The muntjac spindle staining assay: An investigation of the ability of this assay to detect potential aneuploidy inducing agents

Abstract

An assay using muntjac fibroblasts in conjunction with a differential spindle and chromosome stain was developed and evaluated for its ability to detect aneuploidy inducing agents. The effects of the spindle poison colcemid and the human carcinogen diethylstilboestrol were compared with those obtained after treatment of the cells with a number of compounds each believed to impair mitotic spindle function via a different mechanism. Vinblastine and nocodazole were both shown to have colcemid like effects suggesting a common mode of action and potential aneugenic activity. p-Fluorophenylalanine affected the spindle morphology resulting in an accumulation of atypical metaphases but without significant chomosome loss. Hydroquinone induced metaphase arrest and significant spindle damage but at doses very close to the toxic limits for this compound. Lastly, acenaphthene appeared to have no significant effect on mitosis at doses up to toxic levels. The ability of the muntjac fibroblasts to express aneuploid elements in recovery cell populations following treatment with colcemid and p-fluorophenylalanine was also examined. The results were inconclusive although an induction of endoreduplication was - indicated. Although both the spindle staining assay and recovery protocol require further refinement and validation, it was concluded that the assay was able to detect aneugenic agents especially those which act via the spindle.