Mitosis‐to‐G1 phase transition of HTC cells obtained with nocodazole (R 17934; NSC 238159)

Müller, Robert N.; Haverbeke, Y. Van; Blave, A.; Aguilera, Andrés; Michel, Nico; Miller-Faurès, A.; Miller, A. O. A.

doi:10.1016/0014-5793(80)81121-5

Cited by 3 publications

(3 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The degree of synchrony during the S phase can be increased markedly by addition of Aphidicolin early in GI, allowing sufficient time (12 hr) for the cells to accumulate at the Gl/S border. Nocodazole and Aphidicolin alone have been successfully used to accumulate cells of various origin in metaphase or at the G,/S interphase, respectively (Muller et al, 1980;Zieve et al, 1980;Pedrali-Noy et al, 1980). Our results show that they can be used in combination to obtain reasonably good synchrony throughout the phases of the cell cycle.…”

Section: Discussionmentioning

confidence: 71%

Laser Flow Cytofluorometric Analysis of Htc Cells Synchronized With Hydroxyurea, Nocodazole and Aphidicolin

et al. 1981

Self Cite

View full text Add to dashboard Cite

The technique of laser flow cytofluorometry has been used to monitor the arrival in G1 and the subsequent progression through the cell cycle of HTC cells accumulated in metaphase with colcemid alone or after treatment with hydroxyurea and Nocodazole. Under the experimental conditions used in this study, the latter procedure gives much better results, avoiding in particular the extensive formation of micronucleated cells. Aphidicolin, an inhibitor of DNA polymerase, in combination with Nocodazole, provides a useful method to tightly synchronize these cells at the G1/S border.

show abstract

Section: Discussionmentioning

confidence: 71%

Laser Flow Cytofluorometric Analysis of Htc Cells Synchronized With Hydroxyurea, Nocodazole and Aphidicolin

et al. 1981

Self Cite

View full text Add to dashboard Cite

show abstract

“…Since nocodazole presum-ably blocks cells in M phase by preventing microtubule polymerization, it may be possible that some permanent damage was sustained in the cell to prevent or reduce subsequent chromosome decondensation as the cells moved from M to GI phase. This would be an alternate explanation for the lack of change in proton NMR relaxation times between M and G 1 phase in the study by Muller (2). Since colcemid or N20 gas was used in the HeLa experiments ( 1 ), it is necessary to assume that neither of these treatments would have caused effects similar to those of nocodazole.…”

Section: Comparison With Similar Studiesmentioning

confidence: 93%

“…The explanation put forward by Muller et al (2) for the lack of T1 dependency on the cell cycle was that this T 1 variation throughout the cell cycle is cell-line dependent. Since a rather high centrifugation force ( 3000g) was used by Wheatley et al in their cell pellet preparations, it may be speculated that this may have caused the "flat" water content and T 1 for cells across the cell cycle.…”

Section: Comparison With Similar Studiesmentioning

confidence: 97%

The dependence of proton longitudinal and transvese relaxation times on cell‐cycle phase: Mouse MCA‐transformed 10T½ 5 TCL‐15 cells

Belfi

Medendorp²,

Ngo

1991

Magnetic Resonance in Med

View full text Add to dashboard Cite

Attempts to determine proton NMR longitudinal relaxation times (T1) as a function of cell-cycle stage using cells synchronized by chemical methods have yielded conflicting results (P. T. Beall, C. F. Hazlewood, and P. N. Rao, Science 192, 904 (1976); R. N. Muller et al., FEBS Lett. 114, 231 (1980); D. N. Wheatley, et al., J. Cell Sci. 88, 13 (1987]. This has raised the question whether a true dependence of T1 on cell-cycle phase exists. In the present study, the centrifugal elutriation technique was used to obtain relatively pure, synchronized cell populations of TCL-15 cells (a methylcholanthrene-transformed line of mouse 10T1/2 cells) for measurement of proton NMR relaxation rates. This technique provides a means to procure synchronized cell populations without the use of chemical agents as in the above-cited investigations and therefore avoid possible effects caused by the chemical agents of the NMR relaxation processes. Both T1 and the transverse relaxation time, T2, of water protons in synchronized-cell pellets obtained in this study, exhibited a dependence on cell-cycle phase at least for the first half of the cell cycle (G1 to S). Cells in G1 phase exhibited quantitatively higher T1 and T2 relaxation times compared to those measured for cells in mid S phase. Such changes were found to correlate with changes in water content. The distribution of cell-cycle phases of each cell population was determined by the DNA histogram using flow cytometric methods. Possible relaxation mechanisms which may contribute to the cell-cycle-specific phenomena of the intracellular T1 and T2 times are discussed.

show abstract

Hydration, volume changes and nuclear magnetic resonance proton relaxation times of HeLa S-3 cells in M-phase and the subsequent cell cycle

Wheatley¹,

Inglis²,

Foster

et al. 1987

Journal of Cell Science

View full text Add to dashboard Cite

Most mitotic HeLa cells divided into two daughter cells with half the volume of the parent; no additional reduction in volume was detectable during late telophase or the early part of G1. Synchronous growth throughout the next generation cycle was exponential, without an additional and sudden rise in volume being detectable as cells entered M-phase. Since the wet weight/dry weight ratio and the protein per unit mass of cells remained constant throughout the cycle, measurements based on volume changes accurately reflected growth parameters and cell water content. Nuclear magnetic resonance (n.m.r.) t1 (spin-lattice) proton relaxation times for intracellular water for M-phase cells measured at both 32 MHz (25 degrees C) and 80 MHz (37 degrees C) in a Bruker spectrometer had the same values as normal interphase cells. Furthermore, cells arrested in metaphase by nitrous oxide or alkaloids gave the same t1 values. Artificial manipulation of cell hydration by adjustment of the tonicity of the external medium led to t1 values that correlated well with the level of intracellular water. On this basis, a 40% increase in hydration raised t1 values by a factor of 1.29 at 32 MHz and 1.5 at 80 MHz. This is considerably less than the increase by a factor of 1.9 in t1 time reported with the 40% rise in cell water accompanying mitosis, measured at 30 MHz and 25 degrees C under isotonic conditions by others. The protein-synthesizing ability of hydrated cells was reduced to only half the normal level after a doubling of intracellular water. The data from these several different analyses, taken together, strongly indicate that the water content of mitotic cells is very similar to that of interphase cells, and that certain features unique to the mitotic phase of the cell cycle cannot be ascribed to an elevated (free) water content.

show abstract

Mitosis‐to‐G1 phase transition of HTC cells obtained with nocodazole (R 17934; NSC 238159)

Cited by 3 publications

References 5 publications

Laser Flow Cytofluorometric Analysis of Htc Cells Synchronized With Hydroxyurea, Nocodazole and Aphidicolin

Laser Flow Cytofluorometric Analysis of Htc Cells Synchronized With Hydroxyurea, Nocodazole and Aphidicolin

The dependence of proton longitudinal and transvese relaxation times on cell‐cycle phase: Mouse MCA‐transformed 10T½ 5 TCL‐15 cells

Hydration, volume changes and nuclear magnetic resonance proton relaxation times of HeLa S-3 cells in M-phase and the subsequent cell cycle

Contact Info

Product

Resources

About