Secretory activity of bovine ovarian granulosa cells transfected with sense and antisense insulin-like growth factor (IGF) binding protein-3 and the response to IGF-I, GH, LH, oxytocin and oestradiol.
The aim of our in vitro experiments was to examine if IGF binding protein (IGFBP)-3 is involved in control of bovine ovarian secretory activity. For this purpose we performed the transfection of bovine granulosa cells with cDNA sense and antisense constructs increasing or inhibiting IGFBP-3 synthesis. The release of IGFBP-3, progesterone, oxytocin, IGF-I and prostaglandins F (PGF) and E (PGE) by control and transfected cells was compared. The transfected ovarian cells were cultured with and without bLH (100 ng/ml), bGH (100 ng/ml), IGF-I (10 ng/ml), oxytocin (10 ng/ml) and oestradiol-17beta (100 ng/ml). The concentration of IGFBP-3 produced was assessed using ligand and western blotting and secretion of progesterone, oxytocin, IGF-I, PGF and PGE was evaluated using RIA/IRMA techniques. Transfection of cells with the sense IGFBP-3 cDNA construct resulted in the expected increase in IGFBP-3 release, whereas the antisense IGFBP-3 construct induced the expected reduction in IGFBP-3 output. The granulosa cells transfected to overexpress IGFBP-3 had an increase in IGF-I, PGF and PGE release, and a decrease in basal and hormone- or growth factor-induced accumulation of progesterone and oxytocin. The granulosa cells transfected to have reduced IGFBP-3 expression gave primarily significant opposite findings. The present results suggest the involvement of IGFBP-3 in control of bovine ovarian steroid, peptide hormone, growth factor and prostaglandin release. IGFBP-3 is a physiological stimulator of IGF-I and prostaglandin release and an inhibitor of steroid and peptide hormone output.
Sirotkin
A V
AV
Laboratory of Endocrinology, Research Institute of Animal Production, Hlohovská 949 92 Nitra, Slovak Republic. sirotkin@vuzv.sk
Makarevich
A V
AV
Corkins
M R
MR
Kotwica
J
J
Kwon
H B
HB
Bulla
J
J
Hetényi
L
L
D000818:Animals; D002417:Cattle; D016373:DNA, Antisense / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D006107:Granulosa Cells / Q000187:drug effects* / Q000378:metabolism*; D013006:Growth Hormone / Q000494:pharmacology; D006728:Hormones / Q000494:pharmacology*; D066298:In Vitro Techniques; D018972:Insulin-Like Growth Factor Binding Protein 3 / Q000235:genetics* / Q000378:metabolism; D007334:Insulin-Like Growth Factor I / Q000378:metabolism / Q000494:pharmacology; D007986:Luteinizing Hormone / Q000494:pharmacology; D010121:Oxytocin / Q000378:metabolism / Q000494:pharmacology; D011374:Progesterone / Q000378:metabolism; D011458:Prostaglandins E / Q000378:metabolism; D011460:Prostaglandins F / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D016373:DNA, Antisense; D018076:DNA, Complementary; D006728:Hormones; D018972:Insulin-Like Growth Factor Binding Protein 3; D011458:Prostaglandins E; D011460:Prostaglandins F; D011994:Recombinant Proteins; D011374:Progesterone; D004958:Estradiol; D010121:Oxytocin; D007334:Insulin-Like Growth Factor I; D007986:Luteinizing Hormone; D013006:Growth Hormone