Effect of microtubule inhibitors on the expansion of hypoblast and margin of overgrowth of chick blastoderms

Abstract

The effect of the microtubule inhibitors, Nocodazole and taxol, was studied on the expansion of fragments of chick hypoblast (8 to 10 h incubation) and of margin of overgrowth (24 h incubation) cultured on artificial substrata and on the epiboly in intact blastoderms (24 h incubation). Immunocytochemical staining of these cells with antiserum against tubulin showed that 1 micrograms Nocodazole/ml caused disassembly of microtubules, and that 1 microgram taxol/ml led to increased but unordered assembly. The solvent dimethylsulphoxide had no effect. At these concentrations both microtubule inhibitors led to rapid arrest of the expansion of fragments of hypoblast and of margin of overgrowth in culture, and of the epiboly in intact blastoderms. Time-lapse films showed that inhibition of expansion in both situations was reversible within 2 h after removal of the drugs. Phase-contrast microscopy showed remarkably little difference between the morphology of treated as compared to untreated cultures. Measurements of the height of the cells on sectioned fragments of margin of overgrowth showed no differences between treated and untreated cultures. These results suggest that the cytoplasmic microtubule complex is important both for epiboly and for the migration of hypoblast cells in the chick blastoderm. The mechanisms of this microtubule-related migration are not understood.