Response of the phiX174 am3, cs70 transgene to acute and chronic ENU exposure: implications for protocol design.
Studies of other transgenic assays have shown that time after treatment is a very important variable in the analysis of mutation frequencies but that eventually a plateau frequency is reached, indicating that the mutations are neutral. This neutrality is very important for the design of both experiments and testing protocols. Here we show that the phiX174 am3, cs70 transgene gives qualitatively similar results to the other transgenes studied after exposure of the mice to N-ethyl-N-nitrosourea. In the small intestine, the mutant frequency induced by an acute dose did not change significantly from 10 to 70 days post-treatment, indicating that the mutations induced are, indeed, neutral. Likewise, the mutant frequency increased linearly with duration of exposure to ENU at a constant rate. Mutant frequencies obtained were 10 times higher from the chronic exposure than produced by a nearly lethal acute dose. As in previous comparisons of a transgene and the endogenous Dlb-1 locus in the small intestine, the chronic exposure was much more effective at increasing the sensitivity of the transgene than of the endogenous gene. The Dlb-1 locus shows more complex kinetics in this strain, as in others, with mutations initially accumulating at a slower rate, indicating a differential repair of genetic damage.
Lawrence Livermore National Laboratory, Biology and Biotechnology Research Program, 94550 Livermore, CA, USA. cosentino1@llnl.gov
Lidia
L
Cosentino
Heinrich V
HV
Malling
John A
JA
Heddle
D000818:Animals; D010584:Bacteriophage phi X 174 / Q000187:drug effects* / Q000235:genetics; D005038:Ethylnitrosourea / Q000633:toxicity*; D005814:Genes, Viral / Q000187:drug effects* / Q000235:genetics; D007421:Intestine, Small / Q000187:drug effects / Q000378:metabolism; D037102:Lectins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D008817:Mice, Mutant Strains; D008822:Mice, Transgenic; D009153:Mutagens / Q000633:toxicity*; D009154:Mutation; D037121:Plant Lectins; D013997:Time Factors; D018675:Toxicity Tests / Q000379:methods*; D019076:Transgenes / Q000187:drug effects*
D037102:Lectins; D009153:Mutagens; D037121:Plant Lectins; C036554:dolichos biflorus agglutinin; D005038:Ethylnitrosourea