Baseline Sister Chromatid Exchanges Research Articles

Chromosomal Methods in Population Studies

A brief description of chromosome aberrations and sister chromatid exchange (SCE) as cytogenetic endpoints for evaluation of DNA damaging agents is presented. Problems associated with the use of cytogenetic assays as population monitors of radiation and chemical exposures are discussed. Adequate cell sample size requirements and accurate assessment of cummulative exposure effects with increasing age are stressed as important considerations for reliable cost-benefit analysis of population studies involving low level exposures. Examples of population studies using SCE as an indicator of specific chemical exposures are cited, and factors contributing to variations in control baseline SCE levels are discussed. Possible implications of population cytogenetic data on general public health are suggested.

Chromosomal methods in population studies.

A brief description of chromosome aberrations and sister chromatid exchange (SCE) as cytogenetic endpoints for evaluation of DNA damaging agents is presented. Problems associated with the use of cytogenetic assays as population monitors of radiation and chemical exposures are discussed. Adequate cell sample size requirements and accurate assessment of cummulative exposure effects with increasing age are stressed as important considerations for reliable cost-benefit analysis of population studies involving low level exposures. Examples of population studies using SCE as an indicator of specific chemical exposures are cited, and factors contributing to variations in control baseline SCE levels are discussed. Possible implications of population cytogenetic data on general public health are suggested.

Correlations between sister chromatid exchange frequencies and replicon sizes: A model for the mechanism of SCE production

Baseline sister chromatid exchanges (SCEs) increase as a function of average replicon size in a variety of human and Chinese hamster cell lines. This observation is the basis for a model in which SCEs are generated by errors in the unravelling of daughter double helices by topo isomerases. These errors cause exchanges behind the replication fork, not at the fork as several current models assume. This model also provides an explanation for SCEs induced when residual DNA damage that has been replicated interferes with the normal processes of unravelling the daughter strands.

Cytogenetics of Werner's syndrome cultured skin fibroblasts: variegated translocation mosaicism.

Skin fibroblast-like (FL) cells from patients with Werner's syndrome (adult progeria) regularly demonstrate frequent pseudodiploidy involving variable structural rearrangements that are clonal: variegated translocation mosaicism (VTM). Ninety-two percent of 1,538 metaphases from 29 independent strains derived from five patients with Werner's syndrome demonstrated this cytogenetic abnormality. In contrast, only eight (8.4%) of 95 non-Werner's syndrome FL cell cultures demonstrated VTM: seven with low-grade VTM (approximately 5% of 300 metaphases), and one with VTM affecting 90-100% of metaphases. Unlike the cytogenetic abnormalities observed in the terminal stages of normal FL cell cultures, VTM occurs throughout the entire lifespan of Werner's syndrome cultures. Ten of the identifiable break points in 1,005 banded metaphases accounted for 27% of all definable rearrangements. Baseline sister chromatid exchanges were not increased. Cocultivation of Werner's syndrome and normal strains did not induce VTM in the normal strain. The relationship between VTM and the reduced growth potential of Werner's syndrome FL cells is not yet understood, nor is the relationship between these in vitro abnormalities and the presumptive single gene defect that causes the progeroid clinical manifestations of Werner's syndrome.

Characterization of a rat lymphocyte culture system for assessing sister chromatid exchange after in vivo exposure to genotoxic agents.

Lymphocyte culture systems have the major advantage of permitting the analysis of in vivo cytogenetic damage with minimal injury to the animal under study. This paper describes a rat lymphocyte culture system designed for the study of sister chromatid exchange (SCE) induced by in vivo exposure to genotoxic agents. A standard protocol was established in which 1 to 2 ml of blood are removed from rats by cardiac puncture, washed three times with phosphate-buffered saline (pH 7.4), and grown in RPMI 1640. 5-Bromodeoxyuridine (BrdU) (1.0 microM) is added after 24 hr, and cells are harvested after 56 hr of culture. Critical steps for successful blast transformation include washing the blood in buffered saline and the adding of 2.0 to 4.0 microgram phytohemagglutinin/ml to the culture medium. The use of low concentrations of BrdU (less than or equal to 5.0 microM) is recommended to maintain low baseline SCE levels and to avoid cytotoxicity. The mutagenic carcinogen, ethyl methanesulfonate (EMS), was used as a positive control agent and at a dose of 300 mg/kg caused a fourfold increase in SCE frequency. Twenty-eight days after EMS administration (30, 100, 300 mg/kg), lymphocytes from treated animals still displayed SCE levels at least 50% above baseline. This system provides a reliable means of investigating chemically induced SCE in the rat.

Effects of a tumor promoter and an anti-promoter on spontaneous and UV-induced 6-thioguanine-resistant mutations and sister-chromatid exchanges in V79 Chinese hamster cells

The effects of a tumor promoter 12- O-tetradecanoylphorbol-13-acetate (TPA) and/or an anti-promotor antipain (protease inhibitor) on spontaneous and ultraviolet-induced sister-chromatid exchanges (SCEs) and 6-thioguanine- resistant (6TG r) recessive mutations were examined in V79 Chinese hamster cells in culture. TPA and/or antipain neither significantly altered base-line and UV-induced immediate SCE frequencies, nor decreased the level of delayed SCEs which persisted 6–7 days after irradiation. TPA and/or antipain appeared to enhance the recovery of UV-induced 6TG r colonies at the plateau expression phase despite non-mutagenicity by themselves and unaltered metabolic co- operation. Thus, the results conceivably imply that the 6TG r-recessive mutation expression, but not fixation, can be modulated at the cell level by the TPA and/or antipain. Our results, together with the recent results of Loveday and Latt, may argue against the notion that TPA enhances the antipain-suppressible SCEs as an index of mitotic recombination in relevance with a tumor-promotion mechanism.

Tissue‐specific induction of sister chromatid exchanges by ethyl carbamate in mice

Sister chromatid exchange (SCE) techniques were used to analyze the genetic effects of ethyl carbamate (urethane) in cultured mouse-bone marrow cells, and in several different mouse tissues in vivo. Ethyl carbamate concentrations up to 5.0 mg/ml were ineffective in causing a significant elevation of SCE in vitro. After in vivo drug administration, bone marrow, liver and spermatogonial cells all revealed significant dose-related increases in SCE. Baseline and relative incremental levels of SCE were somatic vs germ tissue-specific. Regenerating liver cells exhibited significantly greater absolute SCE values than all other tissues examined. Marrow cells revealed intermediate values, while germ cells were the least sensitive in SCE responsiveness. Spermatogonia required a fourfold higher dose, over that effective in somatic tissues, to promote an approximate doubling of the baseline SCE level. In vivo SCE analysis affords sensitive risk assessments for different tissues. Thus, this approach; should be generally useful for studying compounds with questionable mutagenic potential, and/or those exerting target organ specificities of related biological activity (eg, toxicity, carcinogenesis).

Spontaneous and mitomycin-C induced sister-chromatid exchanges: Comparison of in vivo and in vitro systems

Frequencies of sister-chromatid exchanges (SCE) were measured in vitro in mouse fibroblasts and in vivo in mouse bone-marrow cells. SCE levels in these cell systems were measured in response to varying concentrations of bromodeoxyuridine (BrdU) and mitomycin-C (MMC). Although BrdU was found to induce SCE in both cellular systems, baseline SCE levels were 2- to 3-fold higher in vitro than in vivo. SCE induction was found to be a linear function of MMC concentration in vivo and in vitro; however the slope of the in vivo curve was 5-fold higher. The interaction of BrdU substituted DNA and MMC was examined by administering a fixed dose of MMC with increasing concentrations of BrdU. The induced SCE frequencies appeared to be additive. In addition to measuring drug-induced SCE, the BrdU differential staining technique allows concomitant measurement of the inhibition of cellular replication by the test drugs.

The effect of aging on sister chromatid exchange

The advent of the bromodeoxyuridine(BrdU)-differential staining techniques has greatly facilitated the detection of sister chromatid exchanges (SCE). These SCE have been demonstrated to be an accurate reflection of DNA damage both in vitro in cultured cells and in vivo in mouse and rat bone marrow and spleen cells. In this review, we examine the effect of cellular aging on both baseline and mutagen-induced SCE levels. In all systems examined, aging did not appear to significantly affect the baseline levels of SCE. However, in human fibroblast cultures we have found a significant decrease in the levels of mutagen-induced SCE as a function of both in vitro passage level ( in vitro aging) and the age of the cell culture donor ( in vivo aging). In addition we have found a similar decrease in mutagen-induced SCE levels in both mouse and rat bone marrow cells and mouse spleen cells where examinations were performed entirely in vivo. Diminished mutagen-induced SCE levels were obtained with a wide variety of agents including mitomycin-C, cyclophosphamide, adriamycin, ethyl methanesulfonate and N-acetyl-2-acetoxyaminofluorene. These decreased SCE levels were accompanied by increased frequencies of chromosomal aberrations in the older cell populations. If SCE represents a form of DNA repair as has been suggested by several investigators, our findings would indicate impaired DNA repair occurring in old cells.

Aging and sister chromatid exchange: II. The effect of the in vitro passage level of human fetal lung fibroblasts on baseline and mutagen-induced sister chromatid exchange frequencies

The frequencies of baseline and mutagen-induced sister chromatid exchanges (SCE) were examined in human fetal lung fibroblasts (IMR-90, WI38) as a function of in vitro serial passage (in vitro aging). Although baseline SCE levels remained relatively constant throughout the in vitro lifespan of these cell cultures, a significant decline was observed at middle and late passage in the levels of SCE induced by mitomycin-C, ethyl methane-sulfonate and N-acetoxy-2-acetylaminofluorene. These findings indicate that cellular aging results in an altered response to certain types of induced DNA damage.