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a larger target for point mutation, especially for point mu-
tation at G:C basepairs [discussed in Chen et al., 1998].
As demonstrated in this study, the major adduct produced
by N-OH-AAF is dG-C8-AF, and a large number of in vitro
studies indicate that this adduct primarily induces G 3 T
transversion [Heflich and Neft, 1994]. Comparison of in-
duced mutant frequencies in the spleen indicates that the
four-dose treatment with N-OH-AAF induced 2.7-fold more
lacI mutants than hprt mutants. This value of 2.7 is consis-
tent with what we have previously found for lacI and hprt
lymphocyte mutant frequencies induced by other mutagenic
carcinogens in the Big Blue rat. 7,12-Dimethylbenz[a]an-
thracene, which forms somewhat more bulky adducts with
dA than with dG, induced 1.7-fold more lacI mutants than
hprt mutants [Manjanatha et al., 1998], whereas thiotepa,
which like N-OH-AAF produces mainly dG adducts, in-
duced 2.8-fold more lacI than hprt mutants [Chen et al.,
1998].
A strength of transgenic shuttle vector systems for de-
tecting in vivo mutation is that mutations can be measured
in any tissue from which sufficient DNA can be recovered.
Thus, the transgenic assays may be useful for demonstrating
the tissue-specificity of in vivo mutation, which could be
extremely useful in evaluating the tissue-specific carcino-
genicity of chemical agents. The results of this study are
consistent with the expectation that the hepatocarcinogen
N-OH-AAF should produce a higher mutant frequency in
the target liver than the nontarget tissues like spleen lym-
phocytes. Comparison of the liver lacI response in the rat
with published liver lacI mutant frequencies for mice
treated with 2-AAF or N-OH-AAF also suggests a species-
specificity to the response. Whereas the N-OH-AAF mutant
response reached 16-fold the background frequency in Big
Blue rats, the reported maximum mutant frequencies for
2-AAF– or N-OH-AAF–treated transgenic mice range from
less than twofold background to about fivefold background
[Gunz et al., 1993; Shephard et al., 1993; Brooks et al.,
1995; Frijhoff et al., 1998; Ross and Leavitt, 1998]. Al-
though none of the mouse studies used treatments directly
comparable to our rat treatment, these mutagenicity results
are generally consistent with the greater carcinogenic po-
tency of 2-AAF in rats than in mice [Heflich and Neft, 1994]
and suggest the potential of the Big Blue system for
making interspecies comparisons.
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