Immunocytochemical localization of prostaglandin synthase in the ovine uterus during the oestrous cycle and in early pregnancy

1990 ◽  
Vol 2 (4) ◽  
pp. 311 ◽  
Author(s):  
LA Salamonsen ◽  
JK Findlay
Keyword(s):  
Epithelial Cells ◽  
Early Pregnancy ◽  
Stromal Cells ◽  
Cellular Localization ◽  
In Vitro Release ◽  
Oestrous Cycle ◽  
Intense Staining ◽  
Endometrial Cells ◽  
Ovine Uterus

Prostaglandin (PG) synthase has been localized by immunocytochemistry within the ovine uterus throughout the oestrous cycle and in early pregnancy. On Day 4 of the cycle, PG synthase was located primarily in the stromal cells in caruncular and intercaruncular tissue with little staining in the epithelium. On Days 14 through to 16, the most intense staining was in the luminal epithelial cells (caruncular and intercaruncular) and in epithelial cells of glands close to the uterine lumen. PG synthase was also located in the intercaruncular stromal cells, particularly close to the myometrium. Staining for the enzyme on Day 10 was intermediate between that of Day 4 and Day 14. On Day 15 of pregnancy, the pattern of staining was identical to that on Day 15 of the cycle, with no detectable difference in intensity. When endometrial cells (cycle, Day 14) were cultured with and without ovine trophoblast protein-1 (3 ng mL-1) in vitro, release of PGE and PGF2 alpha was attenuated (54% and 47% of control respectively) but no differences were observed in the intensity of staining for PG synthase in the cells. These results demonstrate marked cyclical changes in the endometrial cell types producing PGs, suggesting differential regulation of PG synthase. In addition, it appears that conceptus-induced changes in PGF2 alpha release do not occur via changes in the concentration or cellular localization of PG synthase, but rather that the activity of the enzyme is modified.

Localization of neutral endopeptidase in the ovine uterus and conceptus during the oestrous cycle and early pregnancy

1995 ◽  
Vol 7 (1) ◽  
pp. 27 ◽  
Author(s):  
SC Riley ◽  
E Wong ◽  
JK Findlay ◽  
LA Salamonsen
Keyword(s):  
Smooth Muscle ◽  
Epithelial Cells ◽  
Smooth Muscle Cells ◽  
Early Pregnancy ◽  
Stromal Cells ◽  
Immunohistochemical Staining ◽  
Muscle Cells ◽  
Neutral Endopeptidase ◽  
Oestrous Cycle ◽  
Ovine Uterus

Neutral endopeptidase (NEP; EC 3.4.24.11), an enzyme which metabolizes several peptides (including oxytocin and endothelins) implicated in the control of uterine function, was found to be localized in the ovine uterus throughout the oestrous cycle and in the uterus and conceptus during early pregnancy, using immunohistochemical techniques. Positive NEP immunoreactivity was found in the endometrium principally in stromal cells, in the vasculature in endothelial and vascular smooth muscle cells, and also weakly in some glandular epithelial cells. In a layer of stromal fibroblasts several cells in thickness underlying the luminal epithelium, staining was much weaker than that in the deeper stromal cells throughout the period examined. NEP staining was also present in smooth muscle cells of the myometrium at all times, and was most intense in the layer of cells adjacent to the endometrium. In the conceptus, NEP immunohistochemical staining was found in uninucleate cells, but not in binucleate trophoblast cells, in epithelial cells of the allantois and amnion, and in the heart and brain of the Day-20 embryo. In ovariectomized ewes treated with oestrogen or progesterone separately or remaining untreated, immunohistochemical staining of NEP was stronger when compared with intact ewes, in caruncular and intercaruncular stroma and epithelia, in glands, in the vasculature and in myometrium. The staining was less intense in all cell types in ewes receiving oestrogen plus progesterone. The expression of NEP and its specific regionalization within the uterus indicate a mechanism by which the availability of biologically important peptides involved in the regulation of the oestrous cycle and implantation, including oxytocin and endothelin, can be controlled by regulation of their metabolism.

scholarly journals Ovarian steroids affect prostaglandin production in equine endometrial cells in vitro

2014 ◽  
Vol 220 (3) ◽  
pp. 263-276 ◽  
Author(s):  
Anna Z Szóstek ◽  
António M Galvão ◽  
Graça M Ferreira-Dias ◽  
Dariusz J Skarzynski
Keyword(s):  
Cell Proliferation ◽  
Epithelial Cells ◽  
Protein Expression ◽  
Stromal Cells ◽  
Positive Control ◽  
Dependent Manner ◽  
Ovarian Steroids ◽  
Endometrial Cells ◽  
Prostaglandin Endoperoxide Synthase

This study aimed to evaluate the influence of ovarian steroids on equine endometrial epithelial and stromal cells, specifically i) prostaglandin (PG) production in a time-dependent manner, ii) specific PG synthases mRNA transcription and protein expression, and iii) cell proliferation. After passage I, cells were exposed to vehicle, oxytocin (OT, positive control, 10−7M), progesterone (P4, 10−7M), 17β estradiol (E2, 10−9M), or P4+E2for 12, 24, 48, or 72 h. Following treatment, PG concentration was determined using the direct enzyme immunoassay (EIA) method. Alterations inPGsynthases mRNA transcriptions,PGsynthases protein expression, and cell proliferation in response to the treatments were determined after 24 h using real-time PCR, western blot, or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide respectively. After 24 h, E2and P4+E2increased PGE2and PGF2αsecretion as well as specific prostaglandin-endoperoxide synthase-2 (PTGS2), PGE2synthases (PGES), and PGF2αsynthases (PGFS) expression in the epithelial cells (P<0.05). Additionally, E2and P4+E2increased PTGS2 expression in stromal cells after 24 h (P<0.05). In stromal cells, P4+E2increased PGE2production as well as PGES expression after 24 h (P<0.05). Both E2and P4+E2increased PGF2αproduction by stromal cells after 24 h (P<0.05). Ovarian steroids affected proliferation of stromal and epithelial cells during the 24-h incubation period (P<0.05). We provide evidence that ovarian steroids affect PG production in equine endometrial cells, upregulating PTGS2, PGES, and PGFS expression. Ovarian steroid-stimulated PG production could be an important mechanism occurring in the equine endometrium that is involved in the regulation of the estrous cycle and early pregnancy.

scholarly journals Effect of LH on prostaglandin F2α and prostaglandin E2 secretion by cultured porcine endometrial cells

Reproduction ◽  
2005 ◽  
Vol 130 (1) ◽  
pp. 105-112 ◽  
Author(s):  
Agnieszka Blitek ◽  
Adam J Ziecik
Keyword(s):  
Epithelial Cells ◽  
Prostaglandin E2 ◽  
Stromal Cells ◽  
Prostaglandin F2α ◽  
Cell Types ◽  
Oestrous Cycle ◽  
Time Dependent ◽  
Dependent Effect ◽  
Endometrial Cells ◽  
Time Dependent Effect

LH appears to be a potent stimulator of the release of endometrial prostaglandins (PGs) in the pig. The aim of the present studies was to examine the effect of LH on PGF2αand PGE2secretion by cultured porcine endometrial cells on days 10–12 and 14–16 of the oestrous cycle and to compare its action with oxytocin. A time-dependent effect of LH (10 ng/ml) on PGF2αrelease from luminal epithelial and stromal cells on days 10–12 was observed (experiment 1). The highest increase in PGF2αsecretion in response to LH was detected in stromal cells after 6 h of incubation (P< 0.001). Epithelial cells responded to LH after a longer exposure time (P< 0.01). A concentration-dependent effect of LH (0.1–100 ng/ml) on PGF2αrelease from stromal cells was examined after 6 h and from epithelial cells after 12 h (experiment 2). Effective concentrations of LH were 10 and 100 ng/ml. LH (10 ng/ml) and oxytocin (100 nmol/l) affected PGF2αand PGE2secretion from endometrial cells on days 10–12 and 14–16 of the oestrous cycle (experiment 3). LH stimulated PGF2αsecretion from both cell types and its action was more potent on days 10–12. LH induced PGE2release, especially in epithelial cells on days 14–16. A stimulatory effect of oxytocin on PGF2αwas confirmed in stromal cells, but this hormone was also shown to enhance PGE2output. These results indicated that LH, like oxytocin, a very effective stimulator of PGF2αrelease, could play an important role in the induction of luteolysis.

scholarly journals Endometrium on-a-chip reveals insulin- and glucose-induced alterations in the transcriptome and proteomic secretome

Endocrinology ◽  
2021 ◽  
Author(s):  
Tiago H C De Bem ◽  
Haidee Tinning ◽  
Elton J R Vasconcelos ◽  
Dapeng Wang ◽  
Niamh Forde
Keyword(s):  
Early Pregnancy ◽  
Stromal Cells ◽  
Protein Coding ◽  
Endometrial Cells ◽  
In Utero Environment ◽  
Lower Chamber ◽  
Transcriptional Changes ◽  
First Time ◽  
Dose Dependent

Abstract The molecular interactions between the maternal environment and the developing embryo that are key for early pregnancy success and are influenced by factors such as maternal metabolic status. Our understanding of the mechanism(s) through which these individual nutritional stressors alter endometrial function and the in utero environment for early pregnancy success is, however, limited. Here we report, for the first time, the use of an endometrium-on-a-chip microfluidics approach to produce a multi-cellular endometrium in vitro. Isolated endometrial cells (epithelial and stromal) from the uteri of non-pregnant cows in the early-luteal phase (Day 4-7), were seeded in the upper chamber of the device (epithelial cells; 4-6 10 4 cells/mL) and stromal cells seeded in the lower chamber (1.5-2 10 4 cells/mL). Exposure of cells to different concentrations of glucose (0.5, 5.0 or 50 mM) or insulin (Vehicle, 1 or 10 ng/mL) were performed at a flow rate of 1µL/min for 72 hr. Quantitative differences in the cellular transcriptome and the secreted proteome of in vitro-derived uterine luminal fluid (ULF) were determined by RNA-sequencing and Tandem Mass Tagging Mass Spectrometry (TMT-MS), respectively. High glucose concentrations altered 21 and 191 protein-coding genes in epithelial and stromal cells, respectively (p&lt;0.05), with a dose-dependent quantitative change in the protein secretome (1 and 23 proteins). Altering insulin concentrations resulted in limited transcriptional changes including transcripts for insulin-like binding proteins that were cell specific but altered the quantitative secretion of 196 proteins. These findings highlight one potential mechanism by which changes to maternal glucose and insulin alter uterine function.

Changes in oestrogen receptor protein, mRNA expression and localization in the endometrium of cyclic and pregnant gilts

1993 ◽  
Vol 5 (3) ◽  
pp. 247 ◽  
Author(s):  
RD Geisert ◽  
RM Brenner ◽  
RJ Moffatt ◽  
JP Harney ◽  
T Yellin ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Mrna Expression ◽  
Oestrogen Receptor ◽  
Cellular Localization ◽  
Oestrous Cycle ◽  
Glandular Epithelium ◽  
Receptor Protein ◽  
Immunocytochemical Staining ◽  
Positive Staining ◽  
Intense Staining

Conceptus secretion of oestrogen on Day 11 of gestation is involved with establishment of pregnancy in the pig. Changes in oestrogen receptor (ER) protein, mRNA and cellular localization in the endometrium were evaluated during the oestrous cycle and early pregnancy of the gilt. In nonpregnant gilts, concentration of nuclear ER in the endometrium increased from Days 0 to 12 followed by a decline on Day 15 of the oestrous cycle. In pregnant gilts, changes in endometrial nuclear ER during Days 10, 12, 15 and 18 were similar to that in cyclic pigs. Analysis of endometrial ER mRNA expression did not detect any difference between cyclic and pregnant pigs between Days 10 and 15 postoestrus. Expression of ER mRNA in endometrium of cyclic and pregnant gilts was greatest on Day 10 followed by a decline on Day 15. Endometrial ER mRNA increased on Day 18 of the oestrous cycle, but remained low during pregnancy. Immunocytochemical localization of ER in the endometria of cyclic and pregnant gilts indicated that there was intense staining for ER in stromal cells and moderate to strong staining in surface and glandular epithelial cells during oestrus (Day 0) and Day 18 of the oestrous cycle. However, stromal ER staining was absent from Days 5 to 15 of the oestrous cycle and continued to be suppressed on Day 18 of pregnancy. Immunocytochemical staining of ER in the surface and glandular epithelium was readily detectable from Days 0 to 12 of the oestrous cycle and during pregnancy. Intensity of staining for ER declined in surface epithelial cells on Day 15 in both cyclic and pregnant pigs whereas positive staining for ER in glandular epithelium was absent. Staining for ER on uterine surface epithelial cells increased during pro-estrus (Day 18) of cyclic gilts but remained similar to Day 15 in pregnant gilts. Changes in endometrial ER protein, mRNA and localization in surface epithelium are consistent with a physiological role for conceptus oestrogen secretion in uterine function and maternal recognition of pregnancy in the pig.

scholarly journals Temporal expression and cellular localization of a gastrin-releasing peptide-related gene in ovine uterus during the oestrous cycle and pregnancy

1998 ◽  
Vol 157 (1) ◽  
pp. 139-148 ◽  
Author(s):  
JC Whitley ◽  
A Shulkes ◽  
LA Salamonsen ◽  
D Vogiagis ◽  
M Familari ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Cellular Localization ◽  
Major Product ◽  
Oestrous Cycle ◽  
Endometrial Tissue ◽  
Cycle Duration ◽  
Gastrin Releasing Peptide ◽  
Stromal Tissue ◽  
Blastocyst Implantation ◽  
Ovine Uterus

Synthesis of both mRNA and peptide for gastrin-releasing peptide (GRP) has been demonstrated in the pregnant endometrium of sheep and women. However, it is not known whether GRP is synthesized in the sheep uterus during the oestrous cycle. Furthermore the cellular site of GRP mRNA synthesis in the uterus has not been determined. Therefore we examined the synthesis of GRP and determined the cellular location of GRP peptide and mRNA in sheep uterus taken at different times during the oestrous cycle (duration 17 days) and pregnancy (duration 145 days). Northern blot analysis of RNA isolated from ovine endometrium revealed low or no GRP mRNA at days 4, 10, 12 and 14 of the oestrous cycle and a 24-fold rise in GRP mRNA (normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA) between days 14 and 16. A similar pattern was observed during early pregnancy, with a 12-fold rise in GRP mRNA:GAPDH mRNA between days 17 and 20 of pregnancy. Levels of GRP peptide were determined by RIA and found to be low in endometrium isolated at days 4, 10, 12 and 14 of the oestrous cycle (1.0-1.6 pmol/g) and 4 to 5-fold higher at day 16. In situ hybridization localized GRP synthesis to the epithelial cells of the uterine glands at day 16 of the oestrous cycle and at days 17, 20, 40 and 50 of pregnancy. At day 140 of pregnancy diffuse hybridization to cells of the myometrium was also observed. Immunohistochemistry localized GRP peptide to the apical cytoplasm of uterine glandular epithelial cells at day 16 of the oestrous cycle. For samples obtained at day 20 of pregnancy, the area surrounding the glands also showed moderately strong staining. Further staining in the glandular lumen and the stromal tissue surrounding the glands was apparent at day 140 of pregnancy. No GRP immunoreactivity could be detected in the peripheral plasma during the oestrous cycle or the first 20 days of pregnancy. Sizing chromatography of GRP immunoreactivity extracted from endometrial tissue taken at day 10 of the oestrous cycle revealed two peaks that co-eluted with GRP(1-27) and GRP(18-27). However, during luteolysis and oestrus the major peak of GRP immunoreactivity extracted from endometrial tissue was larger than GRP(1-27) and similar to that seen previously in the gravid ovine endometrium. These studies demonstrate that a peptide similar to, but larger than, GRP is a major product of the glandular epithelium of the ovine uterus during the luteal regression phase of the oestrous cycle and post-blastocyst implantation in pregnancy and provide further evidence that GRP-related peptides have important regulatory roles in uterine function.

Immunocytochemical localization and changes in endometrial progestin receptor protein during the porcine oestrous cycle and early pregnancy

1994 ◽  
Vol 6 (6) ◽  
pp. 749 ◽  
Author(s):  
RD Geisert ◽  
TN Pratt ◽  
FW Bazer ◽  
JS Mayes ◽  
GH Watson
Keyword(s):  
Progesterone Receptor ◽  
Early Pregnancy ◽  
Cellular Localization ◽  
Oestrous Cycle ◽  
Receptor Protein ◽  
Surface Epithelium ◽  
Immunocytochemical Localization ◽  
Intense Staining ◽  
Pr Protein ◽  
Progestin Receptor

Changes of progesterone receptor (PR) protein and cellular localization in the endometrium were evaluated during the oestrous cycle and early pregnancy of the gilt. During the oestrous cycle, the concentration of total PR protein within the endometrium was highest on Days 0-5 and decreased on Day 10. The endometrial concentration of PR reached a nadir on Day 12 and this level was maintained throughout the remainder of the oestrous cycle (Day 18). In pregnant gilts, the concentration of endometrial PR protein from Day 10 to Day 18 was similar to that in cyclic gilts. Western blot analysis with antiserum specific for the A and B isoforms of PR indicated that porcine endometrium expresses both isoforms of PR. Immunostaining was detectable for both the A and B isoforms of PR from Day 0 to Day 12 of the oestrous cycle. However, no staining was observed on Day 15 and Day 18 of the oestrous cycle or pregnancy Immunocytochemical localization of PR in the endometrium of cyclic gilts and pregnant gilts indicated that there was intense staining for PR in surface epithelium and glandular epithelium during oestrus (Day 0) and on Day 5. However, the staining was less intense on Day 7 and Day 10 of the oestrous cycle and no epithelial staining was observed after Day 12. PRs were present in the stroma and myometrium throughout the oestrous cycle and early pregnancy. The presence of conceptuses during pregnancy did not affect the loss of PR from the uterine epithelium after Day 10 of gestation. Down-regulation of epithelial PR might be one factor involved in the timing of luteolysis during the oestrous cycle as well as conceptus growth and placentation during early pregnancy.

scholarly journals 249.Activin A upregulates endometrial metalloproteases: potential mechanisms for promotion of decidualisation and implantation

2004 ◽  
Vol 16 (9) ◽  
pp. 249
Author(s):  
R. L. Jones ◽  
P. Paiva ◽  
T. J. Kaitu'u ◽  
L. A. Salamonsen
Keyword(s):  
Epithelial Cells ◽  
Stromal Cells ◽  
Transforming Growth Factor ◽  
Embryo Implantation ◽  
Activin A ◽  
Matrix Metalloproteases ◽  
Trophoblast Invasion ◽  
Endometrial Cells ◽  
Mmp Inhibitor

Activin and inhibin subunits are co-expressed by human endometrial epithelial and decidualised stromal cells. Activin A is a potent stimulator of decidualisation in vitro, but the mechanisms are unknown. Matrix metalloproteases (MMPs) are known to be important during decidualisation, as administration of a broad spectrum MMP inhibitor in the rat results in reduced decidualisation. Transforming Growth Factor(TGF)-βs are closely related to activins and inhibit MMP production in endometrial epithelial cells. We hypothesised that activins regulate MMP production during decidualisation and/or trophoblast invasion. Epithelial and stromal cells were isolated from human endometrium and treated for 24 h with activin, inhibin, activin/inhibin, and follistatin. Media were collected and subjected to gelatin and caesin zymography. In epithelial cells, activin A stimulated the expression of latent forms of MMPs-1, -2, -7 and -9, and increased formation of active forms of MMPs-2 and -7. Cotreatment with inhibin prevented this stimulation, whilst inhibin alone completely inhibited MMP production. Treatment with follistatin treatment reduced MMP levels. Similar regulation was seen in stromal cells for MMPs-1, -2 and -9. These data show that activin stimulates the production and activation of MMPs in both endometrial cells, and that inhibin is a potent inhibitor. It is interesting that activin is acting in an opposing manner to TGF-β, indicating that these two closely related proteins have divergent signalling pathways in endometrial cells. Further, this is the first demonstration of a role for inhibin in regulating MMPs and indeed for inhibin action in the endometrium. These findings are of potential importance in understanding regulation of MMPs in the peri-implantation endometrium. Activin is the predominant dimer produced by decidual and epithelial cells, where it may be promoting decidualisation though enhancing MMP production and activation. Furthermore, activin secretion by invasive cytotrophoblasts may stimulate focal decidual MMP production promoting their invasion during embryo implantation.

177 ISOLATION AND CULTURE OF MOUSE ENDOMETRIAL CELLS IN A THREE-DIMENSIONAL CULTURE SYSTEM

2007 ◽  
Vol 19 (1) ◽  
pp. 205
Author(s):  
T.-Y. Fu ◽  
P.-C. Tang ◽  
J.-C. Ju
Keyword(s):  
Epithelial Cells ◽  
Stromal Cells ◽  
Culture System ◽  
Type I Collagen ◽  
Early Embryo ◽  
Type I ◽  
Cytokeratin 18 ◽  
Basal Nucleus ◽  
Endometrial Cells

Implantation of mammalian embryos occurs only during a restricted narrow window. The endometrium becomes highly receptive for the embryos during this period of time. The objective of this study was to establish an in vitro culture system for pre- and post-implantation mouse embryos. In Experiment 1, mouse uterine horns were excised at Day 3.5 post-coitus. After being washed with Dulbecco's phosphate-buffered solution (DPBS), the uterine horns were cut open and incubated with 0.05% trypsin in DPBS at 4&deg;C for 2 h. After trypsinization, the tissues were incubated at 37&deg;C for an additional 30 min. The solution containing epithelial cell suspension was recovered after 30 s of vortexing. The trypsinized uterine horns were then cut into 1-mm3 pieces and digested with collagenase (type I, 1 mg mL-1 in DPBS) at 37&deg;C for 3 h with vigorous shaking. At the end of digestion, the solution was filtered through 40-&micro;m nylon mesh and the flowthrough containing stromal cells was collected. The isolated epithelial and stromal cells were characterized by their morphology and immunocytochemistry. Both types of cells showed positive immunocytochemical reaction with desmin antibody. The cultured epithelial cells formed polyhedral shapes, and more than 95% expressed epithelium-specific protein, cytokeratin-18. On the other hand, most of the spindle-like stromal cells had no signal for cytokeratin-18 expression, although a few scattered cells were positively labeled. In Experiment 2, for construction of the 3-dimensional culture system, epithelial cells obtained by the method described in Experiment 1 were seeded on an artificial basal membrane (ECMatrixTM; Millipore/Upstate/Chemicon, Temecula, CA, USA) with underlying stromal cells embedded in the type I collagen matrix. The whole system was settled in a Millicell&reg; (Millipore) hanging in a 24-well culture plate, and immersed in DMEM medium supplemented with 10% fetal bovine serum, 20 ng mL-1 epidermal growth factor, 63.5 nmol progesterone, and 7.14 nmol 17β-estradiol. The morphology of epithelial cells on the matrix became cuboidal after 48 h of culture. Additionally, the columnar cells with a basal nucleus were observed on the paraffin wax sections. In Experiment 3, mouse E3.5 embryos were recovered and cultured in this established culture system. Normal hatching and/or attachment of the blastocysts were observed after 2 days of culture. In conclusion, our results showed that epithelial cells formed morphologically columnar monolayers and apparently interacted with blastocyst embryos. Successful construction of this model system would facilitate the study of early embryo development through the implantation stage.

scholarly journals Interleukins Affect Equine Endometrial Cell Function: Modulatory Action of Ovarian Steroids

2014 ◽  
Vol 2014 ◽  
pp. 1-11 ◽  
Author(s):  
Anna Z. Szóstek ◽  
Antonio M. Galvão ◽  
Takuo Hojo ◽  
Kiyoshi Okuda ◽  
Dariusz J. Skarzynski
Keyword(s):  
Cell Proliferation ◽  
Early Pregnancy ◽  
Stromal Cells ◽  
Real Time Pcr ◽  
Cell Function ◽  
Positive Control ◽  
Endometrial Cell ◽  
Ovarian Steroids ◽  
Endometrial Cells

The aim of the present study was to investigate the interaction between ovarian steroids, interleukins and prostaglandins (PG) in equine epithelial and stromal cells in vitro. In Experiment 1, cells were exposed to IL-1α(10 ng/mL), IL-1β(10 ng/mL) or IL-6 (10 ng/mL) for 24 h and cell proliferation was determined using MTT. In Experiment 2, cells were exposed to progesterone (P4; 10−7 M); 17-βestradiol (E2; 10−9 M) or P4+E2for 24 h and later medium was replaced with a fresh one treated with IL-1α, IL-1βor IL-6 (10 ng/mL, each) for 24 h. The oxytocin (OT; 10−7 M) was used as a positive control. In Experiment 3, cells were exposed to P4(10−7 M), E2(10−9 M) or P4+E2for 24 h and theIL receptormRNAs transcription was determined using Real-time PCR. Prostaglandins concentration was determined using the direct enzyme immunoassay (EIA) method. Our findings reveal a functional linking between ovarian steroids and IL-stimulated PG secretion by equine endometrial cells. This interaction could be one of the mechanisms responsible for endometrial local orchestrating events during the estrous cycle and early pregnancy.

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