Abstract
To visualize characteristic chromatin distortions we have distinguished first among regularly occurring intermediates of chromatin structures in mammalian (Indian muntjac, CHO, murine preB, rat liver, rat myeloid leukemia, K562 human erythroid leukemia) and Drosophila nuclei. Fluorescence microscopy of chromatin structures isolated from nuclei of reversibly permeable cells revealed a common pathway of chromatin condensation in mammalian cells. Different intermediates in mammalian and Drosophila cells indicate alternative mechanisms of chromosome condensation. Genotoxic agents such as irradiation (alpha, gamma, UV-B) and heavy metals (Cd, Pb, Ni, Hg, Ag) caused alterations in chromatin structures leading to apoptosis. Injury-specific chromatin changes manifested at significantly lower concentrations than non-specific signs of cellular toxicity, suggesting that preapoptotic events are useful indicators of genotoxicity.
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References
Banfalvi G (2009) Apoptotic chromatin changes. Springer, Dordrecht, Fig. 1.10 p. 17, Fig. 4
Bernard J, Malawista SE (1995) Remembrance of professor Marcel Bessis (1917–1994). Blood Cell Mol Dis 21:152–155
Kerr JFR (1971) Shrinkage necrosis: a distinct mode of cellular death. J Pathol 105:13–20
Susin SA, Daugas E, Ravagnan L, Samejima K, Zamzami N, Loeffler M, Costantini P, Ferri KF, Irinopoulou T, Prévost MC, Brothers G, Mak TW, Penninger J, Earnshaw WC, Kroemer G (2000) Two distinct pathways leading to nuclear apoptosis. J Exp Med 192:571–580
Banfalvi G, Klaisz M, Ujvarosi K, Trencsenyi G, Rozsa D, Nagy G (2007) Gamma irradiation induced apoptotic changes in the chromatin structure of human erythroleukemia K562 cells. Apoptosis 12:2271–2283
Humke EW (2000) Web Alert Apoptosis. Chem Biol 7(2):R48–R49
Nagy G, Pinter G, Kohut G, Adam A, Trencsenyi G, Hornok L, Banfalvi G (2010) Time-lapse analysis of cell death in mammalian and fungal cells. DNA Cell Biol 29:249–259
Paulson JR, Laemmli UK (1977) The structure of histone depleted chromosomes. Cell 12:817–828
Adolph KW (1980) Isolation and structural organization of human mitotic chromosomes. Chromosoma 76:23–33
Rattner JB, Lin CC (1985) Radial loops and helical coils coexist in metaphase chromosomes. Cell 42:291–296
Boy De La Tour E, Laemmli UK (1988) The metaphase scaffold is helically folded: sister chromatids have predominantly opposite helical handedness. Cell 55:937–944
Kireeva N, Lakonishok M, Kireev I, Hirano T, Belmont AS (2004) Visualization of early chromosomal condensation: a hierarchical folding, axial glue model of chromosome structure. J Cell Biol 166:775–785
Banfalvi G (1993) Fluorescent analysis of replication and intermediates of chromatin folding in nuclei of mammalian cells. In: Bach PH, Reynolds CH, Clark JM, Mottley J, Poole PL (eds) Biotechnology applications of microinjection, microscopic imaging, and fluorescence. Plenum Press, New York, pp 111–119
Hepperger C, Otten S, von Hase J, Dietzel S (2007) Preservation of large-scale chromatin structure in FISH experiments. Chromosoma 116:117–133
Kobliakova I, Zatsepina O, Stefanova V, Polyakov V, Kireev I (2005) The topology of early- and late-replicating chromatin in differentially decondensed chromosomes. Chromosome Res 13:169–181
Banfalvi G, Sooki-Toth A, Sarkar N, Csuzi S, Antoni F (1984) Nascent DNA chains synthesized in recersibly permeable cells of mouse thymocytes. Eur J Biochem 139:553–559
Flemming W (1882) Zellsubstanz, Kern- und Zelltheilung. Vogel, Leipzig
Balbiani EG (1881) Sur la structure du noyau des cellules salivaires chez les larves de Chironomus. Zool Anz 4(637–641):662–666
Banfalvi G (2008) Chromatin fiber structure and plectonemic model of chromosome condensation in Drosophila cells. DNA Cell Biol 27:65–70
Trencsenyi G, Nagy G, Bako F, Kertai P, Banfalvi G (2012) Incomplete chromatin condensation in enlarged rat myelocytic leukemia cells. DNA Cell Biol 31:470–478
Paniagua P, Nistal M, Amat P, Rodriguez MC, Alonso JR (1986) Quantitative differences between variants of a spermatogonia in man. J Reprod Fertil 77:669–673
Nagy G, Kiraly G, Turani M, Banfalvi G (2013) Cell trivision of hyperploid cells. DNA Cell Biol 32:676–684
Müller I, Boyle S, Singer RH, Bickmore WA, Chubb JR (2010) Stable morphology, but dynamic internal reorganisation, of interphase human chromosomes in living cells. PLoS One 13(5):e11560. doi:10.1371/journal.pone.0011560
Bjorkroth B, Ericsson C, Lamb MM, Danehol B (1988) Structure of the chromatin axis during transcription. Chromosoma 96:333–340
Belmont AS, Braunfeld MB, Sedat JW, Agard DA (1989) Large-scale chromatin structural domains within mitotic and interphase chromosomes in vivo and in vitro. Chromosoma 98:129–143
Lemke J, Claussen J, Michel S, Chudoba I, Mühlig P, Westermann M, Sperling K, Rubtsov N, Grummt UW, Ullmann P, Kromeyer-Hauschild K, Liehr T, Claussen U (2002) The DNA-based structure of human chromosome 5 in interphase. Am J Hum Genet 71:1051–1059
Lippincott-Schwartz J, Snapp E, Kenworthy A (2001) Studying protein dynamics in living cells. Nat Rev Mol Cell Biol 2:444–456
Pulkinnen JO, Joensuu LH, Martikainen P, Servomaa K, Grénman R (1996) Paclitaxel-induced apoptotic changes followed by time-lapse video microscopy in cell lines established from head and neck cancer. J Cancer Res Clin Oncol 122:214–218
Forrester HB, Albright N, Ling CC, Dewey WC (2000) Computerized video time-lapse analysis of apoptosis of REC:Myc cells X-irradiated in different phases of the cell cycle. Radiat Res 154:625–639
Hoepfner D, Brachat A, Philippsen P (2000) Time-lapse video microscopy analysis reveals astral microtubule detachment in the yeast spindle pole mutant cnm67. Mol Biol Cell 11:1197–1211
Lane JD, Allan VJ, Woodman PG (2005) Active relocation of chromatin and endoplasmic reticulum into blebs in late apoptotic cells. J Cell Sci 118:4059–4071
Rieger MA, Schroeder T (2008) Exploring hematopoiesis at single cell resolution. Cell Tissues Organs 188:139–149
Hinchcliffe EH (2005) Using long-term time-lapse imaging of mammalian cell cycle progression for laboratory instruction and analysis. Cell Biol Educ 4:284–290
Dixit R, Cyr R (2003) Cell damage and reactive oxygen species production induced by fluorescence microscopy: effect on mitosis and guidelines for non-invasive microscopy. Plant J 36:280–290
Gacsi M, Nagy G, Pinter G, Basnakian AG, Banfalvi G (2005) Condensation of interphase chromatin in nuclei of Chinese hamster ovary (CHO-K1) cells. DNA Cell Biol 24:43–53
Banfalvi G (2008) Cell cycle synchronization of animal cells and nuclei by centrifugal elutriation. Nat Protoc 3:663–673
Banfalvi G (2011) Synchronization of mammalian cells and nuclei by centrifugal elutriation. Methods Mol Biol 761:25–45
Banfalvi G, Nagy G, Gacsi M, Roszer T, Basnakian AG (2006) Common pathway of chromosome condensation in mammalian cells. DNA Cell Biol 25:295–301
Henriques V, Orskov SL (1936) Untersuchungen fiber die Schwankungen des Kationgehaltes der roten Blutkörperchen. Skand Arch Physiol 74:63–78
Davson H, Danielli JF (1938) Studies on the permeability of erythrocytes factors in cation permeability. Biochem J 32:991–1001
Joyce CRB, Moore H, Weatherall M (1954) The effects of lead, mercury, and gold on the potassium turnover of rabbit blood cells. Br J Pharmacol Chemother 9:463–470
Passow H, Rothstein A (1960) The binding of mercury by the yeast cell in relation to changes in permeability. J Gen Physiol 43:621–633
Braeckman B, Raes H, Van Hoye D (1997) Heavy-metal toxicity in an insect cell line. Effects of cadmium chloride, mercuric chloride and methylmercuric chloride on cell viability and proliferation in Aedes albopictus cells. Cell Biol Toxicol 13:389–397
McBrien DCH, Hassal KA (1965) The effect of toxic doses of copper upon respiration, photosynthesis and growth of Chlorella vulgaris. Physiol Plant 20:113–117
Halldorson H, Gray DA, Shall S (1978) Poly (ADP-ribose) polymerase activity in nucleotide permeable cells. FEBS Lett 85:349–359
Kucera R, Paulus H (1982) Studies on ribonucleoside-diphosphate reductase in permeable animal cells. II. Catalytic and regulatory properties of the enzyme in mouse L cells. Arch Biochem Biophys 214:114–123
Lesh RE, Somlyo AP, Owens GK, Somlyo AV (1995) Reversible permeabilization: a novel technique for the intracellular introduction of antisense oligodeoxynucleotides into intact smooth muscle. Circ Res 77:220–230
Kobayashi S, Kitazawa T, Somlyo AV, Somlyo AP (1989) Cytosolic heparin inhibits muscarinic and alpha-adrenergic Ca2+ release in smooth muscle. Physiological role of inositol 1,4,5-trisphosphate in pharmacomechanical coupling. J Biol Chem 264:17997–18004
Demis DJ, Rothstein A (1955) Relationship of the cell surface to metabolism. XII. Effect of mercury and copper on glucose uptake and respiration of rat diaphragm. Am J Physiol 180:566–574
Jacobs MH, Glassman HN, Parpart AK (1950) Hemolysis and zoological relationship. Comparative studies with four penetrating non-electrolytes. J Exp Zool 113(2):277–300
Wilbrandt W (1941) Die Wirkung von Schwermetallsalzen auf die Erythrocyten- permeabilität für Glycerin. Pflüg Arch Ges Physiol 244(5):637–643
Myrbäck K (1957) Inhibition of yeast invertase (saccharase) by metal ions. V. Inhibition by mercury compounds. Arkiv Kemi 11:471–479
Banfalvi G, Littlefield N, Hass B, Mikhailova M, Csuka I, Szepessy E, Chou WM (2000) Effect of cadmium on the relationship between replicative and repair DNA synthesis in synchronized cho cells. Eur J Biochem 267:6580–6585
Banfalvi G, Gacsi M, Nagy G, Kiss BZ, Basnakian AG (2005) Cadmium induced apoptotic changes in chromatin structure and subphases of nuclear growth during the cell cycle in CHO cells. Apoptosis 10:631–642
Banfalvi G, Ujvarosi K, Trencsenyi G, Somogyi C, Nagy G, Basnakian AG (2007) Cell culture density dependent toxicity and chromatin changes upon cadmium treatment in murine pre-B cells. Apoptosis 12:1219–1228
Banfalvi G, Sarvari A, Nagy G (2012) Chromatin changes induced by Pb and Cd in human cells. Toxicol In Vitro 26:1064–1071
Trencsenyi G, Kertai P, Somogyi C, Nagy G, Dombradi Z, Gacsi M, Banfalvi G (2007) Chemically induced carcinogenesis affecting chromatin structure in rat hepatocarcinoma cells. DNA Cell Biol 26:649–655
Yedjou CG, Milner JN, Howard CB, Tchounwou TB (2010) Basic apoptotic mechanisms of lead toxicity in human leukemia (HI-60) cells. Int J Environ Res Publ Health 7:2008–2017
Columbano A, Ledda-Columbano GM, Coni PP, Faa G, Liguori C, Santa Cruz G, Pani P (1985) Occurrence of cell death (apoptosis) during the involution of liver hyperplasia. Lab Investig 52:670–675
Xu J, Ji L-D, Xu L-H (2006) Lead-induced apoptosis on PC 12 cells: involvement of p53, Bcl-2 family and caspase-3. Toxicl Lett 166:160–167
Agarwal S, Roy S, Ray A, Mazumbder S, Bhattacharya S (2009) Arsenic trioxide and lead acetate induce apoptosis in adult rat stem cells. Cell Biol Toxicol 25:403–413
Nagy G, Laza D, Ujvarosi K, Banfalvi G (2011) Chromatin toxicity of Ni(II) ions in K562 erythroleukemia cells. In: Banfalvi G (ed) Cellular effects of heavy metals. Springer, New York, pp 163–178
Farkas E, Ujvarosi K, Nagy G, Posta J, Banfalvi G (2010) Apoptogenic and necrogenic effects of mercuric acetate on the chromatin structure of K562 human erythroleukemia cells. Toxicol In Vitro 24:267–275
Kasprzak KS, Sunderman FW Jr, Salnikow K (2003) Nickel carcinogenesis. Mutat Res 533:67–97
Nagy G, Turani M, Kovacs KE, Banfalvi G (2011) Chromatin changes upon silver nitrate treatment in human keratocyte HaCaT and K562 erythroleukemia cells. In: Banfalvi G (ed) Cellular effects of heavy metals. Springer, New York, pp 195–217
Nagy G, Gacsi M, Rehak M, Basnakian AG, Klaisz M, Banfalvi G (2004) Gamma irradiation-induced apoptosis in murine pre-B cells prevents the condensation of fibrillar chromatin in early S phase. Apoptosis 9:765–776
Furuya R, Kumagai H, Hishida A (1997) Acquired resistance to rechallenge injury with uranyl acetate in LLC-PK1 cells. J Lab Clin Med 129:347–355
Ujvarosi K, Hunyadi J, Nagy G, Pocsi I, Banfalvi G (2007) Preapoptotic chromatin changes induced by ultraviolet B irradiation in human erythroleukemia K562 cells. Apoptosis 12:2089–2099
Acknowledgments
This work was supported by Hungarian Scientific Research Fund (OTKA grant) T42762 grant to G.B. Transmission electron microscopic images of Helix lucorum chromosomes made in 2005 and provided by former PhD students, T. Roszer and G. Nagy are gratefully acknowledged.
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Banfalvi, G. Apoptotic agents inducing genotoxicity-specific chromatin changes. Apoptosis 19, 1301–1316 (2014). https://doi.org/10.1007/s10495-014-1018-8
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DOI: https://doi.org/10.1007/s10495-014-1018-8