Hulspas R, Houtsmuller A B, Krijtenburg P J, Bauman J G, Nanninga N
Institute for Applied Radiobiology and Immunology TNO, Department of Molecular Pathology, Rijswijk, The Netherlands.
Chromosoma. 1994 Jul;103(4):286-92. doi: 10.1007/BF00352253.
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.
通过共聚焦显微镜分析了G0期(未刺激)和G1期[植物血凝素(PHA)刺激]人淋巴细胞中11号染色体着丝粒周围DNA的核拓扑结构。在三维(3D)分析中,除了核中心外,中心体被用作第二个参考点。使用荧光原位杂交(FISH)和免疫荧光相结合的方法对11号染色体的着丝粒周围DNA和中心体进行标记。为了保留细胞的3D形态,这些技术在悬浮的全细胞上进行。用最近开发的3D软件程序(三维轴和位置的交互式测量)分析细胞的三维图像。当以核中心为参考点时,11号染色体着丝粒在G0期的分布似乎是随机的,但在G1期明显是非随机的。对G1期细胞的进一步统计分析表明,着丝粒随机分布在核膜下方的一个壳层中。在细胞周期的G0期和G1期,中心体和着丝粒之间似乎不存在拓扑关系。