Fisheries Science

, Volume 84, Issue 6, pp 1073–1079 | Cite as

Molecular mechanism of the suppression of larval skeleton by polycyclic aromatic hydrocarbons in early development of sea urchin Hemicentrotus pulcherrimus

  • Toshio Sekiguchi
  • Koji Yachiguchi
  • Masato Kiyomoto
  • Shouzo Ogiso
  • Shuichi Wada
  • Yoshiaki Tabuchi
  • Chun-Sang Hong
  • Ajai K. Srivastav
  • Stephen D. J. Archer
  • Stephen B. Pointing
  • Kazuichi Hayakawa
  • Nobuo SuzukiEmail author
Original Article Environment


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.


Polycyclic 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>.


  1. Adomako-Ankomah A, Ettensohn CA (2013) Growth factor-mediated mesodermal cell guidance and skeletogenesis during sea urchin gastrulation. Development 140:4214–4225CrossRefPubMedGoogle Scholar
  2. Akasaka K, Ueda T, Higashinakagawa T, Yamada K, Shimada H (1990) Spatial patterns of arylsulfatase mRNA expression in sea urchin embryo. Dev Growth Differ 32:9–13CrossRefGoogle Scholar
  3. Alexander FJ, King CK, Reichelt-Brushett AJ, Harrison PL (2017) Fuel oil and dispersant toxicity to the Antarctic sea urchin (Sterechinus neumayeri). Environ Toxicol Chem 36:1563–1571CrossRefPubMedGoogle Scholar
  4. Ball A, Truskewycz A (2013) Polyaromatic hydrocarbon exposure: an ecological impact ambiguity. Environ Sci Pollut Res Int 20:4311–4326CrossRefPubMedGoogle Scholar
  5. Banger K, Toor GS, Chirenje T, Ma L (2010) Polycyclic aromatic hydrocarbons in urban soils of different land uses in Miami, Florida. Soil Sediment Contam 19:231–243CrossRefGoogle Scholar
  6. Beiras R, Vazquez E, Bellas J, Lorenzo J, Fernandez N, Macho G, Marino J, Casas L (2001) Sea-urchin embryo bioassay for in situ evaluation of the biological quality of coastal seawater. Estuar Coast Shelf Sci 52:29–32CrossRefGoogle Scholar
  7. Bellas J, Nieto Ó, Beiras R (2011) Integrative assessment of coastal pollution: development and evaluation of sediment quality criteria from chemical contamination and ecotoxicological data. Cont Shelf Res 31:448–456CrossRefGoogle Scholar
  8. Bue BG, Sharr S, Seeb JE (1998) Evidence of damage to pink salmon populations inhabiting Prince William Sound, Alaska, two generations after the Exxon Valdez Oil Spill. Trans Am Fish Soc 127:35–43CrossRefGoogle Scholar
  9. Charles GD, Bartels MJ, Zacharewski TR, Gollapudi BB, Freshour NL, Carney EW (2000) Activity of benzo[a]pyrene and its hydroxylated metabolites in an estrogen receptor-alpha reporter gene assay. Toxicol Sci 55:320–326CrossRefPubMedGoogle Scholar
  10. 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
  11. 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
  12. Dinnel PA, Stober QJ, DiJulio DH (1981) Sea urchin sperm bioassay for sewage and chlorinated seawater and its relation to fish bioassays. Mar Environ Res 5:29–39CrossRefGoogle Scholar
  13. Duloquin L, Lhomond G, Gache C (2007) Localized VEGF signaling from ectoderm to mesenchyme cells controls morphogenesis of the sea urchin embryo skeleton. Development 134:2293–2302CrossRefPubMedGoogle Scholar
  14. Fedato R, Simonato J, Martinez C, Sofia S (2010) Genetic damage in the bivalve mollusk Corbicula fluminea induced by the water-soluble fraction of gasoline. Mutat Res Genet Toxicol Environ Mutagen 700:80–85CrossRefGoogle Scholar
  15. Fujita K, Takechi E, Sakamoto N, Sumiyoshi N, Izumi S, Miyamoto T, Matsuura S, Tsurugaya T, Akasaka K, Yamamoto T (2010) HpSulf, a heparan sulfate 6-O-endosulfatase, is involved in the regulation of VEGF signaling during sea urchin development. Mech Dev 127:235–245CrossRefPubMedGoogle Scholar
  16. Hannam ML, Bamber SD, Galloway TS, Moody AJ, Jones MB (2010) Effects of the model PAH phenanthrene on immune function and oxidative stress in the haemolymph of the temperate scallop Pecten maximus. Chemosphere 78:779–784CrossRefPubMedGoogle Scholar
  17. Hayakawa K, Nomura M, Nakagawa T, Oguri S, Kawanishi T, Toriba A, Kizu R, Sakaguchi T, Tamiya E (2006) Damage to and recovery of coastlines polluted with C-heavy oil spilled from the Nakhodka. Water Res 40:981–989CrossRefPubMedGoogle Scholar
  18. Hayakawa K, Onoda Y, Tachikawa C, Hosoi S, Yoshita M, Chung SW, Kizu R, Toriba A, Kameda T, Tang N (2007) Estrogenic/antiestrogenic activities of polycyclic aromatic hydrocarbons and their monohydroxylated derivatives by yeast two-hybrid assay. J Health Sci 53:562–570CrossRefGoogle Scholar
  19. Jaruchotikamol A, Jarukamjorn K, Sirisangtrakul W, Sakuma T, Kawasaki Y, Nemoto N (2007) Strong synergistic induction of CYP1A1 expression by andrographolide plus typical CYP1A inducers in mouse hepatocytes. Toxicol Appl Pharmacol 224:156–162CrossRefPubMedGoogle Scholar
  20. Kawamura K (1993) Uni Zouyoushoku to Kakou, Ryutsu. Sapporo Hokkai Suisan Company, Supporo, p 82Google Scholar
  21. Kitajima T, Matsuda R (1982) Specific protein synthesis of sea urchin micromeres during differentiation. Zool Sci 91:200–205Google Scholar
  22. Kobayashi N (1990) Marine pollution bioassay by sea urchin eggs, an attempt to enhance sensitivity. Publ Seto Mar Biol Lab 34:225–237CrossRefGoogle Scholar
  23. Kobayashi N (1991) Marine pollution bioassay by using sea urchin eggs in the Tanabe Bay, Wakayama Prefecture, Japan, 1970–1987. Mar Pollut Bull 23:709–713CrossRefGoogle Scholar
  24. McClay DR (2011) Evolutionary crossroads in developmental biology: sea urchins. Development 138:2639–2648CrossRefPubMedPubMedCentralGoogle Scholar
  25. Mitsunaga-Nakatsubo K, Akimoto Y, Kawakami H, Akasaka K (2009) Sea urchin arylsulfatase, an extracellular matrix component, is involved in gastrulation during embryogenesis. Dev Genes Evol 219:281–288CrossRefPubMedGoogle Scholar
  26. 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
  27. Pillai MC, Vines CA, Wikramanayake AH, Cherr GN (2003) Polycyclic aromatic hydrocarbons disrupt axial development in sea urchin embryos through a beta-catenin dependent pathway. Toxicology 186:93–108CrossRefPubMedGoogle Scholar
  28. Ransick A, Ernst S, Britten RJ, Davidson EH (1993) Whole mount in situ hybridization shows Endo 16 to be a marker for the vegetal plate territory in sea urchin embryos. Mech Dev 42:117–124CrossRefPubMedGoogle Scholar
  29. Romano LA, Wray GA (2006) Endo16 is required for gastrulation in the sea urchin Lytechinus variegatus. Dev Growth Differ 48:487–497CrossRefPubMedGoogle Scholar
  30. Suess MJ (1976) The environmental load and cycle of polycyclic aromatic hydrocarbons. Sci Total Environ 6:239–250CrossRefGoogle Scholar
  31. 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
  32. Unuma T, Sakai Y, Agatsuma Y, Kayaba T (2015) Sea urchin aquaculture in Japan. In: Brown NP, Eddy SD (eds) Echinoderm aquaculture. Wiley, New Jersey, pp 75–126CrossRefGoogle Scholar
  33. 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

Copyright information

© Japanese Society of Fisheries Science 2018

Authors and Affiliations

  • Toshio Sekiguchi
    • 1
  • Koji Yachiguchi
    • 1
  • Masato Kiyomoto
    • 2
  • Shouzo Ogiso
    • 1
  • Shuichi Wada
    • 3
  • Yoshiaki Tabuchi
    • 4
  • Chun-Sang Hong
    • 5
  • Ajai K. Srivastav
    • 6
  • Stephen D. J. Archer
    • 7
  • Stephen B. Pointing
    • 7
    • 8
  • Kazuichi Hayakawa
    • 9
  • Nobuo Suzuki
    • 1
    Email author
  1. 1.Noto Marine Laboratory, Institute of Nature and Environmental TechnologyKanazawa UniversityHousu-gunJapan
  2. 2.Marine and Coastal Research CenterOchanomizu UniversityTateyamaJapan
  3. 3.Department of Animal Bioscience, Faculty of BioscienceNagahama Institute of Bio-Science and TechnologyNagahamaJapan
  4. 4.Division of Molecular Genetics Research, Life Science Research CenterUniversity of ToyamaToyamaJapan
  5. 5.Research and Business Foundation, Hankuk University of Foreign StudiesYongin-siRepublic of Korea
  6. 6.Department of ZoologyD.D.U. Gorakhpur UniversityGorakhpurIndia
  7. 7.Institute for Applied Ecology New ZealandAuckland University of TechnologyAucklandNew Zealand
  8. 8.Yale-NUS CollegeNational University of SingaporeSingaporeSingapore
  9. 9.Institute of Nature and Environmental TechnologyKanazawa UniversityKanazawaJapan

Personalised recommendations