Induction by beta-estradiol of vitellogenin in striped bass (Morone saxatilis): characterization and quantification in plasma and mucus.
Striped bass (Morone saxatilis) were implanted with beta-estradiol to induce the production of vitellogenin, the egg yolk precursor produced by the liver. Electrophoretic analysis revealed that beta-estradiol caused marked production of a plasma protein of apparent molecular mass 170 kDa. Size exclusion chromatography suggested that the estradiol-induced protein circulated as a dimer. This protein was purified from the plasma of estradiol-treated fish by DEAE-agarose column chromatography and used to induce antibodies in rabbits and goats. Western blots revealed that the antiserum bound to the putative vitellogenin in plasma from estradiol-treated fish and adult females, but not with any proteins in male plasma. Western blot of ovarian extract revealed several smaller immunoreactive protein bands and supported the identity of the purified protein as vitellogenin. A competitive ELISA was developed with sensitivity in a range from 8 to 1000 ng/ml. Plasma concentrations of adult females during their spawning migration ranged from 100 to 600 micrograms/ml. Western blot of mucus extract revealed the presence of a 170-kDa protein in vitellogenic female fish along with several minor bands ranging from 50 to 110 kDa. Positive immunoreactivity was present in the surface mucus of all females and in none of the males collected during a spawning migration in the Hudson River.
Kishida
M
M
Department of Zoology, University of Rhode Island, Kingston 02881.
Anderson
T R
TR
Specker
J L
JL
D000818:Animals; D000918:Antibody Specificity; D001492:Bass / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism; D015153:Blotting, Western; D002848:Chromatography, DEAE-Cellulose; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D008297:Male; D008970:Molecular Weight; D009093:Mucus / Q000737:chemistry / Q000378:metabolism*; D010053:Ovary / Q000737:chemistry / Q000378:metabolism; D014819:Vitellogenins / Q000096:biosynthesis* / Q000097:blood / Q000737:chemistry
D001798:Blood Proteins; D014819:Vitellogenins; D004958:Estradiol