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Characterisation of nuclear architectural alterations during in vitro differentiation of human stem cells of myogenic origin.

Rozwadowska N, Kolanowski T, Wiland E, Siatkowski M, Pawlak P, Malcher A, Mietkiewski T, Olszewska M, Kurpisz M - PLoS ONE (2013)

Bottom Line: Additionally, almost all of the chromosomal centromeres in the differentiated cells preferentially localised near the nuclear periphery when compared to the undifferentiated cells.The exceptions were chromosomes 7 and 11, in which we were unable to confirm the centromere repositioning.Based on these data we can conclude that the myogenic differentiation with global gene expression changes is accompanied by the spatial repositioning of chromosomes and chromatin remodelling, which are important processes that regulate cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Reproductive Biology and Stem Cells, Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland.

ABSTRACT
Cell differentiation is based on a synchronised orchestra of complex pathways of intrinsic and extrinsic signals that manifest in the induced expression of specific transcription factors and pivotal genes within the nucleus. One cannot ignore the epigenetic status of differentiating cells, comprising not only histones and DNA modifications but also the spatial and temporal intranuclear chromatin organisation, which is an important regulator of nuclear processes. In the present study, we investigated the nuclear architecture of human primary myoblasts and myocytes in an in vitro culture, with reference to global changes in genomic expression. Repositioning of the chromosomal centromeres, along with alterations in the nuclear shape and volume, was observed as a consequence of myotube formation. Moreover, the microarray data showed that during in vitro myogenesis cells tend to silence rather than induce gene expression. The creation of a chromosome map marked with gene expression changes that were at least 2-fold confirmed the observation. Additionally, almost all of the chromosomal centromeres in the differentiated cells preferentially localised near the nuclear periphery when compared to the undifferentiated cells. The exceptions were chromosomes 7 and 11, in which we were unable to confirm the centromere repositioning. In our opinion, this is the first reported observation of the movement of chromosomal centromeres along differentiating myogenic cells. Based on these data we can conclude that the myogenic differentiation with global gene expression changes is accompanied by the spatial repositioning of chromosomes and chromatin remodelling, which are important processes that regulate cell differentiation.

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Schematic view of expression changes (2-fold) obtained by microarray analysis.Global expression found in all chromosomes showed an increase in gene silencing during in vitro myogenesis (132 transcripts down-regulated vs. 117 transcripts up-regulated). The up-regulated transcripts are in red and the down-regulated transcripts are in blue. The chromosomes subjected to 3D FISH evaluation are shown in schematic ideograms.
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pone-0073231-g007: Schematic view of expression changes (2-fold) obtained by microarray analysis.Global expression found in all chromosomes showed an increase in gene silencing during in vitro myogenesis (132 transcripts down-regulated vs. 117 transcripts up-regulated). The up-regulated transcripts are in red and the down-regulated transcripts are in blue. The chromosomes subjected to 3D FISH evaluation are shown in schematic ideograms.

Mentions: To study changes in the global transcriptome and to correlate the observed centromere repositioning with genes being switched-on/off, we used microarray gene expression analysis. Among the investigated genes, there were 249 transcripts with a 2-fold change in expression in the differentiated myogenic cells compared to the myoblasts. Up-regulation of expression was observed for 117 transcripts, while 132 were silenced (Figure S2 and Table S2). Subsequently, the marked genes were put on chromosome ideograms, and the evaluation was conducted only on chromosomes investigated in the 3D FISH experiments. The observed pattern of up- and down-regulated genes was surprisingly uniform. The chromosomes 1, 3, 11, 12, 17 and X contained an almost equal number of up-regulated genes (from 8 to 6), and one more gene was found to be consistently silenced (from 9 to 7). Chromosome 7 did not follow the rules in which the coding sequences for 8 active and 5 silenced genes were marked. Figure 7 (Table S1) summarises the data that were obtained.


Characterisation of nuclear architectural alterations during in vitro differentiation of human stem cells of myogenic origin.

Rozwadowska N, Kolanowski T, Wiland E, Siatkowski M, Pawlak P, Malcher A, Mietkiewski T, Olszewska M, Kurpisz M - PLoS ONE (2013)

Schematic view of expression changes (2-fold) obtained by microarray analysis.Global expression found in all chromosomes showed an increase in gene silencing during in vitro myogenesis (132 transcripts down-regulated vs. 117 transcripts up-regulated). The up-regulated transcripts are in red and the down-regulated transcripts are in blue. The chromosomes subjected to 3D FISH evaluation are shown in schematic ideograms.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3760906&req=5

pone-0073231-g007: Schematic view of expression changes (2-fold) obtained by microarray analysis.Global expression found in all chromosomes showed an increase in gene silencing during in vitro myogenesis (132 transcripts down-regulated vs. 117 transcripts up-regulated). The up-regulated transcripts are in red and the down-regulated transcripts are in blue. The chromosomes subjected to 3D FISH evaluation are shown in schematic ideograms.
Mentions: To study changes in the global transcriptome and to correlate the observed centromere repositioning with genes being switched-on/off, we used microarray gene expression analysis. Among the investigated genes, there were 249 transcripts with a 2-fold change in expression in the differentiated myogenic cells compared to the myoblasts. Up-regulation of expression was observed for 117 transcripts, while 132 were silenced (Figure S2 and Table S2). Subsequently, the marked genes were put on chromosome ideograms, and the evaluation was conducted only on chromosomes investigated in the 3D FISH experiments. The observed pattern of up- and down-regulated genes was surprisingly uniform. The chromosomes 1, 3, 11, 12, 17 and X contained an almost equal number of up-regulated genes (from 8 to 6), and one more gene was found to be consistently silenced (from 9 to 7). Chromosome 7 did not follow the rules in which the coding sequences for 8 active and 5 silenced genes were marked. Figure 7 (Table S1) summarises the data that were obtained.

Bottom Line: Additionally, almost all of the chromosomal centromeres in the differentiated cells preferentially localised near the nuclear periphery when compared to the undifferentiated cells.The exceptions were chromosomes 7 and 11, in which we were unable to confirm the centromere repositioning.Based on these data we can conclude that the myogenic differentiation with global gene expression changes is accompanied by the spatial repositioning of chromosomes and chromatin remodelling, which are important processes that regulate cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Reproductive Biology and Stem Cells, Institute of Human Genetics, Polish Academy of Sciences, Poznan, Poland.

ABSTRACT
Cell differentiation is based on a synchronised orchestra of complex pathways of intrinsic and extrinsic signals that manifest in the induced expression of specific transcription factors and pivotal genes within the nucleus. One cannot ignore the epigenetic status of differentiating cells, comprising not only histones and DNA modifications but also the spatial and temporal intranuclear chromatin organisation, which is an important regulator of nuclear processes. In the present study, we investigated the nuclear architecture of human primary myoblasts and myocytes in an in vitro culture, with reference to global changes in genomic expression. Repositioning of the chromosomal centromeres, along with alterations in the nuclear shape and volume, was observed as a consequence of myotube formation. Moreover, the microarray data showed that during in vitro myogenesis cells tend to silence rather than induce gene expression. The creation of a chromosome map marked with gene expression changes that were at least 2-fold confirmed the observation. Additionally, almost all of the chromosomal centromeres in the differentiated cells preferentially localised near the nuclear periphery when compared to the undifferentiated cells. The exceptions were chromosomes 7 and 11, in which we were unable to confirm the centromere repositioning. In our opinion, this is the first reported observation of the movement of chromosomal centromeres along differentiating myogenic cells. Based on these data we can conclude that the myogenic differentiation with global gene expression changes is accompanied by the spatial repositioning of chromosomes and chromatin remodelling, which are important processes that regulate cell differentiation.

Show MeSH
Related in: MedlinePlus