title: Comparison of in vivo mutagenesis in the endogenous Hprt gene and the lacI transgene of Big Blue(R) rats treated with 7, 12-dimethylbenz[a]anthracene.
abstract: The lacI transgene of Big Blue(R) (BB) rats was evaluated as a reporter of in vivo mutation by comparing mutant frequencies (MFs) in it and in the endogenous Hprt gene. Seven-week old female BB rats were given single doses of 0, 20, 75 and 130 mg/kg of 7, 12-dimethylbenz(a)anthracene (DMBA) by gavage, and Hprt and lacI MFs in splenic lymphocytes were measured over a period of 18 weeks. The Hprt MFs in treated rats increased for 10 weeks and then declined; 130 mg/kg of DMBA produced a maximum Hprt MF of 168+/-11.4x10-6 clonable lymphocytes, while the MF in control rats was 7.4+/-1. 5x10-6. DMBA exposure of generic F344 rats resulted in a similar time-course of mutant induction but produced about 50% higher Hprt MFs with the 75 and 130 mg/kg doses. In contrast, the lacI MFs increased for 6 weeks and then remained relatively constant; 130 mg/kg of DMBA produced a maximum increase in lacI MF of 341+/-83x10-6 PFU compared with 25+/-5x10-6 PFU in control rats. The Hprt mutant frequencies in DMBA-treated BB and F344 rats were significantly increased over control values for every dose-time combination examined, while only the 130 mg/kg dose consistently produced lacI MFs that were significantly above the controls. In addition, the fold-increase in MF for treated vs. control rats was two times higher for the Hprt gene than the lacI gene due to the higher MFs in the lacI gene of control rats. Differences between the lacI and Hprt genes in the kinetics of mutant induction, in the frequency of induced mutants, and in the sensitivity of mutant detection could be explained at least partially by the properties of these two genes.
authors.affiliation: Department of Health and Human Services, Food and Drug Administration, National Center for Toxicological Research, Division of Genetic Toxicology, Jefferson, AR 72079, USA. mmanjanatha@nctr.fda.gov
authors.forename: M G
authors.initials: MG
authors.lastname: Manjanatha
authors.forename: S D
authors.initials: SD
authors.lastname: Shelton
authors.forename: A
authors.initials: A
authors.lastname: Aidoo
authors.forename: L E
authors.initials: LE
authors.lastname: Lyn-Cook
authors.forename: D A
authors.initials: DA
authors.lastname: Casciano
mesh_terms: D015127:9,10-Dimethyl-1,2-benzanthracene / Q000633:toxicity*; D000818:Animals; D030801:Animals, Genetically Modified; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics*; D002273:Carcinogens / Q000633:toxicity*; D002999:Clone Cells; D004305:Dose-Response Relationship, Drug; D029968:Escherichia coli Proteins; D005260:Female; D007041:Hypoxanthine Phosphoribosyltransferase / Q000096:biosynthesis / Q000235:genetics*; D007700:Kinetics; D056968:Lac Repressors; D008214:Lymphocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D016296:Mutagenesis; D051381:Rats; D011916:Rats, Inbred F344; D011919:Rats, Inbred Strains; D012097:Repressor Proteins / Q000096:biosynthesis / Q000235:genetics*; D013154:Spleen; D013997:Time Factors
chemical_list: D001426:Bacterial Proteins; D002273:Carcinogens; D029968:Escherichia coli Proteins; D056968:Lac Repressors; D012097:Repressor Proteins; D015127:9,10-Dimethyl-1,2-benzanthracene; D007041:Hypoxanthine Phosphoribosyltransferase