Ovine allantoic fluid contains high concentrations of activin A: partial dissociation of immunoactivity and bioactivity.
In a preliminary study, allantoic fluid collected from pregnant sheep across gestational ages of 20-124 days contained significantly higher levels of activin bioactivity (189 +/- 74 ng/ml, mean +/- SE) than did amniotic fluid (3.2 +/- 0.6 ng/ml). Using a combination of chromatography steps, we isolated from 5 L of allantoic fluid approximately 612 microg of immunoactive activin, which eluted over 10 fractions from a C8 reversed-phase column. When these fractions were assayed in a rat pituitary cell culture bioassay, in a specific RIA, and in an activin A two-site ELISA, the RIA activity was skewed to the less hydrophobic side of the activin profile, while the bioactivity was skewed to the more hydrophobic forms. The activity measured in the two-site ELISA more closely matched the mass of activin as determined by laser densitometry. Amino-terminal sequencing of fractions containing either peak immunoactivity or bioactivity showed each to be identical to activin A. This was confirmed by internal sequences from a fraction that eluted in the area of overlapping immunoactivity and bioactivity. A peptide containing at least 18 amino acids at its amino terminus, which were identical to the conserved region of the acute-phase protein serum amyloid A, was identified in the most immunoactive activin fractions.
Institute of Reproduction and Development, Monash University, Clayton, Victoria 3168, Australia.
L M
LM
Foulds
D M
DM
de Kretser
P
P
Farnworth
D
D
Buttress
G
G
Jenkin
N P
NP
Groome
J R
JR
McFarlane
D028341:Activins; D000465:Algorithms; D000481:Allantoin / Q000378:metabolism*; D000653:Amniotic Fluid / Q000378:metabolism; D000818:Animals; D001681:Biological Assay; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005314:Embryonic and Fetal Development / Q000502:physiology; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005865:Gestational Age; D007265:Inhibins / Q000378:metabolism*; D010902:Pituitary Gland / Q000166:cytology / Q000187:drug effects; D011247:Pregnancy; D011863:Radioimmunoassay; D051381:Rats; D012756:Sheep; D013056:Spectrophotometry, Ultraviolet
D028341:Activins; D000481:Allantoin; D007265:Inhibins