Anaphase‐telophase analysis of chromosomal damage induced by chemicals

Dulout, Fernando Noel; Olivero, Ofelia A.

doi:10.1002/em.2860060306

Cited by 15 publications

(10 citation statements)

References 20 publications

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“…Lagging fragments are clearly related with clastogenic events. Chromatin bridges are a consequence of spontaneous or induced chromosome stickiness [Olivero and Dulout, 1984] that results in chromosome fragments (clastogenic effect) or daughter chromatids with more or less DNA than normal. This cannot be considered an aneugenic event because the number of chromosomes does not change.…”

Section: Discussionmentioning

confidence: 99%

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Malsegregation as a possible mechanism of aneuploidy induction by metal salts in MRC‐5 human cells

Seoane¹,

Güerci²,

Dulout³

2002

Environ and Mol Mutagen

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Many aneugenic compounds are known to affect one or more components of the mitotic apparatus leading to an erroneous migration of chromosomes. Malsegregation occurs when a chromosome (or a chromatid) fails to migrate and remains at the metaphase plate. Nondisjunction implies the lack of dissociation between sister chromatids and the migration of both together to the same pole. The aim of the present study was to provide evidence that the aneugenic effect of some metal salts is the consequence of malsegregation at anaphase and that it is not caused by nondisjunction mechanisms. The frequencies of lagging chromosomes at anaphase-telophase of mitosis, hypoploid metaphases, and kinetochore-positive micronuclei induced by cadmium chloride, potassium dichromate, and cacodilic acid (dimethylarsinic acid) in MRC-5 human cells were compared. The data indicate that all the tested compounds are able to induce aneuploidy in MRC-5 human cells. Positive, statistically significant correlations were found when kinetochore-positive micronuclei, hypoploidy, and lagging chromosome frequencies were compared. The results suggest that malsegregation is the main mechanism involved in the induction of aneuploidy by metal salts in MRC-5 cells.

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Section: Discussionmentioning

confidence: 99%

“…Chromosome counting and anaphase-telophase alterations were scored according to the criteria of Dulout and Natarajan [1987] and Dulout and Olivero [1984], respectively. For the analysis of CREST-stained micronuclei, an Olympus BX-40 microscope with an HBO 103 W/2 mercuric light was used.…”

Section: Scoring Proceduresmentioning

confidence: 99%

Malsegregation as a possible mechanism of aneuploidy induction by metal salts in MRC‐5 human cells

Seoane¹,

Güerci²,

Dulout³

2002

Environ and Mol Mutagen

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“…Staining kinetochores in the cytokinesis-blocked micronucleus assay (Eastmond and Tucker, 1989;Lynch and Parry, 1993;Kirsch-Volders et al, 1997;Thompson and Perry, 1988) or in situ hybridization with centromere specific DNA probes, followed by immunofluorescent staining (Eastmond and Pinkel, 1990;Farooqi et al, 1993) are useful to discriminate between clastogens and aneuploidogens. Anaphase-telophase analysis, an ancilliary alternative test system (Nichols et al, 1972;Dulout and Olivero, 1984) has been used to evaluate aneugenic damage by counting lagging chromosomes (Seoane and Dulout, 1994;Seoane, 1999).…”

Section: Introductionmentioning

confidence: 99%

Mechanisms involved in the induction of aneuploidy: the significance of chromosome loss

2000

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The induction of aneuploidy by physical and chemical agents using different test systems was evaluated. The effect of X-rays, caffeine, acetaldehyde, ethanol, diethylstilbestrol, propionaldehyde, and chloral hydrate was studied by chromosome counting in Chinese hamster embryonic diploid cells. Aneugenic ability of cadmium chloride, cadmium sulfate, potassium dichromate, chromium chloride, nickel chloride, and nickel sulfate was assessed by means of anaphase-telophase analysis in Chinese hamster ovary cells. Chromosome counting in human fibroblasts (MRC-5 cell line) was employed to evaluate the effect of cacodilic acid, cadmium chloride, cadmium sulfate, and potassium dichromate. Finally, the induction of kinetochore-positive and kinetochore negative micronuclei by cadmium chloride, cadmium sulfate, potassium dichromate, chromium chloride, and nickel chloride was studied using CREST antibodies. When the effect of different agents was determined by chromosome counting, an increase of hypoploid but not of hyperploid cells was observed. Anaphase-telophase analysis showed that metal salts increased the frequency of lagging chromosomes. This finding has been confirmed by the increment of kinetochore-positive micronuclei using CREST antibodies. Therefore, chromosome loss could be considered as the main cause of induced aneuploidy.
A indução de aneuploidia por agentes físicos e químicos usando diferentes sistemas de teste foi avaliada. O efeito de raios-X, cafeína, acetaldeído, etanol, dietilestilbestrol, propionaldeído e hidrato de cloral foi estudado por contagem cromossômica em células diplóides embriônicas de hamster chinês. A habilidade aneugênica de cloreto de cádmio, sulfato de cádmio, dicromato de potássio, cloreto de crômio, cloreto de níquel e sulfato de níquel foi avaliada por meio de análise de anáfase-telófase em células de ovário de hamster chinês. A contagem cromossômica em fibroblastos humanos (linhagem celular MRC-5) foi empregada para avaliar o efeito de ácido cacodílico, cloreto de cádmio, sulfato de cádmio e dicromato de potássio. Finalmente, a indução de micronúcleos positivos e negativos para cinetocoro por cloreto de cádmio, sulfato de cádmio, dicromato de potássio, cloreto de crômio e cloreto de níquel foi estudada usando anticorpos CREST. Quando o efeito de agentes diferentes foi determinado por contagem cromossômica, observou-se um aumento de células hipoplóides mas não de hiperplóides. A análise anáfase-telófase mostrou que sais metálicos aumentaram a freqüência de cromossomos "lagging". Este achado foi confirmado pelo aumento de micronúcleos positivos para cinetocoro usando anticorpos CREST. Portanto, a perda cromossômica poderia ser considerada a principal causa de aneuploidia induzida

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“…Although accurate, FISH requires special equipment and costly reagents that are not available in most laboratories. An alternative ancillary test system is the analysis of alterations in the anaphasetelophase of cultured mitotic cells (Dulout and Olivero, 1984;Seoane and Dulout, 1994). This test is very simple and can be performed in any cytogenetic laboratory with cell culture facilities.…”

Section: Introductionmentioning

confidence: 99%

“…Lagging fragments arising from chromosome breaks and chromatin bridges are considered indicators of exchangetype aberrations. Lagging chromosomes are presumably produced by alterations at the level of the kinetochore (Dulout and Olivero, 1984;Seoane and Dulout, 1994).…”

Section: Introductionmentioning

confidence: 99%

Contribution to the validation of the anaphase-telophase test: aneugenic and clastogenic effects of cadmium sulfate, potassium dichromate and nickel chloride in Chinese hamster ovary cells

Seoane

Dulout

1999

Genet. Mol. Biol.

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There is increasing evidence that aneuploidy during mitosis may be a factor in the etiology of somatic malignancy. The analysis of alterations in anaphase-telophase of mitosis is a useful test for evaluating the aneuploidogenic and clastogenic ability of chemicals. Several metals have been found to be carcinogenic to humans and animals. However, the underlying mechanisms remain unclear. In the present study the aneugenic and clastogenic abilities of cadmium sulfate, potassium dichromate and nickel chloride were analyzed using the anaphase-telophase test. Chinese hamster ovary (CHO) cells cultured for two cycles were treated with the desired compound for 8 h before cell harvesting. The frequency of cells with chromatin bridges, lagging chromosomes and lagging chromosomal fragments was scored. The mitotic index was determined by counting the number of mitotic cells per 1,000 cells on each coverslip and was expressed as a percentage of the number of mitotic plates. Statistical comparisons were done using the "G" method. Correlation and regression analyses were performed to evaluate variations of the mitotic index. Chromium and cadmium were clastogenic and aneugenic and increased the frequencies of the three types of aberrations scored; nickel had only aneugenic activity because it increased the frequency of lagging chromosomes. These results indicate that the anaphase-telophase test is sufficiently sensitive to detect dose-response relationships that can distinguish clastogenic and/or aneugenic activities and that the results obtained using the anaphase-telophase test were similar to those obtained by chromosome counting.
As evidências de que a aneuploidia durante a mitose pode ser um fator na etiologia de malignidades somáticas estão cada vez mais fortes. A análise de alterações em anáfase-telófase da mitose é um teste útil para a avaliação da capacidade aneuploidogênica e clastogênica de substâncias químicas. Vários metais têm sido identificados como carcinogênicos para o homem e para animais. Contudo, os mecanismos de ação permanecem obscuros. No presente estudo, a capacidade aneugênica e clastogênica do sulfato de cádmio, do dicromato de potássio e do cloreto de níquel foi analisada usando o teste anáfase-telófase. Células do ovário do hamster chinês cultivadas por dois ciclos foram tratadas com o composto desejado por 8 horas antes da colheita das células. Foram quantificadas as freqüências de células com pontes de cromatina, lagging cromossomos e lagging fragmentos cromossômicos. O índice mitótico foi determinado pela contagem do número de células em mitose por 1000 células em cada lamínula e foi expresso como uma porcentagem do número de placas mitóticas. A análise estatística foi feita usando o método "G". Análises de correlação e de regressão foram realizadas para avaliar as variações do índice mitótico. O crômio e o cádmio foram clastogênicos e aneugênicos e aumentaram as freqüências dos três tipos de aberrações avaliadas; o níquel teve apenas atividade aneugênica porque ele aumentou a freqüência de lag...

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Anaphase‐telophase analysis of chromosomal damage induced by chemicals

Cited by 15 publications

References 20 publications

Malsegregation as a possible mechanism of aneuploidy induction by metal salts in MRC‐5 human cells

Malsegregation as a possible mechanism of aneuploidy induction by metal salts in MRC‐5 human cells

Mechanisms involved in the induction of aneuploidy: the significance of chromosome loss

Contribution to the validation of the anaphase-telophase test: aneugenic and clastogenic effects of cadmium sulfate, potassium dichromate and nickel chloride in Chinese hamster ovary cells

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