Pattern of condensation of mouse and Chinese hamster chromosomes in G2 and mitosis of 33258-Hoechst-treated cells

Marcus, Menashe; Nielsén, Karin; Goitein, Ruth; Gropp, A.

doi:10.1016/0014-4827(79)90574-3

Cited by 36 publications

(16 citation statements)

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“…The cells were treated with the fluorochrome 33258-Hoechst during the G 2 phase in order to inhibit the condensation of these normally short c-heterochromatic regions (Marcus et al 1979) and thereby increase the resolving power. As shown Fig.…”

Section: Dnase I Sensitivity Of C-heterochromatin In M Agrestis and mentioning

confidence: 99%

DNase I sensitivity in facultative and constitutive heterochromatin

et al. 1985

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In situ nick translation allows the detection of DNase I sensitive and insensitive regions in fixed mammalian mitotic chromosomes. We have determined the difference in DNase I sensitivity between the active and inactive X chromosomes in Microtus agrestis (rodent) cells, along both their euchromatic and constitutive heterochromatic regions. In addition, we analysed the DNase I sensitivity of the constitutive heterochromatic regions in mouse chromosomes. In Microtus agrestis female cells the active X chromosome is sensitive to DNase I along its euchromatic region while the inactive X chromosome is insensitive except for an early replicating region at its distal end. The late replicating constitutive heterochromatic regions, however, in both the active and inactive X chromosome are sensitive to DNase I. In mouse cells on the other hand, the constitutive heterochromatin is insensitive to DNase I both in mitotic chromosomes and interphase nuclei.

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Section: Dnase I Sensitivity Of C-heterochromatin In M Agrestis and mentioning

confidence: 99%

DNase I sensitivity in facultative and constitutive heterochromatin

et al. 1985

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“…Additionally, the ability of TOP II inhibitors to alter the rate of cell progression through G2 phase has likewise been demonstrated [11,16,24]. Hoechst 33342 is a fluorochrome that binds to the outside of DNA within the minor groove with preference for A+T rich regions of the genome [10,19,33] and it inhibits Top II-mediated DNA relaxation and decatenation reactions [33] by a process other than stabilization of the cleavable complex [13,33].…”

Section: Accumulation Of Cho Cells In G 2 Phase With Hoechst 33342mentioning

confidence: 99%

Reversible synchronization of cultured rodent and human diploid fibroblast cells in G2 phase

Crissman

Oishi²,

Tobey

1996

Methods Cell Sci

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This paper describes a three-step procedure for induction of reversible cell synchrony in the G2 phase of the cell cycle of Chinese hamster ovary (line CHO) cells and non-transformed, human skin fibroblast (HSF) cells. The CHO cells are presynchronized in early S phase by isoleucine deficiency and hydroxyurea blockades. The culture is transferred to medium supplemented with the DNA topoisomerase II inhibitor, Hoechst 33342 for 12 hours after which 95% of the cells are arrested in G2 phase. When G2 synchronized cells are transferred to Hoechst-free, complete medium, they divide as a highly synchronous cohort within 3 hours. The HSF cells are initially cultured in low serum to arrest them in Go and then transferred to complete medium containing aphidicolin to arrest them in early S phase. The culture is then transferred to aphidicolinfree, complete medium with Hoechst 33342 (0.I p.g/ml) for 10 hours after which up to 85% of the cells arrest in G2 phase. Synchronous cell division occurs 3.5 hours after transfer of cells to complete, drug-free medium. The synchrony techniques are useful for studying GriM biochemical events in mammalian cells.Abbreviations: ~x-MEM = minimum essential medium alpha medium; BCS --bovine calf serum; DMSO = dimethyl sulfoxide; EDTA = ethylenediaminetetraacetic acid; FCM = flow cytometry; PBS --phosphate buffered saline; Top II = DNA topoisomerase II

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“…On the basis of the finding that condensationdecondensation cycles relationed with the phosphorylation and de-acetylation of histones produce changes in chromosomal morphology during the mitotic cycle (Marcus et al, 1979), Goyanes and M6ndez (1981) developed a high resolution Rbanding method by cell treatment with 5-bromodeoxyuridine (5-BrdU) and 33258-Hoechst. 33258-Hoechst is a fluorescent benzimidazol compound which shows a specific binding to A-T rich regions in DNA (Kucherlapati et al, 1975;Goyanes & M6ndez, 1981) and produces a bright fluorescence in these areas (Rocchi et al, 1976); Hayman & Sharp, 1981).…”

Section: Introductionmentioning

confidence: 99%

“…The addition of 33258-Hoechst fluorochrome to a cell culture interferes partially with chromosomal condensation during G 2 phase and early mitosis (prophase and metaphase), and inhibits normal metaphase condensation. This phenomenon produces the appearance of metaphasic chromosomes which are longer than chromosomes isolated from untreated cultures (Marcus et al, 1979). Inspite of this effect, 33258-Hoechst does not interfere with cell division (Marcus et al, 1979).…”

Section: Introductionmentioning

confidence: 99%

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The R-banding pattern of the chinese hamster Don cell line

Martínez¹,

Pasantes²,

González³

et al. 1986

Genetica

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Chinese hamster cells (Don line) were treated in vivo with 5-BrdU and 33258-Hoechst fluorochrome for obtaining the partial inhibition of condensation that causes the R-banding pattern. Untreated chromosomes were stained by a standard G-banding method. Statistical measurements show significant differences in the band numbers between the two treatments. The Don cell line in the authors' laboratory presents some karyotypical differences from Don cell lines studied by other authors.

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Pattern of condensation of mouse and Chinese hamster chromosomes in G2 and mitosis of 33258-Hoechst-treated cells

Cited by 36 publications

References 28 publications

DNase I sensitivity in facultative and constitutive heterochromatin

DNase I sensitivity in facultative and constitutive heterochromatin

Reversible synchronization of cultured rodent and human diploid fibroblast cells in G2 phase

The R-banding pattern of the chinese hamster Don cell line

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