Analysis of restriction enzyme‐induced chromosomal aberrations by fluorescence in situ hybridization

Columna, Elma Abella; Giaccia, Amato J.; Evans, James; Yates, Barbara L.; Morgan, William F.

doi:10.1002/em.2850220106

Cited by 16 publications

(2 citation statements)

References 26 publications

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“…to detect DNA adducts, to measure repair in single genes with exquisite sensitivity, to use fluorescent in situ hybridization to see aberrations that go undetected by conventional methods, such as highly complex aberrations that result in pieces of a chromosome being distributed widely among the genome [t:.g., Columna et al, 1993;Hoffmann et al, 19931. we still do not understand exactly how some lesions lead to strand breaks and what happens between strands of DNA to form aberrations.…”

Section: Philosophy Of Testingmentioning

confidence: 99%

“…Despite all the recent advances that allow us to induce DNA double strand breaks with different types of ends by introducing restriction endonucleases [reviewed by Bryant and Johnston, 19931 to study effects of chemicals or heat on replication forks and chain elongation [Painter, 1977;Wong et al, 19931, to detect cell cycle perturbation by flow cytometry, to measure cyclins involved in cell cycle regulations. to detect DNA adducts, to measure repair in single genes with exquisite sensitivity, to use fluorescent in situ hybridization to see aberrations that go undetected by conventional methods, such as highly complex aberrations that result in pieces of a chromosome being distributed widely among the genome [t:.g., Columna et al, 1993;Hoffmann et al, 19931. we still do not understand exactly how some lesions lead to strand breaks and what happens between strands of DNA to form aberrations.…”

Section: Philosophy Of Testingmentioning

confidence: 99%

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Chromosome aberrations induced in vitro: Mechanisms, delayed expression, and intriguing questions

Galloway¹

1994

Environ. Mol. Mutagen.

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Chromosome aberrations, including breakage and rearrangement and numerical changes, are important in carcinogenesis, heritable mutations, embryonic loss, and developmental abnormalities. We can detect DNA reactive agents in in-vitro chromosome aberrations assays, but aberrations are also induced by chemical that do not directly interact with DNA. This article discusses briefly some important aspects of using aberrations in genetic toxicology testing but concentrates on highlights of recent research on aberrations, in particular two areas: (1) persistence through multiple cell cycles of changes that lead to chromosome aberrations, and (2) the relations among DNA synthesis inhibition, DNA damage, cell cycle regulation, and genomic instability, expressed as chromosome breakage, gene amplification, and aneuploidy. An understanding of these mechanisms not only may lead to insights into carcinogenesis but ultimately may help us to interpret results of chromosome aberration tests and to develop a rational assessment of the degree of human risk implied by a positive aberration test.

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Section: Philosophy Of Testingmentioning

confidence: 99%

Section: Philosophy Of Testingmentioning

confidence: 99%

Chromosome aberrations induced in vitro: Mechanisms, delayed expression, and intriguing questions

Galloway¹

1994

Environ. Mol. Mutagen.

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Use of multicolour chromosome painting to identify chromosomal rearrangements in human lymphocytes exposed to bleomycin: A comparison with conventional cytogenetic analysis of giemsa‐stained chromosomes

Ellard

Parry

1995

Environ and Mol Mutagen

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Exchange aberrations induced by bleomycin were identified by multicolour fluorescence in situ hybridisation (FISH) with probes for chromosomes 1, 2, and 3. The frequency and distribution of aberration types were compared to conventional metaphase analysis of Giemsa-stained chromosomes from the same human lymphocyte cultures. The total percentage of exchanges detectable by painting three pairs of chromosomes with separate colours was calculated as 40%. Giemsa staining revealed predominantly asymmetric chromosome exchanges, which are expected to comprise 50% of the total induced exchanges. Genomic exchange frequencies were, therefore, determined by multiplying the observed frequencies from FISH analysis by 2.5 and the number of asymmetric exchanges identified in Giemsa-stained slides by 2.0. By these calculations, the genomic exchange frequency calculated from chromosome painting exceeded that estimated by Giemsa-staining. This difference was due to the identification by chromosome painting of a unique class of cells in which chromosomes had undergone complex exchanges (nonreciprocal exchanges involving multiple mutual sites). The percentage of cells exhibiting exchanges was similar for both methods.

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Chromosomal aberrations induced by defined DNA double-strand breaks: The origin of achromatic lesions

Harvey

Costa

Savage

et al. 1997

Somat Cell Mol Genet

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The mechanisms of formation of chromosomal aberrations are poorly understood, despite the common use of aberrations as a measure of the genetic effects of physical and chemical agents. We have used restriction endonucleases to introduce defined DNA double-strand breaks into mammalian cells, and measured chromosomal aberration formation relative to the activity of the endonuclease. The endonucleases AluI and Sau3AI remain active for a relatively short time under simulated cellular conditions and induce achromatic lesions ('gaps') in chromatids only within the first hour or two following treatment. In contrast, the endonuclease MboI (an isoschizomer of Sau3AI) is active for an extremely long time and continues to produce chromatid gaps during the whole 12 hr sampling period. This observation strongly suggests that the aberrations classified as gaps are a manifestation of unrejoined DNA double-strand breaks. The formation of gaps may relate to the opportunities for repair of DNA breaks in relation to cell-cycle position. It is more difficult to relate the formation of structural chromatid aberrations to the endonuclease activity, although at relatively low concentrations all 3 endonucleases gave similar levels of structural aberrations.

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Analysis of restriction enzyme‐induced chromosomal aberrations by fluorescence in situ hybridization

Cited by 16 publications

References 26 publications

Chromosome aberrations induced in vitro: Mechanisms, delayed expression, and intriguing questions

Chromosome aberrations induced in vitro: Mechanisms, delayed expression, and intriguing questions

Use of multicolour chromosome painting to identify chromosomal rearrangements in human lymphocytes exposed to bleomycin: A comparison with conventional cytogenetic analysis of giemsa‐stained chromosomes

Chromosomal aberrations induced by defined DNA double-strand breaks: The origin of achromatic lesions

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