Detection of aneuploidy by multicolor FISH in mouse sperm after in vivo treatment with acrylamide, colchicine, diazepam or thiabendazole

Multicolor fluorescence in situ hybridization (FISH) was used to investigat e the induction of aneuploidy during meiosis in young adult male mice treat ed with chemicals chosen for the EU sponsored aneuploidy project (acrylamid e, colchicine, diazepam and thiabendazole), The aim of the present study wa s to evaluate the frequency of aneuploid sperm induced by each of these che micals by sperm FISH. Male (102/ElxC3H/El)F-1 mice were treated with acryla mide (120 and 60 mg/kg single dose i.p.), colchicine (1.5 and 3 mg/kg singl e dose, i.p.), diazepam (300, 150 and 75 mg/kg single dose by oral intubati on) or thiabendazole (100 and 300 mg/kg daily for 11 days by oral intubatio n). At 22 days after the last treatment, sperm were collected from the caud a epididymis, Three chromosome FISH was applied to determine hyperhaploid a nd diploid sperm with DNA probes specific for the chromosomes X, Y and 8, F ive animals were treated per dose group and sperm aneuploidy was evaluated in 10 000 sperm per animal, We found significant increases in the frequency of total hyperhaploidy for the males treated with 3.0 mg/kg colchicine (0. 092 versus 0.056%, P < 0.05) and with 1.5 mg/kg colchicine (0.082 versus 0. 050%, P < 0.05), as well for the males treated with 300 mg/kg diazepam (0.0 81 versus 0.050%, P < 0.05), indicating that colchicine and diazepam each i nduced germ cell aneuploidy, We also found significant increases in the fre quency of total diploidy for the males treated with 300 mg/kg diazepam (P < 0.05) and with 300 mg/kg thiabendazole (P < 0.05), No significant effects were found for 120 and 60 mg/kg acrylamide or for the other doses of diazep am and thiabendazole, These first results indicate that the multicolor FISH method is useful to determine aneuploidy induction in sperm of mice.