Biomedical subjects

F W Bazer

Publications and source records attributed to F W Bazer.

At least 235 records · Page 13Linked to original sources

PubMed1985

The carbohydrate structure of porcine uteroferrin and the role of the high mannose chains in promoting uptake by the reticuloendothelial cells of the fetal liver.

Uteroferrin, the iron-containing, progesterone-induced phosphatase of the porcine uterus, is a glycoprotein carrying a single oligosaccharide chain. Most of the uteroferrin isolated from either uterine secretions or allantoic fluid has endoglycosidase H-sensitive carbohydrate chains with either five or six mannose residues. As determined by 1H-NMR spectroscopy, the Man6 oligosaccharide has the following structure. (Formula: see text) The Man5 species lacks the terminal alpha 1,2-linked residue. Uteroferrin is transported across the pig placenta and has been proposed to be involved in iron transfer to the fetus (see Buhi, W. C., Ducsay, C. A., Bazer, F. W., and Roberts, R. M. (1982) J. Biol. Chem. 257, 1712-1721). Injection of 125I-labeled uteroferrin into the umbilical vein of midpregnant fetuses resulted in incorporation of label into the liver, the major site of fetal erythropoiesis. Light and electron microscope autoradiography revealed that the primary sites of uteroferrin uptake were the reticuloendothelial cells lining the liver sinusoids. Reticuloendothelial cells isolated from either fetal pig or adult rat livers were shown to accumulate uteroferrin when cultured in vitro. Uptake was inhibited by yeast mannan and by glycopeptides isolated from either ovalbumin or uteroferrin. Rat cells did not accumulate uteroferrin whose high mannose chains had been removed using endoglycosidase H. Moreover, the K uptake values (3 X 10(-7) M), specific competition by D-mannose and L-fucose bovine serum albumin, and inhibition by EDTA are consistent with an uptake mechanism involving a receptor for high-mannose oligosaccharides on the liver sinusoidal cells. It is suggested that one function of this receptor in the fetal pig is to remove maternally derived uterine glycoproteins from the fetal circulation. In the case of uteroferrin this process provides iron to the fetal liver.

Acid Phosphatase↗

PubMed1985

Localization of the iron transport glycoprotein, uteroferrin, in the porcine endometrium and placenta by using immunocolloidal gold.

Uteroferrin, a glycoprotein implicated in transplacental iron transport in the pig, has been localized within the porcine uterus during mid pregnancy by using immunocolloidal gold labeling of sectioned material. In the endometrium where uteroferrin is synthesized, it was located exclusively in the non-ciliated cells of the glandular epithelium where it appeared to follow a common route of secretory glycoprotein from the cell (i.e., rough endoplasmic reticulum----Golgi----condensing vacuoles----secretory granules). Uteroferrin was present in the lumen of the glands and overlying placental areolae and in the large absorptive chorionic epithelial cells of the areolae. The latter cells contained numerous small vesicles and tubules as well as large endocytotic vacuoles. All these structures contained uteroferrin and are probably involved in translocating the glycoprotein in intact form from the uterine lumen to the blood capillaries draining the placenta.

Acid Phosphatase↗

PubMed1985

Characterization of proteins produced in vitro by periattachment bovine conceptuses.

Bovine conceptuses from Days 16 (n = 4), 19 (n = 6), 22 (n = 3), and 24 (n = 4), and chorion from Day 69 (estrus/mating = Day 0) were cultured for 24 h in modified minimum essential medium (MEM) in the presence of radioactive L-leucine [( 3H] leucine) to characterize de novo synthesis and release of proteins. Proteins released into MEM were identified by two-dimensional polyacrylamide gel electrophoresis, fluorography, and gel and ion exchange chromatography. Major polypeptides identified in MEM were different from those identified in conceptus and chorionic tissues. Both uptake of [3H] leucine and quality of polypeptides produced de novo and released into MEM were related to stage of conceptus development. Percent retention of [3H] leucine in MEM was lowest (P less than 0.01) in Day 16 cultures (1.2 +/- 4.1%), increased in Days 19 (16.8 +/- 3.7%) and 22 cultures (20.9 +/- 5.8%), and decreased (P less than 0.07) in Day 24 cultures (6.9 +/- 4.1%). Complexity of polypeptides increased after Day 16. Days 16, 19, 22 and 24 conceptus culture MEM was enriched in low-Mr, acidic polypeptides (Mr/isoelectric point ranges: 22K-26K/6.5-5.6, 20K-26K/5.5-5.4, and 16K-20K/5.0-4.5), which were not prominent products of Day 29 and 69 tissues. A high-Mr (Mr +/- SEM; 735K +/- 22K) glycoprotein was produced by all conceptus and chorionic tissues. The transient nature of production of low-Mr polypeptides suggests that they may be required during the periattachment period.

Amino Acids↗

PubMed1985

Characterization of proteins produced in vitro by bovine endometrial explants.

Endometrial tissues were obtained from 17 pregnant (P, estrus/mating = Day 0; Day 16, n = 4; Day 19, n = 6; Day 22, n = 3; Day 24, n = 4) and six nonpregnant (NP; Day 16, n = 4; Day 19, n = 2) cattle, as well as from one cyclic (nonbred) cow (Day 4), one later-pregnant cow (Day 69), and both ligated and pregnant uterine horns of three unilaterally pregnant cattle (UP; Day 270). Tissues (approximately equal to 500 mg wet tissue/explant) were cultured for 24 or 48 h in modified minimal essential medium (MEM), in the presence of radioactive amino acid and/or amino sugar substrate(s) (L-[3H] leucine, L-[14C] leucine, and D-[3H] glucosamine), in order to characterize substrate uptake and de novo synthesis and release of proteins and polypeptides in vitro. Endometrial explants from all cattle produced proteins de novo from radiolabeled substrates. Chromatographic (gel filtration, cation, and anion exchange) and two-dimensional polyacrylamide gel electrophoretic analyses revealed complex patterns of primarily acidic radiolabeled polypeptides in dialyzed MEM, which were absent from endometrial tissue homogenates. No qualitative differences were noted in the array of proteins released into MEM associated with either pregnancy status (P vs. NP, UP) or stage of gestation (Days 16, 19, 22, 24, 69, and 270). Medium from all endometrial explants was enriched in polypeptides in four Mr (X 10(3)/pH classes (I, approximately equal to 14/greater than 7.2; II, 19-24/5.4-6.3; III, 28-31/6.9-7.3; and IV, greater than or equal to 150/less than or equal to 5.1).

Amino Acids↗

PubMed1985

Differential patterns of secretory protein localization within the pig uterine endometrium.

We have compared the distribution of two of the major secreted proteins of the porcine uterus within the endometrium of ovariectomized pigs which had received hormone replacement therapy for 30 days. The proteins studied, plasmin/trypsin inhibitor (PI) and uteroferrin (Uf), an iron-containing acid phosphatase, were both secreted into the uterine lumen by ovariectomized gilts given progesterone (P4) or P4 and 17 beta-estradiol but not by animals given 17 beta-estradiol alone or corn oil. The two proteins were localized immunocytochemically within the endometrium using an immunoperoxidase procedure. The results confirmed that production of PI and Uf was P4-dependent and demonstrated that the primary site of synthesis of PI was the surface and upper glandular epithelium, while Uf synthesis was confined to the glandular epithelium. A similar localization of PI and of Uf was found in endometrial tissue from pigs at day 13 (late luteal phase) of the estrous cycle. These results suggest that the uterine epithelium of the pig is regionally differentiated with regard to the production of P4-induced proteins.

Acid Phosphatase↗

PubMed1985

In vitro synthesis and secretion of ovine trophoblast protein-1 during the period of maternal recognition of pregnancy.

The trophoblast of the sheep conceptus secretes a low mol wt, acidic polypeptide called ovine trophoblast protein-1 (oTP-1) from days 13-21 of pregnancy. This protein has been proposed to have an antiluteolytic role during pregnancy, since infusion of oTP-1 into the uterine lumen of nonpregnant ewes lengthened their estrous cycles. In the present study, mRNA from day 16 conceptuses was translated using a cell-free wheat germ lysate, and oTP-1 was immunoprecipitated from the translation mixture by means of a specific rabbit antiserum. The predominant protein synthesized under these conditions was oTP-1, which had a mol wt (21,000) higher than that of oTP-1 secreted by intact conceptuses into culture medium (mol wt, 17,000). In a separate experiment, in vitro secretion of oTP-1 from conceptuses on days 13, 17, and 21 of pregnancy was examined. Conceptuses were cultured for 24 h in the presence of [3H]leucine, and oTP-1 was immunoprecipitated from the culture medium. Incorporation of [3H]leucine into secreted macromolecules (mol wt, greater than 3500) increased 12-fold from days 13 to 21, while release of immunoprecipitable 3H-labeled oTP-1 increased 27-fold from days 13 to 17 and declined by over 50% from days 17 to 21. When results were expressed per microgram DNA, secretion of total macromolecules declined from days 13 to 17 and then remained constant until day 21. The pattern of oTP-1 secretion held whether results were expressed per conceptus or per unit DNA. The interestrous intervals for the donor ewes were shorter for ewes which had their conceptuses removed on day 13 than for ewes flushed on day 17 or 21. Thus, the period of oTP-1 synthesis corresponds closely to the time at which the conceptus acts to extend luteal lifespan.

Animals↗

PubMed1985

Appearance of beta-hexosaminidase and other lysosomal-like enzymes in the uterine lumen of gilts, ewes and mares in response to progesterone and oestrogens.

In one experiment, ovariectomized gilts were treated with corn oil (vehicle), progesterone, oestradiol-17 beta or both steroids. While oestradiol treatment did not stimulate enzyme activity in uterine flushings relative to vehicle-treated animals, gilts treated with progesterone had elevated amounts of all enzymes measured. Progesterone was less effective when co-administered with oestradiol-17 beta. Enzymes were not equally stimulated by progesterone. For example, there was a 909-fold increase in acid phosphatase activity in uterine flushings and a 304-fold increase in beta-N-acetylglucosaminidase, but only a 10-fold increase in beta-glucosidase. Endometrial explants from gilts synthesized and secreted radiolabelled beta-N-acetylglucosaminidase, suggesting that at least some lysosomal enzymes enter the uterus through secretory processes. In other experiments, changes in beta-N-acetyglucosaminidase in uterine fluids of mares and ewes treated with hormonal regimens similar to those given to the gilts were evaluated. Treatment with the combination of progesterone and oestrogen stimulated accumulation of the enzyme relative to that in vehicle-treated animals. The biochemical properties of porcine beta-N-acetylglucosaminidase were examined in detail. Properties of the uterine enzyme were similar to reported values for lysosomal hexosaminidase. These included molecular weight (82 000-89 000), pH optimum (pH 4.4), presence of two isomers (isoelectric points of 5.5 and 8.0) and ability to hydrolyse substrates for glucosaminidase and galactosaminidase. We conclude that steroids induce the accumulation of lysosomal enzymes in the uterine lumen. The degree of stimulation differed between enzymes, suggesting that those enzymes stimulated to the greatest extent may play an important role in pregnancy.

Acetylglucosaminidase↗

PubMed1985

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

Primary cell cultures were established from cells derived from dissociated Day-14 and -16 sheep and pig blastocysts. The appearance of cells in culture from both species was similar. Cultures contained a variety of cells with distinct morphologies, some were small and compact and formed clumps and multiple layers while others were large, flat and formed a monolayer. Within 4 h of culturing small floating fluid-filled spheres of cells were observed in the medium; some of these increased in size to greater than 1 cm diameter over 1-2 weeks. In addition, fluid-filled domes of cells arose from the underlying monolayer. Contractile cells became evident after about 8 days and some became organized into large patches of contracting tissue. Two-dimensional polyacrylamide gel electrophoresis and fluorography were performed on proteins released into the medium by confluent monolayers, floating spheres and floating cells that failed to attach during the first 24 h. All cultures produced as major products proteins with electrophoretic mobilities identical to certain fetal plasma proteins. In general, cultures did not produce proteins characteristic of short-term cultures of whole conceptuses harvested at Days 14-16. In cultures established from sheep blastocysts only the cells that failed to attach produced ovine trophoblast protein-1, a major polypeptide produced by the trophectoderm of the sheep conceptus between Days 13 and 21 of pregnancy.

Animals↗

PubMed1985

Steroid metabolism by endometrial and conceptus tissues during early pregnancy and pseudopregnancy in gilts.

Endometrial and conceptus tissues were obtained on Days 10.5, 11, 12, 16 and 25 of pregnancy and Day 25 of pseudopregnancy of gilts and incubated for 6 h in Minimal Essential Medium (5 ml) containing 35 ng [3H]progesterone. Metabolism of [3H]progesterone to oestrone, oestradiol and oestriol was determined by gas and high-pressure liquid chromatography and successive recrystallizations with unlabelled standards. Conceptuses collected between Days 10.5 and 12 were spherical, tubular or filamentous and incubated with 500 mg endometrium and [3H]progesterone. Production of oestrone by spherical conceptuses was not detected, but was 44-47 pg/tubular conceptus and 21 pg/filamentous conceptus. A similar trend was observed for oestradiol. Conceptus tissues from Days 16 and 25 (chorion) were most active in producing oestrone (123 and 520 pg/mg tissue, respectively) and oestradiol (277 and 876 pg/mg tissue, respectively). Endometrial oestrogen production was less than that for conceptus tissue for oestrone and oestradiol on Days 16 and 25 of gestation. Coincubations of endometrium and conceptus tissues had lower oestrogen production than conceptus alone. Endometrium from Day 25 of pseudopregnancy metabolized [3H]progesterone to several non-polar metabolites, but no oestrogens were detected. An unidentified phenolic metabolite of [3H]progesterone was detected in higher quantities than either oestrone or oestradiol; 445 to 461 pg/conceptus at the tubular stage. These results indicate temporal changes in the conversion of [3H]progesterone to oestrogens by conceptus and endometrial tissue from pregnant gilts, but not endometrium from pseudopregnant gilts.

Animals↗

PubMed1985

Adipose tissue fatty acid metabolism during pregnancy in swine.

In vitro adipose tissue fatty acid pool size (POOL), fatty acid release (FAR) and esterification (EST) were measured in peritoneal (PFP) and subcutaneous mammary (MFP) fat pads of swine at d 15, 30, 45, 60, 75, 90, 105 and 112 of pregnancy. Plasma free fatty acids (FFA) and triglycerides (TG) were not altered by stage of pregnancy. Basal EST in PFP was generally constant across pregnancy with a peak at d 75. Basal EST in MFP was elevated at d 30, 75 and 112. Esterification in response to norepinephrine stimulus (NE) was lower than basal rates in both fat depots. Basal FAR was constant throughout pregnancy in PFP, but elevated at d 75 and 90 in MFP. Fatty acid release in response to NE was biphasic with peaks at d 30 and in late pregnancy (in MFP, micromolar FAR in response to NE was 69.3% greater on d 75 to 112 than on d 45 to 60). Basal POOL was constant throughout pregnancy in both depots and lower than NE-stimulated POOL. All responses to NE were greater in MFP than in PFP, indicating that adipose tissue surrounding the developing mammary gland had higher metabolic activity and a greater response to NE than peritoneal adipose. Changes in fatty acid metabolism during pregnancy in swine are temporally related to published values for plasma steroids, fetal growth and mammary development. Metabolic adaptations in adipose and mannary epithelial tissue occur in synchrony with changing plasma estrogen concentrations, redirecting energy flow from maternal adipose tissue toward developing mammary and fetal tissue.

Adipose Tissue↗

PubMed1985

Influence of environmental temperature on prolificacy of pigs.

Exposure of male and female pigs to elevated ambient temperatures can result in reduced reproductive efficiency. When boars and gilts are exposed to heat stress, respiratory rates increase to enhance evaporative cooling because minimal sweating occurs. During early pregnancy, gilts are especially susceptible to heat stress. Decreased conception rates and reduced litter size occur when gilts are exposed to elevated ambient temperature during Days 0 to 16 after mating. Concentrations of progesterone in peripheral plasma were reduced during Days 13-19 after mating and luteal function was extended to Day 25 in heat-stressed gilts that did not conceive. Increased concentrations of oestradiol during Day 10 to 12 of heat stress may interfere with normal maternal recognition of pregnancy. Heat stress reduced the amount of embryonic tissue present at Day 16 of pregnancy but the protein synthetic activity of the tissue was not altered. Thus some embryos may be lost and a reduction in litter size may occur. The production of oestrogen by the conceptus and uterus is not altered on Day 16, after exposure to heat stress for the previous 8 days. These studies suggest that heat stress during early pregnancy can alter the reproductive endocrine system, especially the control of luteal function. In addition, heat stress may have a direct effect on embryo or conceptus development. Exposure of boars to elevated ambient temperatures causes reductions in semen quality, sperm output and fertility. About 5 weeks are required for boars to recover from the detrimental effects of heat stress and to produce semen with potential for maximal fertility. Increased temperature has an inhibitory effect on spermatid maturation and on testicular androgen biosynthesis. Improvements in reproductive performance can be achieved by increasing evaporative cooling of boars.

Androgens↗

PubMed1984

Effect of exogenous gonadal steroids and pregnancy on uterine luminal prostaglandin F in mares.

Two experiments were conducted to assess the effect of exogenous hormone treatment on uterine luminal prostaglandin F (PGF). In the first experiment ovariectomized pony mares received either corn oil (21 days, n = 3), estradiol valerate (21 days, n = 3), progesterone (21 days, n = 3) or estradiol valerate (7 days) followed by progesterone (14 days, n = 4). Progesterone treated mares had higher (P less than .01) uterine luminal PGF compared with all other groups, and no differences were detected between other treatment comparisons. In Experiment II, uterine fluid was collected from 4 ovariectomized horse mares before and after treatment with estradiol valerate (7 days) followed by progesterone (50 days). Pretreatment uterine luminal PGF levels were lower (P less than .001) than post-treatment levels (.03 vs 76.80 ng/ml). In a third experiment PGF was measured in uterine fluid of pony mares on days 8, 12, 14, 16, 18 and 20 of the estrous cycle and pregnancy. In nonpregnant mares a day effect (P less than .03) was observed in which uterine fluid PGF increased during the late luteal phase and declined thereafter. In contrast, no day effect was observed in pregnant animals and uterine luminal PGF was lower (P less than .001) than in cycling animals. These studies indicate that exogenous progesterone administration results in a large increase in uterine luminal PGF, whereas, pregnancy results in suppression. Taken collectively with previous work from our laboratory, these results suggest that while the endometrium of pregnant mares is capable of producing large amounts of PGF, the presence of a conceptus impedes its synthesis and/or release which allows for luteal maintenance.

Animals↗

PubMed1984

Uteroferrin has N-asparagine-linked high-mannose-type oligosaccharides that contain mannose 6-phosphate.

Uteroferrin is an iron-containing, progesterone-induced, acid phosphatase that is secreted in large amounts by the uterine endometrium of pigs. During pregnancy, it transports iron across the chorioallantois (placenta) for use in fetal hematopoiesis. In this paper, it is reported that uteroferrin synthesized by cultured endometrial explants possesses N-linked, high-mannose, oligosaccharide chains that contain 6- phosphomannose units. The latter is regarded as a possible recognition marker whereby acid hydrolases are targeted to the lysosome. On uteroferrin, however, the majority of the phosphate is in single diester linkages between the mannose and a covering N-acetylglucosamine. It is suggested that uteroferrin is a lysosomal enzyme that has assumed a role in iron transport and metabolism and is secreted because the covering N-acetylglucosamine is not removed.

Acid Phosphatase↗

PubMed1984

Response of plasma glucose, fructose and insulin to dietary glucose and fructose in the lactating sow.

Twenty-two Hampshire-Yorkshire X Large White sows of second and third parity were allotted randomly to one of three dietary treatments. Five sows were fed 6.0 kg/day of a corn-soybean meal lactation diet (control diet). Twelve sows were fed the control diet in which 24% of the composition was supplied by corn syrup containing 72% fructose on a dry matter basis (fructose diet) and five sows were fed the control diet in which 24% of the composition was supplied by powdered dextrose (glucose diet). All diets were fed from days 1 through 21 of lactation. Blood was collected from all sows immediately prior to feeding and hourly for 6 hours postprandial via jugular vein cannulae following a single feeding on seven separate but nonconsecutive days during the 21-day period. Fructose was absorbed from the digestive tract of sows as evidenced by elevated (P less than 0.01) conventions of fructose in plasma. Sows fed the fructose diet also had higher (P less than 0.01) plasma glucose concentrations than did those fed the glucose and control diets. The concomitant elevated glucose concentration following ingestion of the fructose diet was not associated (P greater than 0.10) with increased insulin concentration. Fructose in plasma was associated with a slight but significant increase in insulin although the mean concentration of insulin in plasma was only one-third that measured in sows fed the glucose and control diets. These data suggest that fructose in vivo has a glucose-sparing effect presumably mediated through a physiological mechanism that lowers insulin concentration.

Animals↗

PubMed1984

Suppression of phytohemagglutinin-stimulated lymphocyte blastogenesis by ovine uterine milk protein.

Two basic glycoproteins (UTM-P) with molecular weights of 57,000 and 59,000 were purified from ovine uterine milk collected on Days 125 and 130 of pregnancy. The UTM-P were evaluated for immunosuppressive activity in phytohemagglutinin (PHA)-treated, mixed lymphocyte (MLC) and resting lymphocyte (RLC) cultures. For PHA and RLC cultures, UTM-P (2.5 to 800 micrograms UTM-P/ml) were added to 1 X 10(6) lymphocytes and 0.8 micrograms of PHA (for PHA cultures only), while for the MLC, UTM-P (50 to 1600 micrograms UTM-P/ml) were added to 5 X 10(5) lymphocytes combined from each of two ewes. Following [3H] thymidine addition, cells were later harvested for determination of thymidine incorporation. Lymphocyte blastogenesis was suppressed by UTM-P in PHA (R2 = 0.32 to 0.92, P less than 0.01 to 0.001), MLC (R2 = 0.8, P less than 0.001) and RLC (R2 = 0.65, P les than 0.01) experiments. To determine reversibility, PHA-treated lymphocytes were incubated with UTM-P for 6, 12 or 24 h, then washed to remove surface UTM-P. Incubation was continued in the presence of PHA as with other experiments. Exposure of lymphocytes to UTM-P for 6 or 12 h did not result in suppression of blastogenesis, whereas exposure for 24 h was sufficient for suppression (P less than 0.01). In an additional experiment, UTM-P were added to PHA-treated cultures at 0, 6, 12 or 24 h. Suppression (P less than 0.01) of blastogenesis was observed for each time period. Immunosuppressive activity was not mediated by overall cytotoxicity and was not affected by routine handling and storage of UTM-P. Data from these experiments provide one explanation for tolerance of the conceptus allograft during defined stages of ovine pregnancy.

Animals↗

PubMed1984

Ovine trophoblast protein 1, an early secreted blastocyst protein, binds specifically to uterine endometrium and affects protein synthesis.

Ovine trophoblastic protein-1 (oTP-1), an early secretory protein of the sheep blastocyst, was purified after culturing day 14-16 conceptuses for 24 h in vitro. The localization of oTP-1 in the pregnant day 16 sheep uterus was determined immunocytochemically. The protein was associated with trophectoderm cells of the elongated blastocyst and with the surface and upper glandular epithelium of the maternal uterus. Receptors that bound oTP-1 with high affinity (Kd = approximately 2 X 10(-10) M) were present in crude membrane preparations derived from homogenates of endometria from day 12 nonpregnant and anestrous ewes. Uterine infusion of 125I-labeled oTP-1 into day 12 nonpregnant ewes showed that the majority of the radioactivity was retained in the uterus, and only very small amounts of intact protein appeared to enter the maternal vasculature. There was no significant association with the corpora lutea, ovaries, or other tissues tested. oTP-1 failed to compete with ovine PRL for rabbit mammary gland receptors or with hCG or bovine LH for sheep luteal cell receptors, and the oTP-1 did not stimulate progesterone production by dispersed luteal cells from day 12 cycling ewes. Incubation of endometrial explants from day 12 nonpregnant ewes with 5 micrograms/ml oTP-1 resulted in increased rates of protein release into the medium. Two-dimensional polyacrylamide gel electrophoresis revealed that the synthesis of six polypeptides was stimulated selectively by the presence of oTP-1. Together these data suggest that oTP-1 acts on the maternal endometrium. It is suggested that the interaction of oTP-1 with uterine endometrium may elicit maternal responses which contribute to the maintenance of pregnancy in the sheep.

Animals↗

PubMed1984

Endocrine aspects of early pregnancy in pony mares: a comparison of uterine luminal and peripheral plasma levels of steroids during the estrous cycle and early pregnancy.

Comparisons of estrone, 17 beta-estradiol, and plasma progestin concentrations were made in uterine fluid and peripheral blood of nonpregnant and pregnant pony mares. Concentrations of these steroids were also measured within yolk sac fluid from blastocysts on days 12, 14, 16, and 18 of pregnancy to obtain more complete analyses of the uterine environment (uterine fluid plus yolk sac fluid) of early pregnancy. Thirty mares were randomly assigned to six treatment groups (n = 5/group), and uterine fluid and peripheral blood samples were obtained on days 8, 12, 14, 16, 18, and 20 postovulation. After a recovery period of one estrous cycle, mares were bred at their next estrus. Animals were hysterectomized on the same treatment day to which they had previously been assigned in the nonpregnant phase of this study. Using this design, uterine fluid and peripheral blood samples were collected from each mare on equivalent days of the estrous cycle and pregnancy. Significant differences in day trends were found between nonpregnant and pregnant animals for estrogens and progestins in both uterine fluid and peripheral plasma. Furthermore, these data demonstrate that large increases in estrogens occur after day 12 of pregnancy in uterine and yolk sac fluids, with estrone becoming the predominant estrogen by days 18 and 20 in yolk sac and uterine fluids, respectively. These changes were not detected in peripheral plasma, which indicates that changes occurring within the uterine environment are not discernible in the systemic circulation during early pregnancy. These results indicate that the large amounts of estrogens appearing in uterine fluids during early pregnancy are of conceptus origin and may be an important factor in regulating the environment in which the conceptus develops.

Animals↗

PubMed1984

Proteins released by cultured Day 15-16 conceptuses prolong luteal maintenance when introduced into the uterine lumen of cyclic ewes.

Experiments were performed to determine whether proteins, produced and released into the incubation medium by Day 15-16 sheep conceptuses cultured for 24-48 h, could prolong the functional lifespan of the corpus luteum (CL) when infused into the uterine lumen of cyclic ewes. Beginning on Day 12 (oestrus = Day 0) either a concentrated (2 ml) solution of total conceptus culture medium protein (2.2 mg) or diluted sheep serum (2.2 mg protein) was introduced daily via an indwelling catheter into the uterine lumen of 3 ewes for 7 days (Days 12-18). Peripheral blood samples were collected daily for 14 days (Days 12-25). On Day 25 all ewes were laparotomized and ovaries observed to determine whether CL previously marked with India ink were maintained. All controls had ovulated and formed new CL. By contrast none of the conceptus protein-treated ewes had ovulated, and their peripheral progesterone levels remained elevated. One ewe maintained a functional CL until Day 52 when she was hysterectomized. Light microscopy of histological sections prepared from the endometrium revealed glandular development comparable to that in the endometrium of cyclic animals during late dioestrus. The cells of the CL were similar to those from cyclic animals during mid- to late dioestrus. Ovine trophoblast protein ( oTP -1), a major protein secreted by the sheep conceptus between Days 13 and 21 of pregnancy, was purified from conceptus incubation medium and injected (0.2 mg protein/day) into the uterine lumen of 3 animals. Plasma progesterone concentrations indicated that oTP -1-treated animals maintained luteal function 4 days longer than did control animals.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗