Analysis of mutations in the Pig‐a gene of spleen T‐cells from N‐ethyl‐N‐nitrosourea‐treated fisher 344 rats
- 3 May 2011
- journal article
- research article
- Published by Wiley in Environmental and Molecular Mutagenesis
- Vol. 52 (5), 419-423
- https://doi.org/10.1002/em.20654
Abstract
A rapid in vivo somatic cell gene mutation assay is being developed that measures mutation in the endogenous X-linked phosphatidylinositol glycan, class A gene (Pig-a). The assay detects Pig-a mutants by flow cytometric identification of cells deficient in glycosylphosphatidyl inositol (GPI) anchor synthesis. GPI-deficient, presumed Pig-a mutant cells also can be detected in a cloning assay that uses proaerolysin (ProAER) selection. Previously, we demonstrated that ProAER-resistant (ProAERr) rat spleen T-cells have mutations in the Pig-a gene. In the present study, we report on a more complete analysis of ProAERr rat spleen T-cell mutants and describe a mutation spectrum for mutants isolated from rats 4 weeks after treatment with three consecutive doses of 35.6 mg/kg N-ethyl-N-nitrosourea (ENU). We identified a total of 55 independent mutations, with the largest percentage (69%) involving basepair substitution at A:T. The overall spectrum of Pig-a gene mutations was consistent with the types of DNA adducts formed by ENU and was very similar to what has been described for in vivo ENU-induced mutation spectra in other rodent reporter genes (e.g., in the endogenous Hprt gene and transgenic shuttle vectors). These data are consistent with the rat Pig-a assay detecting test-agent-induced mutational responses. Environ. Mol. Mutagen., 2011. Published 2011 Wiley-Liss, Inc.Keywords
This publication has 18 references indexed in Scilit:
- The in vivo pig‐a gene mutation assay, a potential tool for regulatory safety assessmentEnvironmental and Molecular Mutagenesis, 2010
- Accumulation and persistence of Pig-A mutant peripheral red blood cells following treatment of rats with single and split doses of N-ethyl-N-nitrosoureaMutation Research/Genetic Toxicology and Environmental Mutagenesis, 2009
- Development of an in vivo gene mutation assay using the endogenous Pig‐A gene: II. Selection of Pig‐A mutant rat spleen T‐cells with proaerolysin and sequencing Pig‐A cDNA from the mutantsEnvironmental and Molecular Mutagenesis, 2008
- Development of an in vivo gene mutation assay using the endogenous Pig‐A gene: I. Flow cytometric detection of CD59‐negative peripheral red blood cells and CD48‐negative spleen T‐cells from the ratEnvironmental and Molecular Mutagenesis, 2008
- In vivo mutation assay based on the endogenous Pig‐a locusEnvironmental and Molecular Mutagenesis, 2008
- The effect of time after treatment, treatment schedule and animal age on the frequency of 6-thioguanine-resistant T-lymphocytes induced in Fischer 344 rats by N-ethyl-N-nitrosoureaMutation Research/Genetic Toxicology, 1993
- Mutational spectrum at the Hprt locus in splenic T cells of B6C3F1 mice exposed to N-ethyl-N-nitrosourea.Proceedings of the National Academy of Sciences of the United States of America, 1992
- Statistical test for the comparison of samples from mutational spectraJournal of Molecular Biology, 1987
- O4-Methyl, -ethyl, or -isopropyl substituents on thymidine in poly(dA-dT) all lead to transitions upon replication.Proceedings of the National Academy of Sciences of the United States of America, 1986