Abstract
Cellular division is a fundamental process of cellular growth. First, cells replicate their DNA in S phase and then undergo mitosis which, under normal conditions, leads to complete cell division. Moreover, mitotic activity correlates to cellular growth activity. The simplest and classical method to measure mitotic activity (mitotic index (MI)), is the manual counting of mitotic cells among a given cell population of interest. The latter can be accomplished via phase contrast microscope observation. However, Giemsa staining may improve accuracy and consistency. Fluorescence immunostaining targeting specific phosphorylations of proteins at critical cell cycle steps will provide further improved analysis via high-throughput capacity of flow or imaging cytometer. Finally, time lapse image analysis provides quantitative and qualitative metrics delineating the process of cellular division including timing of division, duration of mitosis, and failure to procced through or complete mitosis.
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References
Howard A, Pelc S (1953) Synthesis of deoxyribonucleic acid in normal and irradiated cells and its relation to chromosome breakage. Heredity 6(supple):261–273
Hoffman JG (1947) Wright's hypothesis: its relation to volume growth of tissue cells and mitotic index. Science 106(2754):343–344. https://doi.org/10.1126/science.106.2754.343-a
Hoffman JG (1949) Theory of the mitotic index and its application to tissue growth measurement. Bull Math Biophys 11(2):139–144. https://doi.org/10.1007/BF02477499
Kim JY, Jeong HS, Chung T, Kim M, Lee JH, Jung WH, Koo JS (2017) The value of phosphohistone H3 as a proliferation marker for evaluating invasive breast cancers: a comparative study with Ki67. Oncotarget. 8(39):65064–65076. https://doi.org/10.18632/oncotarget.17775
Medri L, Volpi A, Nanni O, Vecci AM, Mangia A, Schittulli F, Padovani F, Giunchi DC, Zito A, Amadori D, Paradiso A, Silvestrini R (2003) Prognostic relevance of mitotic activity in patients with node-negative breast cancer. Mod Pathol 16(11):1067–1075. https://doi.org/10.1097/01.MP.0000093625.20366.9D
Wilson L (1975) Action of drugs on microtubules. Life Sci 17(3):303–309. https://doi.org/10.1016/0024-3205(75)90476-2
Eigsti OJ (1938) A cytological study of colchicine effects in the induction of polyploidy in plants. Proceedings of the National Academy of Sciences of the United States of America. 24:56–63. https://doi.org/10.1073/pnas.24.2.56
Davidson D, Macleod RD (1966) Changes in mitotic indices in roots of vicia faba I. Antagonistic Effects of Colchicine and Iaa. Chromosoma. 18(3):421. https://doi.org/10.1007/Bf00332546
Dewey WC, Miller HH (1969) X-ray induction of chromatid exchanges in mitotic and G1 Chinese hamster cells pretreated with Colcemid. Exp Cell Res 57(1):63–70
Alderton GK, Joenje H, Varon R, Borglum AD, Jeggo PA, O'Driscoll M (2004) Seckel syndrome exhibits cellular features demonstrating defects in the ATR-signalling pathway 22. HumMolGenet 13(24):3127–3138
Rossi L, Laas E, Mallon P, Vincent-Salomon A, Guinebretiere JM, Lerebours F, Rouzier R, Pierga JY, Reyal F (2015) Prognostic impact of discrepant Ki67 and mitotic index on hormone receptor-positive, HER2-negative breast carcinoma. Br J Cancer 113(7):996–1002. https://doi.org/10.1038/bjc.2015.239
Chopra DP, Wilkoff LJ (1977) Induction of hyperplasia and anaplasia by carcinogens in organ cultures of mouse prostate. In Vitro 13(4):260–267. https://doi.org/10.1007/BF02615084
Goto H, Tomono Y, Ajiro K, Kosako H, Fujita M, Sakurai M, Okawa K, Iwamatsu A, Okigaki T, Takahashi T, Inagaki M (1999) Identification of a novel phosphorylation site on histone H3 coupled with mitotic chromosome condensation. J Biol Chem 274(36):25543–25549. https://doi.org/10.1074/jbc.274.36.25543
Hendzel MJ, Wei Y, Mancini MA, Van Hooser A, Ranalli T, Brinkley BR, Bazett-Jones DP, Allis CD (1997) Mitosis-specific phosphorylation of histone H3 initiates primarily within pericentromeric heterochromatin during G2 and spreads in an ordered fashion coincident with mitotic chromosome condensation. Chromosoma 106(6):348–360. https://doi.org/10.1007/s004120050256
Paulson JR, Taylor SS (1982) Phosphorylation of histones 1 and 3 and nonhistone high mobility group 14 by an endogenous kinase in HeLa metaphase chromosomes. J Biol Chem 257(11):6064–6072
Gurley LR, D'Anna JA, Barham SS, Deaven LL, Tobey RA (1978) Histone phosphorylation and chromatin structure during mitosis in Chinese hamster cells. Eur J Biochem 84(1):1–15. https://doi.org/10.1111/j.1432-1033.1978.tb12135.x
Wei Y, Mizzen CA, Cook RG, Gorovsky MA, Allis CD (1998) Phosphorylation of histone H3 at serine 10 is correlated with chromosome condensation during mitosis and meiosis in Tetrahymena. Proc Natl Acad Sci U S A 95(13):7480–7484. https://doi.org/10.1073/pnas.95.13.7480
Sawicka A, Seiser C (2012) Histone H3 phosphorylation - a versatile chromatin modification for different occasions. Biochimie 94(11):2193–2201. https://doi.org/10.1016/j.biochi.2012.04.018
Forrester HB, Vidair CA, Albright N, Ling CC, Dewey WC (1999) Using computerized video time lapse for quantifying cell death of X-irradiated rat embryo cells transfected with c-myc or c-ha-ras. Cancer Res 59(4):931–939
Lynch AE, Triajianto J, Routledge E (2014) Low-cost motility tracking system (LOCOMOTIS) for time-lapse microscopy applications and cell visualisation. PLoS One 9(8):e103547. https://doi.org/10.1371/journal.pone.0103547
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Kato, T.A., Haskins, J.S. (2023). Mitotic Index Analysis. 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_3
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DOI: https://doi.org/10.1007/978-1-0716-2433-3_3
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