Browsing by Subject "pregnancy"
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Item Analysis of some novel uterine extracellular matrix proteins and a growth factor(2009-05-15) Al Ramadan, Saeed YaseenThis dissertation focused on two classes of molecules implicated in processes of implantation and placentation in sheep and pigs. Study one examined the temporal/spatial distribution of several Small Integrin-Binding Ligand, N-Linked Glycoprotein (SIBLING) family members in cyclic and pregnant ovine uterus. Studies two and three evaluated the relationships between progesterone (P4) and estrogen (E2) and their receptors (PGR and ESR1, respectively) on FGF7 mRNA expression within the endometrium and placenta of pigs. Study one showed that dentin sialophosphoprotein (DSPP) was first detected in luminal epithelium (LE) of Day 15 cyclic and pregnant sheep. Stromal expression of DSPP was first detected on Day 20 of pregnancy in stratum compactum and remained prominent in stroma through Day 120. Stromal DSPP protein was positively influenced by the conceptus based upon analysis of a unilaterally pregnant ewe model system. Immunoreactive dentin matrix protein 1 (DMP1), matrix extracellular phospoglycoprotein (MEPE) were localized to the stroma of cyclic and pregnant sheep, however, these proteins appeared to be constitutively expressed. BSP was not detected in ovine endometrium. Study two determined the effects of E2, P4, P4+E2, P4+the PGR antagonist (ZK137, 316), and P4+E2+ZK on FGF7 mRNA expression in uterine LE of ovariectomized pigs. Results indicate that P4 is permissive to FGF7 mRNA expression by down-regulating PGR in LE; P4 stimulates PGR-positive uterine stromal cells to release an as yet unidentified progestamedin that induces FGF7 mRNA expression by LE; E2 and P4 can induce FGF7 mRNA in the absence of PGR rendered nonfunctional by ZK. Study three showed the expression of ESR1, PGR and FGF7 in the uterine and placental tissue of pregnant pigs from Day 20 through 85. Results reveal a positive correlation between stromal cell expression of PGR and FGF7 mRNA which suggests that P4 is permissive to FGF7 mRNA expression by down-regulating PGR in LE. FGF7 mRNA in later pregnancy is maintained by the release of progestamedin from PGR-positive stromal cells. A novel finding was the presence of ESR1 in porcine placenta on Days 20 through Day 85 of pregnancy suggesting that E2 may play important roles in the placental biology of the pig.Item Anterior Gradient Homolog 2 and Its Potential Roles In Ovine Pregnancy(2014-08-13) Del Curto, Hannah MargaretAnterior gradient homolog 2 (AGR2) is a proto-oncogene that encodes a secreted protein with biological roles in cell migration, differentiation, and growth, and is also implicated in epithelial barrier function and integrity. AGR2 is up-regulated in prostate cancer, breast cancer, lung cancer, renal carcinomas and endometrial carcinomas, and it contributes to the survival of cells undergoing physiological stress. We previously reported that AGR2 expression in the ovine endometrium increased between days 9 and 12 post-mating, was stimulated by progesterone, and associated with conceptus elongation. In situ hybridization studies localized AGR2 mRNA to the luminal epithelium (LE) of the ovine endometrium. Further, AGR2 mRNA expression is greater from Days 14 to 16 of pregnancy than in the cycle, suggesting a role for conceptus-derived products in regulation of AGR2 expression in the endometrium. Using osmotic pumps to infuse pregnancy levels of hormones into the uterus of cyclic ewes, we found that interferon tau (IFNT) along with cortisol and prostaglandins (PGE2, PGF2?, and PGI2) or both cortisol and prostaglandins increased (P<0.05) expression of AGR2 mRNA in the endometrium compared to controls receiving vehicle and ewes receiving IFNT, cortisol, or prostaglandins alone. A role for AGR2 in placental function during later pregnancy has not been described for any species. Using a model of maternal nutrient restriction, we utilized natural population variance in fetal growth to identify novel genes associated with the development of either intrauterine growth restricted (IUGR) or non-IUGR pregnancies in sheep. Interestingly, microarray analysis of placentomes from IUGR and non-IUGR pregnancies at Day 125 of pregnancy found AGR2 was up-regulated (P<0.05) in IUGR pregnancies. Quantitative real-time PCR analysis confirmed that AGR2 mRNA was 2.7-fold higher in placentomes from IUGR pregnancies. Collectively, results suggest that AGR2 is involved in growth and development of the early conceptus. Additionally, elevated AGR2 expression may be associated with increased levels of physiological stress in the IUGR pregnancy. During late pregnancy, a reduction in the proliferative actions of AGR2 may be necessary for enhanced placental function in the nutritionally restricted ewe.Item Buprenorphine metabolism by preterm placentas: A pilot study(2007-10-31) Rachel McRee Kaufman; Irina Pikuleva; Patricia Shinnick-Gallagher; Mary Treinen-Moslen; Kenneth M. JohnsonThe focus of this investigation is to attempt to identify and characterize the major enzyme responsible for metabolism of buprenorphine to nor-buprenorphine in human preterm placentas and to elucidate whether the development/gestational age of the placenta affects metabolism of buprenorphine. The estimated apparent Km did not appear to change with gestational age suggesting the affinity of buprenorphine does not change throughout gestation. On the other hand, the estimated Vmax did appear to increase with gestational age suggesting the activity increases with gestational age. Our attempt to identify the enzyme responsible for metabolism in earlier gestational age placentas revealed that CYP19 remains the major metabolizing enzyme of buprenorphine but suggests additional enzymes may be involved, namely, CYP3A4, 1A1, 2E1, 2D6 and 2C19. Further studies must be conducted for conclusions to be drawn but we hope this report can contribute to the design of future studies on preterm placental metabolism of drugs.Item Comparison of perceived stress, allostatic load and racial discrimination in different cultural groups of pregnant black women(2008-07-07) Jennifer L. Musgrave-Kelly; R. Jeanne Ruiz, PhD, RN; Sheryl Bishop, PhD; Kay Avant, PhD; Carolyn Phillips, PhD, RN; Alice Hill, PhD, RNOver 26 billion healthcare dollars are spent annually in the United States on the management of pre-term labor and the care of infants born prematurely. Studies have found that elevated levels of stress and anxiety during pregnancy significantly increase women’s risks for poor perinatal outcomes. Previous research studies also suggest differences in stress response may exist between black women born in the U.S. and foreign-born. The purpose of this study was to compare racial discrimination and different measurements of stress, including perceived stress and allostatic load score, in two different cultural groups of African American women. \r\n The specific aims of the research project were to: 1) determine if differences exist between African American women born in the United States and those who are foreign-born on perceived stress, measures of allostatic load and racial discrimination; 2) examine the relationships between racial discrimination, perceived stress, age, income, number of hours worked weekly, gestational age, total allostatic load score and measures of allostatic load in a sample of pregnant African American women; 3) determine the best model from the study variable set that predict each of the study variables. \r\nThe findings suggest the only variable that was statistically different between the two groups of African American women was the mean BMI (U.S.-born mean BMI 31.99 vs foreign-born mean BMI 25.58; p<0.02). Also, clinically significant differences were noted, such as a difference in ages (U.S.-born mean age 23.81 vs foreign-born mean age 30.25), income (U.S.-born mean income $2,385 vs foreign-born mean income $3,108) and measurements of stress (U.S.-born mean PSS score 18.11 vs foreign-born mean PSS score 16.20) and racial discrimination (U.S.-born mean RDS scores 24.01 vs foreign-born mean RDS score 17.25). When examining the entire sample there was a positive correlation between income and perceived stress (r = -0.43; p < 0.019) and between perceived stress scores and racial discrimination scores (r = 0.34; p < 0.063) . Regression models revealed income and racial discrimination predicted perceived stress scale scores (p < 0.023).\r\n The findings provided supporting evidence in the identification of perceived stress and racial discrimination in pregnancy. Additional support was provided for the differences that exist between different groups of women within the same ethnic group. Future research is needed to understand how socio-demographic and psychological variables place a mother and her baby at risk.\r\nItem Early pregnancy diagnosis and embryo/fetus mortality in cattle(Texas A&M University, 2006-04-12) Romano, Juan EduardoPregnancy diagnosis by transrectal ultrasonography (using a 5 MHZ linear probe) presented the maximum sensitivity and negative predictive values at day 26 and day 29 after estrus in heifers and cows, respectively. Palpation per rectum using the fetal membrane slip for pregnancy diagnosis did not increase embryo/fetus mortality when compared with a positive control group of non-palpated females. The use of a controlled randomized block design was a useful approach to study this problem. Blocking for category and number of embryos allowed us to remove these confounding factors. Factors that affected pregnancy loss during the first four months of pregnancy were: period of pregnancy, age of the animal, number of previous lactations and number of embryos. Pregnancy loss was higher during the embryonic than fetal periods. Spontaneous embryo/fetal mortality increased with the age of the animal and lactation number. The risk of spontaneous embryo/fetus mortality was higher in twin than in single pregnancies. Two types of embryo/fetus mortality were noted: Type I and Type II. Type I was characterized by presence of positive fetal membrane slip by palpation per rectum, signs of degeneration by transrectal ultrasonography and persistence of a functional corpus luteum. The uterus took approximately 3 weeks to be noted clean by transrectal ultrasonography and the animals showed estrus one month after the conceptus was diagnosed dead. Type II was characterized by absence of positive signs of pregnancy by palpation per rectum, absence of signs of degeneration by transrectal ultrasonography and absence of a functional corpus luteum. Pregnancy loss in nuclear transfer derived embryos was higher compared to in vivo derived embryos produced by artificial insemination. Pregnancy loss occurred mainly during the transition from the embryonic to the fetal period. Embryo/fetus mortality detected was Type I. Progesterone produced by the corpus luteum was noted at pregnancy levels for approximately two weeks after embryo/fetus death. Protein B, a hormonal placental marker, was maintained at pregnancy levels for approximately 3 weeks after embryo/fetus death. No differences in the levels of the two hormones were noted when comparing females with dead or live conceptuses.Item Effects of Parental Notification and Consent Laws on Teenage Births and Abortions in Texas(2014-08-04) Wills, Laura JeanTeen pregnancy and teen abortions are major public health concerns in the United States. The more than 300,000 births to teens each year often involve increased risk to the health of the mother and the baby. Teenage pregnancy and births also raise a variety of related political, clinical, social, and economic concerns. To make effective recommendations regarding contraception, teenage pregnancy, abortion, and sexual education programs, researchers must produce empirical evidence, which accurately evaluates policy options for persons involved in public health policy and legislation. This study investigated the effect of parental notification and consent laws on teen births and abortions in Texas. This research examined health data of females aged 13 - 21 from Texas for 1995 through 2009. In January 2000, Texas enacted a law requiring a medical provider to notify the parents of a minor female seeking an abortion, before performing the abortion. In 2005, the law changed, requiring notarized parental consent in addition to notification for an unemancipated minor to receive an abortion. Teenage birth and abortion rates, as well as other health outcomes related to pregnancy were analyzed. The data included years before notification laws (1995 ? 1999), the years following enactment of the notification law (2000 ? 2004), and the most current data following the more rigid consent law (2005 ? 2009). The analysis used a time-series approach, specifically Auto Regressive Integrated Moving Average (ARIMA) model-fitting processes, to identify any changes in the patterns of the dependent variables resulting from this legislation. Overall, parental notification and consent laws did seem to have an effect on birth and abortion rates for minors in Texas. Specifically, notification laws seemed to have the greatest impact on 16 ? 17 year old females, reducing birth and abortion rates. Results were mixed in terms of the effect of the more stringent consent laws and the overall impact of both laws on health outcomes. Findings did differ by race/ethnicity category.Item Exploring oral health among pregnant and parenting adolescent women: a mixed methods study(2010-05) Murphey, Christina Leigh; Rew, Lynn; Cappelli, David; Fowles, Eileen; Garcia, Alexandra; Walker, LorraineDespite growing interest in maternal oral health, research aimed at this population is scant. To date, no qualitative studies of adolescent maternal oral health exist. Therefore, the purpose of this descriptive, exploratory, concurrent, mixed-methods study was to explore oral health status, beliefs, and practices, and pregnancy and parenting outcomes in this population by triangulating both quantitative and qualitative data. A non-probability, convenience sample of 46 pregnant and parenting adolescents was recruited. Five questionnaires were administered and visual oral examinations were conducted. Twenty-four of these 46 participants also participated in the qualitative component of the study. Adolescents in this study were both pregnant (n = 20; 43.5%) and parenting (n = 26; 56.5%), and primarily of Hispanic decent (n = 38; 83%). Of the 20 adolescents who were pregnant, four had been told by a nurse or physician that they had a pregnancy complication(s). Among the parenting adolescents, the most common past pregnancy complications were self-reported as prematurity (n = 6; 35%) and high blood pressure (n = 3; 18%). Thirty-three (72%) participants reported ever having dental insurance. While 16 (35%) participants had seen a dentist in the past 6 months, another 15 (33%) did not recall their last dental visit. One adolescent reported never having been to a dentist. Associations among visual oral health status and selected contextual variables were non-significant, which may be attributed to the small sample size. However, moderate significant correlations were found between social connectedness and oral-health-related quality of life, as well as between visual oral health status and measures of self-reported dental health. For the qualitative component, six themes related to oral health value and well-being, oral health knowledge, practices, myths, and barriers to accessing oral health services emerged. Triangulation of the quantitative and qualitative data did not produce statistical significance; however, discrepancies were found between the overall objective, visual oral health status, and the subjective perception of oral health status, which supports the overall findings. Future research should focus on larger studies to further explore associations between social connectedness, oral-health-related quality of life, and objective and subjective measures of oral health status and behaviors.Item Expression and regulation of select interferon stimulated genes in porcine endometrium during pregnancy(2009-05-15) Joyce, Margaret MaryCoordinated signals between the maternal endometrium and conceptus during the peri-implantation period are essential for the establishment and maintenance of pregnancy. In pigs, this involves estrogen secretion from conceptuses as the signal for maternal recognition of pregnancy. Pig conceptuses also secrete interferons (IFN) delta (IFND) and IFN gamma (IFNG). The uterine effects of pig IFNs are not known, although ruminant conceptuses secrete IFN tau (IFNT) for pregnancy recognition, and this increases the expression of IFN-stimulated genes (ISGs) in the endometrium. Therefore, studies were conducted to identify and characterize ISGs in the pig endometrium during pregnancy and to evaluate their regulation by estrogen and conceptus secretory proteins (CSPs) that contain IFNs. In the first study, four classical ISGs, including interferon regulatory factor 1 (IRF1) and signal transducer and activator of transcription 2 (STAT2), were detected in the pig endometrium and increased after Day 12 of pregnancy, specifically in stroma. IRF2, a transcriptional repressor of ISGs, increased in luminal epithelium (LE) by Day 12 of pregnancy. The increase of IRF2 was due to estrogen while the stromal increase of IRF1 was due to IFN-containing CSP infusion. In the second study, the ISG STAT1 increased in LE after Day 12 of pregnancy and estrogen resulted in a similar increase. After Day 15 of pregnancy, STAT1 increased in stroma. Infusion of IFN-containing CSPs resulted in a similar stromal increase. In the third study, the ISGs swine leukocyte antigen (SLA) class I and beta-2microglobulin (B2M) increased in LE between Days 5 and 9 of the estrous cycle and pregnancy and decreased between Days 15 and 20 of pregnancy. By Day 15 of pregnancy, SLAs and B2M increased in stroma where they remained through Day 40. Progesterone increased SLA and B2M in LE, and a progesterone receptor antagonist ablated the upregulation while infusion of IFN-containing CSP increased SLA and B2M in stroma. Collectively, these studies identify ISGs expressed in the pig endometrium during pregnancy. These genes may be involved in protecting the fetal semiallograft from immune rejection, limiting conceptus invasion through the uterine wall, and/or establishing a vascular supply to the conceptus. The interactions of estrogen, progesterone and IFNs to regulate cell-type specific expression of ISGs highlight the complex interplay between endometrium and conceptus for pregnancy recognition and implantation.Item Hope as a Strategy for Improving Student Achievement and Dissuading Repeat Pregnancy in Pregnant and Parenting Adolescents(2011-08-08) McNeill, Elisa HutsonThis dissertation examines the construct of hope and its ability to be taught to pregnant and parenting adolescents as a strategy to improve academic achievement and dissuade repeat adolescent pregnancy. A systemic review of the literature examined fourteen (n=14) empirical studies to ascertain if a relationship existed between achievement and the construct of hope. With 92% of the studies reporting a positive correlation between hope and achievement, one can answer yes to the question of an existing relationship between hope and achievement. A manuscript is presented to summarize the development and implementation of a curriculum, designed to develop requisite skills among adolescent mothers to elevate their levels of hope. The Helping Optimize Planning Efforts (HOPE) curriculum presents specific methods for adolescent mothers to enhance skills related to goal setting, goal attainment and the use of positive self-talk as a mechanism for developing hope or increasing existing levels of hope. The study attempted to assess the ability of the HOPE curriculum to enhance the adolescent mother?s level of hope with the underlying assumption that increasing levels of hope might decrease the probability of a repeat adolescent pregnancy. Findings suggested there was a significant difference in the scores for the Dispositional Trait Hope Scale (DTHS) pre-test (M = 68.5, SD = 7.0) and the DTHS post-test scores (M = 73.2, SD = 5.61) (t(11) = 3.18, p = .009) indicating an increase in global hope. A significant difference was also found in between the State Hope Scale (SHS) pre-test scores (M = 39.0, SD = 4.84) and SHS post-test scores (M = 41.5, SD = 5.21) (t(11) = 4.19, p = .002) indicating an increase in the students point in time level of hope. The evaluation of the curriculum indicated that adolescent mothers can be taught to increase their levels of hope using the HOPE curriculum. Data collected during the evaluation of the HOPE curriculum was further analyzed to identify the constructs that contribute to the building of hope in adolescent mothers. The data suggests that two distinct components, agency and pathway, contribute to increase the level of hope. Examination of the subscales within the DTHS, showed there was a significant difference between the pre-test agency (M = 25.42, SD = 5.81) and the post-test agency scores (M = 27.85, SD = 3.65) (t(11) = 2.83, p = .017). Also, among the DTHS pathway subscale, a significant difference was found between the scores on the pre-test (M = 25.08, SD = 3.58) and the post-test score (M = 26.67, SD = 2.35) (t(11) = 2.22, p = .048). These findings suggest that the increased level of hope on the DTHS for was a reflection of the improvements in components, agency and pathway. These findings suggests that adolescent mothers can be taught to be more hopeful when the components of agency and pathway are developed.Item Interactive Roles of Gonadotropin-Releasing Hormone and RF-Amide Related Peptide 3 in Adenohypophyseal Physiology and Reporduction in the Mare(2013-01-28) Thorson, Jennifer FrancesThe seasonal termination of ovarian cycles in mares initiated near the time of the autumnal equinox is a significant managerial issue for horse breeders world-wide. Studies presented herein had two over-arching aims. In Aim I, objectives were to develop the principals needed to apply gonadotropin-releasing hormone (GnRH) therapeutics for routinely establishing pregnancies in the winter anovulatory mare. We first tested the hypothesis that continuous administration of native GnRH, beginning in either early February or March, would induce ovulation without reversion to an anovulatory state following treatment withdrawal. Continuous 28-d treatments elevated circulating luteinizing hormone (LH) and stimulated spontaneous ovulation much earlier than controls. However, mares treated only in February ceased ovarian cycles at termination of treatment. In contrast, mares administered GnRH in March continued to exhibit estrous cycles. Thus, we concluded that GnRH treatment must continue through March to ensure continued escape from winter anovulation. We then tested the hypothesis that the Julian day of conception could be accelerated in winter anovulatory mares treated continuously with native GnRH for 56 d beginning on February 1. Indeed, GnRH treatment caused a marked increase in the frequency of pregnancy compared to controls. Data illustrated that continuous administration of native GnRH is a practical and highly efficient option for managing seasonal anovulation. In Aim II, we examined hypothalamic distribution, adenohyphyseal receptor gene expression, and ligand functionality of RFRP3 in the mare during the breeding and non-breeding seasons. Hypothalamic RFRP3 mRNA was detected in the mare; however, neither hypothalamic expression of RFRP3 nor its anterior pituitary receptor differed between reproductive states. We then used equine adenohypophyseal cell culture to test the hypothesis that RFRP3 reduces the responsiveness of the equine gonadotrope to GnRH. Addition of RFRP3 to cell culture failed to counter the effects of GnRH. Finally, the effects of a RFRP3 receptor-signaling antagonist (RF9) were examined in winter anovulatory mares. A robust increase in circulating concentrations of LH relative to controls was observed in response to RF9 treatments, but treatments had no effect on adenohypophyseal responsiveness to GnRH. Data provide indirect evidence that antagonism of the RFRP3 system by RF9 may be at the GnRH neuronal level.Item Methadone metabolism in placentas from preterm pregnancies(2005-07-20) Todd Lewis Hieronymus; Dr. Mahmoud Ahmed; Dr. Joel Gallagher; Dr. Gary HankinsThe aim of this investigation is to identify and characterize the enzyme system responsible for the N-demethylation of methadone in human preterm placentas. The metabolism of methadone revealed typical Michaelis-Menten saturation kinetics. Methadone was N-demethylated to EDDP, only, by human placental aromatase. This was confirmed through the use of chemical inhibitors and monoclonal antibodies raised against specific CYP isoforms. The affinity of methadone to CYP19 remained relatively unchanged throughout gestation. However, the activity of the enzyme increased as gestation progressed, but showed wide variations between individual placentas. Taken together, it was shown that aromatase is the major enzyme responsible for the biotransformation of methadone throughout pregnancy. The variability in activity should affect the concentration of the drug within the fetal circulation. Accordingly, this might be an important factor affecting the occurrence and intensity of neonatal abstinence syndrome.Item Motherhood, Media and Reality: Analyzing Female Audience Reception of Celebrity Parenthood as News(2012-10-19) Hatfield, Elizabeth FishThe growing cultural commodity of celebrity news and its increasing focus on celebrities' families is examined by this project to determine what consequence communications about celebrity pregnancy and parenthood have on readers most likely to identify with the stories ? new mothers. While gossip magazines are not meant to provide parenting advice, their editorial focus on parenting may position celebrity parents as role models for audiences. Guided by theories of media effects, this project sought to understand why and how that might happen. Using narrative thematic analysis, two complementary data sets were analyzed: 36 issues sampled from the leading gossip magazines, People and Us Weekly, during 2007-2009, and five focus groups with recent mothers. Gossip magazines positively framed celebrity family life, idealizing the experience by avoiding talk of parenting's daily challenges. Resources such as nannies and personal trainers define celebrity parenting by affording celebrities, especially women, the ability to continue work while maintaining the identity of primary caregivers. A gendered act, consumption was intrinsically part of good celebrity parenting. Expectations for celebrity postpartum weight loss communicated that bigger bodies are a work-in-progress rather than an acceptable new body type. Fathers were visually depicted more often than in conventional parenting media, though these images similarly showed parents performing normative, gendered behaviors. Participants reported escapism as their main reason for reading gossip magazines and parasocial relationships existed with both liked and disliked celebrities. For liked celebrities, a parasocial dialectical tension emerged defining role models as both special and ordinary. For disliked celebrities, negative frames portrayed their parenting behavior as unacceptable and served as the strongest form of social learning from gossip magazines as readers internalized media criticism. Celebrity role models were selected based on feeling similar, serving as fantasy role models whose parenting lifestyles were simultaneously interpreted as aspirational and unattainable. Participants' social comparisons usually evaluated their own parenting experience as preferred to the demands and media environment faced by celebrities. Situations interpreted as incomparable attributed celebrities' success to external factors rather than internal characteristics. Overall, gossip magazines do provide parenting information that expands and impacts the real experience of mothers.Item Progesterone and interferon tau regulated genes in the endometrium of the ovine uterus and expression of interferon stimulated genes in the corpus luteum during early pregnancy in sheep(Texas A&M University, 2008-10-10) Ahn, Hyo WonDuring early pregnancy in ruminants, progesterone (P4) from the corpus luteum (CL) and interferon tau (IFNT) from the conceptus act on the endometrium to regulate genes including interferon stimulated genes (ISGs) that are hypothesized to be important for uterine receptivity and conceptus growth. Previous custom ovine cDNA array analyses identified candidate genes that were regulated by pregnancy, P4 and/or IFNT in the ovine uterus. The first study validated pregnancy, P4 and/or IFNT regulated genes identified by previous custom ovine cDNA microarray analyses. ACTA2, COL3A1, POSTN, SPARC, S100A2, STAT5A and TAGLN were examined. POSTN was upregulated by P4 and S100A2 was downregulated by IFNT. Moreover, functional studies showed that POSTN stimulated attachment of ovine trophectoderm cells. However, neither COL3A1, SPARC, ACTA2 nor TAGLN was regulated by pregnancy, P4 or IFNT in the ovine endometrium. Collectively, these results confirmed that POSTN and S100A2 are P4 and IFNT regulated, respectively, and likely involved in uterine receptivity to conceptus implantation during early pregnancy. The second study determined expression of ISGs in the CL of pregnant ewes. MX1, MX2, ISG15, OAS1, and RSAD2 mRNAs were increased on Day 14 of pregnancy and maintained to Day 18, indicating that IFNT induced expression of ISGs in the CL. These results confirmed that locally produced IFNT has paracrine effects and also endocrine effects on reproductive organs other than the uterine endometrium and maternal immune system.Item Progesterone regulation of endometrial factors supporting conceptus growth and development in the ovine uterus(Texas A&M University, 2008-10-10) Satterfield, Michael CareyProgesterone is unequivocally required for the establishment and maintenance of pregnancy in all mammals studied. Its known functions are complex and encompass global changes in gene expression. Therefore, studies were conducted to characterize the effects of progesterone on expression of genes for endometrial factors having roles in conceptus growth, implantation and establishment of pregnancy. The first study characterized the effect of an artificially induced early increase in circulating progesterone on conceptus growth and development and regulation of expression of galectin-15 (LGALS15), a recently identified protein secreted by the ovine uterine luminal epithelium (LE). Exogenous progesterone beginning on Day 1.5 post-mating accelerated conceptus development on Days 9 and 12. On Day 12 the conceptus was functionally and morphologically advanced to produce greater quantities of interferon tau (IFNT) than blastocysts from control ewes. Further, the endometrium responded to early progesterone and IFNT with early expression of cathepsin L (CTSL), radical S-adenosyl methionine domain containing 2 (RSAD2), and LGALS15 within the endometrium. The second study identifed structural changes within the luminal epithelium which could alter the flux of factors into and out of the uterine lumen to maintain appropriate fetal/maternal communication. In this study, progesterone reduced quantities of proteins associated with both tight and adherens junctions during the elongation period. IFNT subsequently increased these proteins after conceptus elongation. The third and fourth studies identified progesterone-regulated genes which have been implicated as having importance to implantation in sheep, mouse, and human. WNT signaling was transiently downregulated by progesterone, while members of several growth factor families are upregulated including insulin-like growth factor binding proteins (IGFBPs) 1 and 3, hepatocyte growth factor (HGF) and fibroblast growth factor 7 (FGF7), which may enhance conceptus growth. Collectively, these studies assess the role of progesterone in altering gene uterine expression to establish a favorable environment for conceptus development. The long-term goals of these studies are to establish biomarkers of receptivity to conceptus development and implantation, enhance our understanding of gene and pathway regulation in early pregnancy loss, and identify genes which may be targeted in therapeutic strategies to improve reproductive success in humans and animals.Item Progesterone Regulation of Endometrial Gene Expression in the Early Pregnant Ovine Uterus(2012-10-19) Minten, Megan A.Establishment of pregnancy in ruminants requires blastocyst development to form an elongated filamentous conceptus that produces interferon tau (IFNT), the pregnancy recognition signal, and initiate implantation. Blastocyst growth and development is dependent upon secretions from the uterine endometrium. An early increase in post-ovulatory circulating levels of progesterone (P4) stimulates blastocyst growth and conceptus elongation in ruminants. Microarray analysis was used to identify candidate P4-regulated genes and regulatory networks in the endometrium that govern peri-implantation blastocyst/conceptus growth and development. The first study was conducted to validate effects of P4 and/or pregnancy on expression of candidate genes identified by microarray analysis. The genes included: ANGPTL3, CHGA, CLEC4E, CXCL14, EFNA1, EFNB1, FABP3, IFNG, IL6, LGALS3, PTH, RBP4, SLIT2, SLIT3, and VWF. Early P4 treatment up-regulated CXCL14 gene expression in Day 9 ovine endometrium compared to control endometrium, and FABP3, IFNG, IL6 and LGALS3 in Day 12 early P4-treated ovine endometrium. Expression of ANGPTL3, CHGA, CXCL14, EFNA1, EFNB1, LGALS3 and RBP4 was affected by day of pregnancy. Treatment of ewes with P4+RU486 (P4 receptor antagonist) reduced expression of ANGPTL3, CHGA, EFNA1, EFNB1, FABP3, IFNG, IL6, LGALS3, RBP4, and SLIT2, SLIT3 and VWF in comparison to Day 12 P4-treated endometrium. The second study evaluated expression of genes identified by microarray analysis in endometrium from pregnant and cyclic ewes. Genes evaluated included those from the first study. ANGPTL3, CHGA, CXCL14, EFNA1, EFNB1, IFNG, LGALS3, PTH, RBP4, SLIT2, SLIT3 and VWF were affected by day, status and/or their interaction between Days 10 and 16. Of note, FABP3 increased 21-fold between Days 14 to 18 of pregnancy, and IL6 increased 37-fold between Days 14 to 20 of pregnancy. In situ hybridization analysis detected FABP3 mRNA in both luminal and superficial glandular epithelia of pregnant ewes and trophectoderm, whereas IL6 mRNA was detected in immune cells within uterine luminal epithelium and glandular epithelium and trophectoderm. Collectively, these results identify candidate genes encoding for biologically active molecules that regulate growth and development of the ovine conceptus during the peri-implantation period of pregnancy.Item Transcriptional Regulation of Galectin 15 (LGALS15): An Implantation-Related Galectin Uniquely Expressed in the Uteri of Sheep and Goats(2010-10-12) Lewis, Shaye K.Galectins are a family of secreted animal lectins with a high affinity to betagalactosides commonly involved in cellular functions such as apoptosis, adhesion and migration. Galectin 15 (LGALS15), a newest member of the galectin superfamily, has a unique C-terminal RGD sequence and participates in integrin-mediated ovine trophectoderm cell attachment and migration. In the ovine uterus, LGALS15 is expressed only by the endometrial luminal (LE) and superficial glandular (sGE) epithelia, induced by progesterone between Days 10 and 12 of the cycle and pregnancy, and then stimulated by interferon tau (IFNT) from the conceptus after Day 14 of pregnancy. During early pregnancy, the canonical janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway is not active in the endometrial LE/sGE. Therefore, IFNT may utilizes a non-canonical signaling pathway to increase transcription of genes, including CST3, CTSL, HIF2A, LGALS15, and WNT7A, specifically in the endometrial LE/sGE. Alternatively, IFNT and progesterone could indirectly affect epithelial gene expression by influencing gene expression in the stroma, which then communicates with the epithelium. Although the LGALS15 gene is present in ovine, caprine and bovine species, it is only expressed in uteri of sheep and goats. Available data shows a tissue- and speciesspecific expression pattern for LGALS15, likely involving multiple layers of transcription regulation in the ruminant endometrium. Further analysis of the LGALS15 5? promoter/enhancer region revealed similar predicted transcription factor binding sites in all three species, including; PU.1, Ets-1, AP1, Sp1, and GRE or PRE sites. Interestingly, the proximal promoter region of the LGALS15 gene in all three species exhibited a conserved Sp1 binding site upstream of an AP1 binding site on both sense and antisense strands, and with similar spacing between binding sites. Sequence analysis revealed key differences in LGALS15 gene structure between ruminant species including the proximity of repetitive DNA sequences to the transcription start site (+1). Bovine LGALS15 has repetitive DNA sequences start at - 145 whereas in ovine or caprine LGALS15 it starts at about -300. The length of the repetitive DNA sequence is similar (~1.2 kb) in the 5' promoter/enhancer region of LGALS15 in all three species. Transient transfection analyses found that repetitive DNA sequences reduced basal promoter activity and responsiveness to treatments. None of the promoter construct showed responsiveness to interferon tau (IFNT). The bovine LGALS15 gene promoter showed no activity under any experimental conditions. The current studies indicate that uterine LGALS15 is expressed in ovine and caprine but not bovine species. Additionally, repetitive DNA sequences found in the promoter region may contribute to modulating the LGALS15 gene expression. Therefore, the ruminant LGALS15 gene, like other galectins, is under tight transcriptional control involving hormones, requisite transcription factors and potentially chromatin remodeling complexes working synergistically for LGALS15 promoter transactivation.