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First published online November 10, 2004
doi: 10.1242/10.1242/jcs.01508

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Alteration of chromosome positioning during adipocyte differentiation

¹ Department of Pathology, Tokyo Medical University, 6-1-1 Shinjuku, Shinjuku-ku, Tokyo, 160-8402, Japan
² CREST Research Project, Japan Science and Technology Corporation, 4-1-6 Kawaguchi, Saitama, 332-0012, Japan
³ Shinanomachi Research Park, Keio University, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan
⁴ Department of Biosystems Science, School of Advanced Sciences, The Graduate University for Advanced Studies (Sokendai), Shonan Village, Hayama, Kanagawa 240-0193, Japan
⁵ Bioinformatics Business Promotion Department, NEC Corporation, 7-1 Shiba 5-chome, Minato-ku, Tokyo 108-8001, Japan
⁶ Fundamental and Environmental Research Laboratories, NEC Corporation, 34 Miyukigaoka, Tsukuba, Ibaraki 305-8501, Japan
⁷ Cell bank laboratory, Division of Genetics and Mutagenesis, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan

View larger version (56K): [in a new window]   Fig. 1. Induction of adipocyte differentiation in primary human preadipocyte cells. Phase-contrast images (20x) of human preadipocyte cells (A) and matured adipocyte cells (2 weeks after differentiation) (B). (C) A preadipocyte at metaphase in which the DNA has been hybridized with human whole chromosome painting probes for chromosomes 12 (green) and 16 (red).

View larger version (75K): [in a new window]   Fig. 2. Visualization of the human chromosome-12 and chromosome-16 territories (CT12 and CT16, respectively). Gallery of 200 nm serial optical sections (every third section is shown: 0 µm, 0.6 µm, 1.2 µm, 1.8 µm, 2.4 µm, 3.0 µm) through a preadipocyte (A) and a mature adipocyte (C) nucleus after 3D-FISH with chromosome painting probes for chromosomes 12 (green) and 16 (red). A DNA counterstain is shown in blue (scale bar, 5 µm). (B,D) 3D reconstructed images of the nuclei presented in A and C, respectively, with outlines of the painted CTs and the nuclear DNA. The adipocyte nucleus displays a proximal association of one CT12 and one CT16 (C,D), whereas the preadipocyte nucleus shows no association between CT12 and CT16 (A,B).

View larger version (27K): [in a new window]   Fig. 3. Topological deformation method for the standardization of cell nuclear shape. (A) Coordinate system of image. The z-axis runs out from the x-y plane towards the reader. (B) Standardization of the nuclear shape. The broken and solid lines indicate actual and standardized nuclear shapes, respectively. (C) Calculation of chromosomal positions. (D) Quasi-3D visualization of the standardized nucleus shape. Deformation was performed in the projected 2D space; transformed x-y positions and the original z position were used. The cylindrical nucleus was constructed using the standardized 2D nucleus shape.

View larger version (16K): [in a new window]   Fig. 4. Relative positions of human chromosome-12 and chromosome-16 territories (CT12 and CT16, respectively). (A) The definition of a representative peak point for a CT. Green dots indicate the fluorescence peak center (FPC). The location of the CT is represented by the position of its FPC. (B) Schema of the minimum FPC distance. We measured the pixel-to-pixel distance between the FPCs of CT12 and CT16, and then recorded the shortest of the four distances obtained from the four possible pairings as the minimum FPC distance. (C) Quantitative evaluation of the minimum FPC distance between CT12 and CT16. The distances are normalized using the radius of the standardized 2D nucleus. The significance of the difference between minimum FPC distances in preadipocytes and adipocytes was evaluated using the Welch's t test (P=0.017).

View larger version (28K): [in a new window]   Fig. 5. Radial positions of human chromosome-12 and chromosome-16 territories (CT12 and CT16, respectively). Total radial distributions of CT12 and CT16 over the standardized 2D nucleus (n=76=38x2 for preadipocyte, and n=82=41x2 for adipocyte). The mean values (<r>) and standard errors of all distributions are also shown.

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