Pregnancy-associated glycoprotein family (PAG)--as chorionic signaling ligands for gonadotropin receptors of cyclic animals.

The chorionic pregnancy-associated glycoprotein (PAG) family was identified in pigs, cattle and other eutherian mammals. The objective of this study was to examine whether secretory chorionic proteins (including PAGs), produced in vitro by explants of porcine and bovine placental membranes, may interact with other proteins, i.e. gonadal and extragonadal binding sites. Trophoblast (TRF) and trophectoderm (TRD) explants of pigs (n=38; 14-61 dpc-day post coitum) or cotyledons (CT) of cows (n=5; 40-110 dpc) were long-term cultured. Released chorionic proteins were ultra-fractionated from media (>10 kDa) or precipitated [20-75% of (NH(4))(2)SO(4)]. The PAGs were monitored by Western/PAGE (30-73 kDa). Secretory TRF/TRD/CT (+PAG) proteins (0.78-25 microg/ligand) were examined by radioreceptor assay (RRA) with iodinated hCG ((125)I-hCG) for binding-effectiveness by gonadotropin receptors of cyclic pigs and cows (cRc). Gonadal and extragonadal cRc isolated from luteal-phase corpora lutea and uteri (cCLRc, cMYORc and cENDRc) were tested with positive control ligands: porcine LH and hCG (0.39-50 ng/ml). Control proteins produced in vitro by endometrial (END) explants of cyclic (cEND), pseudopregnant (PsEND) and pregnant (pEND) gilts were utilised as negative ligands (0.78-25 microg/ligand). Positive control ligands competed with (125)I-hCG for binding by cCLRc, cMYORc and cENDRc (18-61%/B(0) for hCG and 27-57%/B(0) for LH). Negative ligands (cEND, PsEND and pEND) did not show cRc bindings. This is the first RRA report indicating that in vitro produced porcine TRF/TRD proteins (+PAG) competed (P< or =0.05) with (125)I-hCG for binding by cCLRc, cMYORc and cENDRc in a concentration- and pregnancy stage-dependent manner. The highest competition with (125)I-hCG (up to P< or =0.001) was found for ultra-fractionated TRF/TRD proteins (>10 kDa) during early pregnancy (<22 dpc). The greatest competition (P< or =0.05) of precipitated porcine TRD proteins (>30 dpc) was detected for fractions obtained by saturation with use of 20% of (NH(4))(2)SO(4). Bovine CT proteins revealed lower competition of (125)I-hCG for bovine cCLRc (during 45 dpc only) that was more efficient with CT (up to 71%) than with non-labelled hCG (82%). The PAG proteins may play a role as potential "signal molecules", because they were able to interact with gonadotropin receptors of luteal-phase animals. It seems that the pPAG proteins may be luteoprotective chorionic-origin signals during implantation and placentation, according to binding-effectiveness of the chorionic ligands that was comparable to LH/hCG ligands with gonadal and extragonadal receptors of cyclic animals.