Analysis of chemically induced spindle aberrations in male mouse germ cells: comparison of differential and immunofluorescent staining procedures.

Differential staining of spindle and chromatin was adapted to meiotic cells of male mice. Immunostaining with antitubulin antibodies was developed. Both methods were used to test acrylamide, diazepam and vinblastine for their potential to cause spindle disturbances in male mouse germ cells. The analyses were performed after in vivo treatment, comparing the classical safranin/brilliant blue spindle staining to an immunofluorescent assay. The two staining methods complemented each other. Differential staining seems to be more sensitive to the detection of misplaced chromatin whereas the immunostaining is superior when scoring for spindle structure aberrations. All three chemicals showed spindle activity. Acrylamide mainly caused multipolar spindles which possibly develop from a separation of mother and daughter centrioles during an unspecific meiotic block. Diazepam elevated the level of monopolar spindles and induced the loss of single chromosomes, suggesting an effect on motor proteins. Vinblastine was the strongest spindle poison by far. It produced high numbers of shortened and monopolar spindles as well as multiple chromosome loss.