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• W2735962502 endingPage "493" @default.
• W2735962502 startingPage "485" @default.
• W2735962502 abstract "The Pig-a assay, a recently developed in vivo somatic gene mutation assay, is based on the identification of mutant erythrocytes that have an altered repertoire of glycosylphosphatidylinositol (GPI)-anchored cell surface markers. We hypothesized that the erythrocyte Pig-a assay concept could be applied to rat cauda epididymal spermatozoa (sperm) for germ cell mutagenicity evaluation. We used GPI-anchored CD59 as the Pig-a mutation marker and examined the frequency of CD59-negative sperm using flow cytometry. A reconstruction experiment that spiked un-labeled sperm (mutant–mimic) into labeled sperm at specific ratios yielded good agreement between the detected and expected frequencies of mutant–mimic sperm, demonstrating the analytical ability for CD59-negative sperm detection. Furthermore, this methodology was assessed in F344/DuCrl rats administered N-ethyl-N-nitrosourea (ENU), a prototypical mutagen, or clofibrate, a lipid-lowering drug. Rats treated with 1, 10, or 20 mg/kg body weight/day (mkd) ENU via daily oral garage for five consecutive days showed a dose-dependent increase in the frequency of CD59-negative sperm on study day 63 (i.e., 58 days after the last ENU dose). This ENU dosing regimen also increased the frequency of CD59-negative erythrocytes. In rats treated with 300 mkd clofibrate via daily oral garage for consecutive 28 days, no treatment-related changes were detected in the frequency of CD59-negative sperm on study day 85 (i.e., 57 days after the last dose) or in the frequency of CD59-negative erythrocytes on study day 29. In conclusion, these data suggest that the epidiymal sperm Pig-a assay in rats is a promising method for evaluating germ cell mutagenicity. Environ. Mol. Mutagen. 58:485–493, 2017. © 2017 Wiley Periodicals, Inc." @default.
• W2735962502 created "2017-07-21" @default.
• W2735962502 creator A5047460454 @default.
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• W2735962502 date "2017-07-17" @default.
• W2735962502 modified "2023-10-16" @default.
• W2735962502 title "Applying the erythrocyte <i>Pig-a</i> assay concept to rat epididymal sperm for germ cell mutagenicity evaluation" @default.
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• W2735962502 doi "https://doi.org/10.1002/em.22109" @default.
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