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Reversible Changes of Chromosome Structure upon Different Concentrations of Divalent Cations

Part of: Micrographia Collection

Published online by Cambridge University Press:  17 April 2019

Astari Dwiranti Open the ORCID record for Astari Dwiranti [Opens in a new window] ,
Hideaki Takata Open the ORCID record for Hideaki Takata [Opens in a new window]  and
Kiichi Fukui Open the ORCID record for Kiichi Fukui [Opens in a new window]
Astari Dwiranti
Affiliation:
Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Indonesia, Depok Campus, 16404 West Java, Indonesia
Hideaki Takata
Affiliation:
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Midorigaoka, Ikeda, Osaka 563-8577, Japan
Kiichi Fukui*
Affiliation:
Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka 565-0871, Japan
*
*Author for correspondence: Kiichi Fukui, E-mail: [email protected]
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Abstract

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+.

Keywords

calcium ion (Ca2+)chromosomeEDTAmagnesium ion (Mg2+)reversibilityscanning electron microscopy
Type
Micrographia
Information
Microscopy and Microanalysis , Volume 25 , Issue 3 , June 2019 , pp. 817 - 821
Copyright
Copyright © Microscopy Society of America 2019 

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References

Adolph, KW, Kreisman, LR & Kuehn, RL (1986). Assembly of chromatin fibers into metaphase chromosomes analyzed by transmission electron microscopy and scanning electron microscopy. Biophys J 49, 221–231.Google Scholar
Albiez, H, Cremer, M, Tiberi, C, Vecchio, L, Schermelleh, L, Dittrich, S, Küpper, K, Joffe, B, Thormeyer, T, von Hase, J, Yang, S, Rohr, K, Leonhardt, H, Solovei, I, Cremer, C, Fakan, S & Cremer, T (2006). Chromatin domains and the interchromatin compartment form structurally defined and functionally interacting nuclear networks. Chrom Res 14, 707–733.Google Scholar
Bloomfield, VA (1997). DNA condensation by multivalent cations. Biopolymers 44, 269–282.Google Scholar
Caravaca, JM, Caño, S, Gállego, I & Daban, J (2005). Structural elements of bulk chromatin within metaphase chromosomes. Chrom Res 13, 725–743.Google Scholar
Dwiranti, A, Hamano, T, Takata, H, Nagano, S, Guo, H, Onishi, K, Wako, T, Uchiyama, S & Fukui, K (2014). The effect of magnesium ions on chromosome structure as observed by helium ion. Microscopy Microsc 20, 184–188.Google Scholar
Dwiranti, A, Takata, H, Uchiyama, S & Fukui, K (2016). The effect of magnesium ions on chromosome structure as observed by scanning electron microscopy (SEM) and scanning electron microscope (STEM) tomography. Chrom Sci 19, 19–23.Google Scholar
Eltsov, M, Maclellan, KM, Maeshima, K, Frangakis, AS & Dubochet, J (2008) Analysis of cryo-electron microscopy images does not support the existence of 30-nm chromatin fibers in mitotic chromosomes in situ. Proc Natl Acad Sci 105, 19732–19737.Google Scholar
Engelhardt, M (2004). Condensation of chromatin in situ by cation-dependent charge shielding and aggregation. Biochem Biophys Res Com 324, 1210–1214.Google Scholar
Grigoryev, SA & Woodcock, CL (2012). Chromatin organization—the 30 nm fiber. Exp Cell Res 318, 1448–1455.Google Scholar
Hayashihara, K, Uchiyama, S, Kobayashi, S, Yanagisawa, M, Matsunaga, S & Fukui, K (2008). Isolation method for human metaphase chromosomes. Protoc Exchange doi:10.1038/nprot.2008.166.Google Scholar
Herzog, M & Soyer, M (1983). The native structure of dinoflagellate chromosomes and their stabilization by Ca2+ and Mg2+ cations. Eur J Cell Biol 30, 33–41.Google Scholar
Levi-Setti, R, Gavrilova, KL & Rizzo, PJ (2008). Divalent cation distribution in dinoflagellate chromosomes imaged by high-resolution ion probe mass spectrometry. Eur J Cell Biol 87, 963–976.Google Scholar
Nishino, Y, Eltsov, M, Joti, Y, Ito, K, Takata, H, Takahashi, Y, Hihara, S, Frangakis, AS, Imamoto, N, Ishikawa, T & Maeshima, K (2012). Human mitotic chromosomes consist predominantly of irregularly folded nucleosome fibres without a 30-nm chromatin structure. EMBO J 31, 1644–1653.Google Scholar
Phengchat, R, Takata, H, Morii, K, Inada, N, Murakoshi, H, Uchiyama, S & Fukui, K (2016). Calcium ions function as a booster of chromosome condensation. Sci Rep 6, 38281. DOI: 10.1038/srep38281.Google Scholar
Poirier, MG, Monhait, T & Marko, JF (2002). Reversible hypercondensation and decondensation of mitotic chromosomes studied using combined chemical-micromechanical techniques. J Cell Biochem 85, 422–434.Google Scholar
Strick, R, Strissel, PL, Gavrilov, K & Levi-Setti, R (2001). Cation-chromatin binding as shown by ion microscopy is essential for the structural integrity of chromosomes. J Cell Biol 155, 899–910.Google Scholar
Uchiyama, S, Kobayashi, S, Takata, H, Ishihara, T, Sone, T, Matsunaga, S & Fukui, K (2004). Protein composition of human metaphase chromosomes analyzed by two-dimensional electrophoreses. Cytogenet Genome Res 107, 49–54.Google Scholar
Uchiyama, S, Kobayashi, S, Takata, H, Ishihara, T, Hori, N, Higashi, T, Hayashihara, K, Sone, T, Higo, D, Nirasawa, T, Takao, T, Matsunaga, S & Fukui, K (2005). Proteome analysis of human metaphase chromosomes. J Biol Chem 280, 16994–17004.Google Scholar
Visvanathan, A, Ahmed, K, Even-Faitelson, L, Lleres, D, Bazett-Jones, DP & Lamond, A (2013). Modulation of higher order chromatin conformation in mammalian cell nuclei can be mediated by polyamines and divalent cations. PLoS ONE 8, e67689.Google Scholar