Journal of Aquaculture (한국양식학회지)

The Korean Society of Fisheries and Aquatic Science (한국양식학회)
권호 표지

Use of the Comet Assay to Assess DNA Damage in Hemocytes and Gill of Oyster(Crassostrea gigas) Exposed to Pyrene and Benzo(a)pyrene

Pyrene과 Benzo(a)pyrene에 노출된 굴의 혈구세포과 아가미 세포에서의 DNA손상 측정을 위한 Comet assay의 이용

  • 김기범 (경상대학교 해양환경공학과, 해양산업연구소) ;
  • 배세진 (한국해양연구원 생태환경연구본부)
  • Published : 2003.08.01
Download PDF
⟨ Previous Next ⟩

Abstract

Sessile organisms such as the oyster Crassostrea gigas have been given much attention as a potential biomonitoring indicator to assess the impact of toxicants on aquatic organism. In this study, we exposed cells isolated from gill of oyster (Crassostrea gigas) to hydrogen peroxide in vitro. In addition oysters were in vivo exposed to pyrene and benzo(a)pyrene at various concentrations for 2 weeks. Comet assay was used to detect DNA single strand breaks and to investigate the application of this technique as a tool for aquatic biomonitoring. Hydrogen peroxide increased DNA single strand break with increasing concentration after 30 minutes exposure in vitro. Pyrene and benzo(a)pyrene caused DNA damage only at very high concentration (100 $\mu\textrm{g}$/L or 1000 $\mu\textrm{g}$/L) at two week exposure in vivo. DNA damage was relatively higher at hemocyte than at gill. It suggested that metabolized PAHs are transferred to hemolymph from digestive gland which have a relatively high enzyme activity, and attacked the DNA of hemocyte, while gill accumulated PAHs without degrading them to their metabolites due to low enzyme activity at gill. Both in vitro and in vivo exposure experiments showed that the comet assay is an effective tool on screening whether the organism are exposed to genotoxic contaminants.

Keywords

References

  1. Environ., Res. v.74 Effect of in vitro exposure to tributyltin on generation of oxygen metabolites by oyster hemocytes Anderson,R.S.;L.L.Brubacher;L.M.R.Calvo;E.M.Burreson;M.A.Unger https://doi.org/10.1006/enrs.1997.3751
  2. Ecotoxicol. Environ. Safety v.33 The effects of near ultraviolet radiatio on the toxic effects of polycyclic aromatic hydrocarbons in animals and plants: A review Arfsten,A.P.;D.J.Schaeffer;D.C.Mulveny https://doi.org/10.1006/eesa.1996.0001
  3. Toxicological profile polycyclic aromatic hydrocarbons(PAHs) ATSDR(Agency for Toxic Substances and Disease Registry)
  4. J. Aquaculture v.15 Hematological characteristics of olive flounder (Paralichthys olivaceus) in culture farm influenced by heated effluent water from a power plant in summer Chang,Y.J.;J.W.Hur;P.Chin
  5. Mar. Pollut. Bull. v.44 Detecting genotoxicity using the comet assay following chronic exposure of Manila clam Tapes semidecus satus to polluted estuarine sediments Coughlan,B.M.;M.G.J.Hartl;S.J.O'Reilly;D.Sheehan;C.Morthersill;F.N.A.M.van Pelt;J.O'Halloran;N.M.O'Brien https://doi.org/10.1016/S0025-326X(02)00254-0
  6. Environ. Sci. Technol. v.17 US Mussel Watch 1976-1978: An overview of the trace metal, DDE, PCB, hydrocarbon and artificial radionuclide data Farrington,J.W.;E.D.Goldberg;R.W.Risebrough;J.H.Martin;V.T.Bowen https://doi.org/10.1021/es00114a010
  7. Estuar. Coast. Mar. Sci. v.1 Petroleum hydrocarbons in Narragansett Bay. I. Survey of hydrocarbons in sediments and clams (Mercenaria mercenaria) Farrington,J.W.;J.G.Quinn https://doi.org/10.1016/0302-3524(73)90059-5
  8. Aquat. Toxicol. v.59 Effects of exposure of Mya arenaria and Mactromeris polynyma to contaminated marine sediments on phargocytic activity of hemocytes Fournier,M.;J.Pellerin;M.Lebeuf;P.Brousseau;Y.Morin;D.Cyr https://doi.org/10.1016/S0166-445X(01)00240-5
  9. FEBS Lett. v.281 DNA damage by oxygen derived species. Its mechanism and measurement in mammalian system Halliwell,B.;O.I.Aruma https://doi.org/10.1016/0014-5793(91)80347-6
  10. Mar. Environ. Res. v.54 Use o the Comet assay to assess DNA damage in hemocytes and diestive gland cells of mussels and clams exposed to water contaminated with petroleum hydrocarbons Hamoutene,D.;J.F.Payne;A.Rahimtula;K.Lee https://doi.org/10.1016/S0141-1136(02)00162-9
  11. Science v.240 DNA damage and oxygen radical toxicity Imlay,J.A.;S.Linn https://doi.org/10.1126/science.3287616
  12. Environ. Pollut. v.83 Polynuclear aromatic hydrocarbon contaminants in oysters from the Gulf of Mexico Jackson,T.J.;T.L.Wade;T.J.McDonald;D.L.Wilkinson;J.M.Brooks https://doi.org/10.1016/0269-7491(94)90150-3
  13. Korean J. Food Sci. Technol. v.32 Discrimination of irradiated beef using comet assay Jeong,S.K.;J.H.Park;S.T.i;K.J.Park;H.H.Kim;C.K.Hyun;H.K.Shin
  14. J. Korean Fish Soc. Application of single cell gel electrophoresis for detection of DNA single strand breaks in DNA of fish blood cell Kim,G.B.;R.F.Lee;K.Maruya
  15. Mar. Pollut. Bull. v.38 Disribution and source of polycyclic aromatic hydrocarbons in the vicinity of Incheon Harbor, Korea Kim,G.B.;K.A.Maruya;R.F.Lee;J.H.Lee;C.H.Koh;S.Tanabe https://doi.org/10.1016/S0025-326X(98)00077-0
  16. J. Korean Asso. Radiat. Prot. v.24 Evaluation of protective effect of peach kernel extracs on radiation induced DNA damage in human blood lymphocytes in the single cell gel electrophoresis assay Kim,J.K.;T.W.Park;C.J.Lee;Y.G.Chai
  17. Mar. Environ. Res. v.35 The genotoxic disease syndrome Kurelec,B. https://doi.org/10.1016/0141-1136(93)90101-5
  18. Mar. Environ. Rea. v.54 Different levels of mussel (Mytilus edulis) DNA strand breaks following chronic field and acute laboratory exposure to polycyclic aromatic hydrocarbons Large,A.T.;J.P.Shaw;L.D.Peters;A.D.McIntosh;L.Webster;A.Mally;J.K.Chipman https://doi.org/10.1016/S0141-1136(02)00139-3
  19. J. Korean Soc. Food Sci. Nutr. v.30 Dectection characteristics of TL, ESR and DNA comet for irradiated peanuts by origins Lee,E.;J.Jung;D.Jo;J.H.Kwon
  20. Mar. Environ. Res. v.54 Comet assays to assess DNA damage and repair in grass shrimp embryos exposed to phototoxicants Lee,R.;G.B.Kim https://doi.org/10.1016/S0141-1136(02)00128-9
  21. Mutat. Res. Use of the single cell gel electrophoresis/comet assay for detecting DNA damage in aquatic (marine and freshwater) animals Lee,R.;S.Steinert
  22. Mar. Environ. Res. v.54 Phototoxicity of pyrene and benzo(a)pyrene to embryo-larval stages of the pacific oyser Crassostrea gigas Lyons,B.P.;C.K.Pasoe;I.R.B.McFadzen https://doi.org/10.1016/S0141-1136(02)00124-1
  23. Aquat. Toxicol. v.41 Detection of DNA strand breaks in brown trout (Salmo trutta) hepatocytes and blood cells using the single cell gel electrophoresis (comet) assay Mitchelmore,C.L.;J.K.Chipman https://doi.org/10.1016/S0166-445X(97)00064-7
  24. Aquat. Toxicol. v.41 Evidence for cytochrome P-450 catalysis and free radical involvement in the reproduction of DNA strand breaks by benzo[a]pyrene and nitroaromatics in mussel (Mytilus edulis L.) digestive gland cells Mitchelmore,C.L.;C.Birmelin;J.K.Chipman;D.R.Livingstone https://doi.org/10.1016/S0166-445X(97)00083-0
  25. Ecotoxicol. Environ. Safety v.41 Detection of DNA strand breaks in isolated mussel (Mytilus edulis L.) digestive gland cells usin the come assay Mitchelmore,C.L.;C.Birmelin;D.R.Livingstone;J.K.Chipman https://doi.org/10.1006/eesa.1998.1666
  26. J. Korean Soc. Oceanol. v.36 Polycyclic aromatic hydrocarbons in sediments from the southeastern coastal areas of Korea Moon,H.B.;H.G.Choi;S.S.Kim;P.J.Kim;P.Y.Lee;G.Ok
  27. Aquat. Toxicol. v.35 Detection of DNA damage in individual cells from marine organism using the single cell gel assay Nacci,D.E.;S.Cayula;E.Jackim https://doi.org/10.1016/0166-445X(96)00016-1
  28. Chemosphere v.50 Distribution of polycyclic aromatic hydrocarbons in agricultural soils in South Korea Nam,J.J.;B.H.Song;K.C.Eom;S.H.Lee;A.Smith https://doi.org/10.1016/S0045-6535(02)00764-6
  29. Biochem. Biophys. Res. Commun. v.123 Microelectrophoretic study of radiation-induced DNA damages in individual mammalian cells Ostling,O.;K.J.Johanson https://doi.org/10.1016/0006-291X(84)90411-X
  30. Nature v.303 Fifty years of benzo(a)pyrene Phillips,D.H. https://doi.org/10.1038/303468a0
  31. Experientia v.51 The effect of chronic heat stress on cortisol levels in the Antarctic fish Pagothenia borchgrevinka Ryan,S.N. https://doi.org/10.1007/BF01922428
  32. Biomarkers of Environmental Contamination Shugart,L.R.
  33. Exp. Cell Res. v.175 A simple technique for quantitation of low levels of DNA damage in individual cells Singh,N.P.;M.T.McCoy;R.R.Tice;E.L.Schneider https://doi.org/10.1016/0014-4827(88)90265-0
  34. Mar. Environ. Res. v.42 Contribuion of apoptosis to observed DNA damage in mussel cells Steinert,S.A. https://doi.org/10.1016/0141-1136(95)00056-9
  35. Prog. Nucleic Acid Res. Mol. Biol. v.35 DNA damage produced by ionizing radiation in mammalian cells: identities, mechanism of formation and reparability Ward,J.F. https://doi.org/10.1016/S0079-6603(08)60611-X
  36. Mutat. Res. v.399 Evaluation of the comet assay as a method for the detection of DNA damage in the cells of a marine invertebrate, Mytilus edulis L. (Mollusca: Pelecypoda) Wilson,J.T.;P.L.Pascoe;J.M.Parry;D.R.Dixon https://doi.org/10.1016/S0027-5107(97)00268-6