Manuelidis L
Proc Natl Acad Sci U S A. 1984 May;81(10):3123-7. doi: 10.1073/pnas.81.10.3123.
Paraformaldehyde-fixed tissue from mouse cerebellum was hybridized with biotin-labeled satellite DNA for identification of centromeres. By using avidin-peroxidase conjugates, it was possible to define the nuclear position of centromeres at the ultrastructural level. Three-dimensional analysis of well-resolved centromere arrays were aided by computer reconstruction of serial sections. Different cell types displayed distinct, nonrandom centromere locations. In Purkinje neurons, the majority of detected sequences were clustered together around the central nucleolus, whereas in granule neurons, more numerous, dispersed centromere clusters were associated with the nuclear membrane. In Purkinje cells, peroxidase-labeled regions corresponded to dense heterochromatic aggregates were detected in Purkinje cells of several different species. These observations suggest that in these highly differentiated cells, the nuclear position of centromeres is maintained in evolution despite species differences in centromeric DNA sequence. Such defined ordering of centromeres may be integral to specific functional capacities.
将来自小鼠小脑的多聚甲醛固定组织与生物素标记的卫星DNA杂交,以鉴定着丝粒。通过使用抗生物素蛋白-过氧化物酶结合物,能够在超微结构水平确定着丝粒的核位置。通过对连续切片进行计算机重建,辅助对清晰分辨的着丝粒阵列进行三维分析。不同细胞类型显示出不同的、非随机的着丝粒位置。在浦肯野神经元中,大多数检测到的序列聚集在中央核仁周围,而在颗粒神经元中,更多分散的着丝粒簇与核膜相关。在浦肯野细胞中,过氧化物酶标记区域对应于在几种不同物种的浦肯野细胞中检测到的致密异染色质聚集体。这些观察结果表明,在这些高度分化的细胞中,尽管着丝粒DNA序列存在物种差异,但着丝粒的核位置在进化过程中得以维持。这种着丝粒的明确排列可能是特定功能能力所必需的。