Intrauterine cortisol, aldosterone, and corticosteroid binding globulin-like activity during early porcine pregnancy and the estrous cycle

Studies were conducted to determine whether the corticosteroids cortisol and aldosterone, and corticosteroid-binding globulin (CBG) were present in the porcine early-embryonic environment. Cortisol was measured in uterine flushings from white crossbred gilts at Days 7, 10, 13, and 16 of the estrous cycle and pregnancy. Total content of cortisol increased (p < 0.01) between Days 13 and 16, and immunoreactive CBG (ir-CBG) increased (p < 0.01) between Days 10 and 13, in both cyclic and pregnant gilts. In a separate study with Chinese Meishan gilts, total cortisol and aldosterone content of uterine flushings increased (p < 0.02) between Days 10 and 15 of the estrous cycle and pregnancy. In another study with white crossbred gilts, CBG-like binding activity in uterine flushings was low at Day 10, then increased over 100-fold at Day 15 (p < 0.01). However, levels of CBG-like binding activity on Day 15 were 100-fold lower than those of ir-CBG measured in the previous study and could bind less than 4% of the uterine luminal cortisol. Differences between ir-CBG and CBG binding might be due to the ability of the CBG antibody to recognize either biologically inactive CBG or structurally similar molecules. CBG-like binding activity, which appeared unrelated to glucocorticoid receptors, was also present in the endometrial cytosol of white crossbred gilts. Concentrations (fmol/mg protein) of endometrial CBG-like activity decreased (p = 0.03) between Days 10 and 15 of the estrous cycle and pregnancy, did not differ with reproductive status, and on Day 15 were comparable to concentrations in uterine flushings but threefold lower (p < 0.01) than those in the serum. Equilibrium dissociation constants for CBG-like binding activities were comparable among the three locations. These studies indicate that corticosteroids are present-primarily in the free form-within the porcine uterine lumen and could influence early porcine conceptus development. Endometrial CBG-like binding activity could mediate actions of cortisol or progesterone on uterine function.     

Conceptus, progesterone, and breed effects on uterine protein secretion in swine

This experiment consisted of the following treatment-breed groups: 1) White crossbred gilts, 2) White crossbred gilts treated with progesterone (200 mg/d in corn oil given on d 2 and 3 after estrus), and 3) Chinese Meishan gilts. Pregnant and nonpregnant gilts (n=3 to 6) from each treatment-breed combination were assigned to be slaughtered on d 10, 11, 12, 13, and 15. At slaughter each uterine horn was flushed with 20 mL of minimal essential medium. Uterine flushings were assayed for total protein, acid phosphatase, uteroferrin, retinol-binding protein, and oxytocin. Uterine flush total protein was increased by progesterone treatment, was unaffected by pregnancy status, and was less in Meishans. Similar patterns were found for retinol binding protein and uteroferrin, except that uteroferrin was greater in pregnant than in nonpregnant gilts. Oxytocin was greater in pregnant than in nonpregnant gilts, was not influenced by progesterone treatment, and was similar in Meishan and in White crossbred gilts. These results indicate that the conceptus does not influence secretion of either total protein or retinol binding protein during pregnancy and that the onset of secretion of these uterine proteins may be controlled by progesterone. The presence of the conceptus is associated with increased uteroferrin and oxytocin production. The decreased secretion of uterine proteins in Meishan gilts may partially explain the slower embryonic development that has been reported for this breed.     

The impact of participation in a study abroad programme on students'' conceptual understanding of community health nursing in a developing country

Porcine uterine tissues were collected from Days 10 to 14 of gestation (peri-implantation period) or corresponding days of the estrous cycle. Results indicated a marked increase in beta transforming growth factors (TGFbeta1, TGFbeta2, and TGFbeta3) and TGFbeta receptor (type I and type II) immunostaining in uterine luminal epithelium (ULE) between Days 10 and 14 of gestation, but there was no increase in ULE immunostaining on the corresponding days of the estrous cycle. Uterine glands and stroma were intensely immunopositive in pregnant gilts for TGFbeta isoforms and their receptors, but immunostaining was weak to undetectable in cycling gilts. No differences were detected in myometrium, in which immunostaining was moderate in both cycling and pregnant gilts. Additionally, TGFbeta2 and TGFbeta receptor (type I and type II) immunostaining was detected in uterine monocyte/macrophage-like cells. Western blotting detected the presence of all three TGFbeta isoforms in uterine luminal flushings. The CCL64 cell TGFbeta bioassay detected bioactive TGFbetas++ in uterine luminal flushings on Days 12, 13, an 14 of gestation. These results strongly indicate that uterine expression of TGFbetas and their receptors is pregnancy specific and that bioactive TGFbetas are present at the conceptus-maternal interface in the peri-implantation period in pigs. Thus TGFbetas are likely to be involved in autocrine-paracrine interactions between the maternal uterus and the conceptus.     

Effect of the conceptus and lack of effect of uterine space on endometrial protein secretion during mid-gestation in swine

The effect of the conceptus and of reduced uterine space on endometrial protein secretion was examined on Days 40, 60, and 80 of gestation in white crossbred gilts. Twenty-nine gilts were checked daily for estrus, and 15 were given 5 mg estradiol valerate daily from Days 11 to 15 (Day 0 = day of estrus) of the estrous cycle to induce pseudopregnancy. The remaining 14 pigs were mated during estrus. All pigs were laparotomized on Day 4, and one uterine horn was ligated to produce one crowded and one roomy uterine environment. Pigs were killed on Days 40, 60, and 80 of pregnancy or pseudopregnancy. The reproductive tracts were collected, and placental tissues from pregnant pigs and endometrial tissues from all pigs were cultured in the presence of 3H-leucine to evaluate protein secretion. Conditioned medium was dialyzed, measured for incorporation of radioactivity into nondialyzable macromolecules, and then subjected to two-dimensional (2D)-PAGE to determine the effect of uterine space and day of pregnancy or pseudopregnancy on overall protein secretion rate and secretion of specific proteins. Fetal survival, fetal weight, and placental weight were decreased (p < 0.01) in the crowded uterine environment compared to the roomy uterine environment. Incorporation of 3H-leucine into nondialyzable macromolecules by endometrial tissue in culture was not affected by uterine space. Secretion of nondialyzable macromolecules by endometrium from pregnant pigs was not different from that by endometrium from pseudopregnant pigs on Day 40 but was greater (p < 0.01) on Days 60 and 80.(ABSTRACT TRUNCATED AT 250 WORDS)     

Unusual abundance of arginine and ornithine in porcine allantoic fluid

The concentrations of free arginine, ornithine, and glutamine in porcine allantoic and amniotic fluids were determined on Days 30,35, 40, and 45 of gestation. Arginine and ornithine were the most abundant amino acids in allantoic fluid on Days 35-40 and 45 of gestation, respectively. Arginine and ornithine nitrogen accounted for 40%, 50%, and 55% of the total free alpha-amino acid nitrogen in allantoic fluid on Days 35, 40, and 45 of gestation, respectively. Glutamine was the most abundant amino acid in amniotic fluid during early gestation and was also abundant in allantoic fluid. On Day 45 of gestation, glutamine nitrogen accounted for 41% of the total free alpha-amino acid nitrogen in amniotic fluid. The unusual abundance of arginine (2.5-4.1 mM) and ornithine (1.08-2.52 mM) in allantoic fluid on Days 35-40 of gestation has not been reported for any other biological fluid. These results are novel and interesting with respect to the role of these two basic amino acids in fetal-placental nutrition and metabolism.     

Endothelin-1 and endothelin receptors are present in the sheep uterus and conceptus at implantation

Previous studies have demonstrated that endothelin is present in the ovine endometrium and increases at around the expected time of implantation. To characterize further uterine endothelin at the time of establishment of pregnancy in sheep, endothelin was measured by radioimmunoassay in uterine flushings obtained during the oestrous cycle and in pregnant ewes up to the time of implantation (day 16). During the oestrous cycle, the highest amounts of endothelin were present in uterine flushings on day 14 (1.1 +/- 0.2 ng endothelin/uterus). During early pregnancy, basal levels of endothelin (0.5-0.6 ng endothelin/uterus) were present in uterine flushings for the first 10 days and then increased on day 14 to levels similar to those found at the equivalent stage of the oestrous cycle. On days 15 and 16 of pregnancy, endothelin content in the uterine lumen increased to significantly (P < 0.05) higher concentrations (2.9 +/- 0.4 ng endothelin/uterus) when compared with the non-fertile cycle. The principal isoform present in flushings at the time of implantation was endothelin-1, as determined by reverse-phase HPLC. Endothelin was released principally by purified endometrial epithelial cells in culture, with barely detectable amounts released by endometrial stromal cells or conceptus tissue, which is consistent with the epithelium being the principal source of endothelin in the uterine lumen. Endothelin binding sites were present in endometrium and myometrium, as demonstrated by specific binding of 125I-labelled endothelin-1, which was saturable and displaced by endothelin-1. Both endothelinA and B sub-types of receptors were present as demonstrated by the biphasic displacement of 125I-labelled endothelin-1 binding by the specific endothelinB agonist BQ3020. These were localised principally on luminal and glandular epithelium and in the vasculature of the endometrium and myometrium as shown by autoradiography. Endothelin receptors were also present on the conceptus obtained at the time of implantation. In the day 20 conceptus, endothelin immunostaining was localised principally in the heart, in trophoblast in uninucleate but not in binucleate cells, and in fetal membranes. This immunostaining of the conceptus may represent binding to receptor sites. It is concluded that endothelin-1 is present in the uterine lumen and may play an important role in the paracrine regulation of the conceptus and endometrium at the time of rapid embryo development, implantation and early placentation.     

Folate status response to controlled folate intake in pregnant women

A metabolic study (84-d) was conducted to investigate the folate status response of pregnant subjects (n = 12) during their second trimester and nonpregnant controls (n = 12) to folate intakes approximating the current (400 microg/d) and former (800 microg/d) recommended dietary allowance (RDA). The overall goal of the study was to provide metabolic data to assist in the interpretation of the current RDA for folate. Subjects were fed a controlled diet containing 120 +/- 15 microg/d (mean +/- SD) folate and either 330 or 730 microg/d synthetic folic acid. Outcome variables between and within supplementation groups were compared at steady state. Serum folate was higher (P 0.05) were detected in serum folate between pregnant and nonpregnant women within the same supplementation group. Urinary 5-methyl-tetrahydrofolate excretion was greater (P 0.05) in 5-methyl-tetrahydrofolate excretion were detected between pregnant and nonpregnant women within supplementation groups. Differences (P 相似文献   

Efficacy of an in-feed formulation of ivermectin against adult worms and somatic larvae of Strongyloides ransomi

The efficacy of an in-feed formulation (IVOMEC premix) containing 0.6% ivermectin was tested against Strongyloides ransomi in swine. The efficacy of ivermectin against patent infections of S. ransomi when given via the feed at 2 ppm for 7 days (Days 0-7) to provide 100 mcg ivermectin kg-1 body weight day-1 was evaluated in a study with 16 3-month-old male castrated piglets. Seven days prior to treatment each piglet was infected subcutaneously with 2500 infective larvae of S. ransomi. Fecal egg counts were carried out on Days -7, 0, 7 and 14, and worm counts on Day 14. Efficacy was 100% in all treated piglets. Two trials involving 40 pregnant gilts were carried out to evaluate the efficacy of ivermectin against the somatic larval stages of S. ransomi when given at a daily dose of 100 mcg kg-1 body weight for 7 days starting on Days 66, 78, 92 or 103 of pregnancy. The gilts were each experimentally infected with three subcutaneous injections of 250,000 infective larvae, with the last infection given between 12 and 30 days prior to commencement of treatment. Gilts were confirmed free of pre-existing intestinal stages of S. ransomi prior to ivermectin treatment. Fecal nematode egg counts were carried out in gilts/sows and piglets subsequently born. The Strongyloides larvae present in sow milk 1, 2 and 7 days post partum were counted. Fourteen days post natum, worm counts were performed in four randomly selected piglets for each litter. IVOMEC premix given to pregnant gilts prevented shedding of larvae in sow milk, egg output in feces and the establishment of S. ransomi in piglets.     

Impairment of the nitric oxide-mediated vasodilator response to mental stress in hypertensive but not in hypercholesterolemic patients

A role for connective tissue growth factor (CTGF) in reproductive function has been suggested from recent studies in the pig. To extend these findings, we have analyzed the immunohistochemical localization of CTGF during the estrous cycle and early pregnancy in mice. During the diestrous and early proestrous stages, CTGF was localized at high levels to both luminal and glandular uterine epithelial cells and at much lower levels in the stroma or myometrium. Epithelial expression of CTGF was considerably reduced at estrus. On Days 1.5-3.5 of pregnancy, CTGF was localized mainly to the uterine epithelial cells, which showed a substantially reduced level of CTGF on Day 4.5. On Days 5.5 and 6.5, CTGF was present at high levels in uterine decidual cells. CTGF was detected in the trophectoderm and inner cell mass of the preimplantation embryo on Day 4.5 and became preferentially localized to embryonic endoderm and mesoderm on Days 5.5-6.5. Multiple mass forms of CTGF (Mr 14 000-38 000) were present in endometrial extracts and uterine luminal flushings. Collectively, these data support a role for CTGF in uterine cell growth, migration, adhesion, and extracellular matrix production during the estrous cycle and early pregnancy, as well as in early development of the embryo.     

Growth and cellular proliferation of antral follicles throughout the follicular phase of the estrous cycle in Meishan gilts

Meishan gilts were ovariectomized 2 h after an i.v. injection of 5'-bromo-2'-deoxyuridine (BrdU, a thymidine analogue; 5 mg/kg body weight) on Days 15-19 of the estrous cycle or 24-30 h after observed estrus (post LH, PLH). All antral follicles > or = 3 mm from one ovary were fixed in Carnoy's solution. Granulosa and thecal cell labeling indexes (LI; percentage of nuclei staining for BrdU) as well as LI of cells within the basal, middle, and antral thirds of the granulosa cell layer were estimated for each follicle. In addition, antral and granulosa cell layer volume, granulosa cell layer thickness, granulosa cell density, number of granulosa cells, and number of S-phase cells per hour were estimated for each follicle. Mean follicular diameter increased linearly (p < 0.01) from Day 15 to PLH, with a growth rate of 0.77 mm/day. Granulosa and thecal cell LI decreased (p < 0.01) from Day 15 to PLH; however, granulosa cell LI was greater (p < 0.01) than thecal cell LI on Days 15 and 16 but less (p < 0.05) than thecal cell LI on Day 19. Follicles collected from PLH gilts contained no labeled granulosa cells. Cells within the basal third of the granulosa cell layer contained fewer (p < 0.01) labeled nuclei than did cells within the middle or antral thirds. In addition, LI within the basal and middle thirds of the granulosa cell layer decreased (p < 0.01) from Days 15 to 18 and from Days 15 to 17, respectively, whereas LI within the antral third remained constant from Days 15 to 18. Granulosa cell layer thickness was greatest (p < 0.01) on Day 15, then decreased (p < 0.01) and was similar from Day 16 to PLH. Granulosa cell density was similar from Days 15 to 19, then decreased (p < 0.01) for PLH gilts. Antral and granulosa cell layer volumes increased linearly (p < 0.01) from Days 15 to 19 and Day 15 to PLH, respectively, resulting in 2.8 and 1.9 volume doublings and doubling times of 1.4 and 2.7 days, respectively. Number of granulosa cells per follicle increased linearly (p < 0.01) from Day 15 to PLH, resulting in 1.5 cell doublings and a doubling time of 3.3 days. Number of S-phase cells per follicle per hour was similar from Days 15 to 18 and then decreased (p > 0.01) from Day 18 to PLH. In summary, the percentages of proliferating granulosa and thecal cells decreased throughout the final stages of antral follicular development. Differentiation of granulosa cells occurred from the basal to the antral area as follicles matured. We proposed that, during the latter stages of follicular development, the rapid increase in follicular diameter resulted primarily from expansion of the antral cavity, whereas increases in the granulosa cell layer volume and number of granulosa cells per follicle maintained a constant granulosa cell layer thickness.     

Quantification of receptors for estradiol-17 beta and progesterone in the pituitary and hypothalamus of prepubertal gilts induced to ovulate with pregnant mare's serum and human chorionic gonadotropin

Nuclear and cytoplasmic exchange assays were developed and validated to quantify receptors for estradiol-17 beta (E217 beta) and progesterone (P4) in hypothalamic and pituitary tissues of gilts before, during, and after treatment with pregnant mare's serum (PMS) and human chorionic gonadotropin (hCG). Prepubertal gilts, 5 months old, were assigned randomly to four treatments. One group of gilts received 500 IU PMS (Day 0) and were sacrificed 2 days later (2 days post-PMS); another group received 500 IU PMS on Days 0 and 2, and were sacrificed 4 days from the initial injection (4 days post-PMS). A third group of gilts received PMS (500 IU) on Days 0 and 2, 1000 IU hCG on Day 4, and were sacrificed 5 days after hCG (5 days post-hCG). Controls were given saline on Days 0, 2 and 4 and sacrificed on Day 6. In pituitary tissues, there were no significant changes in numbers of cytoplasmic E2 17 beta receptors, cytoplasmic P4 receptors, nuclear P4 receptors or nuclear E2 17 beta receptors among the control, 2 days post-PMS, 4 days post-PMS or 5 days post-hCG treatment groups. In hypothalamic tissues, no differences in cytoplasmic E2 17 beta receptors, cytoplasmic P4 receptors or nuclear P4 receptors were found among any of the treatments. Nuclear receptors for E2 17 beta in hypothalamic tissues were greater, however, in gilts 2 days post-PMS (P less than 0.05) than in controls or 5 days post-hCG gilts, but they were not different from gilts 4 days post-PMS. Follicular development and serum concentrations of E2 17 beta followed the expected patterns after PMS; only ovaries from hCG-treated pigs contained corpora lutea. Because the PMS-hCG regimen simulated the onset of puberty, it seems that gilts attain puberty without a significant change in the number of receptors for E2 17 beta and P4 in the pituitary or hypothalamus.     

Determination of porcine endometrial phospholipase A2 activity and detection of immunoreactive cyclooxygenase during the oestrous cycle and early pregnancy

Experiments examined the characteristics and activity of phospholipase A2 (PLA2) and examined the presence of immunoreactive cyclooxygenase in endometrium of pigs during the oestrous cycle and early pregnancy. Endometrial PLA2 was calcium-independent and activity of the enzyme was greatest at a pH of 8.0. Activity of PLA2 on Days 10, 12, 14 and 16 of the oestrous cycle did not differ (P > 0.1) from activity on those days during pregnancy. During oestrus and early metoestrus (Days 0-3), cyclooxygenase was present in both glandular and surface epithelium. After Day 10 of the oestrous cycle or pregnancy, staining for cyclooxygenase was less intense in the lower and middle uterine glands. However, the upper glandular epithelium near the surface epithelium stained intensely. By Day 15 of the oestrous cycle or pregnancy, intense staining for cyclooxygenase appeared restricted to the upper uterine glands. These results indicate changes in localization of immunoreactive cyclooxygenase throughout the oestrous cycle and suggest that these are not related to altered secretion of prostaglandins (PGs) during early pregnancy. The stimulatory effects of porcine conceptus products on secretion of PGs during early pregnancy are apparently not associated with increased activity of endometrial PLA2.     

Secretion of bovine uterine proteins in response to type I interferons

Bovine interferon-tau (bIFN-tau) is secreted by the developing conceptus and initiates antiluteolytic events by interacting with uterine membrane receptors. We have identified three endometrial proteins (approximately 8, 16, and 28 kDa; P8, P16, and P28; respectively) that are secreted in response to recombinant (r) bIFN-tau. The objective of this study was to determine whether or not secretion of these proteins was a unique response to IFN-tau during early pregnancy. Three experiments were designed to examine secretion of endometrial proteins as a function of time in culture (0, 3, 6, 12, 18, 24 h), stage of the estrous cycle and pregnancy (Days 15, 18, 0/21), and dose of Type I IFN (0, 0.5, 5, and 25 nM; rbIFN-tau, rbIFN-alpha, and roIFN-tau). Endometrium was cultured for times specified with L-[3H]leusine to generate radiolabeled proteins. Secreted proteins were quantitated by using one-dimensional (1D)-PAGE, fluorography, and densitometry. Secretion of P8, P16, and P28 increased over time (p < 0.0001) in culture and in response to 25 nM rbIFN-tau (p < 0.05). Secretion of P8 in response to rbIFN-tau was higher (p < 0.05). Secretion of P8 in response to rbIFN-tau was higher (p < 0.0005) in endometrium collected from pregnant than nonpregnant heifers, but did not differ across the days examined. Although secretion of P8 was higher (p < 0.0001) in the presence than in the absence of rbIFN-tau, it was not affected by rbIFN-alpha.(ABSTRACT TRUNCATED AT 250 WORDS)     

Maternal dietary protein deficiency decreases amino acid concentrations in fetal plasma and allantoic fluid of pigs

This study was conducted to test the hypothesis that maternal dietary protein deficiency decreases amino acid availability to the fetus, thereby contributing to retarded fetal growth. Primiparous gilts selected genetically for low or high plasma total cholesterol concentrations (low line and high line, respectively) were mated, and then fed 1.8 kg/d of isocaloric diets containing 13% or 0.5% crude protein. At d 40 or 60 of gestation, they were hysterectomized, and maternal and fetal blood samples as well as amniotic and allantoic fluids were obtained for analyses of amino acids, ammonia and urea. Dietary protein restriction decreased (P < 0.05) the following: 1) maternal plasma concentrations of urea at d 40 and 60 of gestation; 2) fetal plasma concentrations of alanine, arginine, branched-chain amino acids (BCAA), glutamine, glycine, lysine, ornithine, proline, taurine, threonine and urea at d 60 of gestation; 3) amniotic and allantoic fluid concentrations of urea at d 40 and 60 of gestation; and 4) allantoic fluid concentrations of alanine, arginine, BCAA, citrulline, cystine, glycine, histidine, methionine, proline, serine, taurine, threonine and tyrosine at d 40 of gestation, in gilts of both genetic lines. At d 60 of gestation, protein deficiency decreased (P < 0.05) allantoic fluid concentrations of arginine, cystine, glycine, taurine and tyrosine in low line gilts and of cystine, glutamine, ornithine, serine, taurine and tyrosine in high line gilts. Low line and high line gilts also differed remarkably in allantoic fluid concentrations of arginine, glutamine, ornithine and ammonia at d 40 and 60 of gestation. Our results suggest the following: 1) protein-deficient gilts maintain maternal plasma concentrations of amino acids by mobilizing maternal protein stores and decreasing oxidation of amino acids during the first half of gestation; 2) protein deficiency may impair placental transport of amino acids from the maternal to the fetal blood; and 3) low line and high line gilts differ in fetal amino acid metabolism. Decreases in concentrations of the essential and nonessential amino acids in the fetus may be a mechanism whereby maternal dietary protein restriction results in fetal growth retardation.     

Contributions of endogenous inhibin and estradiol to the regulation of follicle-stimulating hormone and luteinizing hormone secretion in the pregnant rat

To examine the contributions of endogenous inhibin and estradiol to the regulation of FSH and LH secretion in the pregnant rat, some rats were passively immunized against inhibin and/or estradiol, and others were ovariectomized, on Days 5, 10, 15, and 20 of pregnancy. Ovarian and uterine venous blood was collected separately to confirm the sources of inhibin and steroid hormones during pregnancy. Immunoreactivity of inhibin in the placenta was also examined by RIA. Levels of inhibin in ovarian venous plasma were significantly higher than those in peripheral plasma during pregnancy. No difference was observed between the levels of inhibin in uterine venous plasma and peripheral plasma. No immunoreactivity of inhibin was detected in placental homogenate from rats at Days 10, 15, and 20. FSH secretion significantly increased after immunoneutralization of inhibin during pregnancy. A marked increase in FSH secretion was noted on Days 5 and 20, and the smallest increase was observed on Day 15. Administration of estradiol antiserum (AS) alone did not induce a significant increase in FSH secretion on any day of pregnancy. However, a synergistic effect of estradiol AS and inhibin AS was observed on Day 20. On Days 5, 10, and 20, administration of inhibin AS or estradiol AS induced a significant increase in LH secretion. A synergistic effect of inhibin AS and estradiol AS on LH secretion was observed on Day 5. On Days 5 and 10, significantly high LH secretion was noted in ovariectomized rats as compared with that in rats treated with both inhibin AS and estradiol AS, indicating that other ovarian hormones such as progesterone may be involved in the suppression of LH secretion in these stages of pregnancy. These data indicate that both inhibin and estradiol, predominantly secreted from the ovary, are involved in the regulation of gonadotropin secretion during pregnancy as during the estrous cycle in the rat.     

Identification of a new MAGE gene with tumor-specific expression by representational difference analysis

Early pregnancy in ruminants, such as the sheep, is characterized by relatively extensive development of the conceptus before attachment to the endometrium. Between the period of blastocyst hatching and initial attachment, the uterus responds to signals from the conceptus and adapts to provide an environment that permits the establishment of pregnancy. We used large-format two-dimensional (2D) PAGE to analyze the dynamic changes in protein composition of uterine luminal fluid (ULF) during this stage of pregnancy, and we determined the contribution of each of the extraembryonic membranes and the endometrium to these changes. The majority of the more than 40 pregnancy-associated proteins in ULF at Day 17 were secreted by the conceptus. By 2D gel map comparison and Western blotting, we identified transferrin, secreted by the yolk sac from Day 15, and cytoplasmic actin, one of the most abundant proteins produced by the trophoblast at Day 17. Apolipoprotein A1 and aldose reductase, whose abundance were markedly increased in pregnancy, were identified by peptide microsequencing. Aldose reductase, an enzyme required for the conversion of glucose to fructose, was shown to be synthesized by the trophoblast, and its detection even before the formation of the placenta suggests that the synthesis of fructose may occur much earlier than previously reported.     

Prostaglandin F2alpha-induced luteolysis of aging corpora lutea in hysterectomized pigs

Prostaglandins primarily of uterine origin play an important role in parturition. Hysterectomy of nongravid pigs early in the luteal phase maintains luteal function until about Day 150, whereas the duration of normal pregnancy is about 114 days. A precisely timed peak release of relaxin and coincident decrease in progesterone secretion in unmated hysterectomized gilts are similar to hormonal changes that occur a few hours before parturition. It is hypothesized that prostaglandin F2alpha (PGF2alpha) in hysterectomized pigs mimics abrupt changes in ovarian and pituitary hormone secretion seen before normal parturition and in early lactation. Unmated Yorkshire gilts were hysterectomized on Days 6-8 of a normal estrous cycle, and at 1200 h on Day 113, they were given an i.m. injection of 30 mg PGF2alpha-trihydroxymethylaminomethane (THAM) salt or PBS. None of these gilts expressed behavioral estrus immediately after PGF2alpha or vehicle treatment. On Day 113, PGF2alpha increased peak relaxin (60 ng/ml) compared with that of controls (34 ng/ml; p < 0.01), whereas progesterone decreased abruptly (4 vs. 16 ng/ml in PGF2alpha and PBS; p < 0.01). Prolactin remained at < 5 ng/ml from Day 98 to 120 in controls but peaked at 33 ng/ml immediately after PGF2alpha treatment on Day 113, and then decreased to levels similar to those of controls on Day 120. Sequential bleeding revealed an acute growth hormone release (4.5 ng/ml) immediately after PGF2alpha injection and return to basal levels (< 0.6 ng/ml) on Days 114-120. PGF2alpha induced abrupt shifts in progesterone, relaxin, prolactin, and growth hormone secretion in hysterectomized gilts that mimicked hormone changes seen in late pregnancy, parturition, and early lactation. These findings provide new insight into the role of PGF2alpha in abruptly changing hormone secretions by aging corpora lutea and the pituitary gland even in the absence of conceptuses or the uterus in the pig.     

Endothelin receptor type A and B gene expression in human nonpregnant, term pregnant, and preeclamptic uterus

OBJECTIVE: The aim of this study was to quantify the gene expression of ETA and ETB receptors within the different uterine segments of nonpregnant, normal pregnant, and preeclamptic women. STUDY DESIGN: Biopsy samples from the cervix, isthmus, and corpus uteri were obtained from eight nonpregnant, nine term pregnant, and seven preeclamptic women. The concentration of ETA and ETB receptor messenger ribonucleic acid were determined by a solution hybridization technique with complementary ribonucleic acid probes. Results are presented in counts per minute per microgram of total nucleic acid as mean +/- SEM. RESULTS: The expression of messenger ribonucleic acid encoding the ETA receptor was generally higher in the upper than in the lower uterine segment in nonpregnant, normal pregnant, and preeclamptic myometrium, whereas the opposite pattern was seen with regard to ETB. During normal pregnancy the concentrations of ETA receptor messenger ribonucleic acid in the corpus and ETB receptor messenger ribonucleic acid in the isthmus were significantly elevated compared with those in nonpregnant women. This enhanced gene expression was, however, not observed in the preeclamptic group. CONCLUSION: Our finding of segmentally differentiated endothelin receptor gene expression is compatible with a role for endothelin-1 in stimulating uterine contractions through ETA receptors during spontaneous labor and suggests a relaxing effect of the ETB receptor on the myometrium.     

Prepartum changes in maternal responsiveness and nest defense in Rattus norvegicus..

Examined 297 female rats for changes in maternal responsiveness during late pregnancy by exposing Ss to foster pups in tests lasting 15–30 min under conditions favoring the rapid initiation of maternal behavior. Ss were divided into 17 groups according to state of pregnancy (nonpregnant, Day 17, Day 20, Day 21, or Day 22); type of nest; and parity. Nulliparous Ss were compared with primiparous and with experienced breeders. Nest defense was observed by introducing unfamiliar males (2-min tests) on Day 22. Results are presented for 3 periods: Days 17–20, when maternal responsiveness was lower than in the nonpregnant condition; Day 21, when maternal responsiveness returned to or rose above nonpregnant levels; and on Day 22 (the 3.5 hrs prior to delivery), during which 90% of Ss almost immediately retrieved, gathered, and tended foster pups and during which 92% attacked the unfamiliar intruders. (Attacks were rare earlier.) Maternally experienced Ss were more responsive to pups than nulliparous Ss when nonpregnant and throughout late pregnancy, but both groups were equally likely to show prepartum aggression. (22 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)