Ocean and Polar Research

Korea Institute of Ocean Science & Technology (한국해양과학기술원)
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Cytochemical Measurement of Lysosomal Responses in the Digestive Cells of Wild Pacific Oyster, Crassostrea gigas on the South Coast of Korea

세포화학적 방법을 이용한 남해안 조간대에 서식하는 참굴(Crassostrea gigas) 소화맹낭 세포 내 리소솜 활동 측정에 관한 연구

  • Jeung, Hee-Do (School of Marine Biomedical Science (POST BK21) and Marine and Environmental Research Institute Jeju National University) ;
  • Lee, Jee-Yeon (School of Marine Biomedical Science (POST BK21) and Marine and Environmental Research Institute Jeju National University) ;
  • Hong, Hyun-Ki (School of Marine Biomedical Science (POST BK21) and Marine and Environmental Research Institute Jeju National University) ;
  • Kang, Hyun-Sil (School of Marine Biomedical Science (POST BK21) and Marine and Environmental Research Institute Jeju National University) ;
  • Kim, Young-Ok (South Sea Research Institute, KIOST) ;
  • Choi, Kwang-Sik (School of Marine Biomedical Science (POST BK21) and Marine and Environmental Research Institute Jeju National University)
  • 정희도 (제주대학교 해양과학대학 해양의생명과학부) ;
  • 이지연 (제주대학교 해양과학대학 해양의생명과학부) ;
  • 홍현기 (제주대학교 해양과학대학 해양의생명과학부) ;
  • 강현실 (제주대학교 해양과학대학 해양의생명과학부) ;
  • 김영옥 (한국해양과학기술원 남해연구소) ;
  • 최광식 (제주대학교 해양과학대학 해양의생명과학부)
  • Received : 2012.03.28
  • Accepted : 2012.05.21
  • Published : 2012.06.30
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Abstract

Digestive cells of the bivalves have a highly developed lysosomal system and the system is known to be sensitive to changes in environmental qualities. In this study, we measured lysosomal responses of the digestive cells in wild oyster, Crassostrea gigas using frozen section. Oysters were collected in June 2010 from intertidal areas in the inner and outer bay of Gwangyang off the south coast of Korea. From the tissue sections, we measured the digestive cell lysosomal membrane stability (LMS), level of neutral lipids (NL), lipofuscin (LF) and the digestive gland atrophy (DGA). The DGA and condition index of oysters from the inner bay were significantly lower (P<0.05). The statistical test indicated that LMS levels of oysters in the inner bay and the outer bay were not significantly different since a shorter activity was displayed by oysters from the inner bay than that of oysters in outer bay. The LF deposition level of the oysters in the inner bay displayed significantly higher levels than the outer bay (P<0.05). In contrast, the NL accumulation measured from oysters in outer bay was significantly higher than the level observed in the inner bay (P<0.05). Different levels of DGA and LF that were observed in the inner and outer bays were thought to be associated to different level of environmental contamination and these two assays are considered to be good biomarkers.

이매패류의 소화맹낭은 고도로 발달된 리소솜 시스템을 지니고 있어 소화작용과 함께 오염물질의 해독과 배출작용을 담당한다. 따라서 소화맹낭의 리소솜 시스템은 환경변화에 민감하게 반응한다고 알려져 있다. 이 연구에서는 동결절편을 이용한 조직학적 방법으로 조간대에 서식하는 참굴 소화맹낭의 리소솜 활동을 측정하였다. 참굴은 2010년 6월 광양만 내만의 초남대교, 차면리와 외만의 신덕리, 평산리 조간대에 서식하는 굴을 채집하였다. 제작된 동결절편을 이용하여 lysosomal membrane stability(LMS) 및 lipofuscin(LF) 축적, neutral lipid(NL) 축적, 소화맹낭 위축도(digestive gland atrophy, DGA)를 평가하였다. 비만도 및 소화맹낭 위축도는 내만에 서식하는 참굴이 다른 지역에 비해 유의적으로 낮았다(P<0.05). LMS는 내만이 외만에 비해 짧은 리소솜 막 불안정화 시간을 보였지만 지역별 통계적인 유의차는 없었다. LF 축적은 내만에 서식하는 참굴이 유의적으로 높은 축적을 보였지만(P<0.05), NL 축적의 경우 외만에 서식하는 참굴이 유의적으로 높은 축적을 보였다(P<0.05). DGA와 LF의 경우 내만과 외만에서 뚜렷한 차이를 보였고, 이는 환경오염의 수준과 관련 있을 것으로 사료된다. 따라서 이 두 가지 분석항목은 연안환경평가에서 좋은 biomarker로 사용될 수 있을 것으로 기대된다.

Keywords

References

  1. 한국해양연구원 (2010) 남해 특별관리해역 생태계 건강지수 개발. 한국해양연구원, BSPE98463-2251-3, 381 p
  2. Bayne BL, Brown DA, Burns K, Dixon DR, Ivanovici A, Livingstone DR, Lowe DM, Moore MN, Stebbing ARD, Widdows J (1984) The effects of stress and pollution on marine animals. Praeger, New York, 384 p
  3. Bianchi TS, Canuel EA (2011) Chemicals biomarkers in aquatic ecosystems. Princeton University Press, Princeton, 396 p
  4. Capuzzo JM, Leavitt DF (1988) Lipid composition of the digestive glands of Mytilus edulis and Carcinus maenas in response to pollutant gradients. Mar Ecol-Prog Ser 46:139-145 https://doi.org/10.3354/meps046139
  5. Cho SM, Jeong WG (2005) Spawning impact on lysosomal stability of the Pacific oyster, Crassostrea gigas. Aquaculture 244:383-387 https://doi.org/10.1016/j.aquaculture.2004.12.013
  6. Chung KW (2003) Ultrastructure of chronic liver disease: Lysosomes and lysosomal storage diseases. Korean J Hepatol 9:344-361
  7. Cuervo AM (2004) Autophagy: In sickness and in health. Trends Cell Biol 14:70-77 https://doi.org/10.1016/j.tcb.2003.12.002
  8. Dimitriadis VK, Domouhtsidou GP, Cajaraville MP (2004) Cytochemical and histochemical aspects of the digestive gland cells of the mussel Mytilus galloprovincialis (L.) in relation to function. J Mol Histo 35:501-509
  9. Eble AF, Scro R (1996) General anatomy. In: Kennedy VS, Newell RIE (eds) The eastern oyster Crassostrea virginica. Maryland Sea Grant College, College Park, pp 19-73
  10. Ferreira A, Dolder H (2003) Cytochemical study of spermiogenesis and mature spermatozoa in the lizard Tropidurus itambere (Reptilia, Squamata). Acta Histochem 105:339-352 https://doi.org/10.1078/0065-1281-00731
  11. Fink S (1986) A new integrated concept for the improved preparation of sections of fresh or frozen tissue for light microscope histochemistry. Histochemistry 86:43-52 https://doi.org/10.1007/BF00492344
  12. Hauton C, Hawkins LE, Hutchinson S (2001) Response of haemocyte lysosomes to bacterial inoculation in the oysters Ostrea edulis L. and Crassostrea gigas (Thunberg) and the scallop Pecten maximus (L.). Fish Shellfish Immun 11:143-153 https://doi.org/10.1006/fsim.2000.0301
  13. Hawkins HK (1980) Reactions of lysosomes to cell injury. In: Trump BF, Arstila AV (eds) Pathobiology of cell membranes. Vol 2. Academic Press, New York, pp 252-285
  14. Hong SH, Kannan N, Yim UH, Choi JW, Shim WJ (2011) Polychlorinated biphenyls (PCBs) in a benthic ecosystem in Gwangyang Bay, South Korea. Mar Pollut Bull 62: 2863-2868 https://doi.org/10.1016/j.marpolbul.2011.09.005
  15. Howard D, Smith C (1983) Histological techniques for marine bivalve mollusks. NOAA technical memorandum NMFS-F/NEC-25. Woods Hole, MA, 97 p
  16. Huggett RJ, Kimerle RA, Mehrle PM, Bergman HL (1992) Biomarkers. Lewis Publishers, Boca Raton, 347 p
  17. Jo QT, Choy EJ, Park DW, Jee YJ, Kim SY, Kim Y (2002) Cellular biomarker of membrane stability and hydrolytic enzyme activity in the hemocytes of benzo (a) pyreneexposed Pacific oyster, Crassotrea gigas. J Fish Sci Tech 5:263-270
  18. Kagley AN, Snider RG, Krishnakumar PK, Casillas E (2003) Assessment of seasonal variability of cytochemical responses to contaminant exposure in the blue mussel Mytilus edulis (Complex). Arch Environ Con Tox 44:43-52 https://doi.org/10.1007/s00244-002-1303-3
  19. Kang DH, Choi KS (1999) Evaluation of methods used in the calculation of condition index using the Mussel, Mtilus coruscus (Gould, 1861) collected from Chuja Island, Cheju, Korea. Korean J Malacol 15:57-62
  20. Kang DH, Chu FE, Yang HS, Lee CH, Koh HB, Choi KS (2010) Growth, reproductive condition, and digestive tubule atrophy of Pacific oyster Crassostrea gigas in Gamakman bay off the southern coast of Korea. J Shellfish Res 29:839-845 https://doi.org/10.2983/035.029.0418
  21. Klionsky DJ, Emr SD (2000) Autophagy as a regulated pathway of cellular degradation. Science 290:1717-1721 https://doi.org/10.1126/science.290.5497.1717
  22. Kramer KJM (1994) Biomonitoring of coastal waters and estuaries. CRC Press, Boca Raton, 327 p
  23. Krishnakumar PK, Casillas E, Varanasi U (1994) Effect of environmental contaminants on the health of Mytilus edulis from Puget Sound, Washington, USA. I. Cytochemical measures of lysosomal responses in the digestive cells using automatic image analysis. Mar Ecol-Prog Ser 106:249-261 https://doi.org/10.3354/meps106249
  24. Li Y, Qin Y, Li H, Wu R, Yan C, Du H (2007) Lysosomal acid lipase over-expression disrupts lamellar body genesis and alveolar structure in the lung. Int J Exp Path 88:427-436 https://doi.org/10.1111/j.1365-2613.2007.00547.x
  25. Livingstone DR (1991) Organic xenobiotics metabolism in marine invertebrates. In: Giles R (ed) Advances in comparative and environmental physiology. Springer-Verlag, Berlin, pp 45-185
  26. Lowe DM (1988) Alterations in the cellular structure of Mytilus edulis resulting from exposure to environmental contaminants under field and experimental conditions. Mar Ecol-Prog Ser 46:91-100 https://doi.org/10.3354/meps046091
  27. Lowe DM, Moore MN, Clarke KE (1981) Effect of oil on digestive cells in mussels: quantitative alterations in cellular and lysosomal structure. Aquat Toxicol 1:213-226 https://doi.org/10.1016/0166-445X(81)90016-3
  28. Lullman-Rauch R (1979) Drug-induced lysosomal storage disorders. In: Dingle JT, Jacques PJ, Shaw IH (eds) Lysosomes in biology and pathology. North-Holland, Aamsterdam, pp 49-130
  29. Moore MN (1976) Cytochemical demonstration of latency of lysosomal hydrolases in digestive cells of the common mussel, Mytilus edulis, and changes induced by thermal stress. Cell Tissue Res 175:279-287
  30. Moore MN (1985) Cellular responses to pollutants. Mar Pollut Bull 16:164-169 https://doi.org/10.1016/0025-326X(85)90008-6
  31. Moore MN (1988) Cytochemical responses of the lysosomal system and NADPH-ferrihemoprotein reductase in molluscs to environmental and experimental exposure to xenobiotics. Mar Ecol-Prog Ser 46:81-89 https://doi.org/10.3354/meps046081
  32. Moore MN, Allen JI, McVeigh A (2006) Environmental prognostics: An integrated model supporting lysosomal stress responses as predictive biomarkers of animal health status. Mar Environ Res 61:278-304 https://doi.org/10.1016/j.marenvres.2005.10.005
  33. Moore MN, Livingstone DR, Widdows J (1989) Hydrocarbons in marine mollusks: biological effects and ecological consequences. In: Varanasi U (ed) Metabolism of polycyclic aromatic hydrocarbons in the aquatic environment. CRC press. Boca Raton, pp 303-310
  34. Moore MN, Lowe DM, Koehler A (2004) Biological effects of contaminants: Measurements of lysosomal membrane stability. ICES Techniques in Marine Environmental Sciences (TIMES), Vol 36. ICES, Copenhagen, 31 p
  35. Moore MN, Viarengo A, Donkin P, Hawkins AJS (2007) Autophagic and lysosomal reactions to stress in the hepatopancreas of blue mussels. Aquat Toxicol 84:80-91 https://doi.org/10.1016/j.aquatox.2007.06.007
  36. O'Connor TP, Lauenstein GG (1989) Ten-year trends in chemical contamination in mussels and oysters. J Shellfish Res 8:452
  37. Petrovic S, Ozretic B, Krajnovic-Ozretic M, Bobinac D (2001) Lysosomal membrane stability and metallothioneins in digestive gland of mussels (Mytilus galloprovincialis Lam.) as biomarker in a field study. Mar Pollut Bull 42:1373-1378 https://doi.org/10.1016/S0025-326X(01)00167-9
  38. Pipe RK (1993) The generation of reactive oxygen metabolites by the haemocytes of the mussel Mytilus edulis. Dev Comp Immunol 16:111-122
  39. Raftopoulou EK, Dailianis S, Dimitriadis VK, Kaloyianni M (2006) Introduction of cAMP and establishment of neutral lipid alterations as pollution biomarkers using the mussel Mytilus galloprovincialis. Correlation with a battery of biomarker. Sci Total Environ 368:597-614 https://doi.org/10.1016/j.scitotenv.2006.04.031
  40. Ringwood AH, Hoguet J, Keppler CJ (2002) Seasonal variation in lysosomal destabilization in oyster, Crassostrea virginica. Mar Environ Res 58:793-797
  41. Shim WJ, Kahng SH, Hong SH, Kim NS, Kim SK, Shim JH (2000) Imposex in the rock shell, Thais clavigera, as evidence of organotin contamination in the marine environment of Korea. Mar Environ Res 49:435-451 https://doi.org/10.1016/S0141-1136(99)00084-7
  42. Terman A, Brunk UT (2004) Molecules in focus lipofuscin. Int J Biochem Cell Biol 36:1400-1404 https://doi.org/10.1016/j.biocel.2003.08.009
  43. Viarengo A, Lowe D, Bolognesi C, Fabbri E, Koehler A (2007) The use of biomarker in biomonitoring: A 2-tier approach assessing the level of pollutant-induced stress syndrome in sentinel organisms. Comp Biochem Phys C 146:281-300
  44. Viarengo A, Moore MN, Mancinelli G, Mazzucotelli A, Pipe RK, Farrar SV (1987) Metallothioneins and lysosomes in metal toxicity and accumulation in marine mussels: the effect of cadmium in the presence and absence of phenathrene. Mar Biol 94:251-257 https://doi.org/10.1007/BF00392937
  45. Viarengo A, Nott J (1993) Mechanisms of heavy metals cation homeostasis in marine invertebrates. Comp Biochem Phys C 104:355-372 https://doi.org/10.1016/0742-8413(93)90001-2
  46. Winstead JT (1995) Digestive tubule atrophy in eastern oyster, Crassotrea virgicina (Gmelin 1791), exposed to salinity and starvation stress. J Shellfish Res 14:105-111
  47. Zweytick D, Athenstaedt K, Daum G (2000) Intracellular lipid particles of eukaryotic cells. Biochem Biophys Acta 1469:101-120 https://doi.org/10.1016/S0005-2736(00)00294-7

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