Artificial parthenogenesis in starfish eggs: behavior of nuclei and chromosomes resulting in tetraploidy of parthenogenotes produced by the suppression of polar body extrusion.

Artificial parthenogenesis in starfish requires the activation of oocytes and the suppression of the polar body (PB) extrusion. To induce parthenogenesis we have employed a combination of calcium ionophore A23187 for activating oocytes and cytochalasin B (CB) for the suppression of PB extrusions. The treatment of activated oocytes with CB during meiosis I produced eggs lacking polar bodies, i.e., during meiosis I produced eggs lacking polar bodies, i.e., 0pb eggs, and treatment during meiosis II induced eggs bearing the first PB only, i.e., 1pb eggs. About 90% of both 0pb and 1pb eggs developed parthenogenetically with strong synchrony of cell division among eggs. After meiotic chromosomes in CB-treated oocytes separated finally into monads, they came together to form a single nucleus. The nucleus was found to be tetraploid in the 0pb eggs and diploid in the 1pb eggs. Regardless of the difference in ploidy, both 0pb and 1pb eggs developed as tetraploid embryos. Observations of the behavior of meiotic chromosomes and nuclei revealed that in the 0pb eggs, the first round of chromosomal replication was followed by the first cleavage, as in normally fertilized eggs. In the 1pb eggs, on the other hand, two rounds of chromosomal replication were found to precede the first cleavage, indicating the occurrence of one (first) round of chromosomal replication that is not accompanied with cytokinesis. In the first round of mitosis, a bipolar spindle did not appear, but only a half spindle was formed, resulting in the failure of both karyokinesis and cytokinesis. Thus, both types of parthenogenetic eggs became tetraploid before the first cleavage. At the first cleavage, both types of eggs formed a bipolar spindle and divided into a pair of blastomeres. Based on these observations, we suggest that the meiotic centrosomes remaining in these eggs by the failure of PB extrusion are diverted into mitosis-organizing centers in the mitotic spindle, and this results in parthenogenetic development.