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The three-dimensional organization of telomeres in the nucleus of mammalian cells.

Chuang TC, Moshir S, Garini Y, Chuang AY, Young IT, Vermolen B, van den Doel R, Mougey V, Perrin M, Braun M, Kerr PD, Fest T, Boukamp P, Mai S - BMC Biol. (2004)

Bottom Line: In tumor cells, the 3D telomere organization is distorted and aggregates are formed.The results emphasize a non-random and dynamic 3D nuclear telomeric organization and its importance to genomic stability.Based on our findings, it appears possible to examine telomeric aggregates suggestive of genomic instability in individual interphase nuclei and tissues without the need to examine metaphases.

View Article: PubMed Central - HTML - PubMed

Affiliation: Manitoba Institute of Cell Biology, CancerCare Manitoba, University of Manitoba, 675 McDermot Avenue, Winnipeg, MB, R3E 0V9, Canada. tcychuang@hotmail.com

ABSTRACT

Background: The observation of multiple genetic markers in situ by optical microscopy and their relevance to the study of three-dimensional (3D) chromosomal organization in the nucleus have been greatly developed in the last decade. These methods are important in cancer research because cancer is characterized by multiple alterations that affect the modulation of gene expression and the stability of the genome. It is, therefore, essential to analyze the 3D genome organization of the interphase nucleus in both normal and cancer cells.

Results: We describe a novel approach to study the distribution of all telomeres inside the nucleus of mammalian cells throughout the cell cycle. It is based on 3D telomere fluorescence in situ hybridization followed by quantitative analysis that determines the telomeres' distribution in the nucleus throughout the cell cycle. This method enables us to determine, for the first time, that telomere organization is cell-cycle dependent, with assembly of telomeres into a telomeric disk in the G2 phase. In tumor cells, the 3D telomere organization is distorted and aggregates are formed.

Conclusions: The results emphasize a non-random and dynamic 3D nuclear telomeric organization and its importance to genomic stability. Based on our findings, it appears possible to examine telomeric aggregates suggestive of genomic instability in individual interphase nuclei and tissues without the need to examine metaphases. Such new avenues of monitoring genomic instability could potentially impact on cancer biology, genetics, diagnostic innovations and surveillance of treatment response in medicine.

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Related in: MedlinePlus

Demonstration of the signal-to-noise and spatial resolution of our measurements. The fluorescence intensity is bright (typical signal-to-noise ratio of 10:1). Two pairs of telomeres are shown, 1200 nm apart (top), which can be easily separated, and 400 nm apart (bottom). The inserts show the actual images.
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Figure 3: Demonstration of the signal-to-noise and spatial resolution of our measurements. The fluorescence intensity is bright (typical signal-to-noise ratio of 10:1). Two pairs of telomeres are shown, 1200 nm apart (top), which can be easily separated, and 400 nm apart (bottom). The inserts show the actual images.

Mentions: The optical resolution and signal-to-noise ratio are presented in Fig. 3. The images of two neighbouring telomeres that are 1200 nm and 400 nm apart, and the corresponding intensity along the line connecting the pair, indicates the smallest telomere distance that can still be unambiguously distinguished (approximately 200 nm).


The three-dimensional organization of telomeres in the nucleus of mammalian cells.

Chuang TC, Moshir S, Garini Y, Chuang AY, Young IT, Vermolen B, van den Doel R, Mougey V, Perrin M, Braun M, Kerr PD, Fest T, Boukamp P, Mai S - BMC Biol. (2004)

Demonstration of the signal-to-noise and spatial resolution of our measurements. The fluorescence intensity is bright (typical signal-to-noise ratio of 10:1). Two pairs of telomeres are shown, 1200 nm apart (top), which can be easily separated, and 400 nm apart (bottom). The inserts show the actual images.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC425602&req=5

Figure 3: Demonstration of the signal-to-noise and spatial resolution of our measurements. The fluorescence intensity is bright (typical signal-to-noise ratio of 10:1). Two pairs of telomeres are shown, 1200 nm apart (top), which can be easily separated, and 400 nm apart (bottom). The inserts show the actual images.
Mentions: The optical resolution and signal-to-noise ratio are presented in Fig. 3. The images of two neighbouring telomeres that are 1200 nm and 400 nm apart, and the corresponding intensity along the line connecting the pair, indicates the smallest telomere distance that can still be unambiguously distinguished (approximately 200 nm).

Bottom Line: In tumor cells, the 3D telomere organization is distorted and aggregates are formed.The results emphasize a non-random and dynamic 3D nuclear telomeric organization and its importance to genomic stability.Based on our findings, it appears possible to examine telomeric aggregates suggestive of genomic instability in individual interphase nuclei and tissues without the need to examine metaphases.

View Article: PubMed Central - HTML - PubMed

Affiliation: Manitoba Institute of Cell Biology, CancerCare Manitoba, University of Manitoba, 675 McDermot Avenue, Winnipeg, MB, R3E 0V9, Canada. tcychuang@hotmail.com

ABSTRACT

Background: The observation of multiple genetic markers in situ by optical microscopy and their relevance to the study of three-dimensional (3D) chromosomal organization in the nucleus have been greatly developed in the last decade. These methods are important in cancer research because cancer is characterized by multiple alterations that affect the modulation of gene expression and the stability of the genome. It is, therefore, essential to analyze the 3D genome organization of the interphase nucleus in both normal and cancer cells.

Results: We describe a novel approach to study the distribution of all telomeres inside the nucleus of mammalian cells throughout the cell cycle. It is based on 3D telomere fluorescence in situ hybridization followed by quantitative analysis that determines the telomeres' distribution in the nucleus throughout the cell cycle. This method enables us to determine, for the first time, that telomere organization is cell-cycle dependent, with assembly of telomeres into a telomeric disk in the G2 phase. In tumor cells, the 3D telomere organization is distorted and aggregates are formed.

Conclusions: The results emphasize a non-random and dynamic 3D nuclear telomeric organization and its importance to genomic stability. Based on our findings, it appears possible to examine telomeric aggregates suggestive of genomic instability in individual interphase nuclei and tissues without the need to examine metaphases. Such new avenues of monitoring genomic instability could potentially impact on cancer biology, genetics, diagnostic innovations and surveillance of treatment response in medicine.

Show MeSH
Related in: MedlinePlus