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Ethylnitrosourea-induced mutations in vivo involving the Dolichos biflorus agglutinin receptor in mouse intestinal epithelium

: O'Sullivan, J.F.; Paul, D.; Schmidt, G.H.

Mutation research 228 (1990), Nr.2, S.149-155
ISSN: 0027-5107
Fraunhofer ITA ( ITEM) ()
dolichos biflorus agglutinin; ethylnitrosourea; intestinal epithelium; mutagenesis

A mutagenesis assay system is intoduced based on the induction of mutations in somatic cells of mouse small intestine using ethylnitrosourea (ENU). F sub 1 mice, heterozygous for the Dolichos biflorus agglutinin (DBA) locus (Dlb-1 high a/Dlb-1 high b) encoding the DBA cell surface receptor, were treated in utero on either day 7, day 9 or day 11 post coitum. Mutant intestinal cell populations of adult mice were visualised in whole-mount preparations by the absence of histochemical staining using peroxidase-labelled Dolichos biflorus agglutinin. Loss of staining is attributed to mutagenesis of the Dlb-1 high b allele in the heterozygote. This system allows one to evaluate mammalian mutagenesis in vivo at a single locus. Mutant cell populations appeared as discrete groups of 'striped' villi, each stripe comprising cells derived from unstained crypt stem cells (cf. Schmidt et al., 1985a). A spontaneous mutation level was noted in untreated controls which was found to differ significantly f rom that recorded in mice treated with the mutagen (P is smaller than 0.01). The mutation scores were highly consistent among mice and a small number of animals (i.e., 16) were sufficient to detect mutagenic effects of ENU. Thus, the advantages which accrue from the assay are (1) the ability to detect small clones of mutant cell populations in the intestine (i.e., cells derived from a single mutated crypt); (2) a small number of tested mice are required to generate a conclusive result, especially when compared to the mammalian spot test (Fahrig, 1978)