Molecular mechanism of the suppression of larval skeleton by polycyclic aromatic hydrocarbons in early development of sea urchin Hemicentrotus pulcherrimus
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.
KeywordsPolycyclic aromatic hydrocarbons Early development Spicule formation Sea urchin
This research was supported by grant to T.S., K.H., and N.S. (Program for Advancing Strategic International Networks to Accelerate the Circulation of Talented Researchers no. G2702 by JSPS). Grants to T. S. (Scientific Research [C] no. 18K06312 by JSPS) and to N.S. (Grant-in-Aid for Scientific Research [C] no. 16K07871 by JSPS) partially support this investigation. This study was conducted as part of the cooperative research program of Institute of Nature and Environmental Technology, Kanazawa University <Accept no. 17021>.
- Chen K, Tsutsumi Y, Yoshitake S, Qiu X, Xu H, Hashiguchi Y, Honda M, Tashiro K, Nakayama K, Hano T, Suzuki N, Hayakawa K, Shimasaki Y, Oshima Y (2017) Alteration of development and gene expression induced by in ovo-nanoinjection of 3-hydroxybenzo[c]phenanthrene into Japanese medaka (Oryzias latipes) embryos. Aquat Toxicol 182:194–204CrossRefPubMedGoogle Scholar
- Collier TK, Anulacion BF, Arkoosh MR, Dietrich JP, Incardona JP, Johnson LL, Ylitalo GM, Myers MS (2013) Effects on fish of polycyclic aromatic hydrocarbons (PAHs) and naphthenic acid exposures. In: Tierney KB et al (eds) Fish physiology: organic chemical toxicology of fishes. Elsevier, London, pp 195–255CrossRefGoogle Scholar
- Kawamura K (1993) Uni Zouyoushoku to Kakou, Ryutsu. Sapporo Hokkai Suisan Company, Supporo, p 82Google Scholar
- Kitajima T, Matsuda R (1982) Specific protein synthesis of sea urchin micromeres during differentiation. Zool Sci 91:200–205Google Scholar
- Pagano G, Cipollaro M, Corsale G, Esposito A, Ragucci E, Giordano G, Trieff N (1986) The sea urchin: bioassay for the assessment of damage from environmental contaminants. In: Cairns J Jr (eds) Community toxicity testing, ASTM STP 920. American Society for Testing and Materials, Philadelphia, pp 66–92CrossRefGoogle Scholar
- Suzuki N, Ogiso S, Yachiguchi K, Kawabe K, Makino F, Toriba A, Kiyomoto M, Sekiguchi T, Tabuchi Y, Kondo T, Kitamura K, Hong CS, Srivastav AK, Oshima Y, Hattori A, Hayakawa K (2015) Monohydroxylated polycyclic aromatic hydrocarbons influence spicule formation in the early development of sea urchins (Hemicentrotus pulcherrimus). Comp Biochem Physiol C Toxicol Pharmacol 171:55–60CrossRefPubMedGoogle Scholar
- Urben S, Nislow C, Spiegel M (1988) The origin of skeleton forming cells in the sea urchin embryo. Dev Genes Evol 197:447–456Google Scholar