Vitellogenin in Zoarces viviparus: purification, quantification by ELISA and induction by estradiol-17 beta and 4-nonylphenol.
Vitellogenin was isolated by gel filtration and ion-exchange chromatography from plasma of estradiol-treated male Zoarces viviparus. The purification of vitellogenin was followed through each step by native polyacrylamide gelelectrophoresis (PAGE). The purified vitellogenin was used to raise anti-vitellogenin antibodies in rabbits. The specificity of the affinity purified antibody raised against vitellogenin was assessed by Western blot analysis. No crossreactivity was observed with plasma from non-induced control males. A direct enzyme-linked immunosorbent assay (ELISA) was developed by use of the raised anti-vitellogenin. The detection limit for the purified standard vitellogenin by the ELISA method was 5 ng ml-1 Vitellogenin levels were quantified in plasma of pregnant female Zoarces viviparus, which were held in aquaria with different concentrations of the estrogenic compound 4-nonylphenol dissolved in the ambient seawater. A marked effect of exposure to ambient 4-nonylphenol was observed by significant dose-dependent increases in the level of plasma vitellogenin of the pregnant fish as well as of embryos exposed in vitro.
Korsgaard
B
B
Institute of Biology, Odense University, Denmark.
Pedersen
K L
KL
D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D005399:Fishes / Q000097:blood* / Q000196:embryology; D008297:Male; D010636:Phenols / Q000494:pharmacology*; D011247:Pregnancy; D011817:Rabbits; D014819:Vitellogenins / Q000096:biosynthesis / Q000097:blood* / Q000302:isolation & purification*
D010636:Phenols; D014819:Vitellogenins; D004958:Estradiol; C041594:4-nonylphenol