Protein production by cultures established from Day-14-16 sheep and pig conceptuses

Abstract

Summary. Primary cell cultures were established from cells derived from dissociated

Chorionic expression of heterogeneous products of the PAG (Pregnancy-Associated Glycoprotein) gene family secreted in vitro throughout embryonic and foetal development in the pig

“…The adsorption-purification procedure appropriately eliminated immunoglobulins that are not porcine pregnancy-specific: several antibodies against antigenic proteins dominantly expressed in uterine endometrial tissue and antigens expressed in both trophectodermal and endometrial tissues, e.g. anti-uteroferrin, anti-serpin, anti-Kunitz-type proteinase inhibitors, anti-lysosyme or anti-retinol binding proteins (see: [1,4,5,[29][30][31][32][33]). The M r of immunoreactive porcine trophectodermal proteins produced in vitro (35-72 kDa), examined with the use of newly produced anti-pPAG sera (specific for pPAG fusion proteins during immunoscreening of cDNA clones), were very similar to forms of pPAG proteins detected initially by antisera against heterologous ruminant PAG antigens.…”

“…The pellets containing trophectodermal proteins were dissolved in 20 mM Tris HCl/2 mM EDTA, dialysed (MWCO 12-14 kDa) to remove (NH 4 ) 2 SO 4 , then the proteins were fractionated and concentrated in Amicon cartridges (MWCO 30 kDa), according to previously identified properties of the highly glycosylated forms of pPAG/BP proteins [1,4,5,[29][30][31][32][33]. The separated porcine proteins were analysed by 1D-SDS-PAGE and monitored by heterogeneous Western blotting with well-defined various anti-ruminant PAG sera, raised either against single purified bovine or ovine native PAG proteins, as immunogens [6,27,35].…”

“…In the pig, conditioned media of cultured trophoblast tissues, recovered from pregnant animals during implantation time, contain several forms of BP -basic proteins that have been identified as major multiple secretory products of conceptuses [29,30]. These BP proteins vary in their M.W.…”

Two-dimensional polyacrylamide gel electrophoresis of proteins synthesized and released by conceptuses and endometria from pony mares

“…Included among these were proteins whose electrophoretic mobilities resembled those of alpha-fetoprotein and transferrin (Fig. 1, Day 16, parentheses) (Buhi et al, 1982;Godkin et al, 1985). Positive identification of these proteins is in progress.…”

“…2. The yolk sac membranes incorporated the radiolabel into proteins that migrated in a manner resembling alpha-fetoprotein and transferrin (parentheses; Buhi et al, 1982;Godkin et al, 1985). These proteins remain to be positively identified.…”