Radioimmunoassay for fibroblast growth factor (FGF): release by the bovine anterior pituitary in vitro.
A radioimmunoassay (RIA) was developed to measure fibroblast growth factor (FGF) using antiserum generated against a synthetic replicate of [Tyr10]FGF(1-10). The antisera, previously shown to be capable of inhibiting the biological action of FGF on bovine aortic arch endothelial cells in vitro, are highly specific for the amino-terminus of FGF. In the RIA, the antisera recognize the decapeptide antigen [Tyr10]FGF(1-10) and the intact mitogen on an equimolar basis and show less than 0.01% cross-reactivity with N-acetyl-[Tyr10]FGF(1-10). Bovine adenohypophysial cells maintained in primary monolayer culture release and ir-FGF which is indistinguishable from the intact mitogen in as much as it is retained on heparin-Sepharose affinity columns and shows a dose-dependent and parallel displacement in RIA. The release of ir-FGF by the bovine adenohypophysis can be increased with forskolin (10(-5) M) or KCl (50 mM). Preincubation of pituitary cells with 17 beta-estradiol has no measurable effects on basal ir-FGF, but increases the release after KCl treatment 2-3-fold. These results show that ir-FGF can be released by the bovine adenohypophysis in vitro and lend credence to the hypothesis that FGF plays a physiological role in the homeostatic mechanisms regulating mesoderm-derived cell growth.
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D005576:Colforsin; D003429:Cross Reactions; D004224:Diterpenes / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology; D005346:Fibroblast Growth Factors / Q000031:analogs & derivatives / Q000032:analysis*; D010903:Pituitary Gland, Anterior / Q000032:analysis*; D011189:Potassium Chloride / Q000494:pharmacology; D011863:Radioimmunoassay
D004224:Diterpenes; D005576:Colforsin; D004958:Estradiol; D005346:Fibroblast Growth Factors; D011189:Potassium Chloride