Background: The zebrafish is emerging as an important genetic system for the study of vertebrate development, and many zygotic mutations affecting embryogenesis have been isolated. The early events in development are under the control of maternal genes but are relatively unexplored. Here, the process of axis specification is investigated.
Results: The vegetal pole of the zygote transiently contains a dense array of parallel microtubules, while microtubules near the equator are disorganized. Irradiation of the zygote with ultraviolet light disrupts the formation of the vegetal microtubule array and causes loss of the axis; brief treatment with nocodazole at this stage also causes defects in the axis. During cleavage stages, yolk cortical microtubules reorganize to form arrays that apparently extend from marginal blastomeres. Prolonged exposure to cold (18 degrees C) or incubation in nocodazole prior to the 32-cell stage disrupts cortical microtubules and causes premature formation of the yolk syncytial layer; these treatments also prevent formation of an axis, as indicated by the absence of goosecoid and forkhead2 expression and of translocation of beta-catenin into nuclei. Cortical microtubule arrays are required for the transport of particles from the vegetal hemisphere into marginal blastomeres, as shown by the movement of polystyrene beads; treatments that prevent axis formation also prevent the entry of beads into blastomeres.
Conclusions: To form an organizer, zebrafish blastomeres appear to require substances which are transported from the vegetal hemisphere of the yolk cell by cortical microtubules. Initial asymmetry appears dependent on an array of parallel microtubules at the vegetal pole.