109 EXPRESSION OF GENES RELATED TO ENDOMETRIAL RECEPTIVITY IN EQUINE ENDOMETRIUM DURING THE ESTROUS CYCLE AND EARLY PREGNANCY

2015 ◽  
Vol 27 (1) ◽  
pp. 147 ◽  
Author(s):  
M. Hitit ◽  
A. Guzeloglu ◽  
C. Ozel ◽  
M. O. Atli ◽  
E. Kurar ◽  
...  
Keyword(s):  
Early Pregnancy ◽  
Reproductive Tract ◽  
Cell Signalling ◽  
Endometrial Receptivity ◽  
Oestrous Cycle ◽  
Mrna Levels ◽  
Expression Levels ◽  
Expression Of Genes ◽  
Relative Mrna Expression ◽  
One Way Anova

A set of genes that display differential expression levels in the reproductive tract could serve as beneficial markers of endometrial receptivity. SERPINA14 is present in the uterus during pregnancy and suppresses lymphocyte accumulation. Osteopontin is the ligand of integrin β3 and enables trophoblast communication during implantation. Leukemia inhibitory factor (LIF) is involved in inflammatory cell signalling and contributes to implantation by regulating immune cells. The objective was to assess the expression of SERPINA14, osteopontin, and LIF mRNAs in the equine endometrium during the oestrous cycle and early pregnancy. Biopsies were obtained from mares on day of ovulation (d 0, n = 4), late diestrus (LD, n = 4, high progesterone [P4]), and after luteolysis at the beginning of oestrus phase (AL, n = 4, <1 ng mL–1 P4) of the cycle. Biopsies were also taken on days 14 (P14, n = 4), 18 (P18, n = 4), and 22 (P22; n = 4) of pregnancy. Relative mRNA expression levels of genes were quantified using real-time quantitative RT-qPCR in duplicate. Data were analysed using one-way ANOVA, and l.s.d. test was applied. Both the oestrous cycle and early pregnancy increased SERPINA14 mRNA levels compared to d0. Expression of LIF mRNA was not significantly regulated except for a decline at AL. Expression of osteopontin mRNA was up-regulated during the oestrous cycle at LD while early pregnancy inhibited this up-regulation. The results suggest that the genes studied related to endometrial receptivity are strictly regulated accordingly to the stage of oestrous cycle, probably by circulating ovarian steroids, specifically progesterone, and pregnancy-associated factors are also involved in this regulation. This project was partially funded by TUBITAK 107O035 to AG and DUBAP 14VF12 to MOA. MH was supported by OYP 2013-090.

Sulfated glycoprotein-1 (SGP-1) expression in ovine endometrium during the oestrous cycle and early pregnancy

1995 ◽  
Vol 7 (5) ◽  
pp. 1053 ◽  
Author(s):  
TE Spencer ◽  
GH Graf ◽  
FW Bazer
Keyword(s):  
Early Pregnancy ◽  
Blot Hybridization ◽  
Immunohistochemical Localization ◽  
Oestrous Cycle ◽  
Northern Hybridization ◽  
Mrna Levels ◽  
Slot Blot ◽  
Slot Blot Hybridization ◽  
Endometrial Epithelium ◽  
Sulfated Glycoprotein

This study determined effects of day of oestrous cycle and early pregnancy on sulfated glycoprotein-1 (SGP-1) expression in ovine endometrium. A 364-bp clone of the ovine SGP-1 mRNA was amplified from reverse transcribed Day-15 cyclic endometrial mRNA using the polymerase chain reaction (PCR) and primers specific for the rat SGP-1 mRNA sequence. Nucleotide sequence of the ovine SGP-1 cDNA shared significant identity with rat SGP-1 and human prosaposin. Ewes (n = 40) were hysterectomized on either Day 1, 6, 11, 13 or 15 of the oestrous cycle or on Day 11, 13, 15, 17 or 25 of early pregnancy. Total cellular RNA was isolated from endometrium and subjected to Northern and slot blot hybridization analyses using an antisense cRNA probe transcribed from the ovine SGP-1 cDNA clone. A single 2.6-kb mRNA transcript was detected by Northern hybridization analyses. Slot blot hybridization analyses indicated that steady-state levels of endometrial SGP-1 mRNA varied during the oestrous cycle (cubic, P < 0.02) and increased between Day 11 and Day 25 of early pregnancy (linear, P < 0.01). On Days 11, 13 and 15, endometrial SGP-1 mRNA levels were greater in pregnant ewes than in cyclic ewes (day x pregnancy status, P < 0.01). Immunohistochemical localization of SGP-1 in uterine tissues with rabbit anti-rat SGP-1 antibody revealed intense immunoreactivity associated primarily with the endometrial epithelium. These results indicate that the ovine endometrium expresses SGP-1, a prosaposin, and that SGP-1 expression varies during the oestrous cycle and is enhanced by the conceptus. The presence of SGP-1 in the endometrium suggests intracellular and extracellular roles for this protein in glycosphingolipid metabolism or transport in the uterine environment.

scholarly journals The expression of the IGF system in the bovine uterus throughout the oestrous cycle and early pregnancy

2000 ◽  
Vol 165 (2) ◽  
pp. 231-243 ◽  
Author(s):  
RS Robinson ◽  
GE Mann ◽  
TS Gadd ◽  
GE Lamming ◽  
DC Wathes
Keyword(s):  
Cross Sections ◽  
Early Pregnancy ◽  
In Situ Hybridisation ◽  
Endometrial Biopsy ◽  
Oestrous Cycle ◽  
Luminal Epithelium ◽  
Mrna Levels ◽  
Igf Binding Proteins ◽  
Igf System ◽  
Igf I

The IGF system is expressed in the uterus during the oestrous cycle and early pregnancy and is likely to play an important role in regulating the development of the embryo and uterus. The IGF peptides (IGF-I and -II) mediate their effects through the type 1 IGF receptor (IGF-1R), while the IGF-binding proteins (IGFBP-1 to -6) modulate their interaction with the receptor. In this study, the expression of the IGF system in the bovine uterus was determined throughout the oestrous cycle and on day 16 of pregnancy. Endometrial biopsy samples were collected from four cows over three cycles such that there were samples for every 2 days from day 0 (oestrus) to day 14 and then every day until day 21. To assess the effect of pregnancy, uterine horn cross-sections were collected on day 16 from 15 pregnant (PREG), five inseminated non-pregnant (INP) and nine uninseminated cyclic controls (CONT). The expression of mRNA for the IGFs, IGF-1R and IGFBP-1 to -5 was determined by in situ hybridisation and the results were quantified by measuring the optical density units from autoradiographs. The main region of IGF-I mRNA expression was the sub-epithelial stroma underlying the luminal epithelium. The expression of IGF-I mRNA was highest at oestrus and lowest during the early and late luteal phases. On day 16, IGF-I mRNA levels were low in all groups, with pregnancy having no effect on the IGF-I mRNA concentrations. The strongest expression of IGF-II mRNA was in the caruncular stroma, with pregnancy having no significant effect in this region. IGF-1R mRNA was also present in the caruncles and was strongly expressed in all epithelial cells both throughout the oestrous cycle and during early pregnancy. The expression of IGFBP-1 mRNA was confined to the luminal epithelium, with the strongest expression seen on day 14 of the cycle. On day 16 the expression of IGFBP-1 mRNA was higher in the PREG group compared with the CONT group. The expression of IGFBP-2 mRNA was localised to the sub-epithelial stroma with more INP than PREG cows showing detectable levels of IGFBP-2. The strongest expression of IGFBP-3 mRNA was in the caruncular stroma; expression in the endometrial stroma was similarly decreased during early pregnancy. IGFBP-5 mRNA was mainly expressed in the inner ring of myometrium and was not affected by pregnancy on day 16. In conclusion, these results show that many components of the uterine IGF system are differentially regulated during the oestrous cycle and early pregnancy and suggest that modulation of the IGF system may influence uterine activity during this period.

scholarly journals Differences in expression of genes related to steroidgenesis in abdominal subcutaneous adipose tissue of pregnant women with and without PCOS; a case control study

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Neda Emami ◽  
Ashraf Moini ◽  
Parichehreh Yaghmaei ◽  
Vahid Akbarinejad ◽  
Maryam Shahhoseini ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Cytochrome P450 ◽  
Pregnant Women ◽  
Polycystic Ovary ◽  
Hydroxysteroid Dehydrogenase ◽  
Mrna Levels ◽  
Gene Expressions ◽  
P450 Monooxygenase ◽  
Expression Levels ◽  
Expression Of Genes

Abstract Background It was reported that steroid-related gene expressions in the adipose tissue (AT) of women differ between women affected with polycystic ovary syndrome (PCOS) and non-PCOS. Although association between PCOS in mother and offspring’s health is a crucial issue, there are few studies focusing on AT of pregnant women suffering from PCOS. Our objectives were to determine the differences between mRNA expression levels of key steroid-converting enzymes in abdominal subcutaneous AT of pregnant women afflicted with PCOS and non-PCOS. Methods Twelve pregnant women with PCOS (case) and thirty six non-PCOS pregnant women (control) (1:3 ratio; age- and BMI-matched) undergoing cesarean section were enrolled for the present study. Expressions of fifteen genes related to steriodogenesis in abdominal subcutaneous AT were investigated using quantitative real-time PCR. Results No significant differences were detected with respect to age, BMI (prior pregnancy and at delivery day), gestational period and parity among pregnant women with PCOS and non-PCOS. Most of the sex steroid-converting genes except 17β-Hydroxysteroid dehydrogenases2 (17BHSD2), were highly expressed on the day of delivery in subcutaneous AT. Women with PCOS showed significantly higher mRNA levels of steroidgenic acute regulator (STAR; P < 0.001), cytochrome P450 monooxygenase (CYP11A1; P < 0.05), 17α-hydroxylase (CYP17A1; P < 0.05), and 11β-Hydroxysteroid dehydrogenase (11BHSD1 and 11BHSD2; P < 0.05). The expression of steroid 21-hydroxylase (CYP21) in non-PCOS was fourfold higher than those of women with PCOS (P < 0.001). There were no significant differences between relative expression of aromatase cytochrome P450 (CYP19A1), 3β-hydroxysteroid dehydrogenase (3BHSD1 and 3BHSD2), and 17BHSD family (1, 3, 5, 7, and 12) between the two groups. Conclusion The expression levels of genes related to sex steroids metabolism were similar to age-matched and BMI- matched pregnant non-PCOS and pregnant women with PCOS at delivery day. However, the alterations in gene expressions involved in glucocorticoids and mineralocorticoids metabolism were shown. It is necessary to point out that further studies regarding functional activity are required. More attention should be given to AT of pregnant women with PCOS that was previously ignored.

scholarly journals Differences in Expression of Genes Related to Steroidgenesis in Abdominal Subcutaneous Adipose Tissue of Pregnant Women with and without PCOS

2020 ◽  
Author(s):  
Neda Emami ◽  
Ashraf Moini ◽  
Parichehreh Yaghmaei ◽  
Vahid Akbarinejad ◽  
Maryam Shahhoseini ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Cytochrome P450 ◽  
Pregnant Women ◽  
Polycystic Ovary ◽  
Mrna Levels ◽  
Gene Expressions ◽  
P450 Monooxygenase ◽  
Expression Levels ◽  
Expression Of Genes ◽  
Subcutaneous Adipose

Abstract Background: It was hypothesized that steroid-related gene expressions in the adipose tissue (AT) of women are different between women with and without polycystic ovary syndrome (PCOS). However, little information exists regarding pregnant women suffering from PCOS when interrelationships among maternal PCOS, patient’s quality of life and offspring’s health is a consideration issue in health and diseases. Our objectives were to compare the mRNA expression levels of key steroid-converting enzymes in subcutaneous AT of PCOS vs. non-PCOS pregnant women.Methods: The subjects were 48 pregnant women (12 PCOS and 36 non–PCOS women) undergoing cesarean section, from whom abdominal subcutaneous AT samples were retrieved. Expressions of fifteen genes related to steriodogenesis were investigated using quantitative real-time PCR. Results: No significant differences were found with respect to age, BMI (before pregnancy and at delivery day), gestational period and parity among non-PCOS and PCOS pregnant women. Most of the sex steroid-converting genes except 17β-Hydroxysteroid dehydrogenases2 (17BHSD2), were highly expressed on the day of delivery in subcutaneous AT. PCOS patients showed significantly higher mRNA levels of steroidgenic acute regulator (STAR; P<0.001), cytochrome P450 monooxygenase (CYP11A1; P<0.05), 17α-hydroxylase (CYP17A1; P<0.05), and 11β-Hydroxysteroid dehydrogenase (11BHSD1-2; P<0.05). The expression of steroid 21-hydroxylase (CYP21) in non-PCOS women was 4-fold higher than for PCOS women (P < 0.001). There were no significant differences between relative expression of aromatase cytochrome P450 (CYP19A1), 3β-hydroxysteroid dehydrogenase1-2 (3BHSD1-2), and 17BHSD family (1, 3, 5, 7, and 12) between the two groups.Conclusion: The expression levels of genes related to sex steroids metabolism were similar in age-matched and BMI- matched non-PCOS and PCOS pregnant women at delivery day. However, the alterations in gene expressions involved in glucocorticoids and mineralocorticoid metabolism was shown which warrants further studies regards functional activity. More attention should be given to AT of PCOS mothers that was previously ignored.

Expression of leukaemia inhibitory factor at the conceptus–maternal interface during preimplantation development and in the endometrium during the oestrous cycle in the mare

2016 ◽  
Vol 28 (10) ◽  
pp. 1642 ◽  
Author(s):  
M. de Ruijter-Villani ◽  
C. Deelen ◽  
T. A. E. Stout
Keyword(s):  
Early Pregnancy ◽  
Critical Role ◽  
Endometrial Receptivity ◽  
Oestrous Cycle ◽  
Inhibitory Factor ◽  
Leukaemia Inhibitory Factor ◽  
Luminal Epithelium ◽  
Signal Transducer ◽  
Blastocyst Development ◽  
Mrna And Protein Expression

Leukaemia inhibitory factor (LIF) plays a critical role in blastocyst development and implantation in several species. The present study investigated mRNA and protein expression for LIF, as well as the low-affinity LIF receptor (LIFR) and interleukin-6 signal transducer (IL6ST), in equine endometrium, trophoblast and histotroph during early pregnancy and in the endometrium during the oestrous cycle. Endometrial LIF mRNA expression was upregulated after Day 21 of pregnancy, whereas LIF immunoreactivity increased in the endometrium on Day 28. Expression of LIF mRNA in the yolk sac membrane increased from Day 21 of pregnancy, whereas LIF immunoreactivity increased from Day 28 in the trophoblast. LIFR and IL6ST mRNA was expressed in the endometrium during both the oestrous cycle and early pregnancy and, although LIFR and IL6ST protein were localised to the glandular epithelium during the cycle and first 14 days of pregnancy, from Day 21 they were located in the luminal epithelium. Trophoblast expression of LIFR and IL6ST increased as pregnancy proceeded. In conclusion, LIF expression increased at the conceptus–maternal interface during capsule attenuation. Because contemporaneous upregulation of both LIFR and IL6ST was also observed in the trophoblast, we propose that LIF plays an important role in the development of endometrial receptivity for trophoblast growth, apposition and adhesion in mares.

scholarly journals Temporal and spatial associations of oestrogen receptor alpha and progesterone receptor in the endometrium of cyclic and early pregnant mares

Reproduction ◽  
2005 ◽  
Vol 130 (2) ◽  
pp. 241-250 ◽  
Author(s):  
L S Hartt ◽  
S J Carling ◽  
M M Joyce ◽  
G A Johnson ◽  
D K Vanderwall ◽  
...  
Keyword(s):  
Steady State ◽  
Early Pregnancy ◽  
Receptor Gene ◽  
Cell Types ◽  
Oestrous Cycle ◽  
Mrna Levels ◽  
Steady State Levels ◽  
Low Levels ◽  
Temporal And Spatial

Uterine function is primarily controlled by the combined actions of oestrogen and progesterone working through their cognate nuclear receptors. The mechanism of establishment of pregnancy in the mare is of interest because it involves prolonged pre-attachment and conceptus migration phases, and both invasive and non-invasive placental cell types, and as such has been an important comparative model. This study characterised regulation of oestrogen (ER) and progesterone (PR) receptors in the endometrium of the mare during the oestrous cycle and early pregnancy. Endometrial tissues collected during the oestrous cycle and early pregnancy were analysed for steady-state levels of ER and PR mRNA and protein. Steady-state levels of ER and PR mRNA were highest on days 0, 17 and 20 in cyclic mares and lowest on days 11 and 14. A day-by-status interaction was detected, indicating that day 17 and day 20 pregnant mares exhibited low levels of ER and PR compared with the corresponding days of the oestrous cycle. In situ hybridisation analyses showed receptor mRNA localisation primarily in the luminal epithelium (LE), glandular epithelium (GE) and stroma around oestrus. During dioestrus and early pregnancy, receptors were not detected in the LE, and were lower in the stroma and deeper GE. Changes in hybridisation intensity in these cell types were consistent with changes in mRNA levels detected by slot-blot hybridisation. ER and PR proteins were detected in the nuclei of LE, GE and stromal cells. Consistent with results from in situ hybridisation, levels of ER and PR immunoreactivity were higher around oestrus, declined to low levels during dioestrus and remained low during early pregnancy. Results described here for temporal and spatial changes in steroid receptor gene expression in mares show the greatest similarities with those described for cattle and sheep.

121 RELATIONSHIP BETWEEN PROSTAGLANDIN E2 METABOLITE, UTERINE ARTERIAL VASODILATORY CHANGES, AND PROGESTERONE DURING THE OESTROUS CYCLE AND EARLY PREGNANCY IN BEEF COWS

2013 ◽  
Vol 25 (1) ◽  
pp. 207
Author(s):  
H. L. Sánchez-Rodríguez ◽  
J. M. Feugang ◽  
R. C. Youngblood ◽  
R. C. Vann ◽  
S. T. Willard ◽  
...  
Keyword(s):  
Prostaglandin E2 ◽  
Early Pregnancy ◽  
Vascular Function ◽  
Reproductive Tract ◽  
Sharp Decrease ◽  
Female Reproductive Tract ◽  
Oestrous Cycle ◽  
Potential Candidate ◽  
Vascular Effects ◽  
Uterine Arteries

Vasodilation of the female reproductive vasculature is essential to supply the increasing demands associated with reproductive processes; including maintenance of a healthy functional luteal structure, and the establishment and continuance of pregnancy. Due to its vasodilatory effects, prostaglandin E2 has been identified as a potential candidate sustaining such vascular effects. Here, the relationship between systemic concentrations of prostaglandin E2 metabolite (PGEM) and progesterone (P4), and uterine arterial vasodilatory changes were evaluated during the oestrous cycle and early pregnancy. Synchronised Angus cows (n = 19) were artificially inseminated (AI) and examined for pregnancy 45 d post-AI. Jugular blood samples were collected on Days 0, 3, 6, 10, 16, 20, 25, and 32 (Day 0 = AI) for serum PGEM and P4 evaluation. Vasodilatory changes of the uterine arteries were also characterised using the B-mode ultrasound luminal circumference (UAC). Retrospective comparisons were carried out between pregnant (n = 10) and nonpregnant (n = 9) animals. The GLM Procedure of SAS (SAS Institute Inc., Cary, NC, USA) was used to evaluate the effects of pregnancy status and sampling day over PGEM, UAC, and P4. Pearson correlations between PGEM, UAC, and P4 were obtained by the CORR Procedure of SAS. All variables were measured in triplicate and thresholds for significance and tendency were fixed at P ≤ 0.05 and 0.06 to 0.10, respectively. From Day 0 to Day 20, the overall PGEM concentration tended to be higher in nonpregnant cows compared to pregnant ones (P = 0.08; 90.97 ± 4.38 v. 78.28 ± 3.01 pg mL–1, respectively), while a shift in favor of pregnant cows was observed at Day 32 (P = 0.29; 89.88 ± 14.23 v. 106.3 ± 7.17 pg mL–1, respectively). Both uterine arteries showed similar UAC (P ≥ 0.05); therefore, they were combined in one average value per animal each sampling day. There was an overall increase of the UAC over time, from 10.49 ± 0.01 mm at Day 0 to 12.17 ± 0.02 mm at Day 20 (P ≤ 0.0001), but no significant differences were found between pregnant and nonpregnant cows during this period (P ≥ 0.05). An overall sharp increase in P4 concentrations was observed from Day 3 to Day 16 (0.60 ± 0.12 to 9.66 ± 0.45 ng mL–1, P ≤ 0.0001); while from Day 16 to Day 32, pregnant cows maintained steady and higher P4 concentrations than nonpregnant ones (9.87 ± 0.31 v. 4.40 ± 1.09 ng mL–1, respectively; P = 0.002). As expected, nonpregnant cows showed a sharp decrease in P4 concentrations from Day 16 to Day 25, followed by a second increase at Day 32 (9.27 ± 0.20, 0.85 ± 0.33, and 6.04 ± 1.67 ng mL–1, respectively; P = 0.0007). The UAC and PGEM concentrations were positively correlated in pregnant (r = 0.48; P = 0.002), but not in nonpregnant animals (r = 0.29; P = 0.21). Similarly, UAC and P4 concentrations were also positively associated in pregnant (r = 0.47; P = 0.002), but not in nonpregnant cows (r = 0.01; P = 0.96). Our findings support the role of PGE2 as a potential mediator of the vascular function in the female reproductive tract and, subsequently, as an essential factor to ensure adequate P4 production able to sustain early pregnancy in the bovine. USDA-ARS Grant#58-6402-3-0120.

121 EXPRESSION OF HYALURONAN SYSTEM COMPONENTS IN THE EARLY PREGNANT OVINE ENDOMETRIUM

2015 ◽  
Vol 27 (1) ◽  
pp. 152
Author(s):  
Z. Kiyma ◽  
M. Hitit ◽  
C. Ozel ◽  
G. Sen ◽  
M. Kose ◽  
...  
Keyword(s):  
Hyaluronic Acid ◽  
Reproductive Tract ◽  
Mrna Level ◽  
Cell Signalling ◽  
Molecular Size ◽  
Cell Surface Receptor ◽  
Mrna Levels ◽  
Tissue Samples ◽  
Surface Receptor ◽  
Almost All

Hyaluronan (HA; hyaluronic acid), a member of glycosaminoglycans (GAG) family, is the main polysaccharide and is expressed in almost all tissues including those of the reproductive tract. Three different hyaluronic acid-synthase (HAS) enzymes, HAS1, HAS2, and HAS3, synthesise HA. The action of HA depends on its molecular size and its cell surface receptor (CD44). Hyaluronidases (HYAL) are a group of enzymes, HYAL-1 and HYAL-2, that degrade HA. Roles of the HA system in reproductive events include oocyte maturation, fertilization, and implantation. It is also known that ovarian steroids, specifically progesterone, regulate the HA system in the endometrium. Moreover, HA-mediated cell signalling participates in embryonic development. The aim of this study was to evaluate expression of the HA system at the mRNA level in the early pregnant ovine endometrium at pre-implantation stage. Therefore, endometrial tissue samples were collected on Day 13 of the oestrous cycle (n = 10) and pregnancy (n = 14). Relative mRNA expression levels of HA system genes were quantified using quantitative RT-qPCR. Data were analysed using one-way ANOVA and l.s.d. test. All of the studied components of the HA system (HAS1, HAS2, HAS3, HYAL1, HYAL2, and CD44) were expressed in the ovine endometrium. Steady-state mRNA levels of HAS1, HYAL1, HYAL2, and CD44 were not significantly different between cyclic and early pregnant ovine endometrium on Day 13. However, expression of HAS2 and HAS3 appeared to be down-regulated in early pregnancy. Considering that both cyclic and pregnant endometrium on Day 13 is under the influence of progesterone, detected differential regulation of some components of HA system in the ovine endometrium may be directly related to the effects of the presence of an embryo. The role of the HA system in ovine endometrium at later stages of pregnancy warrants further investigation. Partially funded by TUBITAK-112R022 to ZK and SUBAP to AG.

111 EXPRESSION OF PHOSPHOLIPASE A2 ISOFORMS IN EQUINE ENDOMETRIUM DURING THE ESTROUS CYCLE AND EARLY PREGNANCY

2015 ◽  
Vol 27 (1) ◽  
pp. 148 ◽  
Author(s):  
C. Ozel ◽  
A. Guzeloglu ◽  
M. Hitit ◽  
M. O. Atli ◽  
E. Kurar ◽  
...  
Keyword(s):  
Arachidonic Acid ◽  
Phospholipase A2 ◽  
Early Pregnancy ◽  
Membrane Phospholipids ◽  
Steady State Levels ◽  
In The Beginning ◽  
Relative Mrna Expression ◽  
One Way Anova ◽  
Mrna Expression Levels ◽  
In Pregnancy

Phospholipase A2 (PLA2) is involved in the synthesis of prostaglandins (PG) as it releases arachidonic acid from the membrane phospholipids to be a precursor for cyclooxygenase enzymes. Therefore, it is critically important during luteolysis and at the time of maternal recognition of pregnancy. The embryo must attenuate endometrial PGF2α production, whereas PGE2 is considered to be luteoprotective. Furthermore, implantation also requires the action of PGs. A balance should be maintained in PG synthesis in the endometrium. The objective of this study was to evaluate the expression of isoforms of PLA2 (cytosolic [cPLA2], secretory [PLA2], and calcium independent [iPLA2]) in equine endometrium during the oestrous cycle and early pregnancy. Biopsies were obtained from mares on the day of ovulation (d0, n = 4), late diestrus (LD, n = 4, high progesterone [P4]), and after luteolysis in the beginning of oestrus phase (AL, n = 4, <1 ng mL–1 P4) of the cycle. Biopsies were also taken on days 14 (P14, n = 4), 18 (P18, n = 4), and 22 (P22; n = 4) of pregnancy. Relative mRNA expression levels of genes were quantified using real-time quantitative RT-qPCR. Data were analysed using one-way ANOVA and l.s.d. test. Compared with the day of oestrus (d0), steady-state levels of cPLA2 and sPLA2 were down-regulated at LD, where P4 was high, and expression of both were up-regulated again at AL. In contrast, iPLA2 expression was higher at LD and then decreased again at AL. Pregnancy decreased expression of cPLA2 and sPLA2 mRNA at P14 and P18 compared with their respective cycle days. Late diestrus elevation in the expression of iPLA2 was suppressed by pregnancy at P14; however, it was up-regulated later in pregnancy at P22. The results suggest that expression of cPLA2 and sPLA2 is negatively correlated with circulating progesterone concentrations. Pregnancy further inhibits their expression in the equine endometrium. However, iPLA2 expression seems to be positively correlated by progesterone presence and its expression increased as equine pregnancy advanced. This project was partially funded by TUBITAK 107O035 and SUBAP to AG.

scholarly journals Characterization of local and peripheral immune system in pregnant and non-pregnant ewes

2021 ◽  
Author(s):  
Laurel D Quirke ◽  
Paul H Maclean ◽  
Neville A Haack ◽  
Sara J Edwards ◽  
Axel Heiser ◽  
...  
Keyword(s):  
Gene Expression ◽  
Lymph Node ◽  
Immune System ◽  
T Cell ◽  
Lymph Nodes ◽  
Early Pregnancy ◽  
Reproductive Tract ◽  
Reproductive Tissue ◽  
Expression Of Genes ◽  
Peripheral Lymph

Abstract Modulation of the immune system is known to be important for successful pregnancy but how immune function might differ between the lymph nodes draining the reproductive tract and peripheral lymph nodes is not well understood. Additionally, if immune system changes in response to the presence of an embryo during early pregnancy, and if this response differs in local versus peripheral immune tissue, has not been well characterized. To address these questions, we examined expression of genes important for immune function using NanoString technology in the ampulla and isthmus of the oviduct, endometrium, lymph nodes draining the reproductive tract (lumbo-aortic and medial iliac) as well as a peripheral lymph node (axillary), the spleen and circulating immune cells from ewes on day 5 of the estrous cycle or pregnancy. Concentrations of estradiol and progesterone in plasma were also determined. Principal component analysis revealed separation of the local from the peripheral lymph nodes (MANOVA P = 3.245e-08, R 2 = 0.3) as well as separation of tissues from pregnant and non-pregnant animals [lymph nodes (MANOVA P = 2.337e-09, R 2 = 0.5), reproductive tissues (MANOVA P = 2.417e-14, R 2 = 0.47)]. Nine genes were differentially (FDR &lt;0.10) expressed between lymph node types, with clear difference in expression of these genes between the lumbo-aortic and axillary lymph nodes. Expression of these genes in the medial iliac lymph node was not consistently different to either the axillary or the lumbo-aortic lymph node. Expression of IL10RB was increased (P &lt; 0.05) by 24% in the reproductive tissue of the pregnant animals comparing to non-pregnant animals. Analysis of gene categories revealed that expression of genes of the T cell receptor pathway in reproductive tract tissues was associated (P &lt; 0.05) with pregnancy status. In conclusion, assessment of gene expression of reproductive and immune tissue provides evidence for a specialization of the local immune system around the reproductive tract potentially important for successful establishment of pregnancy. Additionally, differences in gene expression patterns in reproductive tissue from pregnant and non-pregnant animals could be discerned as early as day 5 of pregnancy. This was found to be associated with expression of genes important for T-cell function and thus highlights the important role of these cells in early pregnancy.

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