Abranches R, Beven A F, Aragón-Alcaide L, Shaw P J
Department of Cell Biology, John Innes Centre, Colney, Norwich NR4 7UH, United Kingdom.
J Cell Biol. 1998 Oct 5;143(1):5-12. doi: 10.1083/jcb.143.1.5.
We have determined the relationship between overall nuclear architecture, chromosome territories, and transcription sites within the nucleus, using three-dimensional confocal microscopy of well preserved tissue sections of wheat roots. Chromosome territories were visualized by GISH using rye genomic probe in wheat/rye translocation and addition lines. The chromosomes appeared as elongated regions and showed a clear centromere-telomere polarization, with the two visualized chromosomes lying approximately parallel to one another across the nucleus. Labeling with probes to telomeres and centromeres confirmed a striking Rabl configuration in all cells, with a clear clustering of the centromeres, and cell files often maintained a common polarity through several division cycles. Transcription sites were detected by BrUTP incorporation in unfixed tissue sections and revealed a pattern of numerous foci uniformly distributed throughout the nucleoplasm, as well as more intensely labeled foci in the nucleoli. It has been suggested that the gene-rich regions in wheat chromosomes are clustered towards the telomeres. However, we found no indication of a difference in concentration of transcription sites between telomere and centromere poles of the nucleus. Neither could we detect any evidence that the transcription sites were preferentially localized with respect to the chromosome territorial boundaries.
我们利用小麦根保存完好的组织切片的三维共聚焦显微镜,确定了细胞核内整体核结构、染色体领地和转录位点之间的关系。在小麦/黑麦易位系和附加系中,使用黑麦基因组探针通过基因组原位杂交(GISH)对染色体领地进行可视化。染色体呈现为细长区域,显示出明显的着丝粒-端粒极化,两条可视化的染色体在细胞核中大致相互平行排列。用端粒和着丝粒探针标记证实,所有细胞中都呈现出显著的拉布尔(Rabl)构型,着丝粒明显聚集,并且细胞列常常在几个分裂周期中保持共同的极性。通过在未固定的组织切片中掺入溴尿嘧啶三磷酸(BrUTP)来检测转录位点,结果显示出众多焦点均匀分布于整个核质中的模式,以及核仁中标记更强的焦点。有人提出小麦染色体中富含基因的区域向端粒聚集。然而,我们没有发现细胞核端粒极和着丝粒极之间转录位点浓度存在差异的迹象。我们也没有检测到任何证据表明转录位点相对于染色体领地边界存在优先定位。