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Titolo:
Development and validation of a homologous zebrafish (Danio rerio Hamilton-Buchanan) vitellogenin enzyme-linked immunosorbent assay (ELISA) and its application for studies on estrogenic chemicals
Autore:
Fenske, M; van Aerle, R; Brack, S; Tyler, CR; Segner, H;
Indirizzi:
Leipzig Halle GmbH, UFZ Ctr Environm Res, Dept Chem Ecotoxicol, D-04318 Leipzig, Germany Leipzig Halle GmbH Leipzig Germany D-04318 col, D-04318 Leipzig, Germany Univ Exeter, Hatherly Labs, Sch Biol Sci, Exeter, Devon, England Univ Exeter Exeter Devon England s, Sch Biol Sci, Exeter, Devon, England Univ Halle, Dept Biol, Halle, Germany Univ Halle Halle GermanyUniv Halle, Dept Biol, Halle, Germany
Titolo Testata:
COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY
fascicolo: 3, volume: 129, anno: 2001,
pagine: 217 - 232
SICI:
1532-0456(200107)129:3<217:DAVOAH>2.0.ZU;2-K
Fonte:
ISI
Lingua:
ENG
Soggetto:
SALMON SALMO-SALAR; VITELLINE ENVELOPE PROTEINS; BASS MORONE-SAXATILIS; RAINBOW-TROUT; PLASMA VITELLOGENIN; ONCORHYNCHUS-MYKISS; GENE-EXPRESSION; REPRODUCTIVE-CYCLE; CYPRINID FISH; INDUCTION;
Keywords:
enzyme-linked immunosorbent assay (ELISA); environmental estrogens; 17 alpha-ethinylestradiol (EE2); homologous anti-body; vitellogenin (VTG); zebrafish;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
51
Recensione:
Indirizzi per estratti:
Indirizzo: Fenske, M Leipzig Halle GmbH, UFZ Ctr Environm Res, Dept Chem Ecotoxicol, Permoserstr 15, D-04318 Leipzig, Germany Leipzig Halle GmbH Permoserstr 15 Leipzig Germany D-04318 rmany
Citazione:
M. Fenske et al., "Development and validation of a homologous zebrafish (Danio rerio Hamilton-Buchanan) vitellogenin enzyme-linked immunosorbent assay (ELISA) and its application for studies on estrogenic chemicals", COMP BIOC C, 129(3), 2001, pp. 217-232

Abstract

Vitellogenin (VTG) was isolated by anion exchange chromatography from plasma of female zebrafish (Danio rerio) induced with 17 alpha -ethinylestradiol (EE2). The purity of the VTG isolate was confirmed by polyacrylamide gel electrophoresis (SDS-PAGE). Purified VTG was used to raise polyclonal antibodies in rabbits and the specificity of the antisera for VTG confirmed by Western blot analysis of plasma proteins separated by SDS-PACE. The antibodies cross-reacted with two proteins in the plasma of female zebrafish, with molecular masses of approximately 142 and 171 kDa. No cross-reactivity was observed with any other plasma proteins. A competitive enzyme-linked immunosorbent assay (ELISA) was developed using the polyclonal zebrafish VTG (z-VTG) antibodies and purified z-VTG as ligand and standard, respectively. Thez-VTG ELISA was sensitive with a detection limit of between 2.0 and 3.0 ngpurified VTG/ml, and a working range between 3 and 500 ng/ml (30-85% binding). The ELISA demonstrated precision, with inter- and intra-assay variations of 7.5 +/- 2.7 and 4.9 +/- 1.4%, respectively. Plasma from adult zebrafish and whole body homogenates from juvenile zebrafish diluted parallel withthe z-VTG standard in the EI-ISA, validating the assay for quantifying z-VTG in both of these tissues. Exposure of adult male zebrafish to EE2 via water induced a concentration-dependent induction of VTG with a lowest observed effect concentration (LOEC) less than or equal to 1.67 ng EE2/1 (for a 21-day exposure). The homologous z-VTG ELISA provides a valuable tool for the study of environmental estrogens in zebrafish. (C) 2001 Elsevier Science Inc. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 02/04/20 alle ore 18:15:27