Roles of ovine trophoblast protein-1 and oestradiol/prolactin in the establishment of pregnancy in sheep and pigs.
Explore the source record for details and available documents.
Animals↗
Biomedical subjects
J P Harney
Publications and source records attributed to J P Harney.
Effects of conceptus and conceptus secretory products on uterine development in the pig.
Pregnant gilts (n = 6) were subjected to surgery on Day 3 of pregnancy and uterine horns, within gilts, were assigned to either (1) ligation 15 cm below the uterotubal junction (REST) or (2) ligation proximal to the uterine body (NONREST). On Day 15 of pregnancy gilts were hysterectomized and length and weight of both uterine horns were recorded. NONREST uterine horns were longer (P less than .05) than those of REST, but weights were similar. Conceptuses from the REST uterine horns were retarded in development. Morphometric analysis of surface (SE) and glandular (GE) epithelial cells detected no effect of treatment on height, width, area, nuclear area or percentage of nucleus. Porcine conceptus secretory proteins (pCSP) were obtained from medium in which pig conceptuses, collected on Day 15 of pregnancy, were cultured for 30 h. Culture medium was pooled, dialysed and concentrated by Amicon ultrafiltration for intrauterine infusion. Serum proteins (SP) were obtained from blood collected from a Day 15 pregnant gilt and diluted for intrauterine infusion. Catheters were placed into both uterine horns or cyclic gilts (n = 8) on Day 10 of the oestrous cycle and uterine horns were ligated proximal to the uterine body. Gilts received either oestradiol valerate (E2) (5 mg) or corn oil (CO) on Days 11 and 12. On Days 12-14 each gilt received twice daily infusions of Day 15 pCSP in one uterine horn and SP in the other uterine horn.(ABSTRACT TRUNCATED AT 250 WORDS)
Analysis of Variance↗
Retinol-binding protein: a major secretory product of the pig conceptus.
Pig conceptuses and endometrial explants recovered from gilts between Days 10 and 15 of pregnancy were cultured in leucine-deficient or methionine-deficient medium supplemented with 3H-leucine or 35S-methionine, respectively, for 30 h. Conceptus and endometrial tissues from Day 15 of pregnancy were fixed in Bouin's fixative for immunocytochemistry and light microscopy. Conceptus culture medium from Day 15 of pregnancy was pooled, dialyzed, and fractionated by anion exchange and gel filtration chromatography. A family of 3-5 low molecular weight (Mr) acidic (Mr = 19,000-22,000; pI = 5.6-6.5) 3H-leu-labeled proteins were isolated and identified by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), electroblotting, and fluorography. The two major proteins (pCSP-1 and pCSP-2) were excised from a polyvinylidene difluoride transfer membrane, and NH2-terminal amino acids were sequenced. One peptide was sequenced through 33 amino acids and the second, which shared 100% homology, was sequenced through 22 amino acids. Analysis of the larger sequence indicated that it shared 93.9% and 90.9% homology with the first 33 amino acids of human and rabbit plasma retinol-binding protein (RBP), respectively. Analyses of culture medium from pig conceptus incubations by 2D-PAGE and immunoprecipitation with rabbit anti-human RBP serum indicated that immunoreactive RBP was produced between Days 10 and 15 of pregnancy and was present in Day 30 allantoic fluid. Western blotting of enriched fractions of Day 15 conceptus RBP followed by immunostaining indicated that five isoforms of radiolabeled RBP were present. Immunoreactive RBP was detected in trophectoderm and yolk sac of conceptuses and endometrial surface and glandular epithelium at Day 15 of pregnancy. Results from this study demonstrate that pig conceptuses secrete RBP prior to onset of conceptus elongation and throughout the peri-implantation period, which suggests that RBP and associated retinoids influence conceptus development.
Amino Acid Sequence↗
Ovine trophoblast protein-one inhibits development of endometrial responsiveness to oxytocin in ewes.
In experiment (Exp) 1, 12 cyclic ewes had catheters placed into each uterine horn on Day 7 (estrus = Day 0). On Days 11-15, 6 ewes received twice-daily intrauterine infusions of 1.5 mg serum protein (SP) into each uterine horn and 6 ewes received infusions of 1.08 mg SP + 0.42 mg ovine conceptus secretory proteins (oCSP) containing 25 micrograms ovine trophoblast protein-one (oTP-1) as determined by radioimmunoassay (25-35% bioactive by antiviral assay). SP-infused and oCSP-infused ewes had similar plasma 13,14-dihydro-15-keto prostaglandin F2 alpha (PGF2 alpha) profiles in response to oxytocin on Day 11, but SP ewes became more responsive (p less than 0.01) to oxytocin on Days 13 and 15 than oCSP ewes. SP ewes also had greater incorporation of [3H]inositol into inositol trisphosphate (IP3) (+3449%, p less than 0.01) and total inositol phosphate (IP) (+760%, p less than 0.08), in response to oxytocin, than did oCSP ewes (+553 and +168% for IP3 and total IP, respectively) in endometrium collected at ovariectomy/hysterectomy on Day 16. Mean CL weights on Day 16 and mean concentrations of progesterone in plasma collected at 12-h intervals on Days 6-16 were not different for SP and oCSP ewes, but concentrations of progesterone were lower (p less than 0.05) in SP ewes on Days 15-16 than for oCSP ewes. These results indicate that oTP-1 may prevent luteolysis by inhibiting development of endometrial responsiveness to oxytocin and, therefore, reduce oxytocin-induced synthesis of IP3 and PGF2 alpha.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Onset of secretion of proteins with antiviral activity by pig conceptuses.
In Exp. 1, antiviral activity was detected in Day-15 pregnant uterine flushings (6222 +/- 2167 units/ml) and in conceptus culture medium collected at 0, 1, 2, 4, 8, 16, 24, 32, 40, and 48 h (95, 375, 650, 1216, 1600, 2100, 2017, 2083, 3500 and 5000 units/ml, respectively; R2 = 0.81, P less than 0.01; y = 190.0 + 252.7x - 11.2x2 + 0.2x3. In Exp. 2, antiviral activity of Day-15 conceptus culture medium was reduced 99% after boiling for 20 min (P less than 0.01) and, after 18 h dialysis (6000-8000 Mr cut-off), 100% of the activity was in the retentate. In Exp. 3, antiviral activity was not detected in cultures of conceptuses from Days 10 and 11 and activity was maximal for Day 14 and Day 15 conceptuses (2100 and 2083 units/ml, respectively). Effects of day were best described by a quadratic regression equation (y = 17,652 - 3263x + 150x2; R2 = 0.55, P less than 0.01). In Exp. 4, changes in antiviral activity detected in uterine flushings from pregnant gilts on Days 8, 10, 11, 12, 14 and 15 (1.3, 0, 6.7, 63.3, 580 and 1663 units/ml, respectively) were described by the equation y = -20,743 + 6189x - 606x2 + 20x3 (R2 = 0.85, P less than 0.01). In Exp. 5, low antiviral activities (5-30 units/ml) were detected in all plasma samples collected from the uterine artery and uterine vein of pregnant and cyclic gilts, but values were not significantly influenced by pregnancy status, day or site of collection.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Influence of the endometrium, protease inhibitors and freezing on antiviral activity of proteins secreted by pig conceptuses.
In Exp. 1, only medium from cultures containing conceptus tissue had antiviral activity (P less than 0.05). Addition of Day-15 pregnant endometrium or Day-14 cyclic uterine flush proteins to cultures containing 200 mg conceptus tissue decreased antiviral activity (conceptus x endometrial protein interaction, P less than 0.06). Effects of endometrium (-54%) and uterine flush proteins (-40%) on antiviral activity of conceptus cultures did not differ from each other (P greater than 0.10). In Exp. 2, antiviral activity was only detected in cultures containing conceptus tissue (P less than 0.06). The amount of antiviral activity in cultures of Day-15 conceptus tissue was not influenced differently (P greater than 0.10) by culture in medium conditioned by endometrium from Day 10 or Day 12 of pregnancy. However, antiviral activity was undetectable in medium conditioned by endometrium from one of the Day-12 gilts. In Exp. 3, antiviral activity was present in medium from only 1 of 3 cultures from Day-12 gilts when assayed unfrozen. Antiviral activity was lower (P less than 0.01) in cultures of conceptuses from Day 12 than Day 14 of pregnancy; however, antiviral activity increased quadratically (P less than 0.05) when cultures contained 0, 0.01, 0.1 and 1.0 units/ml aprotinin, respectively. Freezing and thawing culture medium did not reduce (P greater than 0.10) antiviral activity compared to medium assayed unfrozen (1438 vs 1354 units/ml, respectively). These results suggest a regulatory influence of the endometrium on secretion of antiviral proteins by pig conceptuses in vitro.
Animals↗
Endometrial inositol phosphate turnover in pigs is reduced during pregnancy and estradiol-induced pseudopregnancy.
Three experiments were conducted to examine inositol phosphate (IP) turnover in response to treatments applied in vitro to endometrium from cyclic (CYC), pregnant (PREG) and estradiol-induced pseudopregnant (PSP) gilts. In Exp. 1, treatments (in 25 microliters .1 M NaHCO3) were 1) control (NaHCO3), 2) 125 ng oxytocin, 3) .25 micrograms prolactin, 4) 2.5 micrograms prolactin and 5) 5 micrograms pig conceptus secretory proteins (pCSP). Basal IP turnover on d 14 (estrus = d 0) for CYC was 3.9 to 5.0-fold greater than for PREG gilts and .6 to 1.1-fold greater than for PSP gilts (P less than .05). Oxytocin increased IP turnover 23 to 42% in CYC gilts (P less than .05), but not in PREG or PSP gilts. The treatment x reproductive status interaction (P less than .05) indicated that pCSP increased IP turnover 74 to 140% in PREG gilts but decreased it 18 to 22% in CYC and 17 to 50% in PSP gilts. In Exp. 2, treatments were applied in a 2 x 2 x 2 arrangement: 1) 0 or 125 ng oxytocin; 2) 0 or 2.5 micrograms prolactin and 3) 0 or 5 micrograms pCSP. Basal IP turnover on d 14 was 3.3 to 5.4-fold greater (P less than .05) in CYC than in PSP gilts and was affected by interaction (P less than .05) of pCSP and prolactin. Inositol phosphate turnover was increased by prolactin (12 to 22%) and by pCSP (7 to 34%) but, when combined, the stimulatory effects of each were eliminated.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Effect of porcine conceptus secretory proteins on interestrous interval and uterine secretion of prostaglandins.
Porcine conceptus secretory proteins (pCSP) were obtained from medium in which pig conceptuses, collected on Day 15 of pregnancy, were cultured for 30 h. Culture medium was pooled, dialyzed, and concentrated by Amicon ultrafiltration for intrauterine infusion. Serum proteins (SP) were obtained from blood collected from a Day 15 pregnant gilt and diluted for intrauterine infusion. Catheters were placed into both uterine horns and the inferior vena cava of cyclic (Day 8) gilts. Single blood samples were collected at 0800 h on Days 9, 10, and 11. On Day 11, all gilts received 1 mg estradiol-17 beta (E2) i.m. at 0800 h. Protein infusions commenced on Day 12 and continued through Day 15, twice daily at 0800 h and 2000 h. Protein infusions per uterine horn were (1) 4.0 mg pCSP + 4.0 mg SP (pCSP, 4 gilts) and (2) 8.0 mg SP (SP, 4 gilts). Blood samples were collected every 15 min on Days 12 through 17 between 0805 h and 1100 h. Single blood samples were collected at 0800 h after Day 17 until gilts exhibited estrus. Concentrations of prostaglandin (PG) E, 13,14-dihydro-15-keto-PGF2 alpha (PGFM), and progesterone (P4) were measured by specific radioimmunoassays. Interestrous intervals for pCSP-treated (18.2 days) and SP-treated (18.0 days) gilts were not different (SEM = 0.8 days) and temporal changes in concentrations of P4 in plasma did not differ between pCSP-treated (29.2 ng/ml) and SP-treated (31.2 ng/ml) gilts.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Comparative aspects of maternal recognition of pregnancy between sheep and pigs.
Sheep conceptuses secrete a protein, oTP-1, between Days 10 and 21 of gestation which is responsible for establishment of pregnancy. oTP-1 inhibits uterine production of luteolytic amounts of PGF-2 alpha (PGF) produced in response to oestradiol and oxytocin. oTP-1 does not compete with oxytocin for binding to oxytocin receptors, but may interfere with oxytocin stimulation of the inositol phospholipid system. Pig conceptuses secrete oestrogens between Days 10 and 15 of pregnancy which are essential for establishment of pregnancy. Oestrogens, directly or indirectly, alter secretion of PGF from an endocrine direction (towards uterine vasculature) to an exocrine direction (towards the uterine lumen). PGF sequestered in the uterine lumen is unavailable to exert a luteolytic effect on the CL. Pig conceptus secretory proteins stimulate uterine production of PGF and PGE. Conceptus secretory proteins of sheep and pigs include proteins which have antiviral activity and may be considered interferons. In sheep, oTP-1 has both antiluteolytic and antiviral properties. The specific pig conceptus secretory proteins(s) possessing antiviral activity has not been established. Unlike oTP-1, however, it does not appear to possess antiluteolytic activity.
Animals↗
Prostaglandin secretion by perifused porcine endometrium: further evidence for an endocrine versus exocrine secretion of prostaglandins.
Bilateral perifusion devices were utilized for measurement of prostaglandin secretion by luminal and myometrial surfaces of porcine endometrium. Tissues were collected from Days 10, 12 and 14 pregnant, Day 14 cyclic and Day 14 estrogen-induced pseudopregnant gilts. Each tissue was placed into duplicate perifusion devices and perifused with Krebs-Ringer Bicarbonate solution at 3 ml/10 min for 2 h, fractions collected every 10 min and oxytocin (1 IU/ml) perifused during fractions 6-10 to the luminal side of one chamber and to the myometrial side of the other chamber. Secretion rates of PGF were higher (P less than 0.05) than PGE2 for each status. Secretion rates of PGF and PGE2 were higher (P less than 0.01) from the luminal side for Day 12 pregnant, Day 14 pregnant and Day 14 pseudo-pregnant gilts, whereas secretion was higher from the myometrial side for Day 10 pregnant and Day 14 cyclic gilts. Oxytocin increased (P less than 0.01) prostaglandin secretion from the luminal side regardless of reproductive status. Pregnancy at Day 12 and Day 14, as well as estrogen treatment, were associated with prostaglandin secretion in a luminal (exocrine) orientation versus a myometrial (endocrine) orientation for Day 14 cyclic and Day 10 pregnant gilts. These data indicate an estrogen associated switch between Days 10 and 12 of pregnancy from an endocrine to an exocrine secretion of prostaglandins.
Animals↗
Health manpower project.
Explore the source record for details and available documents.
Education, Nursing, Continuing↗