Fundulus heteroclitus gonadotropins.5: Small scale chromatographic fractionation of pituitary extracts into components with different steroidogenic activities using homologous bioassays

Fractionation and characterization of gonadotropins (GtH) from Fundulus heteroclitus pituitary extracts were carried out using a biocompatible liquid chromatographic procedure (Pharmacia FPLC system). Chromatographic fractions were monitored for gonadotropic activities (induction of oocyte maturatio...

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Bibliographic Details
Published in:Reproductive biology and endocrinology 2004-03, Vol.2 (1), p.14-14, Article 14
Main Authors: Lin, Yu-Wai Peter, Petrino, Teresa R, Wallace, Robin A
Format: Article
Language:English
Subjects:
20-alpha-Dihydroprogesterone - biosynthesis
Animals
Biological Assay - methods
Chromatography - methods
Estradiol - biosynthesis
Female
Fundulidae
Gonadotropins, Pituitary - analysis
Gonadotropins, Pituitary - pharmacology
Male
Ovarian Follicle - drug effects
Ovarian Follicle - metabolism
Pituitary Gland - chemistry
Testosterone - biosynthesis
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Summary:Fractionation and characterization of gonadotropins (GtH) from Fundulus heteroclitus pituitary extracts were carried out using a biocompatible liquid chromatographic procedure (Pharmacia FPLC system). Chromatographic fractions were monitored for gonadotropic activities (induction of oocyte maturation and steroid production) using homologous follicle bioassays in vitro. Size-exclusion chromatography eluted gonadotropic activity in one major protein peak (Mr approximately 30,000). Anion-exchange and hydrophobic-interaction chromatography (HIC) yielded two distinct peaks of 17beta-estradiol (E2)- and 17alpha-hydroxy,20beta-dihydroprogesterone (DHP)-promoting activity with associated oocyte maturation. Two-dimensional chromatography (chromatofocusing followed by HIC) resolved pituitary extracts into two active fractions; both induced E2 synthesis, but one was relatively poor in eliciting DHP and testosterone production. Thus, using homologous bioassays, at least two quantitatively different gonadotropic (steroidogenic) activities: an E2-promoting gonadotropin (GtH I-like) and a DHP-promoting gonadotropin (GtH II-like), which has a lower isoelectric point but greater hydrophobicity than the former, can be distinguished from F. heteroclitus pituitaries by a variety of chromatographic procedures. This study complements previous biochemical and molecular data in F. heteroclitus and substantiates the duality of GtH function in a multiple-spawning teleost.
ISSN:1477-7827
1477-7827
DOI:10.1186/1477-7827-2-14