Abstract
The ability to analyze cell division in both spatial and temporal dimensions within an organism is a key requirement in developmental biology. Specialized cell types within individual organs, such as those within shoot and root apical meristems, have often been identified by differences in their rates of proliferation prior to the characterization of distinguishing molecular markers. Replication-dependent labeling of DNA is a widely used method for assaying cell proliferation. The earliest approaches used radioactive labeling with tritiated thymidine, which were later followed by immunodetection of bromodeoxyuridine (BrdU). A major advance in DNA labeling came with the use of 5-ethynyl-2′deoxyuridine (EdU) which has proven to have multiple advantages over BrdU. Here we describe the methodology for analyzing EdU labeling and retention in whole plants and histological sections of Arabidopsis.
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References
Clowes FAL (1956) Localization of nucleic acid synthesis in root meristems. J Exp Bot 7:307–312
Clowes FAL (1956) Nucleic acids in root apical meristems of Zea. New Phytol 55:29–34
Booth A, Cutter EG, Postlethwait SN, Voeller BR, Wareing PF (1959) Use of radioactive tracers in studies of the shoot apex. Nature 183:1580–1582
Gratzner HG (1982) Monoclonal antibody to 5-bromo- and 5-iododeoxyuridine: a new reagent for detection of DNA replication. Science 218:474–475
Gratzner HG, Leif RC, Ingram DJ, Castro A (1975) Use of antibody specific for bromodeoxyuridine for immunofluorescent determination of DNA-replication in single cells and chromosomes. Exp Cell Res 95:88–94
Salic A, Mitchison TJ (2008) A chemical method for fast and sensitive detection of DNA synthesis in vivo. Proc Natl Acad Sci U S A 105:2415–2420
Kotogany E, Dudits D, Horvath GV, Ayaydin F (2010) A rapid and robust assay for detection of S-phase cell cycle progression in plant cells and tissues by using ethynyl deoxyuridine. Plant Methods 6:5
Perilli S, Perez-Perez JM, Di Mambro R, Peris CL, Diaz-Trivino S, Del Bianco M, Pierdonati E, Moubayidin L, Cruz-Ramirez A, Costantino P, Scheres B, Sabatini S (2013) RETINOBLASTOMA-RELATED protein stimulates cell differentiation in the Arabidopsis root meristem by interacting with cytokinin signaling. Plant Cell 25:4469–4478
Vanstraelen M, Baloban M, Da Ines O, Cultrone A, Lammens T, Boudolf V, Brown SC, De Veylder L, Mergaert P, Kondorosi E (2009) APC/C-CCS52A complexes control meristem maintenance in the Arabidopsis root. Proc Natl Acad Sci U S A 106:11806–11811
Schubert I, Schubert V, Fuchs J (2011) No evidence for “break-induced replication” in a higher plant - but break-induced conversion may occur. Front Plant Sci 2:8
She W, Grimanelli D, Rutowicz K, Whitehead MW, Puzio M, Kotlinski M, Jerzmanowski A, Baroux C (2013) Chromatin reprogramming during the somatic-to-reproductive cell fate transition in plants. Development 140:4008–4019
Zhu Y, Weng M, Yang Y, Zhang C, Li Z, Shen WH, Dong A (2011) Arabidopsis homologues of the histone chaperone ASF1 are crucial for chromatin replication and cell proliferation in plant development. Plant J 66:443–455
Ichihashi Y, Kawade K, Usami T, Horiguchi G, Takahashi T, Tsukaya H (2011) Key proliferative activity in the junction between the leaf blade and leaf petiole of Arabidopsis. Plant Physiol 157:1151–1162
Kakar K, Zhang H, Scheres B, Dhonukshe P (2013) CLASP-mediated cortical microtubule organization guides PIN polarization axis. Nature 495:529–533
Kelliher T, Walbot V (2011) Emergence and patterning of the five cell types of the Zea mays anther locule. Dev Biol 350:32–49
Xiong Y, McCormack M, Li L, Hall Q, Xiang C, Sheen J (2013) Glucose-TOR signalling reprograms the transcriptome and activates meristems. Nature 496:181–186
Xu D, Huang W, Li Y, Wang H, Huang H, Cui X (2012) Elongator complex is critical for cell cycle progression and leaf patterning in Arabidopsis. Plant J 69:792–808
Neef AB, Luedtke NW (2011) Dynamic metabolic labeling of DNA in vivo with arabinosyl nucleosides. Proc Natl Acad Sci U S A 108:20404–20409
Cruz-Ramirez A, Diaz-Trivino S, Wachsman G, Du Y, Arteaga-Vazquez M, Zhang H, Benjamins R, Blilou I, Neef AB, Chandler V, Scheres B (2013) A SCARECROW-RETINOBLASTOMA protein network controls protective quiescence in the Arabidopsis root stem cell organizer. PLoS Biol 11:e1001724
Kerk NM, Ceserani T, Tausta SL, Sussex IM, Nelson TM (2003) Laser capture microdissection of cells from plant tissues. Plant Physiol 132:27–35
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Kazda, A., Akimcheva, S., Watson, J.M., Riha, K. (2016). Cell Proliferation Analysis Using EdU Labeling in Whole Plant and Histological Samples of Arabidopsis. In: Caillaud, MC. (eds) Plant Cell Division. Methods in Molecular Biology, vol 1370. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3142-2_13
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DOI: https://doi.org/10.1007/978-1-4939-3142-2_13
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3141-5
Online ISBN: 978-1-4939-3142-2
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