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Title: Distribution of chromosome 18 and X centric heterochromatin in the interphase nucleus of cultured human cells

Abstract

In situ hybridization of human chromosome 18 and X-specific alphoid DNA-probes was performed in combination with three dimensional (3D) and two dimensional (2D) image analysis to study the interphase distribution of the centric heterochromatin (18c and Xc) of these chromosomes in cultured human cells. 3D analyses of 18c targets using confocal laser scanning microscopy indicated a nonrandom disposition in 73 amniotic fluid cell nuclei. In agreement with the 3D observations 18c targets were found significantly closer to the center of the 2D nuclear image (CNI) and to each other in all these cultures compared to a random distribution derived from corresponding ellipsoid or cylinder model nuclei. For comparison, a chromosome X-specific alphoid DNA probe was used to investigate the 2D distribution of chromosome X centric heterochromatin in the same cell types. Two dimensional Xc-Xc and Xc-CNI distances fit a random distribution in diploid normal and Bloom's syndrome nuclei, as well as in nuclei with trisomy X. The different distributions of 18c and Xc targets were confirmed by the simultaneous staining of these targets in different colors within individual nuclei using a double in situ hybridization approach.

Authors:
; ; ; ; ;  [1];  [2]
  1. (Univ. of Heidelberg (West Germany))
  2. (European Molecular Biology Lab., Heidelberg (West Germany))
Publication Date:
OSTI Identifier:
5377114
Resource Type:
Journal Article
Resource Relation:
Journal Name: Experimental Cell Research; (United States); Journal Volume: 189:1
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; HETEROCHROMATIN; SUBCELLULAR DISTRIBUTION; HETEROCHROMOSOMES; DNA HYBRIDIZATION; HUMAN X CHROMOSOME; CELL CULTURES; CELL CYCLE; FIBROBLASTS; MAN; TWO-DIMENSIONAL ELECTROPHORESIS; ANIMAL CELLS; ANIMALS; CHROMATIN; CHROMOSOMES; CONNECTIVE TISSUE CELLS; DISTRIBUTION; ELECTROPHORESIS; HUMAN CHROMOSOMES; HYBRIDIZATION; MAMMALS; PRIMATES; SOMATIC CELLS; VERTEBRATES; X CHROMOSOME; 550400* - Genetics

Citation Formats

Popp, S., Scholl, H.P., Loos, P., Jauch, A., Cremer, C., Cremer, T., and Stelzer, E. Distribution of chromosome 18 and X centric heterochromatin in the interphase nucleus of cultured human cells. United States: N. p., 1990. Web. doi:10.1016/0014-4827(90)90249-A.
Popp, S., Scholl, H.P., Loos, P., Jauch, A., Cremer, C., Cremer, T., & Stelzer, E. Distribution of chromosome 18 and X centric heterochromatin in the interphase nucleus of cultured human cells. United States. doi:10.1016/0014-4827(90)90249-A.
Popp, S., Scholl, H.P., Loos, P., Jauch, A., Cremer, C., Cremer, T., and Stelzer, E. Sun . "Distribution of chromosome 18 and X centric heterochromatin in the interphase nucleus of cultured human cells". United States. doi:10.1016/0014-4827(90)90249-A.
@article{osti_5377114,
title = {Distribution of chromosome 18 and X centric heterochromatin in the interphase nucleus of cultured human cells},
author = {Popp, S. and Scholl, H.P. and Loos, P. and Jauch, A. and Cremer, C. and Cremer, T. and Stelzer, E.},
abstractNote = {In situ hybridization of human chromosome 18 and X-specific alphoid DNA-probes was performed in combination with three dimensional (3D) and two dimensional (2D) image analysis to study the interphase distribution of the centric heterochromatin (18c and Xc) of these chromosomes in cultured human cells. 3D analyses of 18c targets using confocal laser scanning microscopy indicated a nonrandom disposition in 73 amniotic fluid cell nuclei. In agreement with the 3D observations 18c targets were found significantly closer to the center of the 2D nuclear image (CNI) and to each other in all these cultures compared to a random distribution derived from corresponding ellipsoid or cylinder model nuclei. For comparison, a chromosome X-specific alphoid DNA probe was used to investigate the 2D distribution of chromosome X centric heterochromatin in the same cell types. Two dimensional Xc-Xc and Xc-CNI distances fit a random distribution in diploid normal and Bloom's syndrome nuclei, as well as in nuclei with trisomy X. The different distributions of 18c and Xc targets were confirmed by the simultaneous staining of these targets in different colors within individual nuclei using a double in situ hybridization approach.},
doi = {10.1016/0014-4827(90)90249-A},
journal = {Experimental Cell Research; (United States)},
number = ,
volume = 189:1,
place = {United States},
year = {Sun Jul 01 00:00:00 EDT 1990},
month = {Sun Jul 01 00:00:00 EDT 1990}
}
  • The aim of this work was to measure simultaneously and in a quantitative manner double-strand breaks (DSBs), interphase chromosome breaks and cell lethality either immediately after irradiation, or at various times thereafter (up to 24 h), in cells of three nontransformed human fibroblast cell lines of widely different intrinsic radiosensitivity. We wished to assess initial damage, repair kinetics and residual damage at the DNA and the chromosome level, and to correlate these parameters with cell killings. We employed HF19 cells, a normal fibroblast cell line, AT2 cells, a radiosensitive cell line from a patient suffering from ataxia telangiectasia (AT), andmore » 180BR cells, a radiosensitive cell line from a patient with no clinical symptoms of AT. AT2 and 180BR cells, in addition to being radiosensitive, also display a reduced ability to repair potentially lethal damage compared to HF19 cells. The yield of DSBs, as measured by pulsed-field gel electrophoresis, is similar in all three cell lines (slopes correspond to 1.6-1.7% Gy{sup -1} of DNA-associated radioactivity released from the gel well into the lane). In contrast, residual DSBs measured 24 h after irradiation are almost zero for HF19 cells (0.1% confidence interval=0-1.4%), but are 12.5% ({plus_minus}2.3%) and 43.8% ({plus_minus}1.2%) of those measured immediately after irradiation in HF19, AT2 and 180BR cells, respectively. Neither the initial yield of DSBs nor that of excess interphase chromosomes breaks can explain the differences in radiosensitivity between the three cell lines; however, there is a correlation between residual DSBs, rate of DSB rejoining at 24 h, residual interphase chromosome breaks on the one hand and cell survival on the other hand. 74 refs., 6 figs., 4 tabs.« less
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