Clastogenicity of diepoxybutane in bone marrow cells and male germ cells of mice

Abstract

The bifunctional metabolite of 1,3-butadiene, 1,2:3,4-diepoxybutane (DEB), was tested in the mouse bone marrow micronucleus assay and in male mouse germ cell tests, namely the analysis of first cleavage divisions and the dominant lethal assay. All experiments were performed with single intraperitoneal treatment of the animals. In the micronucleus test, DEB doses of 4.5, 9.0, 18.0 and 36.0 mg/kg body weight were tested at a sampling interval of 24 h for bone marrow. The dose response for the induction of micronuclei in polychromatic erythrocytes was linear with the lowest effective dose of 9.0 mg/kg body weight. No sensitivity difference was observed between male and female mice. the cytogenetic analysis of first cleavage division chromosomes was performed after treatment of male mice with 17, 26, 34, 43 and 52 mg/kg body weight of DEB and mating the males to hormonally stimulated females on days 7, 14, 21 and 28 after treatment. The two higher doses caused general toxicity evidenced by the poor mating behavior of the males. Only 13 and 20% of the mated females were fertilized on day 7 after treatment of the males with 43 and 52 mg/kg body weight of DEB, respectively. An increased number of unfertilized oocytes was obtained from fertilized females on day 7 after treatment of the males with 34 mg/kg body weight of DEB. With a dose of 26 mg/kg body weight, it was demonstrated that chromosomal aberrations were only induced in spermatozoa (mating on day 7 after treatment) while spermatids (mating on days 14 and 21) and spermatocytes (mating on day 28) were not susceptible to the clastogenic effect of DEB. The response in spermatozoa in the dose range 17-34 mg/kg body weight was linear up to 26 mg/kg body weight and reached a plateau thereafter. The results of the dominant lethal experiments performed in the dose range 18-54 mg/kg body weight gave results similar to the cytogenetic study. With the highest dose tested, the toxicity and cytotoxicity during the first 8 mating days after treatment dramatically reduced the number of pregnant females and, consequently, the total implantations, so that no significant dominant lethal effect could be assessed. During mating days 9-12 (treated late spermatids), a significant dominant lethal effect was observed. With the two lower doses (18 and 36 mg/kg body weight), the dominant lethal effect was restricted to spermatozoa. The good correlation of the chromosomal aberrations with dominant lethal mutations confirms the chromosomal origin of dominant lethal effects. The clastogenic effect of DEB in somatic cells and in germ cells of mice was of the same order of magnitude.