Chemerin Affects P4 and E2 Synthesis in the Porcine Endometrium during Early Pregnancy

Chemerin, belonging to the adipokine family, exhibits pleiotropic activity. We hypothesised that the adipokine could be involved in the regulation of steroidogenesis in the porcine endometrium. Thus, the aim of this study was to determine the effect of chemerin on the key steroidogenic enzyme proteins’ abundance (Western blot), as well as on P4 and E2 secretion (radioimmunoassay) by the porcine endometrium during early pregnancy and the mid-luteal phase of the oestrous cycle. Moreover, we investigated the hormone impact on Erk and Akt signalling pathway activation (Western blot). Chemerin stimulated E2 production on days 10 to 11 of pregnancy. On days 10 to 11 and 15 to 16 of gestation, and on days 10 to 11 of the cycle, chemerin enhanced the expression of StAR and all steroidogenic enzyme proteins. On days 12 to 13 of pregnancy, chemerin decreased StAR and most of the steroidogenic enzyme proteins’ abundance, whereas the P450C17 abundance was increased. On days 27 to 28 of pregnancy, chemerin increased StAR and P450C17 protein contents and decreased 3βHSD protein amounts. It was noted that the adipokine inhibited Erk1/2 and stimulated Akt phosphorylation. The obtained results indicate that chemerin affected P4 and E2 synthesis through the Erk1/2 and Akt signalling pathways.

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110 Temporal Changes in Endometrial Gene Expression Between Ipsi- and Contralateral Uterine Horns in Cattle

Reproduction Fertility and Development ◽

10.1071/rdv30n1ab110 ◽

2018 ◽

Vol 30 (1) ◽

pp. 194

Author(s):

J. M. Sánchez ◽

C. Passaro ◽

N. Forde ◽

S. Behura ◽

J. A. Browne ◽

...

Keyword(s):

Gene Expression ◽

Stem Cells ◽

Luteal Phase ◽

Expression Patterns ◽

Uterine Horn ◽

Oestrous Cycle ◽

Signalling Pathway ◽

Signalling Pathways ◽

Robust Analysis ◽

The Impact

The transfer of an embryo into the uterine horn contralateral to the ovary bearing the corpus luteum has been associated with a decreased pregnancy rate in cattle compared with transfer into the ipsilateral horn. These findings suggest that the environment in the contralateral horn is less conducive to supporting conceptus development than that of the ipsilateral horn. Therefore, this study compared the endometrial transcriptome of the ipsi- and contralateral uterine horns during the luteal phase. Endometrial samples from the ipsi- (IPSI) and contralateral (CONTRA) horns were collected from synchronized nonpregnant beef heifers on Days 5, 7, 13 or 16 post-oestrus (n = 5 heifers per time point). Total RNA was isolated and sequenced. Differences in the transcriptome were determined by edgeR-robust analysis. Principal component analysis found that IPSI and CONTRA have distinct patterns of gene expression on each day, with Day 5 exhibiting the most variation and Day 16 being least variable. Further, the 2 uterine horns had distinct expression patterns on Day 5, with IPSI exhibiting significantly higher variation in gene expression compared twitho CONTRA. EdgeR-robust analysis found 217 (201 up- and 16 down-regulated), 54 (44 up- and 10 down-regulated), 14 (13 up- and 1 down-regulated), and 18 (14 up- and 4 down-regulated) differentially expressed genes (DEG; >2-fold change, false discovery rate P < 0.05) between IPSI and CONTRA endometria on Days 5, 7, 13, and 16 of the oestrous cycle, respectively. The top 5 canonical pathways associated with DEG between IPSI and CONTRA during the luteal phase of the oestrous cycle were involved in signalling pathways regulating pluripotency of stem cells (73/138), progesterone-mediated oocyte maturation (55/89), endometrial cancer (31/51), ErbB signalling pathway (50/87), and mTOR signalling pathway (36/61). The impact of DEG on signalling pathways was assessed using a pathway perturbation algorithm called Signalling Pathway Impact Analysis (SPIA). This topology-based pathway analysis was conducted using the Bioconductor ToPAseq package (https://bioconductor.org/packages/release/bioc/html/ToPASeq.html) and revealed that signalling pathways regulating pluripotency of stem cells showed the highest perturbation score when IPSI was compared with CONTRA irrespective of day. Discovering and cataloguing which pathways are perturbed in each uterine horn throughout the oestrous cycle may contribute to our understanding of the mechanisms underlying early embryonic loss. Ths study was supported by Science Foundation Ireland (13/IA/1983) and the Irish Department of Agriculture, Food and The Marine (13S528).

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Expression of inter-alpha-trypsin inhibitor heavy chains in endometrium of cyclic and pregnant gilts

Reproduction ◽

10.1530/rep.0.1260621 ◽

2003 ◽

pp. 621-627 ◽

Cited By ~ 15

Author(s):

RD Geisert ◽

MD Ashworth ◽

Keyword(s):

Gene Expression ◽

Extracellular Matrix ◽

Western Blot ◽

Protease Inhibitor ◽

Early Pregnancy ◽

Trypsin Inhibitor ◽

Serine Protease Inhibitor ◽

Oestrous Cycle ◽

Heavy Chains ◽

Formation Of Complexes

Attachment of the placenta to the uterus in pigs involves extracellular interaction between the expanding trophoblastic membrane and the thick glycocalyx present on the uterine epithelial microvilli. Formation of complexes between members of inter-alpha-trypsin inhibitor family may function in the maintenance of the extracellular matrix. This study investigated the change in the inter-alpha-trypsin inhibitor heavy chains (ITIH1, ITIH2, ITIH3 and ITIH4) during the oestrous cycle and early pregnancy in pigs. Gene expression of ITIH1, ITIH2, ITIH3 and ITIH4 was detected in the endometrium of cyclic and pregnant gilts; however, gene expression of ITIH was not altered throughout the oestrous cycle or early pregnancy. Western blot analysis with an ITIH antiserum identified the possible linkage forms of ITIH with the serine protease inhibitor, bikunin. Pregnancy altered the release of the various inter-alpha-inhibitor forms from the endometrium during the period of trophoblastic attachment. The results from this study indicate that the inter-alpha-trypsin inhibitor family plays an important role in maintenance of the uterine surface glycocalyx during placental attachment in pigs.

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Early Pregnancy in the Ewe: Effects of Oestradiol and Progesterone on Uterine Metabolism and on Embryo Survival

Australian Journal of Biological Sciences ◽

10.1071/bi9770279 ◽

1977 ◽

Vol 30 (4) ◽

pp. 279 ◽

Cited By ~ 35

Author(s):

BG Miller ◽

NW Moore ◽

Leigh Murphy ◽

GM Stone

Keyword(s):

Embryo Transfer ◽

Embryo Development ◽

Early Pregnancy ◽

Protein Metabolism ◽

Hormonal Regulation ◽

Luteal Phase ◽

Progesterone Receptors ◽

Oestrous Cycle ◽

Embryo Survival ◽

Day Of Embryo Transfer

The hormonal regulation of embryo development during early pregnancy in the ewe has been examined. Ovariectomized ewes received injections of oestradiol (E2) and progesterone (P) according to schedules designed to simulate endogenous ovarian secretion during the luteal phase of the previous oestrous cycle (priming P), around the time of oestrus (oestrous E2 ) and during early pregnancy (maintenance P, maintenance E2)' Embryos were transferred to the ewes on the 4th day after induced oestrus, and ewes were killed at 6 or 13 days after transfer to assess embryo development. Cytosol concentrations of oestradiol 'and progesterone receptors and RNA and protein metabolism in the endometrium and amounts of protein in uterine flushings were examined on the day of embryo transfer and 6 days after transfer.

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Circulating concentrations of oxytocin during pregnancy in ewes

Acta Endocrinologica ◽

10.1530/acta.0.1060277 ◽

1984 ◽

Vol 106 (2) ◽

pp. 277-281 ◽

Cited By ~ 5

Author(s):

D. Schams ◽

A. Lahlou-Kassi

Keyword(s):

Early Pregnancy ◽

Luteal Phase ◽

Oestrous Cycle ◽

Slight Effect ◽

Unknown Mechanism ◽

Maternal Recognition Of Pregnancy ◽

Time Courses

Abstract. Oxytocin and progesterone were measured radioimmunologically during a control cycle, early pregnancy and during several time courses until term in 4 ewes of a local Maroccan breed of sheep. Oxytocin concentrations increased in a similar matter after day 3–4 of the control cycle or after mating and decreased on days 13–15 of early pregnancy about 2 days earlier than during the oestrous cycle. After days 18–19 of pregnancy oxytocin concentrations remained low until term at concentrations as observed during the non-luteal phase of the oestrous cycle. Contrary progesterone concentrations remained high throughout gestation. Maternal recognition of pregnancy seems to have only a slight effect on the secretion pattern of oxytocin for few days after day 12. The results indicate a similar unknown mechanism during the cycle and early pregnancy for the vanishing oxytocin secretion and perhaps synthesis.

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Characterization and proteolytic activity of a cathepsin L-like polypeptide in endometrium and uterine flushings of cycling, pregnant and steroid-treated ovariectomized gilts

Reproduction Fertility and Development ◽

10.1071/r96106 ◽

1997 ◽

Vol 9 (4) ◽

pp. 395 ◽

Cited By ~ 12

Author(s):

Rodney D. Geisert ◽

Robert M. Blair ◽

Thea Pratt ◽

Michael T. Zavy

Keyword(s):

Western Blot ◽

Proteolytic Activity ◽

Early Pregnancy ◽

Cellular Localization ◽

Cathepsin L ◽

Oestrous Cycle ◽

Glandular Epithelium ◽

Specific Substrate ◽

Substrate Metabolism ◽

Uterine Proteins

Cathepsin L has been proposed to be involved with the endothelial–chorial type of placentation in the cat. Little information concerning the presence and secretion of cathepsin L is available for a species with noninvasive epitheliochorial placentation such as the pig. Cathepsin L activity in uterine flushings and endometrium from gilts during different days of the oestrous cycle and early pregnancy was analysed through specific substrate metabolism and Western blot analyses with antiserum against cat endometrial cathepsin L. This antiserum was utilized to determine the cellular localization of the enzyme within porcine endometrium. Cathepsin L activity within uterine flushings was elevated on Day 15 of the oestrous cycle and early pregnancy, with activity declining on Day 18. Cat cathepsin L antiserum cross-reacted with a group of 46, 40 and 38 kDa uterine proteins and detected a product within the surface and glandular epithelium of the endometrium. The appearance of the 40 kDa protein was first detected on Day 10 of the oestrous cycle with the 38 kDa proteins appearing on Day 15 and 18 of pregnancy. The 40 and 38 kDa uterine proteins appear to be steroid regulated as 12 days of progesterone administration is necessary to detect the proteins and cathepsin L activity.

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Expression of chemerin receptors CMKLR1, GPR1 and CCRL2 in the porcine pituitary during the oestrous cycle and early pregnancy and the effect of chemerin on MAPK/Erk1/2, Akt and AMPK signalling pathways

Theriogenology ◽

10.1016/j.theriogenology.2020.07.032 ◽

2020 ◽

Vol 157 ◽

pp. 181-198

Author(s):

Katarzyna Kisielewska ◽

Edyta Rytelewska ◽

Marlena Gudelska ◽

Marta Kiezun ◽

Kamil Dobrzyn ◽

...

Keyword(s):

Early Pregnancy ◽

Oestrous Cycle ◽

Signalling Pathways

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Localization of oestradiol, progesterone and oxytocin receptors in the uterus during the oestrous cycle and early pregnancy of the ewe

Journal of Endocrinology ◽

10.1677/joe.0.1380479 ◽

1993 ◽

Vol 138 (3) ◽

pp. 479-NP ◽

Cited By ~ 90

Author(s):

D. C. Wathes ◽

M. Hamon

Keyword(s):

Progesterone Receptor ◽

Early Pregnancy ◽

Luteal Phase ◽

Specific Binding ◽

Progesterone Receptors ◽

Oestrous Cycle ◽

Luminal Epithelium ◽

Paracrine Factors ◽

Plasma Progesterone ◽

Oxytocin Receptors

ABSTRACT Uterine tissue samples were collected from 47 ewes at various stages of the oestrous cycle and early pregnancy (until day 21) and during seasonal anoestrus. Cryostat sections were immunostained to determine the localization of oestradiol and progesterone receptors using specific monoclonal antibodies. Oxytocin receptors were localized by autoradiography in sections from the same ewes using the 125I-labelled oxytocin antagonist d(CH2)5[Tyr(Me)2,Thr4,Tyr-NH29]- vasotocin. Plasma progesterone measurements were made during the preceding cycle up to the time of slaughter. Oestradiol receptor concentrations were maximal in all regions of the tract at oestrus. Immunostaining of the luminal epithelium, superficial glandular epithelium, stroma and myometrium decreased in the early luteal phase but was maintained for longer in the deep glands. Progesterone receptor immunostaining in the luminal epithelium and superficial glands developed in the early luteal phase (days 1–2) with a somewhat later appearance in the deep glands (days 5–7). Progesterone receptor concentrations in the stroma and myometrium also reached a maximum in the early luteal phase. Myometrial staining was clearly maintained throughout the luteal phase whereas stromal staining was variable between ewes. For both oestradiol and progesterone receptors no differences were apparent between pregnant and non-pregnant ewes between days 2 and 12, but pregnant ewes did not show the general increases in oestradiol receptor staining associated with luteolysis on days 14–15. Oxytocin receptors first developed in the luminal epithelium of non-pregnant ewes on day 14 of the cycle and spread to the superficial glands, caruncular stroma, deep glands and myometrium at oestrus before decreasing in reverse order on days 1–2. Specific binding was not detectable on days 5–12 of the cycle or on days 14 or 21 of pregnancy. The appearance of oxytocin receptors in the luminal epithelium on day 14 preceded that of both the oestradiol and progesterone receptors in the epithelial cells and the fall in plasma progesterone. It was followed by the development of oestradiol and oxytocin receptors in the superficial glands, deep glands, caruncular stroma and myometrium, with the two receptor populations showing a significant positive association in these tissues. The loss of oxytocin receptors in all regions occurred as plasma progesterone levels were increasing, but the association between these two variables was only significant in the superficial glands. The development of progesterone receptors in different tissues could not be explained on the basis of either oestradiol receptor content or plasma progesterone. We conclude that all three receptor populations change in a dynamic manner during the oestrous cycle with variations both between days and between different uterine compartments. The complex pattern of receptor formation and loss suggests that, in addition to the circulating steroid hormone concentrations, local paracrine factors are likely to be involved in their regulation. Journal of Endocrinology (1993) 138, 479–491

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Studies of the oestrous cycle, oestrus and pregnancy in the koala (Phascolarctos cinereus)

Reproduction ◽

10.1530/reprod/120.1.49 ◽

2000 ◽

pp. 49-57 ◽

Cited By ~ 19

Author(s):

SD Johnston ◽

MR McGowan ◽

P O'Callaghan ◽

R Cox ◽

V Nicolson

Keyword(s):

Luteal Phase ◽

Post Partum ◽

Plasma Concentrations ◽

External Genitalia ◽

Oestrous Cycle ◽

Phascolarctos Cinereus ◽

Pregnant Females ◽

Peripheral Plasma ◽

The Mean ◽

High Concentrations

As an integral part of the development of an artificial insemination programme in the captive koala, female reproductive physiology and behaviour were studied. The oestrous cycle in non-mated and mated koalas was characterized by means of behavioural oestrus, morphology of external genitalia and changes in the peripheral plasma concentrations of oestradiol and progestogen. The mean (+/- SEM) duration of the non-mated oestrous cycle and duration of oestrus in 12 koalas was 32.9 +/- 1.1 (n = 22) and 10.3 +/- 0.9 (n = 24) days, respectively. Although the commencement of oestrous behaviour was associated with increasing or high concentrations of oestradiol, there were no consistent changes in the morphology or appearance of the clitoris, pericloacal region, pouch or mammary teats that could be used to characterize the non-mated cycle. As progestogen concentrations remained at basal values throughout the interoestrous period, non-mated cycles were considered non-luteal and presumed anovulatory. After mating of the 12 koalas, six females gave birth with a mean (+/- SEM) gestation of 34.8 +/- 0.3 days, whereas the remaining six non-parturient females returned to oestrus 49.5 +/- 1. 0 days later. After mating, oestrous behaviour ceased and the progestogen profile showed a significant increase in both pregnant and non-parturient females, indicating that a luteal phase had been induced by the physical act of mating. Progestogen concentrations throughout the luteal phase of the pregnant females were significantly higher than those of non-parturient females. Parturition was associated with a decreasing concentration of progestogen, which was increased above that of basal concentrations until 7 days post partum.

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P013 The JAK-3 and TYK-2 / STAT pathways are activated in moderate to severe ulcerative colitis

Journal of Crohn s and Colitis ◽

10.1093/ecco-jcc/jjz203.142 ◽

2020 ◽

Vol 14 (Supplement_1) ◽

pp. S136-S137

Author(s):

M Loza ◽

J M Brea ◽

C Calviño-Suarez ◽

I Baston-Rey ◽

R Ferreiro-Iglesias ◽

...

Keyword(s):

Ulcerative Colitis ◽

Animal Models ◽

Inflammatory Response ◽

Western Blot ◽

Signalling Pathways ◽

Target Engagement ◽

Inclusion Criteria ◽

Phosphorylated Proteins ◽

Single Centre Study ◽

Stat Pathway

Abstract Background Ulcerative colitis (UC) is a chronic, progressive and disabling disease with a complex pathology of unknown aetiology influenced by genetic, environmental and microbiota factors that lead to an immunological and inflammatory response in the colon. Janus Activated Kinase (JAK) family plays a key role in modulating the adaptive and innate inflammatory response. The JAK/STAT pathway involvement in UC has been demonstrated in both animal models and human studies. Thus, overexpressed JAK-3 has been detected in the intestine of patients with UC, suggesting a key role in their pathophysiology and the inhibition of TYK-2 in animal models resulted in an improvement of the disease, which would explain its implication in the inflammatory process. We hypothesise here that there could be an activation of JAK-3 and TYK-2 signalling pathways in UC patients. Thus, we aimed to detect the activation of both signalling pathways by means of western-blot studies in UC patient samples Methods A prospective, observational single-centre study was designed. Inclusion criteria were adult patients with endoscopic active UC (more than Mayo-0) confirmed in a programmed colonoscopy. All patients signed informed consent. Samples were obtained from overstock of routine biopsies in the more severe segment affected of the large bowel. Tissues were homogenised and processed in order to obtain cell lysates by employing RIPA buffer and ultrasounds. The degree of activation of the JAK-3 and TYK-2 pathways was measured by detecting the phosphorylation of both targets as well as of STAT1, STAT3, STAT4, STAT5 and STAT6 through western blot by employing specific antibodies for total and phosphorylated proteins. Results 19 UC patients were consecutively included. Mean age was 46 years old. 53% were female, 47% were extensive colitis (E3) and 53% left-side colitis (E2). Regarding endoscopic activity, 26% had Mayo-1, 53% Mayo-2, and 21% Mayo-3. Immunoreactive bands for both phosphorylated JAK-3 and TYK-2 were detected in the biopsies from UC patients, evidencing that colonic inflammation leads to an activation of both targets. The study of STATs phosphorylation showed immunoreactive bands for phosphorylated forms of STAT1, STAT3, STAT4, STAT5 and STAT6 confirming the activation of both signalling-pathways in these patients (Figure 1). Conclusion The developed translational workflows involving basic/clinical research confirm the activation of both JAK-3 and TYK-2-dependent signalling pathways in UC patients, validating both kinases as targets for treating UC. The developed methodology allows studying the target engagement for future JAK-3/ TYK-2 inhibitors employed in clinical trials.

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