Dietzel S, Eils R, Sätzler K, Bornfleth H, Jauch A, Cremer C, Cremer T
Institute of Human Genetics, University of Heidelberg, Germany.
Exp Cell Res. 1998 May 1;240(2):187-96. doi: 10.1006/excr.1998.3934.
Multicolor fluorescence in situ hybridization with a whole chromosome composite probe for the X-chromosome and microdissection probes for the Xp and Xq arms, as well as for the Xp terminal, Xq terminal, and X centromer specific subregional probes, was applied to three-dimensional (3D) preserved human female amniotic fluid cell nuclei. Confocal laser scanning microscopy and three-dimensional image analysis demonstrated distinctly separated Xp arm and Xq arm domains. 3D distance measurements revealed a high variability of intrachromosomal distances between Xpter, Xcen, and Xqter specific probes within both X territories. A 3D distance measurement error of +/- 70 nm was found in control experiments using quartz glass microspheres labeled with different fluorochromes. Our data argue against the hypothesis of Walker et al. (1991, Proc. Natl. Acad. Sci. USA 88, 6191-6195) that a looped structure of the inactive X territory is formed by tight telomere-telomere associations.
采用针对X染色体的全染色体复合探针以及针对Xp和Xq臂、Xp末端、Xq末端和X着丝粒特异性亚区域探针的显微切割探针，对三维（3D）保存的人类女性羊水细胞核进行多色荧光原位杂交。共聚焦激光扫描显微镜和三维图像分析显示Xp臂和Xq臂结构域明显分离。三维距离测量显示，在两个X区域内，Xpter、Xcen和Xqter特异性探针之间的染色体内距离存在高度变异性。在使用不同荧光染料标记的石英玻璃微球的对照实验中，发现三维距离测量误差为±70 nm。我们的数据与Walker等人（1991年，《美国国家科学院院刊》88，6191 - 6195）的假设相悖，该假设认为失活X区域的环状结构是由紧密的端粒 - 端粒关联形成的。
