Dwiranti Astari, Takata Hideaki, Fukui Kiichi
Department of Biology, Faculty of Mathematics and Natural Sciences,Universitas Indonesia,Depok Campus, 16404 West Java,Indonesia.
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology,Midorigaoka, Ikeda, Osaka 563-8577,Japan.
Microsc Microanal. 2019 Jun;25(3):817-821. doi: 10.1017/S1431927619000266. Epub 2019 Apr 17.
The structural details of chromosomes have been of interest to researchers for many years, but how the metaphase chromosome is constructed remains unsolved. Divalent cations have been suggested to be required for the organization of chromosomes. However, detailed information about the role of these cations in chromosome organization is still limited. In the current study, we investigated the effects of Ca2+ and Mg2+ depletion and the reversibility upon re-addition of one of the two ions. Human chromosomes were treated with different concentrations of Ca2+and Mg2+. Depletion of Ca2+ and both Ca2+ and Mg2+ were carried out using 1, 2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid and ethylenediaminetetraacetic acid (EDTA), respectively. Chromosome structure was examined by fluorescence microscopy and scanning electron microscopy. The results indicated that chromosome structures after treatment with a buffer without Mg2+, after Ca2+ depletion, as well as after depletion of both Mg2+, and Ca2+, yielded fewer compact structures with fibrous chromatin than those without cation depletion. Interestingly, the chromatin of EDTA-treated chromosomes reversed to their original granular diameters after re-addition of either Mg2+ or Ca2+ only. These findings signify the importance of divalent cations on the chromosome structure and suggest the interchangeable role of Ca2+ and Mg2+.
多年来,染色体的结构细节一直是研究人员感兴趣的内容,但中期染色体是如何构建的仍然没有得到解决。有人提出二价阳离子是染色体组织所必需的。然而,关于这些阳离子在染色体组织中的作用的详细信息仍然有限。在当前的研究中,我们研究了Ca2+和Mg2+缺失的影响以及重新添加这两种离子之一后的可逆性。用不同浓度的Ca2+和Mg2+处理人类染色体。分别使用1,2 - 双(2 - 氨基苯氧基)乙烷 - N,N,N',N' - 四乙酸和乙二胺四乙酸(EDTA)进行Ca2+以及Ca2+和Mg2+的缺失处理。通过荧光显微镜和扫描电子显微镜检查染色体结构。结果表明,与未进行阳离子缺失处理的染色体相比,用不含Mg2+的缓冲液处理后、Ca2+缺失后以及Mg2+和Ca2+都缺失后的染色体结构产生的致密结构和纤维状染色质较少。有趣的是,仅重新添加Mg2+或Ca2+后,经EDTA处理的染色体的染色质直径恢复到其原始颗粒直径。这些发现表明二价阳离子对染色体结构的重要性,并暗示了Ca2+和Mg2+的可互换作用。