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카드뮴에 노출된 바지락 (Ruditapes philippinarum) 의 세포성 면역 반응

Cellular Immune Responses of the Manila Clam, Ruditapes philippinarum, Exposed to Cadmium Chloride

초록

바지락 (Ruditapes philippinarum) 은 우리나라 조간대에 널리 분포하며 잘 발달된 생체조절 능력을 가지고 있어 환경모니터링 종으로서 가치가 높은 생물이다. 본 연구는 산업활동으로 해양환경에서 그 농도가 높아지고 있으며 생물체에 강한 독성을 가진 카드뮴이 바지락의 세포성 면역에 미치는 영향을 연구함으로써 바지락의 면역 기작을 이해하고 환경 모니터링 기법으로의 적용 가능성을 조사하기 위하여 실시되었다. 실험을 위하여 바지락은 $50\;{\mu}g/l$의 카드뮴에 8일간 노출되었으며, 유세포 분석기를 이용하여 혈구의 미분화 세포의 구성비율, DNA 손상도, 혈구괴사율, 혈구 apoptosis 비율 및 혈구사망율을 측정하였다. 조사 결과 카드뮴에 노출된 바지락은 대조구와 비교하여 미분화세포, DNA 손상도, 세포괴사율, 세 apoptosis 비율 및 세포 사망율이 통계적으로 유의하게 증가하였다. 특히 apoptosis 비율은 세포괴사율보다 높게 증가함에 따라 바지락 혈구의 경우 본 연구에서 사용된 카드뮴의 농도는 세포괴사보다는 apoptosis를 유도하는데 더 효과적인 농도인 것으로 판단된다. 본 연구를 통하여 중금속 오염에 대한 바지락의 세포성 면역반응은 유세포 분석기를 이용하여 정량이 가능하였으며, 세포성 면역반응을 이용한 환경모니터링에 유용하게 사용될 수 있을 것으로 기대된다.

keywords
ruditapes philippinarum, cadmium chloride, flow cytometry, immune parameter, environmental monitoring

Abstract

The Manila clam, Ruditapes philippinarum, has been considered as a sentinel species due to dominant distribution along the coast of Korea and well developed regulatory system. In order to develop and understand immune responses of the Manila clams, clams were exposed to $50\;{\mu}g/L$ of cadmium chloride (Cd) for 8 days and monitored the cellular immune parameters of the hemocytes including blast cell composition, DNA damage, necrosis, apoptosis and hemocyte mortality using a flow cytometer. The results showed that all immune parameters analyzed in the present study increased remarkably compared to the controls and the increases were statistically significant. Apoptosis rate was higher than necrosis rate in the clams exposed to Cd suggesting that apoptosis was preferably induced by the concentration of Cd used in the present study. Our study indicates that the measurement of cellular immune responses of the Manila clam using flow cytometer will be a useful technique for assessment of heavy metal contamination in marine environment.

keywords
ruditapes philippinarum, cadmium chloride, flow cytometry, immune parameter, environmental monitoring

참고문헌

1.

(1995). . Techniques in Fish Immunology and Pathology of Aquatic Invertebrates, 4, 55-64.

2.

Auffret. (1997). . Comparative Biochemistry and Physiology Part A Physiology, 118(3), 705-712. 10.1016/S0300-9629(97)00017-0.

3.

Sabourault, C.;de Sousa, G.;Amichot, M.;Cuany, A.;Rahmani, R.;Salaun, J.-P.;Berge, J.-B.. (1999). Tissue-specific induction and inactivation of cytochrome P450 catalysing lauric acid hydroxylation in the sea bass, Dicentrarchus labrax. Comparative Biochemistry and Physiology Part B Biochemistry and Molecular Biology, 122(2), 253-263. 10.1016/S0305-0491(99)00006-1.

4.

Brousseau, P.;Pellerin, J.;Morin, Y.;Cyr, D.;Blakley, B.;Boermans, H.;Fournier, M.. (1999). Flow cytometry as a tool to monitor the disturbance of phagocytosis in the clam Mya arenaria hemocytes following in vitro exposure to heavy metals.. Toxicology, 142(2), 145-156. 10.1016/S0300-483X(99)00165-1.

5.

(2002). . The Science of The Total Environment, 247, 295-311.

6.

CHENG. (1988). . Journal of Invertebrate Pathology, 51(3), 207-214. 10.1016/0022-2011(88)90027-4.

7.

Coughlan, B.M.;Hartl, M.G.J.;O'Reilly, S.J.;Sheehan, D.;Morthersill, C.;van Pelt, F.N.A.M.. (2002). Detecting genotoxicity using the Comet assay following chronic exposure of Manila clam Tapes semidecussatus to polluted estuarine sediments. Marine Pollution Bulletin, 44(12), 1359-1365. 10.1016/S0025-326X(02)00254-0.

8.

(1996). . Microbiology Review, 60, 641-696.

9.

Allam, B.;Ashton-Alcox, K.A.;Ford, S.E.. (2002). Flow cytometric comparison of haemocytes from three species of bivalve molluscs. Fish & Shellfish Immunology, 13(2), 141-158. 10.1006/fsim.2001.0389.

10.

11.

Dailianis, S.. (2009). Production of superoxides and nitric oxide generation in haemocytes of mussel Mytilus galloprovincialis (Lmk.) after exposure to cadmium: A possible involvement of Na<SUP>+</SUP>/H<SUP>+</SUP> exchang. Fish & Shellfish Immunology, 27(3), 446-453. 10.1016/j.fsi.2009.06.016.

12.

Kim, J.;Soh, J.. (2009). Cadmium-induced apoptosis is mediated by the translocation of AIF to the nucleus in rat testes. Toxicology Letters, 188(1), 45-51. 10.1016/j.toxlet.2009.03.006.

13.

Rittschof, D.;McClellan-Green, P.. (2005). Molluscs as multidisciplinary models in environment toxicology. Marine Pollution Bulletin, 50(4), 369-373. 10.1016/j.marpolbul.2005.02.008.

14.

Tsangaris, G.T.;Tzortzatou-Stathopoulou, F.. (1998). Cadmium induces apoptosis differentially on immune system cell lines. Toxicology, 128(2), 143-150. 10.1016/S0300-483X(98)00032-8.

15.

(2000). . Journal of Inorganic Biochemistry, 79, 241–244.

16.

Waisberg. (2003). . Toxicology, 192(2-3), 95-117. 10.1016/S0300-483X(03)00305-6.

17.

Walsh, G.M.;Dewson, G.;Wardlaw, A.J.;Levi-Schaffer, F.;Moqbel, R.. (1998). A comparative study of different methods for the assessment of apoptosis and necrosis in human eosinophils. Journal of Immunological Methods, 217(1), 153-163. 10.1016/S0022-1759(98)00103-3.

18.

(2003). . Comparative Biochemistry and Physiology Part C, 134, 227-234.

19.

Lau, P.S.;Wong, H.L.. (2003). Effect of size, tissue parts and location on six biochemical markers in the green-lipped mussel, Perna viridis. Marine Pollution Bulletin, 46(12), 1563-1572. 10.1016/S0025-326X(03)00321-7.

20.

Lee. (1999). . Journal of Biological Chemistry, 274(28), 19792-19798. 10.1074/jbc.274.28.19792.

21.

Oceanography and Marine Environment of the Basque Country. (2004). . Elsevier Oceanography Series, 70(70), 335-364. 10.1016/S0422-9894(04)80052-7.

22.

Micic. (2001). . Aquatic Toxicology, 55(1-2), 61-73. 10.1016/S0166-445X(01)00156-4.

23.

O'Connor, T.P.. (2002). National distribution of chemical concentrations in mussels and oysters in the USA. Marine Environmental Research, 53(2), 117-143. 10.1016/S0141-1136(01)00116-7.

24.

(2006). . Journal of Korean Fisheries Society, 39, 123-131.

25.

Pipe, R.K.;Coles, J.A.. (1995). Environmental contaminants influencing immunefunction in marine bivalve molluscs. Fish & Shellfish Immunology, 5(8), 581-595. 10.1016/S1050-4648(95)80043-3.

26.

Rank, J.;Jensen, K.;Jespersen, P.H.. (2005). Monitoring DNA damage in indigenous blue mussels (Mytilus edulis) sampled from coastal sites in Denmark. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 585(1), 33-42. 10.1016/j.mrgentox.2005.04.008.

27.

Risso-de Faverney, C.;Devaux, A.;Lafaurie, M.;Girard, J.P.;Bailly, B.;Rahmani, R.. (2001). Cadmium induces apoptosis and genotoxicity in rainbow trout hepatocytes through generation of reactive oxygene species. Aquatic Toxicology, 53(1), 65-76. 10.1016/S0166-445X(00)00154-5.

28.

Hegaret, H.;Wikfors, G.H.;Soudant, P.. (2003). Flow-cytometric analysis of haemocytes from eastern oysters, Crassostrea virginica, subjected to a sudden temperature elevation - I. Haemocyte types and morphology. Journal of Experimental Marine Biology and Ecology, 293(2), 237-248. 10.1016/S0022-0981(03)00236-3.

29.

Hegaret, H.;Wikfors, G.H.;Soudant, P.. (2003). Flow cytometric analysis of haemocytes from eastern oysters, Crassostrea virginica, subjected to a sudden temperature elevation - II. Haemocyte functions: aggregation, viability, phagocytosis, and res. Journal of Experimental Marine Biology and Ecology, 293(2), 249-265. 10.1016/S0022-0981(03)00235-1.

30.

Ji, J.;Choi, H.J.;Ahn, I.Y.. (2006). Evaluation of Manila clam Ruditapes philippinarum as a sentinel species for metal pollution monitoring in estuarine tidal flats of Korea: Effects of size, sex, and spawning on baseline accumulation. Marine Pollution Bulletin, 52(4), 447-453. 10.1016/j.marpolbul.2005.12.012.

31.

(2000). . Journal of Fish Pathology, 13, 129-136.

32.

Gutierrez, A.M.;Reboredo, G.R.;Mosca, S.M.;Catala, A.. (2007). Non-enzymatic lipid peroxidation of microsomes and mitochondria from liver, heart and brain of the bird Lonchura striata: Relationship with fatty acid composition. Comparative Biochemistry and Physiology - Part A Molecular & Integrative Physiology, 146(3), 415-430. 10.1016/j.cbpa.2006.11.018.

33.

(2007). . Korean Journal of Food Science and Technology, 39, 353-359.

34.

(2003). . Journal of Food Hygiene and. Safety, 18, 125-132.

35.

Lambert, C.;Soudant, P.;Choquet, G.;Paillard, C.. (2003). Measurement of Crassostrea gigas hemocyte oxidative metabolism by flow cytometry and the inhibiting capacity of pathogenic vibrios. Fish & Shellfish Immunology, 15(3), 225-240. 10.1016/S1050-4648(02)00160-2.

36.

(1988). . American Fisheries Society Special Publication, 18, 225-237.

37.

Gielazyn. (2003). . Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 542(1-2), 15-22. 10.1016/j.mrgentox.2003.07.010.

38.

Galan, A.;Garcia-Bermejo, L.;Troyano, A.;Vilaboa, N.E.;Fernandez, C.;de Blas, E.;Aller,. (2001). The role of intracellular oxidation in death induction (apoptosis and necrosis) in human promonocytic cells treated with stress inducers (cadmium, heat, X-rays). European Journal of Cell Biology, 80(4), 312-320. 10.1078/0171-9335-00159.

39.

(2006). . General and Comparative Endocrinololgy, 145, 133–139.

40.

Giamberini, L.;Cajaraville, M.P.. (2005). Lysosomal responses in the digestive gland of the freshwater mussel, Dreissena polymorpha, experimentally exposed to cadmium. Environmental Research, 98(2), 210-214. 10.1016/j.envres.2004.11.003.

41.

Goedken, M.;De Guise, S.. (2004). Flow cytometry as a tool to quantify oyster defence mechanisms. Fish & Shellfish Immunology, 16(4), 539-552. 10.1016/j.fsi.2003.09.009.

42.

Goldberg. (1986). . Environmental Monitoring and Assessment, 7(1), 91-103. 10.1007/BF00398031.

43.

44.

Hegaret, H.;da Silva, P.M.;Wikfors, G.H.;Lambert, C.;De Bettignies, T.;Shumway, S.E.;Sou. (2007). Hemocyte responses of Manila clams, Ruditapes philippinarum, with varying parasite, Perkinsus olseni, severity to toxic-algal exposures. Aquatic Toxicology, 84(4), 469-479. 10.1016/j.aquatox.2007.07.007.

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