The nuclear position of pericentromeric DNA of chromosome 11 appears to be random in GO and non-random in G1 human lymphocytes |
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Authors: | R Hulspas A B Houtsmuller P -J Krijtenburg J G J Bauman N Nanninga |
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Institution: | (1) Department of Molecular Pathology, Institute for Applied Radiobiology and Immunology TNO, P.O. Box 5815, NL-2280 HV Rijswijk, The Netherlands;(2) Section of Molecular Cytology, Institute for Molecular Cell Biology, BioCentrum Amsterdam, University of Amsterdam, Plantage Muidergracht 14, NL 1018 TV Amsterdam, The Netherlands;(3) Present address: UMMC Cancer Center, Two Biotech, suite 202 373 Plantation St., 01605 Worcester, MA, USA |
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Abstract: | The nuclear topography of pericentromeric DNA of chromosome 11 was analyzed in G0 (nonstimulated) and G1 phytohemagglutinin (PHA) stimulated] human lymphocytes by confocal microscopy. In addition to the nuclear center, the centrosome was used as a second point of reference in the three-dimensional (3D) analysis. Pericentromeric DNA of chromosome 11 and the centrosome were labeled using a combination of fluorescent in situ hybridization (FISH) and immunofluorescence. To preserve the 3D morphology of the cells, these techniques were performed on whole cells in suspension. Three-dimensional images of the cells were analyzed with a recently developed 3D software program (Interactive Measurement of Axes and Positioning in 3 Dimensions). The distribution of the chromosome 11 centromeres appeared to be random during the G0 stage but clearly non-random during the G1 stage, when the nuclear center was used as a reference point. Further statistical analysis of the G1 cells revealed that the centromeres were randomly distributed in a shell underlying the nuclear membrane. A topographical relationship between the centrosome and the centromeres appeared to be absent during the G0 and G1 stages of the cell cycle. |
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