplay between epigenetic modification and chromatin compaction is implicated in the regulation of gene expression, and it comprises one of the most fascinating frontiers in cell biology. Although a complete picture is still lacking, it is generally accepted that the differentiation of embryonic stem (ES) cells is accompanied by a selective condensation into heterochromatin with concomitant gene silencing, leaving access only to lineage-specific genes in the euchromatin. ES cells have been reported to have less condensed chromatin, as they are capable of differentiating into any cell type. However, pluripotency itself - even prior to differentiation - is a split state comprising a naïve state and a state in which ES cells prime for differentiation. Here, we show that naïve ES cells decondense their chromatin in the course of downregulating the pluripotency marker Nanog before they initiate lineage commitment. We used fluorescence recovery after photobleaching, and histone modification analysis paired with a novel, to our knowledge, optical stretching method, to show that ES cells in the naïve state have a significantly stiffer nucleus that is coupled to a globally more condensed chromatin state. We link this biophysical phenotype to coinciding epigenetic differences, including histone methylation, and show a strong correlation of chromatin condensation and nuclear stiffness with the expression of Nanog. Besides having implications for transcriptional regulation and embryonic cell sorting and suggesting a putative mechanosensing mechanism, the physical differences point to a system-level regulatory role of chromatin in maintaining pluripotency in embryonic development.
| Original language | English |
|---|---|
| Pages (from-to) | 2060-2070 |
| Number of pages | 11 |
| Journal | Biophysical Journal |
| Volume | 103 |
| Issue number | 10 |
| DOIs | |
| State | Published - Nov 21 2012 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Biophysics
Cite this
Chromatin decondensation and nuclear softening accompany Nanog downregulation in embryonic stem cells. / Chalut, Kevin J.; Höpfler, Markus; Lautenschläger, Franziska; Boyde, Lars; Chan, Chii Jou; Ekpenyong, Andrew; Martinez-Arias, Alfonso; Guck, Jochen.
In: Biophysical Journal, Vol. 103, No. 10, 21.11.2012, p. 2060-2070.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Chromatin decondensation and nuclear softening accompany Nanog downregulation in embryonic stem cells
AU - Chalut, Kevin J.
AU - Höpfler, Markus
AU - Lautenschläger, Franziska
AU - Boyde, Lars
AU - Chan, Chii Jou
AU - Ekpenyong, Andrew
AU - Martinez-Arias, Alfonso
AU - Guck, Jochen
PY - 2012/11/21
Y1 - 2012/11/21
N2 - The interplay between epigenetic modification and chromatin compaction is implicated in the regulation of gene expression, and it comprises one of the most fascinating frontiers in cell biology. Although a complete picture is still lacking, it is generally accepted that the differentiation of embryonic stem (ES) cells is accompanied by a selective condensation into heterochromatin with concomitant gene silencing, leaving access only to lineage-specific genes in the euchromatin. ES cells have been reported to have less condensed chromatin, as they are capable of differentiating into any cell type. However, pluripotency itself - even prior to differentiation - is a split state comprising a naïve state and a state in which ES cells prime for differentiation. Here, we show that naïve ES cells decondense their chromatin in the course of downregulating the pluripotency marker Nanog before they initiate lineage commitment. We used fluorescence recovery after photobleaching, and histone modification analysis paired with a novel, to our knowledge, optical stretching method, to show that ES cells in the naïve state have a significantly stiffer nucleus that is coupled to a globally more condensed chromatin state. We link this biophysical phenotype to coinciding epigenetic differences, including histone methylation, and show a strong correlation of chromatin condensation and nuclear stiffness with the expression of Nanog. Besides having implications for transcriptional regulation and embryonic cell sorting and suggesting a putative mechanosensing mechanism, the physical differences point to a system-level regulatory role of chromatin in maintaining pluripotency in embryonic development.
AB - The interplay between epigenetic modification and chromatin compaction is implicated in the regulation of gene expression, and it comprises one of the most fascinating frontiers in cell biology. Although a complete picture is still lacking, it is generally accepted that the differentiation of embryonic stem (ES) cells is accompanied by a selective condensation into heterochromatin with concomitant gene silencing, leaving access only to lineage-specific genes in the euchromatin. ES cells have been reported to have less condensed chromatin, as they are capable of differentiating into any cell type. However, pluripotency itself - even prior to differentiation - is a split state comprising a naïve state and a state in which ES cells prime for differentiation. Here, we show that naïve ES cells decondense their chromatin in the course of downregulating the pluripotency marker Nanog before they initiate lineage commitment. We used fluorescence recovery after photobleaching, and histone modification analysis paired with a novel, to our knowledge, optical stretching method, to show that ES cells in the naïve state have a significantly stiffer nucleus that is coupled to a globally more condensed chromatin state. We link this biophysical phenotype to coinciding epigenetic differences, including histone methylation, and show a strong correlation of chromatin condensation and nuclear stiffness with the expression of Nanog. Besides having implications for transcriptional regulation and embryonic cell sorting and suggesting a putative mechanosensing mechanism, the physical differences point to a system-level regulatory role of chromatin in maintaining pluripotency in embryonic development.
UR - http://www.scopus.com/inward/record.url?scp=84869406280&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84869406280&partnerID=8YFLogxK
U2 - 10.1016/j.bpj.2012.10.015
DO - 10.1016/j.bpj.2012.10.015
M3 - Article
C2 - 23200040
AN - SCOPUS:84869406280
VL - 103
SP - 2060
EP - 2070
JO - Biophysical Journal
JF - Biophysical Journal
SN - 0006-3495
IS - 10
ER -