Molecular mechanism of the suppression of larval skeleton by polycyclic aromatic hydrocarbons in early development of sea urchin <Emphasis Type="Italic">Hemicentrotus pulcherrimus</Emphasis>

Polycyclic aromatic hydrocarbons including benz[a]anthracene (BaA) are priority pollutants in the aquatic environment. Our previous study revealed that BaA and its metabolite, 4-monohydroxylated BaA (4-OHBaA) inhibit larval skeletogenesis in the sea urchin Hemicentrotus pulcherrimus. Here we report studies to elucidate the target of skeletogenesis inhibition elicited by BaA and 4-OHBaA. First, we performed an in vitro experiment using isolated micromeres which give rise to the larval skeletogenic mesenchyme. However, skeletogenesis was not repressed by BaA and 4-OHBaA, implying that these chemicals indirectly influence on the formation of larval skeleton. Next, we analyzed their influence in vivo using embryos. Vascular endothelial growth factor (VEGF) that is expressed in the ectoderm and induces spicule formation was inhibited by BaA and 4-OHBaA treatment. These chemicals also suppressed the expression of the heparan sulfate 6-O endosulfatase (Sulf) known as a VEGF signaling modulator. We, therefore, propose that BaA and 4-OHBaA effects on larval skeletogenesis via VEGF signaling. Furthermore, we showed that the expression of Endo16 mRNA, an endodermal marker, decreased after BaA and 4-OHBaA exposure, suggesting that these chemicals affect endodermal function together with skeletogenesis. This study demonstrates that BaA and 4-OHBaA exert multiple detrimental effects on the development of H. pulcherrimus.     

Accumulation of a bitter amino acid,pulcherrimine, in sea urchin ovaries and its relevance to the annual reproductive cycle

Hemicentrotus pulcherrimus in the Tohoku area is not utilized as a fishery product because the ovaries have a bitter taste almost all year. To discover the cause, we compared the annual reproductive cycles and ovarian accumulation of a bitter amino acid, pulcherrimine, in H. pulcherrimus collected at Mikuni (Fukui Prefecture), where it is fished in summer, and Iwaki (Fukushima Prefecture), where it is not utilized. At both sites, pulcherrimine increased in maturing ovaries as oogenesis proceeded from autumn to winter, decreased after spawning in spring, but remained in spent and immature ovaries for a few months. In Mikuni, pulcherrimine was scarcely detected in the ovaries for several months in summer. However, in Iwaki, late spawning delayed the disappearance of pulcherrimine in summer, and early initiation of oogenesis advanced the restart of pulcherrimine accumulation in autumn compared to Mikuni. Consequently, the period when the ovaries were palatable because of the absence of pulcherrimine during summer was short in Iwaki. The major cause of such short-term palatability could be the lower seawater temperature in the Tohoku area. Furthermore, pulcherrimine was never detected in other sea urchins, although a bitter taste was found in the mature ovaries of some species.