The spirmatid micronucleus test with the dissection technique detects the germ cell mutagenicity of acrylamide in rat meiotic cells

Abstract

As part of the development and validation of the spermatid micronucleus test (SMNT) in the project ‘Detection of Germ Cell Mutagens’ sponsored by the CEC we studied the mutagenicity of acrylamide (AA) and mitomycin C (MMC). Of two alternative techniques, we used the ‘dissection technique’ based on microdissection of seminiferous tubules offering a narrow window for evaluation of cell state sensitivity, and including DNA-specific staining and scoring. AA given as a single injection of 50 or 100 mg/kg did not significantly increase MN frequencies. When a subchronic treatment (4 × 50 mg/kg) was given, a significant increase over background was observed 18 and 19 days after the last injection, indicating genotoxic activity in preleptotene spermatocytes and late spermatogonial stages. MMC given as single injections of 0.5 or 1.0 mg/kg increased MN frequencies significantly 17, 18, 19 and 20 days after treatment as a result of clastogenicity in S phase cells. DNA flow cytometry did not show cytotoxicity of AA to preleptotene spermatocytes, but a small decrease in the numbers of stem cells. If spindle disturbances are caused by AA, as suggested, they were not detectable by induction of spermatid MN in vivo 1 or 3 days after treatment or by treatment with AA of cultured sgements of a seminiferous tubules undergoing meiotic divisions in vitro. In conclusion, the SMNT with the dissection technique is able to show the germ cell clastogenicity of AA and MMC, AA was observed to have a much weaker MN inducing potency than MMC.