Abstract
The well-established human epidermal keratinocyte (HEK) differentiation model was investigated to determine possible alterations in chromosome territory (CT) association during differentiation. The seven human chromosomes (1, 4, 11, 12, 16, 17, and 18) selected for this analysis are representative of the chromosome size and gene density range of the overall human genome as well as including a majority of genes involved in epidermal development and differentiation (CT1, 12, and 17). Induction with calcium chloride (Ca2+) resulted in morphological changes characteristic of keratinocyte differentiation. Combined multi-fluorescence in situ hybridization (FISH) and computational image analysis on the undifferentiated (0 h) and differentiated (24 h after Ca2+ treatment) HEK revealed that (a) increases in CT volumes correspond to overall nuclear volume increases, (b) radial positioning is gene density-dependent at 0 h but neither gene density- nor size-dependent at 24 h, (c) the average number of interchromosomal associations for each CT is gene density-dependent and similar at both time points, and (d) there are striking differences in the single and multiple pairwise interchromosomal association profiles. Probabilistic network models of the overall interchromosomal associations demonstrate major reorganization of the network during differentiation. Only ~40 % of the CT pairwise connections in the networks are common to both 0 and 24 h HEK. We propose that there is a probabilistic chromosome positional code which can be significantly altered during cell differentiation in coordination with reprogramming of gene expression.
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Acknowledgments
This research was supported by grants from the National Institutes of Health (GM-072131) to R.B, the National Science Foundation (IIS-0713489, IIS-1115220 and IIIS-1422591) to J.X. and R.B., and the University at Buffalo Foundation (9351115726) to R.B. A part of this research was also supported by a Mark Diamond Research Foundation grant to N.S. Dr. Andrew J. Fritz provided advise and valuable discussion. We thank Drs. Satrajit Sinha and Rose Anne Romano, Department of Biochemistry, University at Buffalo for providing the keratin antibodies and valuable advise.
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Sehgal, N., Seifert, B., Ding, H. et al. Reorganization of the interchromosomal network during keratinocyte differentiation. Chromosoma 125, 389–403 (2016). https://doi.org/10.1007/s00412-015-0546-5
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DOI: https://doi.org/10.1007/s00412-015-0546-5