多氯联苯对孔雀鱼卵黄蛋白原的诱导及检测Induction and Determination of Vitellogenin in Male Guppies(Poecilia reticulata)after Exposed to Polychlorinated Biphenyls
詹翠琼;黄絮洁;方展强;马广智;姚静;
摘要(Abstract)：
为探讨利用孔雀鱼(Poecilia reticulata)卵黄蛋白原(Vitellogenin,Vtg)作为生物标志物检测环境雌激素的可行性,采用浸浴法分别使用17β-雌二醇(E2)和多氯联苯(PCBs)对雄性孔雀鱼成鱼进行染毒,30d后测定其性腺系数(GSI)和肝指数(LSI),并将鱼体整体匀浆进行常规聚丙烯酰胺凝胶电泳(Native-PAGE)及磷、脂和糖蛋白特异染色分析.结果表明,PCBs对雄性孔雀鱼具有雌激素效应,和E2均可以诱导雄性孔雀鱼体内产生Vtg,但诱导组雄鱼GSI及LSI与对照组比较均无显著性差异(p>0.05).Native-PAGE及磷、脂、糖蛋白特异性分析表明,孔雀鱼Vtg是一种富含磷、脂、糖的蛋白,具有VtgⅠ、VtgⅡ和VtgⅢ等3种形式,其分子量分别为642kDa、541kDa和441kDa.雄性孔雀鱼Vtg可作为环境雌激素监测的有效生物标记物.
关键词(KeyWords)： 17β-雌二醇;多氯联苯;卵黄蛋白原;常规聚丙烯酰胺凝胶电泳;孔雀鱼
基金项目(Foundation): 广东省科技计划项目(No.2004B40101015;No.2006B36601008); 华南师范大学2004-2005年度学生课外科研项目(No.67)
作者(Author): 詹翠琼;黄絮洁;方展强;马广智;姚静;
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参考文献(References)：
- Anderson M J,Olsen H,Matsumura F,Hinton D E.1996.Invivo modulation of17β-estradiol induced vitellogenin synthesis andestrogen receptor in rainbow trout(Oncorhynchus mykiss)livercells by b-naphthoflavone[J].Toxicol Appl Pharmacol,137:210-218
- Bayley M,Jacob R.Nielsen J R.1999.Guppy sexual behavioras an effect biomarker of estrogen mimics[J].Ecotoxicology andEnvironmental Safety,43:68-73
- Bergeron J M,Crews D,McL achlan J A.1994.PCBs asenvironmental estrogens:turtle sex determination as a biomarker ofenvironmental contamination[J].Environ Health Persp,102:780-781
- Bing X,Ru S G,Isoda H,Jiang M,Wang W,Wang S H.2004.The identification of vitellogenin induction by17β-estradiolin male Carassius auratus[J].Journal of Fisheries of China,28(3):236-240(in Chinese)
- Cardinali M,Maradonna F,Olivotto I,Bortoluzzi G,Mosconi G,Polzonetti-Magni A M,Carnevali O.2004.Temporary impairmentof reproduction in freshwater teleost exposed to nonylphenol[J].Reproductive Toxicology,18:597-604
- Christiansen L B,Pedersen K L,Ko rsgaard B,Bjerregaard B.1998.Estrogenicity of xenobiotics in rainbow trout(Oncorhynchusmykiss)using in vivo synthesis of vitellogenin as a biomarker[J].Mar Environ Res,46:137-140
- Cutting J A,Roth T F.1973.Stain ing of phospho-proteins onacrylamide gel electropherograms[J].Anal Biochem,54:386-394
- Fairbank G,Steck T L,Wallice D F H.1971.Electrophoroeticanalysis of the major polypeptide of the human erythrocytemembrane[J].Biochem J,10:2606-2617
- Folin L,Norrgren L.1998.Physiological and morphological studiesof feral perch before after remediation of a PCB contaminate lake:J-rnsj"n[J].Ambio,27(5):418-423
- Kinnberg K,Toft G.2003.Effects of estrogenic and antiandrogeniccompounds on the testis structure of the adult guppy(Poeciliareticulata)[J].Ecotoxicology and Environmental Safety,54:16-24
- Li K,Zhou Z L,Yu J,Wang M S,Yu X J.2003.Developmentof ELISA for detecting Carassius auratus vitellogenin[J].ChinaEnvironmental Science,23(3):276-280(in Chinese)
- Li M H,Wang Z R.2005.Effect of nonylphenol on plasmavitellogenin of male adult guppies(Poecilia reticulata)[J].Environmental Toxicology,20(1):53-59
- Ohkubo N,Mochida K,Adachi S,Hara A,Hotta K,NakamuraY,Matsubara T.2003.Development of enzyme-linked immunosorbentassays for two forms of vitellogenin in Japanese common goby(Acanthogobius flavimanus)[J].General and ComparativeEndocrinology,3:353-364
- Qin Z F,Zhou J M,Chu S G,Xu X B.2003.Effects of Chinesedomestic polychlorinated biphenyls(PCBs)on gonadal differentiationin Xenopus laevis[J].Environ Persp,114:553-556
- Roub al W T,Lomax D P,Willis M L,Johnson L L.1997.Purification and partial characterization of English sole(Pleuronectesvetulus)vitellogenin[J].Comp Biochem Physiol,118B:613-622
- Sumpter J P,Jobling S.1995.Vitellogenesis as a biomarker forestrogenic contamination of the aquatic environment[J].EnvironHealth Perspect,103(suppl.7):173-178
- Sun X T,Zhang S C.2001.Purification and characterization of aputative vitellogenin from the ovary of amphioxus(Branchiostomabelcheri tsingtaunese)[J].Comp Biochem physical,129B:121-127
- Toft G,Baatrup E.2001.Sexual characteristics are altered by4-tert-octylphenol and17β-estradiol in the adult male guppy(Poeiliareticulata)[J].Ecotoxicology and Environmental Safety,48:76-84
- Toft G,Baatrup E.2003.Altered sexual characteristics in guppies(Poecilia reticulata)exposed to17β-estradiol and4-tert-octylphenolduring sexual development[J].Ecotoxicology and EnvironmentalSafety,56:228-237
- Wen R S,Fang Z Q,Jiang S G,Xu J,Ma G Z.2007.Developmentof ELISA for detecting Xiphophorus helleri vitellogenin[J].Researchof Environmental Sciences,20(4):144-149(in Chinese)
- Wester P W,Canton J H,Bisschop A.1985.Histopathological studyof Poecilia reticulata(guppy)after long-term-hexachlorocyclohexaneexposure[J].Aquat Toxicol,6:271-296
- Yang A G,Mao J F,Yao L B.2001.Experimental Technology ofBiochemistry and Molecule Biology[M].Beijing:High EducationPress.40-42(in Chinese)
- Zhao B,Liu Z T,Xu Z F,Yang Y B.2005.Estrogenic effects ofphenolic compounds on juvenile goldfish(Carassius auratus)[J].Acta Scientiae Circumstantiae,25(9):1259-1264(in Chinese)
- 邴欣,汝少国,Isoda H,姜明,王蔚,王诗红.2004.17β-雌二醇对雄性金鱼卵黄蛋白原的诱导作用[J].水产学报,28(3):236-240
- 李康,周忠良,于静,王明山,余晓君.2003.鲫鱼(Carassiusauratus)卵黄蛋白原的ELISA检测[J].中国环境科学,23(3):276-280
- 温茹淑,方展强,江世贵,徐杰,马广智.2007.剑尾鱼卵黄蛋白原的ELISA检测[J].环境科学研究,20(4):142-147
- 杨安钢,毛积芳,药立波.2001.生物化学与分子生物学实验技术[M].北京:高等教育出版社.40-42
- 赵兵,刘征涛,徐章法,杨永滨.2005.酚类化合物对金鱼幼鱼的雌激素效应研究[J].环境科学学报,25(9):1259-1264