Natural History Sciences, Hokkaido University.
Research Institute for Electronic Science (RIES), Hokkaido University.
Cell Struct Funct. 2021 Nov 20;46(2):95-101. doi: 10.1247/csf.21011. Epub 2021 Sep 25.
Among the inheritance of cellular components during cell division, deoxyribonucleic acid (DNA) and its condensate (chromosome) are conventionally visualized using chemical tag-labeled nucleotide analogs. However, associated mutagenesis with nucleotide analogs in the visualization of chromosomes is cause for concern. This study investigated the efficiency of using stable isotope labels in visualizing the replicating cultured human cell-chromosomes, in the absence of analog labels, at a high spatial resolution of 100 nm. The distinct carbon isotope ratio between sister chromatids reflected the semi-conservative replication of individual DNA strands through cell cycles and suggested the renewal of histone molecules in daughter chromosomes. Thus, this study provides a new, powerful approach to trace and visualize cellular components with stable isotope labeling.Key words: stable isotope, chromosome replication, semi-conservative replication, imaging, mass spectrometry.
在细胞分裂过程中细胞成分的遗传中，脱氧核糖核酸（DNA）及其凝聚物（染色体）通常使用化学标记核苷酸类似物进行可视化。然而，在染色体可视化中使用核苷酸类似物相关的诱变是令人关注的。本研究调查了在没有类似物标记的情况下，使用稳定同位素标记以高空间分辨率 100nm 观察培养的人类细胞染色体的效率。姐妹染色单体之间明显的碳同位素比值反映了单个 DNA 链通过细胞周期的半保留复制，并提示了子染色体中组蛋白分子的更新。因此，本研究提供了一种新的、强大的方法，通过稳定同位素标记来追踪和可视化细胞成分。
稳定同位素、染色体复制、半保留复制、成像、质谱。
