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Analysis of mutations in the Pig‐a gene of spleen T‐cells from N ‐ethyl‐ N ‐nitrosourea‐treated fisher 344 rats
Author(s) -
Miura Daishiro,
Shaddock Joseph G.,
Mittelstaedt Roberta A.,
Dobrovolsky Vasily N.,
Kimoto Takafumi,
Kasahara Yoshinori,
Heflich Robert H.
Publication year - 2011
Publication title -
environmental and molecular mutagenesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1
H-Index - 87
eISSN - 1098-2280
pISSN - 0893-6692
DOI - 10.1002/em.20654
Subject(s) - microbiology and biotechnology , mutant , biology , mutation , mutagenesis , mutagen , somatic cell , gene , spleen , gene mutation , nitrosourea , in vivo , reporter gene , genetics , gene expression , dna , immunology , chemotherapy
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 (ProAER r ) rat spleen T‐cells have mutations in the Pig‐a gene. In the present study, we report on a more complete analysis of ProAER r 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.