4-chloro-o-phenylenediamine induces a dose-related increase in G:C > T:A transversions and one major DNA adduct in the liver of Big Blue mice after 26 weeks in feed treatment.
The monocyclic aromatic amine 4-chloro-o-phenylenediamine (4-C-o-PDA), a known mutagen and mouse hepatocarcinogen, was tested for its in vivo mutagenic potential in the Big Blue transgenic mouse assay system. Genomic DNA was isolated from liver tissue of control and treated animals and lacI mutants were recovered. In an initial 2-week study 4-C-o-PDA was administered daily per os to groups of male and female C57BL/6 Big Blue mice at doses of 0 and 200 mg/kg for 2 weeks (on working days) followed by a treatment free expression time of 10 days. Only a weak increase in the mutant frequencies in females was observed. In a 26-week study, where 4-C-o-PDA was given to groups of male and female Big Blue mice in feed at dietary concentrations of 0, 5,000 and 10,000 ppm, 4-C-o-PDA was found to induce a pronounced dose-dependent increase in mutant frequencies in either sex. In the present work, we analyzed the mutation spectrum by automated DNA sequencing of lacI mutants from both studies. Following the 2-week administration of 4-C-oT:A transversions in both sexes. In addition, upon 26-week treatment with 4-C-o-PDA, one major DNA adduct was detected by 33P postlabelling and subsequent multidimensional thin layer chromatography. It is concluded that 4-C-oT:A transversions after 26 weeks in feed treatment. This result indicates that the sensitivity of the Big Blue transgenic assay system, in detecting a unique chemically induced mutation spectrum, is dependent on experimental parameters, such as treatment time. The data suggest that the formation of one major DNA adduct upon 4-C-o-PDA treatment may be critical for its mutagenicity.
Staedtler
F
F
Novartis Pharma, Preclinical Safety - Toxicology/Pathology, WS-2881. 3.07A, CH-4002, Basel, Switzerland. frank.staedtler@pharma.novartis.com
Crespo-Perez
J
J
Sagelsdorff
P
P
Steiner
S
S
Suter
W
W
D000225:Adenine; D000821:Animal Feed; D000818:Animals; D003596:Cytosine; D018736:DNA Adducts / Q000187:drug effects / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005260:Female; D006147:Guanine; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009153:Mutagens / Q000378:metabolism / Q000494:pharmacology*; D010655:Phenylenediamines / Q000008:administration & dosage / Q000378:metabolism / Q000494:pharmacology*; D017354:Point Mutation; D013941:Thymine
D018736:DNA Adducts; D009153:Mutagens; D010655:Phenylenediamines; D006147:Guanine; C015060:4-chloro-1,2-diaminobenzene; D003596:Cytosine; D000225:Adenine; D013941:Thymine