Transcriptomic analysis of the porcine endometrium during early pregnancy and the estrous cycle (original) (raw)
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Biology of Reproduction, 2006
Porcine trophoblast attachment to the uterine surface is associated with increased conceptus and endometrial production of prostaglandins. Conceptus secretion of estrogen on Day 12 of gestation is important for establishment of pregnancy; however, early (Days 9 and 10) exposure to exogenous estrogens results in embryonic mortality. Present studies established the temporal and spatial pattern of endometrial PTGS1 (prostaglandin-endoperoxide synthase 1) and PTGS2 expression during the estrous cycle and early pregnancy and determined the effect of early estrogen treatment on endometrial PTGS expression in pregnant gilts. Endometrial PTGS1 mRNA expression increased 2-to 3-fold after Day 10 of the estrous cycle and pregnancy, whereas PTGS2 mRNA expression increased 76-fold between Days 5 and 15 of the estrous cycle and pregnancy. Increased expression of the PTGS2 transcript was detected in the lumenal epithelium after Day 10 in both cyclic and pregnant gilts. There was a 10-and 20-fold increase in endometrial PTGS2 protein expression between Days 5 and 18 of the estrous cycle and pregnancy respectively. Administration of estrogen on Days 9 and 10 of gestation increased endometrial PTGS2 mRNA and protein on Day 10, but decreased PTGS2 mRNA and protein in lumenal epithelium (LE) on Day 12 of gestation compared to vehicle-treated gilts. The present study demonstrates that an increase in uterine epithelial PTGS2 expression occurs after Day 10 of the estrous cycle and early pregnancy in the pig. The conceptus-independent increase in the uterine LE indicates that a novel pathway exists for endometrial induction PTGS2 expression before conceptus elongation and attachment to the uterine surface. Epithelial expression of PTGS2 may serve as one of the signals for placental attachment and embryo survival in the pig. Early administration of estrogen on Days 9 and 10 of pregnancy alters endometrial PTGS2 mRNA and protein expression, which may, at least in part, represent a mechanism by which endocrine disruption of pregnancy causes total embryonic loss during implantation in the pig.
Estradiol-17β-Induced Changes in the Porcine Endometrial Transcriptome In Vivo
International Journal of Molecular Sciences
Estradiol-17β (E2) is a key hormone regulating reproductive functions in females. In pigs, E2, as the main conceptus signal, initiates processes resulting in prolonged corpus luteum function, embryo development, and implantation. During early pregnancy the endometrium undergoes morphological and physiological transitions that are tightly related to transcriptome changes. Recently, however, the importance of E2 as a primary conceptus signal in the pig has been questionable. Thus, the aim of the present study was to determine the effects of E2 on the porcine endometrial transcriptome in vivo and to compare these effects with transcriptome profiles on day 12 of pregnancy. Microarray analysis revealed differentially expressed genes (DEGs) in response to E2 with overrepresented functional terms related to secretive functions, extracellular vesicles, cell adhesion, proliferation and differentiation, tissue rearrangements, immune response, lipid metabolism, and many others. Numerous common...
Endocrinology, 2009
Before implantation, the porcine endometrium and trophoblast synthesize elevated amounts of luteoprotective prostaglandin estradiol-17 (E 2) (PGE 2). We hypothesized that embryo signal, E 2 , and PGE 2 modulate expression of key enzymes in PG synthesis: PG-endoperoxide synthase-2 (PTGS2), microsomal PGE synthase (mPGES-1), PGF synthase (PGFS), and PG 9-ketoreductase (CBR1) as well as PGE 2 receptor (PTGER2 and-4) expression and signaling within the endometrium. We determined the site of action of PGE 2 in endometrium during the estrous cycle and pregnancy. Endometrial tissue explants obtained from gilts (n ϭ 6) on d 11-12 of the estrous cycle were treated with vehicle (control), PGE 2 (100 nM), E 2 (1-100 nM), or phorbol 12-myristate 13-acetate (100 nM, positive control). E 2 increased PGE 2 secretion through elevating expression of mPGES-1 mRNA and PTGS2 and mPGES-1 protein in endometrial explants. By contrast, E 2 decreased PGFS and CBR1 protein expression. E 2 also stimulated PTGER2 but not PTGER4 protein content. PGE 2 enhanced mPGES-1 and PTGER2 mRNA as well as PTGS2, mPGES-1, and PTGER2 protein expression. PGE 2 had no effect on PGFS, CBR1, and PTGER4 expression and PGF 2␣ release. Treatment of endometrial tissue with PGE 2 increased cAMP production. Cotreatment with PTGER2 antagonist (AH6809) but not PTGER4 antagonist (GW 627368X) inhibited significantly PGE 2-mediated cAMP production. PTGER2 protein was localized in luminal and glandular epithelium and blood vessels of endometrium and was significantly up-regulated on d 11-12 of pregnancy. Our results suggest that E 2 prevents luteolysis through enzymatic modification of PG synthesis and that E 2 , PGE 2 , and endometrial PTGER2 are involved in a PGE 2 positive feedback loop in porcine endometrium. (Endocrinology 150: 3823-3832, 2009) P orcine embryos secrete enhanced levels of estrogens, mainly estradiol-17 (E 2) between d 10 and 13 after fertilization, just before implantation in the process of the maternal recognition of pregnancy (1-4). It has been demonstrated that systemic estrogen administration between d 11 and 13 of the estrous cycle prevents corpus luteum regression and extends this gland's lifespan in the pig (5). The action of estrogens may be luteoprotective and antiluteolytic (3, 4, 6). However, estrogen alone does not fully exert the effect of the embryo on growth and development of corpus luteum (7). It has been suggested that besides estrogens, prostaglandin (PG) E 2 (PGE 2) could be involved in the maternal recognition of pregnancy as a luteoprotective/antiluteolytic factor (8, 9). Endome-trial PGE 2 , and PGF 2␣ , which has an opposite luteolytic effect, are involved in reproduction processes in many species (10, 11). It was demonstrated that inhibition of PG synthesis before implantation causes pregnancy failure in different species, including the pig (11, 12). PGs are synthesized by PG-endoperoxide synthase (PTGS) and specific terminal PG synthases: PGE synthase (PGES) and PGF synthase (PGFS) (13, 14). Moreover, PGE 2 can be converted into PGF 2␣ by PG 9-ketoreductase/carbonyl reductase (CBR1) (15-17). It was found that highly inducible forms of PTGS and PGES in the porcine endometrium are PTGS2, (known also as PGHS-2 or COX-2) (18, 19) and microsomal PGES-1 (mPGES-1), respectively (14).
Estrogen-Induced Disruption of Neonatal Porcine Uterine Development Alters Adult Uterine Function
Biology of Reproduction, 2002
In the pig, estradiol-17 valerate (EV) exposure from birth (Postnatal Day [PND] 0) disrupts estrogen receptor-␣ (ER)-dependent uterine development and increases embryo mortality in adults. To determine effects of neonatal EV exposure on adult uterine morphology and function, 36 gilts received corn oil (CO) or EV from PND 0 to PND 13. Cyclic and pregnant (PX) adults from each treatment group were hysterectomized on Day 12 after estrus/mating. Treatment and pregnancy effects were determined for uterine weight and horn volume, uterine luminal fluid (ULF) protein and estradiol content, endometrial incorporation of 3 H-leucine (3 H-Leu) into nondialyzable product, and endometrial mRNA levels for ER, progesterone receptor (PR), uteroferrin (UF), retinol-binding protein (RBP), and keratinocyte growth factor (KGF). Adults cycled normally and had similar numbers of corpora lutea. Uteri of PX gilts contained tubular/ filamentous conceptuses, and ULF estradiol content was unaffected by treatment. However, pregnancy increased uterine weight and size only in CO gilts (Treatment ؋ Status, P Ͻ 0.01). Treatment reduced ULF protein content (P Ͻ 0.01), endometrial 3 H-Leu incorporation (P Ͻ 0.05), and the pregnancy-associated increase in ULF protein (Treatment ؋ Status, P Ͻ 0.01). Treatment did not affect endometrial ER or PR mRNA levels but attenuated the pregnancy-associated increase in UF mRNA (Treatment ؋ Status; P Ͻ 0.01), increased RBP (P Ͻ 0.10), and decreased KGF mRNA levels (P Ͻ 0.05). These results establish that transient postnatal estrogen exposure affects porcine uterine responsiveness to potentially embryotrophic signals and that estrogen-sensitive postnatal uterine organizational events are determinants of uterine size and functionality.
Proteomic analysis of the endometrium during early pregnancy in the domestic pig
Reproduction, Fertility and Development, 2017
Reproductive processes in domestic pigs have been studied extensively. Pigs are one of the main sources of meat for human consumption and are an established model for investigations into mammalian, including human, reproductive physiology. Studies of the uterus during early pregnancy will lead to a better understanding of mechanisms governing pregnancy. Proteomics provides the possibility to explore endometrial functions in an unbiased way. The aim of the study was to compare endometrium harvested from Days 12–13 and 15–16 of pregnancy with the corresponding days of the oestrous cycle. We identified endometrial proteins that are unique to the early stages of pregnancy (Days 12–13 and 15–16). Twenty-one proteins were identified that were uniquely expressed on the selected days of pregnancy or the oestrous cycle. Out of 21 identified proteins, 14 referred to the pregnancy periods. Systemic analysis of the identified proteins revealed cell adhesion and cytoskeletal organisation as two ...
Life, 2020
During the early stages of pregnancy, the uterine endometrium undergoes dramatic morphologic and functional changes accompanied with dynamic variation in gene expression. Pregnancy-stage specific differentially expressed gene (DEG)-transcript-probes were investigated and identified by comparing endometrium transcriptome at 9th day (9D), 12th day (12D) and 16th day (16D) of early pregnancy in Polish large-white (PLW) gilts. Endometrium comparisons between 9D-vs-12D, 9D-vs-16D and 12D-vs-16D of early pregnancy identified 6049, 374 and 6034 highly significant DEG-transcript-probes (p < 0.001; >2 FC). GO term enrichment analysis identified commonly shared upregulated endometrial DEG-transcript-probes (p < 0.001; >2 FC), that were regulating the gene functions of anatomic structure development and transport (TG), DNA-binding and methyltransferase activity (ZBTB2), ion-binding and kinase activity (CKM), cell proliferation and apoptosis activity (IL1B). Downregulated DEG-transc...
Bioscientifica Proceedings, 2019
Timing of conceptus growth and attachment to the uterine luminal epithelium is regulated by progesterone secretion from the corpus luteum and by expression of progesterone receptor in the uterine epithelia and stroma. Conceptus growth and uterine attachment are temporally associated with the disappearance of progesterone receptors from uterine epithelia. While the loss of progesterone receptor from the endometrial epithelia on day 10 of the oestrous cycle and pregnancy has been well documented, the factors involved with cell specific down-regulation of progesterone receptor are yet to be established. We propose that several progesterone stimulated factors activate nuclear factor kappa B (NF-kB) within the uterine epithelia, which leads to inhibition of progesterone receptor and concomitant stimulation of endometrial genes expressed during early conceptus development. Although oestrogens secreted by pig conceptuses function to establish pregnancy, timing of endometrial exposure to oestrogen is critical. Early oestrogen administration alters the pattern of gene expression through the NF-kB system desynchronising the uterine environment for conceptus implantation resulting in later embryonic loss.
Biology of Reproduction, 2005
Secreted phosphoprotein 1 (SPP1, commonly referred to as osteopontin and formerly known as bone sialoprotein 1, early Tlymphocyte activation 1) is an extracellular matrix/adhesion molecule that is upregulated in the pregnant uterus of all mammals examined to date. This study focused on the pig, which has true epitheliochorial placentation and exhibits induction of SPP1 mRNA in luminal epithelium (LE) just before conceptus attachment and in glandular epithelium (GE) after Day 30 of pregnancy. The objective of this study was to determine steroid regulation of SPP1 mRNA and protein in porcine uterine epithelium. To examine the effect of estrogen, cyclic gilts were treated daily (Days 11-14) with 5 mg estradiol benzoate (i.m.) and hysterectomized on Day 15. To evaluate the long-term effect of pseudopregnancy, cyclic gilts were given daily injections (Days 11-15) with steroid as above and hysterectomized on Day 90. In situ hybridization showed high expression of SPP1 mRNA only in LE contiguous with apposing conceptus tissue on Day 15 of pregnancy. In contrast, estrogen injection resulted in moderate but uniform SPP1 mRNA in all LE of Day 15 nonpregnant gilts, with expression maintained through Day 90 of pseudopregnancy. SPP1 mRNA also localized to the GE of Day 90 pseudopregnant gilts, similar to expression in late gestation. Consistent with in situ hybridization results, SPP1 protein localized to the apical surface of LE in all estrogentreated gilts and in the GE on Day 90 of pseudopregnancy. We conclude that, in pregnant pigs, SPP1 is induced by conceptus estrogen in uterine LE and is regulated in GE in a manner coincident with CL/placental progesterone production.
Premature Estrogen Exposure Alters Endometrial Gene Expression to Disrupt Pregnancy in the Pig
Endocrinology, 2007
Establishment and maintenance of pregnancy in the pig involve intricate communication between the developing conceptuses and maternal endometrium. Conceptus synthesis and release of estrogen during trophoblastic elongation are essential factors involved with establishing conceptus-uterine communication. The present study identified endometrial changes in gene expression associated with implantation failure and complete pregnancy loss after premature exposure of pregnant gilts to exogenous estrogen. Gilts were treated with either 5 mg estradiol cypionate (EC) or corn oil on d-9 and -10 gestation, which was associated with complete conceptus degeneration by d-17 gestation. Microarray analysis of gene expression revealed that a total of eight, 32, and five genes were up-regulated in the EC endometrium, whereas one, 39, and 16 genes were down-regulated, on d 10, 13, and 15, respectively. Four endometrial genes altered by EC, aldose reductase (AKR1B1), secreted phosphoprotein 1 (SPP1), CD24 antigen (CD24), and neuromedin B (NMB), were evaluated using quantitative RT-PCR and in situ hybridization. In situ hybridization localized gene expression for NMB, CD24, AKR1B1, and SPP1 in the luminal epithelium, and confirmed the expression patterns from RT-PCR analysis. The aberrant expression patterns of endometrial AKR1B1, SPP1, CD24, and NMB 3-4 d after premature estrogen exposure to pregnant gilts may be involved with conceptus attachment failure to the uterine surface epithelium and induction of endometrial responses that disrupt the establishment of a viable pregnancy. (Endocrinology 148: 4761-4773, 2007) First