Subphases of DNA Replication in <i>Drosophila</i> Cells

Rehák, M.; Csuka, I; Szepessy, Edit; Bánfalvi, Gáspár

doi:10.1089/104454900750019362

Cited by 20 publications

(5 citation statements)

References 15 publications

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“…22 These C-values indicate that nuclear growth takes place in several subphases coinciding with the path of chromosome replication referred to as subphases of DNA replication. 22,23 Conclusion Initial stages of apoptosis were detected earlier after 0.5 µM CdCl 2 treatment in CHO cells by a highly sensitive and quantitative method based on random primed oligonucleotide synthesis (ROPS assay). 10 In this paper we describe that apoptotic changes caused by 1 µM cadmium chloride treatment are similar to those observed by others.…”

Section: Discussionmentioning

confidence: 99%

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Cadmium induced apoptotic changes in chromatin structure and subphases of nuclear growth during the cell cycle in CHO cells

et al. 2005

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CHO cells were grown in the presence of 1 mu M CdCl(2) and subjected to ATP-dependent replicative DNA synthesis after permeabilization. By decreasing the density of the cell culture replicative DNA synthesis was diminishing. At higher than 2 x 10(6) cell/ml concentration Cd had virtually no effect on the rate of DNA replication. Growth at higher cell concentrations could be suppressed by increasing Cd concentration. After Cd treatment cells were synchronized by counterflow centrifugal elutriation. Cadmium toxicity on cell growth in early and mid S phase led to the accumulation of enlarged cells in late S phase. Flow cytometry showed increased cellular and nuclear sizes after Cd treatment. As the cells progressed through the S phase, 11 subpopulations of nuclear sizes were distinguished. Apoptotic chromatin changes were visualized by fluorescent microscopy in a cell cycle dependent manner. In the control untreated cells the main transitory forms of chromatin corresponded to those we have published earlier (veil-like, supercoiled chromatin, fibrous, ribboned structures, chromatin strings, elongated prechromosomes, precondensed chromosomes). Cadmium treatment caused: (a) the absence of decondensed veil-like structures and premature chromatin condensation in the form of apoptotic bodies in early S phase (2.2-2.4 average C-value), (b) the absence of fibrous structures, the lack of supercoiled chromatin, the appearance of uncoiled ribboned chromatin and perichromatin semicircles, in early mid S phase (2.5-2.9 C), (c) the presence of perichromatin fibrils and chromatin bodies in mid S phase (2.9-3.2 C), (d) early intra-nuclear inclusions, elongated forms of premature chromosomes, the extrusion and rupture of nuclear membrane later in mid S phase (3.3-3.4 C), (e) the exclusion of chromatin bodies and the formation of clusters of large-sized perichromatin granules in late S phase (3.5-3.8 C) and (f) large extensive disruptions and holes in the nuclear membrane and the clumping of incompletely folded chromosomes (3.8-4. C).

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Section: Discussionmentioning

confidence: 99%

“…The peaks of nuclear growth expressed in C-values coincide with the replication checkpoints 22 which are likely to correspond to the subphases of chromosome replication. 23 …”

Section: Discussionmentioning

confidence: 99%

Cadmium induced apoptotic changes in chromatin structure and subphases of nuclear growth during the cell cycle in CHO cells

et al. 2005

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“…Starting from 94 h (20.2%), PLM steadily increased and peaked at 102 h (40.4%), thereafter the percent of labeled mitoses decreased at 106 to 19.6%. This wavy pattern of the curve is probably related to the phenomenon of DNA synthesis discontinuity in eukaryotic cells [3,4,6,7]. The fact that PLM was minimum from 38 to 94 h (on average, it did not exceed 10%) attests to the beginning of a new wave of PLM related to S period of the second mitotic cycle.…”

Section: Resultsmentioning

confidence: 99%

Effect of Irradiation on DNA Synthetic Period of the Mitotic Cycle in Cells

et al. 2012

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Kinetics of DNA synthesis in mitotic cycle of mouse corneal epithelial cells after single γ-irradiation (4 Gy) at the end of S period was studied by the method of radioautography. Normally, S period of corneal epithelial cells consists of several stages separated by intervals without DNA synthesis. The estimated mean duration of the first (S(1)) and second (S(2)) phases of S period was 16 and 10 h, respectively, and the interval between them was 7 h. Single irradiation at the end of S period changed the duration of mitotic cycle periods: S(2) phase became 2.2-longer than S(1) phase and the duration of g(1) period decreased, because the time for reparation in irradiated cell increases at the expense of g(1) period. Shortening of g(1) period is a factor promoting the appearance of transformed cells.

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“…The JE-6B lowvolume elutriator rotor is excluded from the JE-5.0 elutriation system. The protocol presented here has been developed to enable cell cycle studies in a variety of cell lines (25)(26)(27). The elutriation of 1-2 × 10 8 cells in a JE-6B rotor and a J2-21 centrifuge was used most frequently in our experiments (28).…”

Section: Hardwarementioning

confidence: 99%

Synchronization of Mammalian Cells and Nuclei by Centrifugal Elutriation

Bánfalvi¹

2016

Methods in Molecular Biology

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Synchronized populations of large numbers of cells can be obtained by centrifugal elutriation on the basis of sedimentation properties of small round particles, with minimal perturbation of cellular functions. The physical characteristics of cell size and sedimentation velocity are operative in the technique of centrifugal elutriation also known as counterstreaming centrifugation. The elutriator is an advanced device for increasing the sedimentation rate to yield enhanced resolution of cell separation. A random population of cells is introduced into the elutriation chamber of an elutriator rotor running in a specially designed centrifuge. By increasing step-by-step the flow rate of the elutriation fluid, successive populations of relatively homogeneous cell size can be removed from the elutriation chamber and used as synchronized subpopulations. For cell synchronization by centrifugal elutriation, early log S phase cell populations are most suitable where most of the cells are in G1 and S phase (>80 %). Apoptotic cells can be found in the early elutriation fractions belonging to the sub-Go window. Protocols for the synchronization of nuclei of murine pre-B cells and high-resolution centrifugal elutriation of CHO cells are given. The verification of purity and cell cycle positions of cells in elutriated fractions includes the measurement of DNA synthesis by [H]-thymidine incorporation and DNA content by propidium iodide flow cytometry.

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Subphases of DNA Replication in Drosophila Cells

Cited by 20 publications

References 15 publications

Cadmium induced apoptotic changes in chromatin structure and subphases of nuclear growth during the cell cycle in CHO cells

Cadmium induced apoptotic changes in chromatin structure and subphases of nuclear growth during the cell cycle in CHO cells

Effect of Irradiation on DNA Synthetic Period of the Mitotic Cycle in Cells

Synchronization of Mammalian Cells and Nuclei by Centrifugal Elutriation

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