Abstract
Chromosome analysis is one of most fundamental techniques for cytogenetic studies. Chromosomes are conventionally prepared from mitotic cells arrested by colcemid block protocol. Premature chromosome condensation (PCC) technique is an alternative to obtain chromosomes. It was more than half century ago that the first observation of PCC phenomena reported. Since then, cell-fusion-mediated PCC method has been developed and introduced in many fields of chromosome analysis. More than quarter century ago, novel PCC technique using chemical drug has been developed. Afterwards, this simple and efficient drug-induced PCC technique becomes a standard protocol for preparing chromosomes. Thus, it seems to be the good time to introduce PCC technique protocol for the artisans in the field of cytogenetic studies.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Gotoh E, Durante M (2006) Chromosome condensation outside of mitosis: mechanisms and new tools. J Cell Physiol 209(2):297–304. https://doi.org/10.1002/jcp.20720
Johnson RT, Rao PN (1970) Mammalian cell fusion: induction of premature chromosome condensation in interphase nuclei. Nature 226(247):717–722
Cornforth MN, Bedford JS (1983) X-ray--induced breakage and rejoining of human interphase chromosomes. Science 222(4628):1141–1143
Gotoh E, Asakawa Y, Kosaka H (1995) Inhibition of protein serine/threonine phophatases directly induces premature chromosome condensation in mammalian somatic cells. Biomed Res 16(1):63–68
Rao PN, Johnson RT (1970) Mammalian cell fusion: studies on the regulation of DNA synthesis and mitosis. Nature 225(228):159–164
Rao PN, Johnson RT (1971) Mammalian cell fusion. IV Regulation of chromosome formation from interphase nuclei by various chemical compounds. J Cell Physiol 78(2):217–223
Sperling K, Rao PN (1974) Mammalian cell fusion. V Replication behaviour of heterochromatin as observed by premature chromosome condensation. Chromosoma 45(2):121–131
Hanks SK, Gollin SM, Rao PN, Wray W, Hittelman WN (1983) Cell cycle-specific changes in the ultrastructural organization of prematurely condensed chromosomes. Chromosoma 88(5):333–342
Gotoh E, Asakawa Y (1996) Detection and evaluation of chromosomal aberrations induced by high doses of gamma-irradiation using immunogold-silver painting of prematurely condensed chromosomes. Int J Radiat Biol 70(5):517–520
Asakawa Y, Gotoh E (1997) A method for detecting sister chromatid exchanges using prematurely condensed chromosomes and immunogold-silver staining. Mutagenesis 12(3):175–177
Durante M, Furusawa Y, Gotoh E (1998) A simple method for simultaneous interphase-metaphase chromosome analysis in biodosimetry. Int J Radiat Biol 74(4):457–462
Hittelman WN, Pollard M (1984) Visualization of chromatin events associated with repair of ultraviolet light-induced damage by premature chromosome condensation. Carcinogenesis 5(10):1277–1285
Pantelias GE, Maillie HD (1984) The use of peripheral blood mononuclear cell prematurely condensed chromosomes for biological dosimetry. Radiat Res 99(1):140–150
Hittelman WN (1986) Visualization of chromatin events during DNA excision repair in XP cells: deficiency in localized but not generalized chromatin events. Carcinogenesis 7(12):1975–1980
Pantelias GE (1986) Radiation-induced cytogenetic damage in relation to changes in interphase chromosome conformation. Radiat Res 105(3):341–350
Hittelman WN (1990) Direct measurement of chromosome repair by premature chromosome condensation. Prog Clin Biol Res 340B:337–346
Suzuki M, Watanabe M, Suzuki K, Nakano K, Matsui K (1992) Heavy ion-induced chromosome breakage studied by premature chromosome condensation (PCC) in Syrian hamster embryo cells. Int J Radiat Biol 62(5):581–586
Gotoh E, Tanno Y (2005) Simple biodosimetry method for cases of high-dose radiation exposure using the ratio of the longest/shortest length of Giemsa-stained drug-induced prematurely condensed chromosomes (PCC). Int J Radiat Biol 81(5):379–385
Gotoh E, Tanno Y, Takakura K (2005) Simple biodosimetry method for use in cases of high-dose radiation exposure that scores the chromosome number of Giemsa-stained drug-induced prematurely condensed chromosomes (PCC). Int J Radiat Biol 81(1):33–40
Gotoh E (2007) Visualizing the dynamics of chromosome structure formation coupled with DNA replication. Chromosoma 116(5):453–462. https://doi.org/10.1007/s00412-007-0109-5
Ono T, Yamashita D, Hirano T (2013) Condensin II initiates sister chromatid resolution during S phase. J Cell Biol 200(4):429–441. https://doi.org/10.1083/jcb.201208008
Voisin P (2015) Standards in biological dosimetry: a requirement to perform an appropriate dose assessment. Mutat Res Genet Toxicol Environ Mutagen 793:115–122. https://doi.org/10.1016/j.mrgentox.2015.06.012
Lu X, Zhao H, Feng JB, Zhao XT, Chen DQ, Liu QJ (2016) Dose response of multiple parameters for calyculin A-induced premature chromosome condensation in human peripheral blood lymphocytes exposed to high doses of cobalt-60 gamma-rays. Mutat Res Genet Toxicol Environ Mutagen 807:47–54. https://doi.org/10.1016/j.mrgentox.2016.06.010
Guerrero-Carbajal C, Romero-Aguilera I, Arceo-Maldonado C, Gonzalez-Mesa JE, Cortina-Ramirez GE, Garcia-Lima O (2019) Dose response of prematurely condensed chromosome rings after gamma irradiation. Int J Radiat Biol 95(5):607–610. https://doi.org/10.1080/09553002.2019.1566677
Sun M, Moquet J, Barnard S, Lloyd D, Ainsbury E (2020) A simplified Calyculin A-induced premature chromosome condensation (PCC) protocol for the Biodosimetric analysis of high-dose exposure to gamma radiation. Radiat Res 193(6):560–568. https://doi.org/10.1667/rr15538.1
Cornforth MN, Bedford JS (1983) High-resolution measurement of breaks in prematurely condensed chromosomes by differential staining. Chromosoma 88(4):315–318
Lau YF, Arrighi FE (1980) Studies of mammalian chromosome replication. I BrdU-induced differential staining patterns in interphase and metaphase chromosomes. Cytogenet Cell Genet 27(2–3):176–183
Gotoh E, Durante M (2006) Chromosome condensation outside of mitosis: mechanisms and new tools. J Cell Physiol. 209:297–304. and cover page.
Iliakis GE, Pantelias GE (1990) Production and repair of chromosome damage in an X-ray sensitive CHO mutant visualized and analysed in interphase using the technique of premature chromosome condensation. Int J Radiat Biol 57(6):1213–1223
Pantelias GE, Iliakis GE, Sambani CD, Politis G (1993) Biological dosimetry of absorbed radiation by C-banding of interphase chromosomes in peripheral blood lymphocytes. Int J Radiat Biol 63(3):349–354
Durante M, George K, Wu H, Yang TC (1996) Rejoining and misrejoining of radiation-induced chromatin breaks. I experiments with human lymphocytes. Radiat Res 145(3):274–280
Yamashita K, Yasuda H, Pines J, Yasumoto K, Nishitani H, Ohtsubo M, Hunter T, Sugimura T, Nishimoto T (1990) Okadaic acid, a potent inhibitor of type 1 and type 2A protein phosphatases, activates cdc2/H1 kinase and transiently induces a premature mitosis-like state in BHK21 cells. EMBO J 9(13):4331–4338
Schlegel R, Belinsky GS, Harris MO (1990) Premature mitosis induced in mammalian cells by the protein kinase inhibitors 2-aminopurine and 6-dimethylaminopurine. Cell Growth Differ 1(4):171–178
Ghosh S, Paweletz N, Schroeter D (1992) Failure of kinetochore development and mitotic spindle formation in okadaic acid-induced premature mitosis in HeLa cells. Exp Cell Res 201(2):535–540. https://doi.org/10.1016/0014-4827(92)90307-t
Gotoh E (2007) Visualizing the dynamics of chromosome structure formation coupled with DNA replication. Chromosoma [Epub ahead of printing]
Bryant PE, Mozdarani H (2007) A comparison of G2 phase radiation-induced chromatid break kinetics using calyculin-PCC with those obtained using colcemid block. Mutagenesis [Epub ahead of printing]
Johnson RT, Gotoh E, Mullinger AM, Ryan AJ, Shiloh Y, Ziv Y, Squires S (1999) Targeting double-strand breaks to replicating DNA identifies a subpathway of DSB repair that is defective in ataxia-telangiectasia cells. Biochem Biophys Res Commun 261(2):317–325
Gotoh E, Kawata T, Durante M (1999) Chromatid break rejoining and exchange aberration formation following gamma-ray exposure: analysis in G2 human fibroblasts by chemically induced premature chromosome condensation. Int J Radiat Biol 75(9):1129–1135
IAEA (2001) Cytogenetic analysis for radiation dose assesment. A manual. Technical reports series no. 405. International Atomic Energy Agency, Vienna
Bezrookove V, Smits R, Moeslein G, Fodde R, Tanke HJ, Raap AK, Darroudi F (2003) Premature chromosome condensation revisited: a novel chemical approach permits efficient cytogenetic analysis of cancers. Genes Chromosomes Cancer 38(2):177–186
El Achkar E, Gerbault-Seureau M, Muleris M, Dutrillaux B, Debatisse M (2005) Premature condensation induces breaks at the interface of early and late replicating chromosome bands bearing common fragile sites. Proc Natl Acad Sci U S A 102(50):18069–18074
Srebniak MI, Trapp GG, Wawrzkiewicz AK, Kazmierczak W, Wiczkowski AK (2005) The usefulness of calyculin A for cytogenetic prenatal diagnosis. J Histochem Cytochem 53(3):391–394
Mochida A, Gotoh E, Senpuku H, Harada S, Kitamura R, Takahashi T, Yanagi K (2005) Telomere size and telomerase activity in Epstein-Barr virus (EBV)-positive and EBV-negative Burkitt's lymphoma cell lines. Arch Virol 150(10):2139–2150. https://doi.org/10.1007/s00705-005-0557-2
Deckbar D, Birraux J, Krempler A, Tchouandong L, Beucher A, Walker S, Stiff T, Jeggo P, Lobrich M (2007) Chromosome breakage after G2 checkpoint release. J Cell Biol 176(6):749–755
Kanda T, Kamiya M, Maruo S, Iwakiri D, Takada K (2007) Symmetrical localization of extrachromosomally replicating viral genomes on sister chromatids. J Cell Sci 120(Pt 9):1529–1539
Sun M, Moquet J, Lloyd D, Ainsbury E (2020) A faster and easier biodosimetry method based on calyculin A-induced premature chromosome condensation (PCC) by scoring excess objects. J Radiol Prot 40(3):892–905. https://doi.org/10.1088/1361-6498/aba085
Soni A, Murmann-Konda T, Magin S, Iliakis G (2019) A method for the cell-cycle-specific analysis of radiation-induced chromosome aberrations and breaks. Mutat Res 815:10–19. https://doi.org/10.1016/j.mrfmmm.2019.04.001
Okayasu R, Liu C (2019) G1 premature chromosome condensation (PCC) assay. Methods Mol Biol 1984:31–38. https://doi.org/10.1007/978-1-4939-9432-8_4
Masui Y (1974) A cytostatic factor in amphibian oocytes: its extraction and partial characterization. J Exp Zool 187(1):141–147
Masui Y (2001) From oocyte maturation to the in vitro cell cycle: the history of discoveries of maturation-promoting factor (MPF) and cytostatic factor (CSF). Differentiation 69(1):1–17
Dunphy WG, Brizuela L, Beach D, Newport J (1988) The Xenopus cdc2 protein is a component of MPF, a cytoplasmic regulator of mitosis. Cell 54(3):423–431
Gautier J, Norbury C, Lohka M, Nurse P, Maller J (1988) Purified maturation-promoting factor contains the product of a Xenopus homolog of the fission yeast cell cycle control gene cdc2+. Cell 54(3):433–439
Maller J, Gautier J, Langan TA, Lohka MJ, Shenoy S, Shalloway D, Nurse P (1989) Maturation-promoting factor and the regulation of the cell cycle. J Cell Sci Suppl 12:53–63
Doree M, Galas S (1994) The cyclin-dependent protein kinases and the control of cell division. FASEB J 8(14):1114–1121
Prasanna PG, Escalada ND, Blakely WF (2000) Induction of premature chromosome condensation by a phosphatase inhibitor and a protein kinase in unstimulated human peripheral blood lymphocytes: a simple and rapid technique to study chromosome aberrations using specific whole-chromosome DNA hybridization probes for biological dosimetry. Mutat Res 466(2):131–141
Pantelias GE, Maillie HD (1983) A simple method for premature chromosome condensation induction in primary human and rodent cells using polyethylene glycol. Somatic Cell Genet 9(5):533–547. https://doi.org/10.1007/bf01574257
Prasanna PG, Blakely WF (2005) Premature chromosome condensation in human resting peripheral blood lymphocytes for chromosome aberration analysis using specific whole-chromosome DNA hybridization probes. Methods Mol Biol 291:49–57
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Gotoh, E. (2023). Chemical-Induced Premature Chromosome Condensation Protocol. In: Gotoh, E. (eds) Chromosome Analysis. Methods in Molecular Biology, vol 2519. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2433-3_5
Download citation
DOI: https://doi.org/10.1007/978-1-0716-2433-3_5
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2432-6
Online ISBN: 978-1-0716-2433-3
eBook Packages: Springer Protocols