Abstract

BackgroundCharacterizing the nuclear orientation of chromosomes in the three-dimensional (3D) nucleus by multicolor banding (mBANDing) is a new approach towards understanding nuclear organization of chromosome territories. An mBANDing paint is composed of multiple overlapping subchromosomal probes that represent different regions of a single chromosome. In this study, we used it for the analysis of chromosome orientation in 3D interphase nuclei. We determined whether the nuclear orientation of the two chromosome 11 homologs was random or preferential, and if it was conserved between diploid mouse Pre B lymphocytes of BALB/c origin and primary B lymphocytes of congenic [T38HxBALB/c]N wild-type mice. The chromosome orientation was assessed visually and through a semi-automated quantitative analysis of the radial and angular orientation patterns observed in both B cell types.ResultsOur data indicate that there are different preferential patterns of chromosome 11 orientation, which are not significantly different between both mouse cell types (p > 0.05). In the most common case for both cell types, both copies of chromosome 11 were oriented in parallel with the nuclear border. The second most common pattern in both types of B lymphocytes was with one homolog of chromosome 11 positioned with its telomeric end towards the nuclear center and with its centromeric end towards the periphery, while the other chromosome 11 was found parallel with the nuclear border. In addition to these two most common orientations present in approximately 50% of nuclei from each cell type, other orientations were observed at lower frequencies.ConclusionsWe conclude that there are probabilistic, non-random orientation patterns for mouse chromosome 11 in the mouse B lymphocytes we investigated (p < 0.0001).

Highlights

  • Characterizing the nuclear orientation of chromosomes in the three-dimensional (3D) nucleus by multicolor banding is a new approach towards understanding nuclear organization of chromosome territories

  • The mBANDing technique was used to study the nuclear organization of chromosome 11 in a diploid mouse Pre B lymphocyte line of BALB/c origin [21] and in B lymphocytes of congenic [T38HxBALB/c]N wild-type mice [22]

  • We performed a semi-quantitative analysis of the radial arrangement of individual mBAND probe distributions on a subset of 45 nuclei per cell type using eADS software [10]

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Summary

Introduction

Characterizing the nuclear orientation of chromosomes in the three-dimensional (3D) nucleus by multicolor banding (mBANDing) is a new approach towards understanding nuclear organization of chromosome territories. We used it for the analysis of chromosome orientation in 3D interphase nuclei. The chromosome orientation was assessed visually and through a semi-automated quantitative analysis of the radial and angular orientation patterns observed in both B cell types. Chromosomes occupy specific regions in the threedimensional (3D) interphase nucleus, so-called chromosome territories (CTs) [1]. The radial arrangement of CTs shows cell-type specific differences [1,2]. Changes in the position of CTs have, been observed during cell differentiation, senescence and tumorigenesis. This occurs for example during adipocyte [13] and human epidermal keratinocyte differentiation [14]. Chromosome positions change when a cell becomes quiescent, senescent or when it re-enters the cell cycle [15,16]

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