The spirmatid micronucleus test with the dissection technique detects the germ cell mutagenicity of acrylamide in rat meiotic cells

Lähdetie, Jaana; Suutari, A.; Sjöblom, Tiina

doi:10.1016/0027-5107(94)90100-7

Cited by 30 publications

(11 citation statements)

References 28 publications

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“…AA has also proven to be genotoxic to germ cells [Dearfield et al, 1995]. AA induced micronuclei in mice spermatids, and heritable chromosome translocations and specific locus mutations in postomeiotic sperm and spermatogonia [Lahdetie et al, 1994;Xiao and Tates, 1994]. AA also elevated the frequency of dominant lethal mutations probably accompanying with chromosome aberrations leading to death of embryo [Shelby et al, 1987;Adler et al, 1994].…”

Section: Discussionmentioning

confidence: 96%

Genotoxicity of acrylamide in vitro: Acrylamide is not metabolically activated in standard in vitro systems

Naoki

Yasui

Oda

et al. 2011

Environ and Mol Mutagen

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The recent finding that acrylamide (AA), a genotoxic rodent carcinogen, is formed during the frying or baking of a variety of foods raises human health concerns. AA is known to be metabolized by cytochrome P450 2E1 (CYP2E1) to glycidamide (GA), which is responsible for AA's in vivo genotoxicity and probable carcinogenicity. In in-vitro mammalian cell tests, however, AA genotoxicity is not enhanced by rat liver S9 or a human liver microsomal fraction. In an attempt to demonstrate the in vitro expression of AA genotoxicity, we employed Salmonella strains and human cell lines that overexpress human CYP2E1. In the umu test, however, AA was not genotoxic in the CYP2E1-expressing Salmonella strain or its parental strain. Moreover, a transgenic human lymphoblastoid cell line overexpressing CYP2E1 (h2E1v2) and its parental cell line (AHH-1) both showed equally weak cytotoxic and genotoxic responses to high (>1 mM) AA concentrations. The DNA adduct N7-GA-Gua, which is detected in liver following AA treatment in vivo, was not substantially formed in the in vitro system. These results indicate that AA was not metabolically activated to GA in vitro. Thus, AA is not relevantly genotoxic in vitro, although its in vivo genotoxicity was clearly demonstrated.

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Section: Discussionmentioning

confidence: 96%

Genotoxicity of acrylamide in vitro: Acrylamide is not metabolically activated in standard in vitro systems

Naoki

Yasui

Oda

et al. 2011

Environ and Mol Mutagen

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“…In addition, when the suspension method is used in conjunction with immunofluorescence or molecular cytogenetics to detect centromere-carrying micronuclei, no acrosome staining can be performed and, therefore, round spermatids scored for micronucleus frequencies do not represent a narrow temporal window after the completion of the meiotic divisions. For these reasons, a better detection of clastogenic effects has been achieved by using more than one sampling time or a multiple treatment schedule [ Lähdetie et al, 1994b;Russo et al, 1994;Xiao and Tates, 1994]. Similarly, slight interlaboratory differences in the choice of time intervals from exposure to spermatid preparation may be the cause of some controversial results [Russo and Levis, 1992a,b;Allen et al, 1994;Nutley et al, 1996].…”

Section: Spermatidsmentioning

confidence: 99%

In vivo rodent erythrocyte micronucleus assay. II. Some aspects of protocol design including repeated treatments, integration with toxicity testing, and automated scoring

Hayashi¹,

MacGregor²,

Gatehouse³

et al. 2000

Environ. Mol. Mutagen.

236

107

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An expert working group on the in vivo micronucleus assay, formed as part of the International Workshop on Genotoxicity Test Procedures (IWGTP), discussed protocols for the conduct of established and proposed micronucleus assays at a meeting held March 25–26, 1999 in Washington, DC, in conjunction with the annual meeting of the Environmental Mutagen Society. The working group reached consensus on a number issues, including: (1) protocols using repeated dosing in mice and rats; (2) integration of the (rodent erythrocyte) micronucleus assay into general toxicology studies; (3) the possible omission of concurrently‐treated positive control animals from the assay; (4) automation of micronucleus scoring by flow cytometry or image analysis; (5) criteria for regulatory acceptance; (6) detection of aneuploidy induction in the micronucleus assay; and (7) micronucleus assays in tissues (germ cells, other organs, neonatal tissue) other than bone marrow. This report summarizes the discussions and recommendations of this working group. In the classic rodent erythrocyte assay, treatment schedules using repeated dosing of mice or rats, and integration of assays using such schedules into short‐term toxicology studies, were considered acceptable as long as certain study criteria were met. When the micronucleus assay is integrated into ongoing toxicology studies, relatively short‐term repeated‐dose studies should be used preferentially because there is not yet sufficient data to demonstrate that conservative dose selection in longer term studies (longer than 1 month) does not reduce the sensitivity of the assay. Additional validation data are needed to resolve this point. In studies with mice, either bone marrow or blood was considered acceptable as the tissue for assessing micronucleus induction, provided that the absence of spleen function has been verified in the animal strains used. In studies with rats, the principal endpoint should be the frequency of micronucleated immature erythrocytes in bone marrow, although scoring of peripheral blood samples gives important supplementary data about the time course of micronucleus induction. When dose concentration and stability are verified appropriately, concurrent treatment with a positive control agent is not necessary. Control of staining and scoring procedures can be obtained by including appropriate reference samples that have been obtained from a separate experiment. For studies in rats or mice, treatment/sampling regimens should include treatment at intervals of no more than 24 hr (unless the test article has a half‐life of more than 24 hr) with sampling of bone marrow or blood, respectively, within 24 or 40 hr after the last treatment. The use of a DNA specific stain is recommended for the identification of micronuclei, especially for studies in the rat. In the case of a negative assay result with a non‐toxic test article, it is desirable that systemic exposure to the test article is demonstrated. The group concluded that successful application of automated scoring by...

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“…Acrylamide is a rodent carcinogen and probable human carcinogen [1,2], and a mutagen in mammalian somatic cells in vitro and in vivo, particularly in assays that detect induction of chromosomal damage [1,[3][4][5][6][7]. In addition, acrylamide is a mammalian germ cell mutagen, inducing high frequencies of dominant lethal mutations (generally associated with chromosomal alterations that result in death of embryos around the time of implantation) [8][9][10][11][12], heritable chromosomal translocations [12][13][14], and specific locus mutations [15] in postmeiotic sperm and spermatogonial stem cells [9] of male mice.…”

Section: Introductionmentioning

confidence: 99%

Comparison of Germ Cell Mutagenicity in Male CYP2E1-Null and Wild-Type Mice Treated with Acrylamide: Evidence Supporting a Glycidamide-Mediated Effect

Ghanayem

Witt

El-Hadri

et al. 2005

Biology of Reproduction

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Acrylamide is an animal carcinogen and probable human carcinogen present in appreciable amounts in heated carbohydrate-rich foodstuffs. It is also a germ cell mutagen, inducing dominant lethal mutations and heritable chromosomal translocations in postmeiotic sperm of treated mice. Acrylamide's affinity for male germ cells has sometimes been overlooked in assessing its toxicity and defining human health risks. Previous investigations of acrylamide's germ cell activity in mice showed stronger effects after repeated administration of low doses compared with a single high dose, suggesting the possible involvement of a stable metabolite. A key oxidative metabolite of acrylamide is the epoxide glycidamide, generated by cytochrome P4502E1 (CYP2E1). To explore the role of CYP2E1 metabolism in the germ cell mutagenicity of acrylamide, CYP2E1-null and wild-type male mice were treated by intraperitoneal injection with 0, 12.5, 25, or 50 mg acrylamide (5 ml saline)(-1) kg(-1) day(-1) for 5 consecutive days. At defined times after exposure, males were mated to untreated B6C3F1 females. Females were killed in late gestation and uterine contents were examined. Dose-related increases in resorption moles (chromosomally aberrant embryos) and decreases in the numbers of pregnant females and the proportion of living fetuses were seen in females mated to acrylamide-treated wild-type mice. No changes in any fertility parameters were seen in females mated to acrylamide-treated CYP2E1-null mice. Our results constitute the first unequivocal demonstration that acrylamide-induced germ cell mutations in male mice require CYP2E1-mediated epoxidation of acrylamide. Thus, CYP2E1 polymorphisms in human populations, resulting in variable enzyme metabolic activities, may produce differential susceptibilities to acrylamide toxicities.

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The spirmatid micronucleus test with the dissection technique detects the germ cell mutagenicity of acrylamide in rat meiotic cells

Cited by 30 publications

References 28 publications

Genotoxicity of acrylamide in vitro: Acrylamide is not metabolically activated in standard in vitro systems

Genotoxicity of acrylamide in vitro: Acrylamide is not metabolically activated in standard in vitro systems

In vivo rodent erythrocyte micronucleus assay. II. Some aspects of protocol design including repeated treatments, integration with toxicity testing, and automated scoring

Comparison of Germ Cell Mutagenicity in Male CYP2E1-Null and Wild-Type Mice Treated with Acrylamide: Evidence Supporting a Glycidamide-Mediated Effect

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