Development of enzyme-linked immunosorbent assays for two forms of vitellogenin in Japanese common goby (Acanthogobius flavimanus).
Two vitellogenins (Vgs) were detected in serum from estradiol-17beta (E(2))-injected Japanese common goby (Acanthogobius flavimanus). Vitellogenins with molecular masses of 530 kDa (Vg-530) and 320 kDa (Vg-320) were purified, and used to raise specific antisera in rabbits. Sandwich enzyme-linked immunosorbent assays (ELISAs) for Vg-530 and Vg-320 were developed using the antisera and the isolated Vgs. The sensitivity ranges of these ELISAs were 1.25-160 ng/ml for Vg-530 and 0.26-66 ng/ml for Vg-320, and very low cross-reactivity was found with the alternate Vg in each assay. Treatment of male gobys with E(2) by injection and immersion induced both Vgs in sera in a dose-dependent manner. The mean concentrations of the Vgs increased from 10 ng/L E(2) exposure for three weeks. Serum concentrations of the two Vgs in field-collected maturing females increased in accordance with increment of E(2) level and ovarian development, and the mean concentrations of Vg-530 were higher than those of Vg-320 in maturing female. These results indicate that the sandwich ELISAs for Vg-530 and Vg-320 developed in the present study is useful as an assay system for surveys of estrogenic activity in coastal areas of Japan.
Ohkubo
Nobuyuki
N
Hokkaido National Fisheries Research Institute, Fisheries Research Agency, 116, Katsurakoi, Kushiro, Hokkaido 085-0802, Japan. nobuyuki@fra.affrc.go.jp
Mochida
Kazuhiko
K
Adachi
Shinji
S
Hara
Akihiko
A
Hotta
Komei
K
Nakamura
Yukio
Y
Matsubara
Takahiro
T
D000818:Animals; D015153:Blotting, Western; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D005399:Fishes / Q000378:metabolism*; D007101:Immersion; D007267:Injections; D008297:Male; D014819:Vitellogenins / Q000032:analysis*
D014819:Vitellogenins; D004958:Estradiol