Expression of Na+, K+-ATPase α-Subunit in Animalized and Vegetalized Embryos of the Sea Urchin, Hemicentrotus pulcherrimus.

A marked increase in the Na⁺, K⁺-ATPase activity of sea urchin embryos occurred following an elevation of its mRNA level, revealed by Northern blotting analysis, in developmental period between the swimming blastula and the late gastrula stage. cDNA clone of Na⁺, K⁺-ATPase α-subunit, obtained from γgt10 cDNA library of sea urchin gastrulae, was digested with EcoRl ad Hindlll. The obtained 268 bp cDNA fragment, hybridized to a 4.6 Kb RNA, was used as probe for Northern blotting analysis. The level of Na⁺, K⁺-ATPase mRNA was higher in embryo-wall cell fraction isolated from late gastrulae (ectoderm cells) than the level in the bag fraction, containing mesenchyme cells (mesoderm cells) and archenteron (endoderm cells). The activity of Na⁺, K⁺-ATPase and the level of its mRNA were higher in animalized embryos obtained by pulse treatment with A23187 for 3 hr, starting at the 8–16 cell stage and were considerably lower in vegetalized embryos induced by 3 hr treatment with Li⁺ than that in normal embryos at the post gastrula corresponidng stage. Augmentation of Na⁺, K⁺-ATPase gene expression can be regarded as a marker for ectoderm cell differentiation at the post gastrula stage, which results from determination of cell fate in prehatching period.     

Regulating potential in development of a direct developing echinoid, Peronella japonica

The regulating potential along the animal-vegetal axis of a direct developing echinoid, Peronella japonica, was investigated using LiCl. Animal caps isolated from 16-cell stage P. japonica embryos developed to permanent blastulae with an amniotic cavity. Treatment of animal caps with LiCl induced them to vegetalize with differentiation of the endoderm and subsequently develop into pluteus-like larvae. The larvae derived from the LiCl-treated animal caps were able to metamorphose and establish an adult body plan. A considerable fraction of whole embryos treated with LiCl exogastrulated and/or evaginated an amniotic cavity. The timing of the sensitivity to LiCl-mediated induction of evagination of the amniotic cavity was earlier than that for exogastrulation. Peronella japonica embryos became sensitive to LiCl induction of exogastrulation later than embryos of indirect developers. Some larvae with evaginated archenteron and/or evaginated amniotic cavity had metamorphic potential. These results suggest that LiCl can induce both vegetalization and evagination of invaginating structures. The present study is the first to show the potential of the presumptive ectoderm region to regulate the establishment of the adult body plan without any influence from other blastomeres, revealing that the regulating potential of sea urchin embryos is much larger than previously thought.