Abstract
It has been predicted that nocodazole-inhibited cells are not synchronized because nocodazole-arrested cells with a G2-phase amount of DNA would not have a narrow cell-size range reflecting the cell size of some specific, presumably G2-phase, cell-cycle age. Size measurements of nocodazole-inhibited cells now fully confirm this prediction. Further, release from nocodazole inhibition does not produce cells that move through the cell cycle mimicking the passage of normal unperturbed cells through the cell cycle. Nocodazole, an archetypal whole-culture synchronization method, can inhibit growth to produce cells with a G2-phase amount of DNA, but such cells are not synchronized. Cells produced by a selective (i.e., non-whole-culture) method not only have a specific DNA content, but also have a narrow size distribution. The current view of cell-cycle control that is based on methods that are not suitable for cell-cycle analysis must therefore be reconsidered when results are based on whole-culture synchronization.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Avisar D, Segal M, Sneh B, Zilberstein A (2005) Cell-cycle-dependent resistance to Bacillus thuringiensis Cry1C toxin in Sf9 cells. J Cell Sci 118:3163–3171
Cooper S (1991) Bacterial growth and division. Academic Press, San Diego
Cooper S (1998) Mammalian cells are not synchronized in G1-phase by starvation or inhibition: considerations of the fundamental concept of G1-phase synchronization. Cell Prolif 31:9–16
Cooper S (2002) Reappraisal of G1-phase arrest and synchronization by lovastatin. Cell Biol Int 26:715–727
Cooper S (2003) Rethinking synchronization of mammalian cells for cell-cycle analysis. Cell Mol Life Sci 6:1099–1106
Cooper S (2004a) Is whole-culture synchronization biology’s "perpetual motion machine"? Trends Biotech 26:266–269
Cooper S (2004b) Whole-culture synchronization can not, and does not, synchronize cells. Trends Biotech 22:274–276
Cooper S, Shedden K (2003) Microarray analysis of gene expression during the cell cycle. Cell Chromosom 2:1–12
Eward KL, Van Ert MN, Thornton M, Helmstetter CE (2004) Cyclin mRNA stability does not vary during the cell cycle. Cell Cycle 3:1057–1061
Gong J, Traganos F, Darzynkiewicz Z (1995) Growth imbalance and altered expression of cyclins B1, A, E, and D3 in MOLT-4 cells synchronized in the cell cycle by inhibitors of DNA replication. Cell Growth Differ 6:1485–1493
Harper JV (2005a) Synchronization of cell populations in G1/S and G2/M phases of the cell cycle. Methods Mol Biol 296:157–166
Harper JV (2005b) Synchronization of cell populations in G1/S and G2/M phases of the cell cycle. Methods Mol Biol 296:157–166
Helmstetter CE (1991) Description of a baby machine for Saccharomyces cerevisiae. New Biol 3:1089–1096
Helmstetter C, Cummings D (1963) Bacterial synchronization by selection of cells at division. Proc Natl Acad Sci USA 50:767–774
Helmstetter C, Cummings D (1964) An improved method for the selection of bacterial cells at division. Biochim Biophys Acta 82:608–610
Helmstetter CE, Thornton M, Romero A, Eward KL (2003) Synchrony in human, mouse and bacterial cell cultures—a comparison. Cell Cycle 2:42–45
Jansen-Durr P, Meichle A, Steiner P, Pagano M, Finke K, Botz J, Wessbecher J, Draetta G, Eilers M (1993) Differential modulation of cyclin gene expression by MYC. Proc Natl Acad Sci USA 90:3685–3689
Keyomarsi K, Sandoval L, Band V, Pardee AB (1991) Synchronization of tumor and normal cells from G1 to multiple cell cycles by lovastatin. Cancer Res 51:3602–3609
Kung AL, Sherwood SW, Schimke RT (1990) Cell line-specific differences in the control of cell cycle progression in the absence of mitosis. Proc Natl Acad Sci USA 87:9553–9557
Ludlow JW, Glendening CL, Livingston DM, DeCaprio JA (1993) Specific enzymatic dephosphorylation of the retinoblastoma protein. Mol Cell Biol 13:367–372
Ouyang B, Lan Z, Meadows J, Pan H, Fukasawa K, Li W, Dai W (1998) Human Bub1: a putative spindle checkpoint kinase closely linked to cell proliferation. Cell Growth Differ 9:877–885
Summers MK, Bothos J, Halazonetis TD (2005) The CHFR mitotic checkpoint protein delays cell cycle progression by excluding cyclin B1 from the nucleus. Oncogene 24:2589–2598
Thornton M, Eward KL, Helmstetter CE (2002) Production of minimally disturbed synchronous cultures of hematopoietic cells. Biotechniques 32:1098–1105
Whitfield M, Sherlock G, Saldanha A, Murray JI, Ball CA, Alexnder KE, Matese JC, Perou CM, Hurt MM, Brown PO, Botstein D (2002) Identification of genes periodically expressed in the human cell cycle and their expression in tumors. Mol Biol Cell 13:1977–2000
Author information
Authors and Affiliations
Corresponding author
Additional information
This work was supported by the National Science Foundation (grant MCB–0323346) and (in part) by the National Institutes of Health (University of Michigan’s Cancer Center, support grant 5 P30 CA46592). G.I., M.T., and P. B. are associated with the Undergraduate Research Opportunity Program of the University of Michigan, which also supported this research.
Rights and permissions
About this article
Cite this article
Cooper, S., Iyer, G., Tarquini, M. et al. Nocodazole does not synchronize cells: implications for cell-cycle control and whole-culture synchronization. Cell Tissue Res 324, 237–242 (2006). https://doi.org/10.1007/s00441-005-0118-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00441-005-0118-8