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The spatial repositioning of adipogenesis genes is correlated with their expression status in a porcine mesenchymal stem cell adipogenesis model system

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Abstract

Alterations in the nuclear positioning of chromosomes and specific genes during differentiation and development have suggested strongly the existence of a relationship between non-random organization of the genome and its function. In this study, we have examined the genome organization in interphase nuclei during adipogenesis, using the pig as a model organism. We hypothesized that changes in the gene expression profile and chromatin remodeling which occur during cellular differentiation would elicit repositioning of whole chromosomes, moving specific genes on them to different regions of the nucleus. We established an in vitro adipogenesis differentiation system using mesenchymal stem cells, derived from porcine bone marrow. The nuclear position of seven adipogenesis genes (PPARG, SREBF1, FABP4, CEBPA, CEBPB, CREB, and GATA2), two control genes (SOX9 and MYL1), and six chromosomes carrying these gene loci (SSC4, SSC6, SSC12, SSC13, SSC15, and SSC17) was determined. We found that during adipogenesis, using the in vitro stem cell model system, in contrast to our original hypothesis, the nuclear position of genes involved in adipogenesis was altered radically with the up-regulation of gene expression correlating with these genes becoming more internally located within nuclei. Chromosome territories, containing these genes, were also found to alter their nuclear position during the in vitro adipogenesis model, with the most dramatic repositioning being SSC4 that moved from the nuclear periphery towards the nuclear interior. We found that during in vitro adipogenesis chromosome territories decondensed and the genes were found on loops and projections of chromatin, away from the main body of the chromosomes. From our data, it appears that the temporal repositioning of genes, emanating away from chromosomes, during adipogenesis is correlated with gene activity, supporting models of the involvement of spatial genome repositioning in regulating gene expression and the nuclear interior being an important region of the nucleus for transcription.

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Acknowledgments

We thank Prof. Wendy Bickmore and Dr. Paul Perry (MRC HGU, Edinburgh) for erosion script, Prof. Malcom A. Ferguson-Smith (Cambridge Resource Centre for Comparative Genomics, Cambridge University, UK) for flow-sorted pig chromosome paints, and Prof. Fengtang Yang (Wellcome Trust Sanger Institute, Cambridge, UK) for paint probe for porcine chromosome 12. We also thank Dr. Maciej Szydlowski (Poznan University of Life Sciences, Poland) for helping with statistical analysis. This study was financed by the Polish Ministry of Science and Higher Education, grant N301 3381 33, fellowship 9/MOB/2007/0 and monies awarded to JMB from Brunel University.

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  1. Department of Genetics and Animal Breeding, Poznan University of Life Sciences, Wolynska 33, 60-637, Poznań, Poland

    Izabela Szczerbal

  2. Laboratory of Nuclear and Genomic Health, Centre for Cell & Chromosome Biology, Biosciences, School of Health Sciences and Social Care, Brunel University, West London, UB8 3PH, UK

    Izabela Szczerbal, Helen A. Foster & Joanna M. Bridger

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  1. Izabela Szczerbal
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  2. Helen A. Foster
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  3. Joanna M. Bridger
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Correspondence to Izabela Szczerbal or Joanna M. Bridger.

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Communicated by T. Misteli

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Table S1

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Table S2

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Table S3

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Table S4

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Fig. S1

Ideograms of porcine chromosomes (SSC4, SSC6, SSC12, SSC13, SSC15, SSC17) showing the location of selected genes. (JPG 145 kb)

Fig. S2

Relative positioning of four gene loci—PPARG, SREBF1, CEBPA, and FABP4 with nucleoli. Nucleoli were revealed with anti-fibrillarin (green) and gene probes were delineated in red. a MSC; b AD14. c The graph shows the percentage of total gene signals co-localized with nucleoli (means ± SEM). Scale bar, 10 μm (JPG 234 kb)

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Szczerbal, I., Foster, H.A. & Bridger, J.M. The spatial repositioning of adipogenesis genes is correlated with their expression status in a porcine mesenchymal stem cell adipogenesis model system. Chromosoma 118, 647–663 (2009). https://doi.org/10.1007/s00412-009-0225-5

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