25 2 2005 2 ACTA SCIENTIAE CIRCUMSTANTIAE Vol. 25,No. 2 Feb., 2005,. [J ].,2005,25 (2) :135-140 MA Taowu,WANG Zijian. Fish exprimental model in the screening and testing for the environmental endocrine disruptors[j ]. Acta Scientiae Circumstantiae,2005, 25 (2) :135-140 1,2, 1 3 11, 100085 ; 21, 416000 :.,,.,,. : ; ; ; ; ; :025322468 (2005) 0220135208 :X171 :A Fish exprimental model in the screening and testing for the environmental endocrine disruptors MA Taowu 1,2, WANG Zijian 1 3 11State Key Laboratory of Environmental Aquatic Chemistry, Research Center for Eco2Evironmental Sciences, Chinese Academy of Sciences, Beijing 100085 ; 21College of Biological Resource & Environmental Science, Jishou University, Jishou 416000 Abstract : In recent years, the exploration of methodology for screening and testing the environmental endocrine disruptors has become an important issue for scien2 tific research and environmental management. In the field of aquatic eco2toxicology, development of methodology, such as fish experimental model and its related in vitro or in vivo bioassays is mainly concerned. And this review summarized the scientific principals of testing endocrine disruption effects and its biomarkers associat2 ed with endocrine system of fish, main bioassaying one of which was used currently in laboratories and governmental agencies. It also indicated that it was necessary to establish the local fish experimental model and the assay methodological system for screening and testing endocrine disruptors. in China. Keywords : Endocrine disruption ; Fish experimental model ; Biomarker ; Screening ; Assay ; Chinese rare minnow.,,,,,.,. ( Endo2 crine disruptors, EDs),,.,,,,..,,., ( ) ( ) :2004210208 ; :2004211222 : (20337020) ; : (1968 ),, ; 3
136 25., EDs,.,. 1 111. ( ) [1 ].,,,, ( ),. 2 [2 ], ( 1). 1 Fig. 1 The schematic diagram of the endocrine system and hormone regulation of fish, 2 2., ( GtH).,. ( ).. ( Danio rerio) 5 : [3 ]., (17, 20 2DP).. Leydig, Sertoli, 112 (112KT) [4 ].,.,,. T 4,T 4. T 3, T 3 [5 ],. T 3 T 4.,.,...,., GtH ( 1). GtH, GtH.. GtH.,,.
2 : 137, T 4 T 3 TSH [6 ]. T 4 T 3 TRH, T3 GnRH [7 ]. 112,, ( ) ( (SBPs) )., : EDs ; EDs ( ), ;,, ( TBT) [8 ] ;, TCDD [9 ] ;. Kime (1999) [10 ] EDs, ( PCBs o, p 2DDT ), ( p, p 2DDTΠDDE ), ( TBT ), ( Vinclozolin, Procymidone), ( ), ( PCBs ), ( PCBs ) ( TBT ).,. [11, 12 ],,,., [2 ], 1,. 2 ( Biomarkers) EDs. EDs, ( ), ( )., EDs. EDs mrna. (VTG) ( Zrp) (CHG) ( T,KT,E 2,T 3 T 4 ) EDs, VTG mrna., GtH E 2,E 2 VTG [13 ] ( 2).,, E 2 VTG,.,VTG, 2 VTG Fig. 2 The modulation of VTG production in fish
138 25. Wester [14 ] 2HCH VTG, VTG EDs., VTG. PCBs, [15 ],, VTG, [10 ]. VTG [10 ]. P450, (Ah2 ) EROD. PCBs P450 [16, 17 ]. P450,,..,,.. ( EDs).,., VTG VTG. 2,3,7,82TCDD 2,3,7,82 TCDF, 2,3,4,7,82PCDF 2, PCB, 2 E 2 VTG, [16 ]. 2 Ah,Ah CYP1A1, [18 ], 2 E 2 [16 ], Ah [19 ]. EDs P450,.,, [20 ].,, ( Testi2ova) VTG [21,22 ]. 3 17 2 [23 ]. [21,24 ].,,, ( ). 3 17 2 (a : ;b : ;c : ;d : ) Fig. 3 Gonad alterations of Chinese rare minnow ( Gobiocypris rarus) af2 ter exposed to 17 2ethinylestradiol (a : normal ovaries ( 100) ; b : testi2ova ( 400) ; c :normal mature testis ( 100) ; d : de2 generated ovaries ( 400) ) [24 ] 3 [20, 21 ] [25 ] ( )., Tier1, ; Tier2,
2 : 139., 3, Tier1 14 ; Tier2 ;Tier3 [26 ]. EDs ( In vitro test system) ( In vivo test system)., [27,28 ].,,,. [17,29 ].,,. Tier1 : ( ) ( ) (VTG ) [20 ]. Tier2 : ( ) ( ) (VTG ) [21 ]., EDs. ( Carassius auratus) ( Morone saxatilis) ( Oryzias latipes) ( Oncorhynchus mykiss) ( Pimephales promelas) ( Sparus auratus) ( Mycteroperca microlepis) ( Salvelinus alpinus) 1 [ 30] Table 1 VTG molecular weight of various fishes Π 10 3 140 147 ( Abramis brama) 170 ( Leuciscus idus) 200 ( Anguilla anguilla) 176 180 ( Cichlasoma dimerrus) 180 ( Oreochromis mossambicus) 156 ( Danio rerio) 183 ( Gobiocypris rarus) 158 Π 10 3 154 150 211 120, 180 130,200 142,171 147,170, VTG, VTG,,,.,EDs. 1 ( ) VTG, VTG,., VTG VTG mrna., EDs,.,,. 4. : ; ;, ;,.,. ( Oryzias latipes) ( Pimepe2 hales promelas) ( Danio rerio) ( Cyprin2 odon variegantus) ( Oncorhyrichus mykiss).,,,, ( ),..,., 315 715 cm., 24 25,. ( ) 4,, 3 5 d 50 250. 80 % 100 %, 25 415 6 d, OECD ISO ( ) USEPA. EDs
140 25.,., 4 5 cm,., 28, 31 25..., 70 80 d, 5 10 d, 150 400,,,, 3 d. 30,...,,, 215 510 cm,, 5 35,,.., 3,, 25 30.,.,, 200,. 500, d2rr [31 ],,,,,, [32 ].,, ( XX) (XY ).,..,, [33 ]. EDs,.,., 315 510cm, (0 40 ), 25 30, 60 %..., 2..,24h. 15 30,,,. (40 % 60 %). 28, 5 6 d.,, EDs. 5.,,,,,.,, ;, ( VTG ). [34 ], ( Gobiocypris rarus)..,,.,,,,,, 2 9 ( 415 d) d 100 600 ( 250 ) [34 ], (26 1) 2 d, 95 %,., (26 1) 4 5., 5 6 cm. EDs,. (
2 : 141 ) EDs. 6,,.,,,. : [ 1 ] Thomas M J, Thomas J A. Hormone Assays and Endocrine Function [ A ].ΠΠ Hayes A W. Principles and Methods of Toxicology Fourth Edition [ C]. Philadelphia : TAYLOR & FRANCIS, 2001 : 1383 1413 [ 2 ] Orlando E F, Guillette L J Jr. The Handbook of Environmental Chemistry [M]. Berlin Heidelberg : Springer2Verlag, 2002 [ 3 ] Selman K, Wallace R A, Sarka A, et al. Stages of oocyte develop2 ment in the zebrafish, Brachydanio Rerio [J ]. Journal of Morphol, 1993, 218 : 203 224 [ 4 ] Redding J M, Patino R. Reproductive Physiology[ A ].ΠΠEvans D H. Physiology of Fishes [ C]. Boca Raton, FL : CRC Press, 1993 : 503 534 [ 5 ]. [ M]. :,1999 : 207 208 [ 6 ] Yoshiura Y, Sohn Y C, Munakata A, et al. Molecular cloning of the cdna encoding the beta subunit of thyrotropin and regulation of its gene expression by thyroid hormones in the goldfish, Carassius aura2 tus[j ]. Fish Physiol Biochem, 1999, 21 : 201 210 [ 7 ] Parhar I S, Soga T, Sakuma Y. Thyroid hormone and estrogen regu2 late brain region2specific messenger ribonucleic acids encoding three gonadotropin2releasing hormone genes in sexually immature male fish, Oreochromis niloticus[j ]. Endocrinology, 2000, 141 : 1618 1626 [ 8 ] Ronis MJ J, Mason A Z. The metabolism of testosterone by the peri2 winkle ( Littorina littorea) in vitro and in vivo : Effects of tributyltin [J ]. Mar Environ Res, 1996, 42 : 161 166 [ 9 ] Romkes M, Piskorskapliszczynska J, Safe S. Effects of 2, 3, 7, 82 tetrachlorodibenzo2p2dioxin on hepatic and uterine estrogen receptor levels in rats[j ]. Toxicol Appl Pharmacol,1987, 87 : 306 314 [10 ] Kime D E. Environmentally induced endocrine abnormalities in fish [ A]. ΠΠHester R E, Harrison R M. Endocrine Disrupting Chemicals [ C]. Cambridge, UK: The Royal Society of Chemistry, Thomas Graham House, 1999 [ 11 ] Folmar L C, Denslow N D, Rao V, et al. Vitellogenin induction and reduced serum testosterone concentrations in feral male carp ( Cypri2 nus carpio) captured near a major metropolitan sewage treatment plant [J ]. Environ Health Perspect, 1996, 104 : 1096 1101 [ 12 ] Orlando E F, Denslow N D, Folmar L C, et al. A comparison of the reproductive physiology of largemouth bass, Micropterus salmoides, collected from the Escambia and Blackwater Rivers in Florida [J ]. Environ Health Perspect, 1999, 107 : 199 204 [ 13 ] Tyler C R, Sumpter J P, Kawauchi H, et al. Involvement of gonado2 tropin in the uptake of vitellogenin into vitellogenic oocytes of the rainbow trout, Oncorhynchus mykiss [ J ]. Gen Camp Endocrinol, 1991, 84 :291 299 [14 ] Wester P W, Canton J H, Bisschop A. Histopathological study of Poecilia reticulata (guppy) after long2term 182hexachlorocyclohexane exposure[j ]. Aquat Toxicol, 1985, 6 : 271 296 [15 ] Donohoe R M, Curtis L R. Estrogenic activity of chlordecone, o, and p 2DDT and o, p 2DDE in juvenile rainbow trout : induction of vitellogenesis and interaction with hepatic estrogen binding sites [J ]. Aquat Toxicol, 1996, 36 : 31 52 [16 ] Anderson M J, Miller M R, Hinton D E. In vitro modulation of 17 2 estradiol2induced vitellogenin synthesis : Effects of cytochrome P4501A1 inducing compounds on rainbow trout ( Oncorhynchus rnykiss) liver cells [J ]. Aquat Toxicol, 1996, 34 : 327 350 [ 17 ] Wan X, Ma T, Wu W, et al. EROD activities of a primary cell cul2 ture of grass carp ( Ctenopharyngodon idellus) hepatocytes exposed to polychlorinated aromatic hydrocarbons [J ]. Ecotoxicol Environ Saf, 2004, 58 : 84 89 [18 ] Wang X, Porter W, Krishnan V, et al. Mechanism of 2, 3, 7, 82 tetrachlorodibenzodioxin ( TCDD) mediated decrease of the nuclear estrogen receptor in MCF27 human breast cancer cells [J ]. Mol Cell Endocrinol, 1993, 96 : 159 166 [ 19 ] DeVito MJ, Xinfrang M, Babish J G, et al. Dose2response relation2 ships in mice following subchronic exposure to 2, 3, 7, 82tetrachlo2 rodibenzo2p2dioxin : CYP 1A 1, CYP 1A2, estrogen receptor, and protein tyrosine phosphorylation [J ]. Toxicol Appl Pharmacol, 1994, 124 : 82 90 [20 ] USEPA. Draft Detailed Review Paper on Fish Screening Assays for Endocrine Disruption [ M ]. EPA Contact Number 682W2012022, Washington DC, 2002 [21 ] USEPA. Draft Detailed Review Paper on a Fish Two Generation Tox2 icity Test [ M ]. DC, 2002 EPA Contact Number 682W2012023, Washington [22 ],,,. [J ].,2004, 24 (3) : 487 492 [23 ] Wang Z. Testing endocrine disruption chemicals using fish model [ A ]. Proceedings of International Symposium on Biotechnology for Environmental Pollution Control [ C]. Beijing, China, 2004 : 348 349 [24 ] Zha J M,Wang Z. Estimation the chronic toxicity of effluents from different technology treatment in urban sewage treatment plant based on early stage development effect of Japeness medaka ( Oryzias latipes) [A]. Proceedings of International Symposium on Biotechnol2 ogy for Environmental Pollution Control [ C]. Beijing, China, 2004 : 335 341
142 25 [25 ] OECD. Draft Detailed Review Paper : Appraisal of Test Methods for Sex2Hormone Disrupting Chemicals [ C ]. OECD Environmental Health and Safety Publications, Environment Directorate, Paris, 2001 [26 ] Hutchinson T H, Pickford D B. Ecological risk assessment and test2 ing for endocrine disruption in the aquatic environment [J ]. Toxicolo2 gy, 2002,181 182 :383 387 [27 ] Fent K. Fish cell lines as versatile tools in ecotoxicology : assessment of cytotoxicity, cytochrome P4501A induction potential and estrogenic activity of chemicals and environmental samples [ J ]. Toxicol In Vitro, 2001,15 : 477 488 [ 28 ] Behrens A, Schirmer K, Bols N C, et al. Polycyclic aromatic hydro2 carbons as inducers of cytochrome P4501A enzyme activity in the rainbow trout liver cell line, RTL2W1, and in primary cultures of rainbow trout hepatocytes [J ]. Environ Toxicol Chem, 2001, 20 : 632 643 [29 ] Smeets J M W, Rankouhi T R, Nichols KM, et al. In vitro vitello2 genin production by carp (Cyprinus carpio) hepatocytes as a screen2 ing method for determining ( anti) estrogenic activity of xenobiotics [J ]. Toxicol Appl Pharmacol, 1999, 157 : 68 76 [30 ]. [D]. :,2004 :51 [31 ] Yamamoto T. Medaka ( Killifish) : Biology and Strains [M]. Tokyo : Keigaku Pub. Co., 1975 [32 ] Edmunds J S, McCarthy R A, Ramsdell J S. Permanent and func2 tional male to female sex Reversal in d2rr strain medaka ( Oryzias latipes) following egg microinjection of Health Perspect, 2000, 108 : 219 224 o, p 2DDT [ J ]. Environ [33 ] Knorr S, Braunbeck T. Decline in reproductive success, sex rever2 sal, and developmental alterations in Japanese medaka ( Oryzias latipes) after continuous exposure to octylphenol [J ]. Ecotoxicol En2 viron Saf, 2002, 51 :187 196 [34 ]. [J ]., 1992,16 (2) :165 174