Transcriptomic profile of the porcine oviductal epithelial cells during the peri-conceptional period.

The optimal environment in the oviduct is created by adjusting its ultrastructure and secretory activity to serve the most suitable protection of gametes and to support embryo development. Through gametes/embryo's presence inside the oviduct, the oviductal transcriptomic profile may be altered, and these changes may be caused by DNA methylation. The results of the present study documented that in the epithelial cells of the ampulla and isthmus of the oviducts collected from pigs during the peri-conceptional period, the most differentially expressed genes (DEGs) were down-regulated.

Peri-conceptional nutritional restriction alters transcriptomic profile in the peri-implantation pig embryos.

Restricted nutritional consumption during the peri-conceptional period affects the potential for DNA methylation and alters endometrial transcriptomic profile during the peri-implantation period. The restricted diet fed to females during the peri-conceptional period may affect the transcriptomic profile in peri-implantation embryos. In the present study, the transcriptome of embryos of normal-diet-fed gilts was determined and compared with that in embryos of restricted-diet-fed gilts during the peri-implantation period.

Transcriptomic analysis of the oviduct of pigs during the peri-conceptional period.

The optimal environment in the oviduct is created by adjusting its ultrastructure and secretory capacity to protect gametes and embryos. It was hypothesized that direct contact between the isthmic epithelium and 2- and 4-cell-stage embryos would alter the transcriptomic profile of the isthmus in pigs. Microarray analysis was performed to determine the alterations in gene expression of the isthmus on Days 2-3 of pregnancy in pigs (after natural mating) during embryo presence in the oviduct.

Synthesis of steroid hormones in the porcine oviduct during early pregnancy.

Past studies of the oviducts have documented oviductal steroid production during the oestrous cycle in pigs. The present study examined whether the pig oviducts are the source of steroid hormones during early pregnancy. In the ampulla and isthmus, the expression of 3β-hydroxysteroid dehydrogenase (3βHSD) and aromatase cytochrome P450 (CYP19) mRNA by real-time PCR, cellular localization and quantities of the studied proteins by immunofluorescence and Western blot analysis, and concentration of steroid hormones in oviductal flushings by radioimmunoassay, were studied.

Peri-conceptional under-nutrition alters transcriptomic profile in the endometrium during the peri-implantation period-The study in domestic pigs.

Female under-nutrition during early pregnancy may affect the physiological pattern of the transcriptomic profile in the endometrium. We aimed to determine if restricted diet applied to females during peri-conceptional period, that is, from the onset of the oestrus until day nine of pregnancy, alters transcriptomic profile in the endometrium during the peri-implantation period. The restricted diet gilts were fed forage, in which the dose of proteins and energy had been reduced by 30% compared to normal diet.

Interleukin-1β system in the oviducts of pigs during the oestrous cycle and early pregnancy.

Interleukin-1β (IL-1β) acts throughout the IL-1β system, which contains IL-1β and the IL-1β receptor (IL-1R), accessory protein (IL-1RacP), and receptor antagonist (IL-1Ra). In pigs, the expression of the members of the IL-1β system was documented in uterine tissues during the oestrous cycle and early pregnancy, as well as in embryos harvested during the peri-implantation period. In the oviducts of non-gravid and gravid pigs, the expression of the IL-1β system is unknown.

Expression of 17β-hydroxysteroid dehydrogenase and the effects of LH, FSH and prolactin on oestrone and 17β-oestradiol secretion in the endometrium of pigs during early pregnancy and the oestrous cycle.

The endometrium of pregnant and cyclic pigs is a source of oestrone (E1) and 17β-oestradiol (E2). However, the roles of LH, FSH and prolactin (PRL) as regulators of endometrial steroidogenesis, and the presence of 17β-hydroxysteroid dehydrogenase (17β-HSD) in the porcine endometrium, remain unknown. Therefore, in the present study we examined 17β-HSD expression and the effects of LH, FSH and PRL on E1 and E2 release in vitro in endometrial explants harvested from gravid pigs on Days 10-11 (embryo migration within the uterus), 12-13 (maternal recognition of pregnancy) and 15-16 (beginning of implantation) and compared them with results obtained in non-gravid pigs.

Periconceptional undernutrition affects in utero methyltransferase expression and steroid hormone concentrations in uterine flushings and blood plasma during the peri-implantation period in domestic pigs.

Female undernutrition during early pregnancy may affect the physiological pattern of genomic DNA methylation. We hypothesised that in utero DNA methylation may be impaired in females fed a restrictive diet in early pregnancy. In this study we evaluated whether poor maternal nutritional status, induced by applying a restricted diet during the peri-conceptional period, may influence: (1) the potential for in utero DNA methylation, expressed as changes in the mRNA expression and protein abundance of methyltransferases: DNA methyltransferase 1 (DNMT1) and DNMT3a in the endometrium and the myometrium, (2) the intrauterine microenvironment, measured as oestradiol 17β (E) and progesterone (P) concentrations in uterine flushings and (3) plasma concentration of E and P during the peri-implantation period.

Effect of neonatal or adult heat acclimation on plasma fT3 level, testicular thyroid receptors expression in male rats and testicular steroidogenesis in vitro in response to triiodothyronine treatment.

This study aimed to evaluate the effect of heat acclimation of neonatal and adult rats on their testes response to in vitro treatment with triiodothyronine (T3). Four groups of rats were housed from birth as: 1) control (CR) at 20°C for 90 days, 2) neonatal heat-acclimated (NHA) at 34°C for 90 days, 3) adult heat-acclimated (AHA) at 20°C for 45 days followed by 45 days at 34°C and 4) de-acclimated (DA) at 34°C for 45 days followed by 45 days at 20°C. Blood plasma and both testes were harvested from 90-day old rats.

Distinct testicular steroidogenic response mechanisms between neonatal and adult heat-acclimated male rats.

Background: In comparison to short-term gonad heat exposure, little is known about the molecular mechanisms that regulate testicular steroidogenesis during long-term whole body heat acclimation.

Material And Methods: Testicular slices from neonatal (NHA) and adult (AHA) heat-acclimated Wistar rats were analysed in vitro to assess the mRNA expression and enzymatic activity of steroidogenic enzymes under basal and luteinising hormone (LH) or prolactin (PRL) stimulated conditions compared with control rats (CR). Furthermore, a de-acclimated group (DA) was created by transferring adult NHA rats to control conditions.

Effect of neonatal or adult heat acclimation on testicular and epididymal morphometry and sperm production in rats.

The accessory gland weight, testicular and epididymal morphometry and sperm production were analyzed in four groups of rats housed at 20 or 34°C: (1) control rats (CR) kept at 20°C from birth to day 90; (2) adult heat-acclimated rats (AHA) kept at 20°C from birth to day 45 followed by 34°C to day 90; (3) neonatal heat-acclimated rats (NHA) kept at 34°C from birth to day 90 and (4) de-acclimated rats (DA) kept at 34°C from birth to day 45 followed by 20°C to day 90. In NHA and DA rats, accessory gland weight was higher than in controls. Despite the lack of differences in testicular and epididymal morphometry, curvilinear velocity of spermatozoa was lower in the NHA group compared to controls.

Transcriptomic analysis of the myometrium during peri-implantation period and luteolysis--the study on the pig model.

In pigs, implantation begins with the attachment of embryos to the endometrium. As the process is regulated by the expression of numerous genes, endometrial transcriptomic profiles have been extensively studied in early gravid pigs. However, the myometrium, a secretory tissue, should not be neglected, as it can also participate in the regulation of implantation in early pregnant pigs.

Novel aspects of cytokine action in porcine uterus--endometrial and myometrial production of estrone (E1) in the presence of interleukin 1beta (IL1beta), interleukin 6 (IL6) and tumor necrosis factor (TNFalpha)--in vitro study.

Interleukin 1beta (IL-1beta), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNFalpha) increased (P < 0.05) estrone (E1) release from endometrial explants of pregnant pigs on days 10 to 11 after 12 h of tissue incubation in vitro with cytokines and on days 12 to 13 after 6 h of incubation. After 12 h of incubation on days 12 to 13 and 15 to 16 of pregnancy only IL6 increased E1 release.

The effect of interleukin-1β, interleukin-6, and tumor necrosis factor-α on estradiol-17β release in the myometrium: the in vitro study on the pig model.

Estradiol-17β (E2) is a potent regulator of early pregnancy and the estrous cycle in pigs. Production of E2 occurs in the porcine myometrium, but the factors involved in its regulation are unknown. In this in vitro study, it was investigated whether interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α affect the release of E2 from the porcine myometrium on Days 10 to 11, 12 to 13, and 15 to 16 of pregnancy and the estrous cycle.

Transcriptomic analysis of the porcine endometrium during early pregnancy and the estrous cycle.

The goal of this study was to describe the alterations in the transcriptome of the endometrium in pigs during the beginning of implantation (days 15-16 of pregnancy) compared to cyclic pigs during the onset of luteolysis (days 15-16 of the estrous cycle). The global expression of genes in porcine gravid and non-gravid endometria was investigated using the Porcine (V2) Two-color gene expression microarray, 4 × 44 (Agilent, USA). Analysis of the microarray data showed that, of 589 accurately annotated genes, the expression of 266 genes was up-regulated and expression of 323 was down-regulated in the endometrium harvested during early pregnancy compared with the endometrium during the estrous cycle.

The effect of interleukin 1β and interleukin 6 on estradiol-17β secretion in the endometrium of pig during early pregnancy and the estrous cycle.

Estrogens are produced by porcine embryos during early pregnancy. It was found that the uterus of pigs might be a source of steroid hormones, including estrogens. However, the factors involved in the regulation of endometrial steroidogenesis remain unknown.

The interleukin-1β system in the corpora lutea of pigs during early pregnancy and the estrous cycle.

Expression of mRNAs encoding interleukin-1β (IL-1β), IL-1β receptor I (IL-1RI), IL-1 receptor accessory protein (IL-1RAcP) and IL-1 receptor antagonist (IL-1Ra), as well as synthesis of IL-1β and IL-1RI proteins, were examined in the corpus luteum (CL) during critical stages of CL activity on days 10-16 of pregnancy and 2-16 of the estrous cycle. Luteal cells were cultured in vitro with IL-1β, and the effect on release of steroid hormones was determined. Expression of the IL-1β system in the CL changed significantly during pregnancy and the estrous cycle.

The activity and localization of 3β-hydroxysteroid dehydrogenase/Δ(5)-Δ(4) isomerase and release of androstenedione and progesterone by uterine tissues during early pregnancy and the estrous cycle in pigs.

Steroid hormones are produced by the porcine uterus. We hypothesized that the uterus in pigs possesses active 3β-hydroxysteroid dehydrogenase/Δ(5)-Δ(4) isomerase (3β-HSD) responsible for progesterone and androstenedione production, that uterine steroids may supplement the amount of steroid hormones produced by embryos and corpus luteum and that these steroids are necessary for maintenance of pregnancy. In this study, we examined 1) endometrial and myometrial expression of 3β-HSD mRNA, 2) uterine 3β-HSD protein activity and 3) in vitro production of A(4) and P(4) by uterine slices harvested from pigs on days 10 to 11, 12 to 13 and 15 to 16 of pregnancy and the estrous cycle.

Role of interleukin-1β in the regulation of porcine corpora lutea during the late luteal phase of the cycle and during pregnancy.

Interleukin-1β (IL-1β) may regulate ovarian physiology. In this study, the influence of IL-1β on secretory activity within the corpora lutea (CL) of cyclic and gravid pigs was determined in vitro during different stages of the CL lifespan, e.g.

The effect of tumor necrosis factor α (TNFα), interleukin 1β (IL1β) and interleukin 6 (IL6) on endometrial PGF2α synthesis, metabolism and release in early-pregnant pigs.

Cytokines produced by the porcine uterus and embryos may be involved in the regulation of endometrial prostaglandin synthesis, metabolism, and release. We studied the effect of tumor necrosis factor α (TNFα), interleukin 1β (IL1β) and interleukin 6 (IL6) on: 1) endometrial release of prostaglandin F2α (PGF2α), 2) expression of the terminal enzyme of PGF2α synthesis--PGF synthase mRNA (PGFS mRNA), 3) secretion of PGF(2)α metabolite--13,14-dihydro-15-keto PGF2α (PGFM) by the endometrium and 4) presence and activity of endometrial NAD-dependent 15-hydroxyprostaglandin dehydrogenase (15-PGDH). The effects of cytokines were determined on days 10-11 and days 12-13, e.

Interleukin 1β-induced synthesis and secretion of prostaglandin E₂ in the porcine uterus during various periods of pregnancy and the estrous cycle.

Peri-implantation porcine embryos express interleukin-1β (IL-1β), which could affect uterine activity during early pregnancy. In vitro studies were conducted to determine if IL-1β stimulates secretion of PGE₂ and expression of cyclooxygenase-2 (COX-2) mRNA and microsomal PGE synthase-1 (mPGES-1) mRNA in uterine tissues harvested from pigs on days 10 to 11, 12 to 13 and 15 to 16 of pregnancy and the estrous cycle. IL-1β (10 ng/ml) increased PGE₂ secretion and mPGES-1 mRNA expression in uterine tissues isolated from pigs between days 10 to 13 of pregnancy and the estrous cycle.

The effect of progesterone on oxytocin-stimulated intracellular Ca2+ mobilisation and prostaglandin secretion in porcine endometrium.

We have studied in the porcine endometrium the expression of oxytocin receptor (OTR) mRNA and the effect of progesterone (P4) on oxytocin/oxytocin receptor (OT/OTR) function concerning intracellular Ca2+ mobilisation ([Ca2+]i), prostaglandin F2alpha (PGF2alpha) and E2 (PGE2; PG) secretion. Tissue was taken from cyclic and early pregnant pigs (days 14-16). A higher expression of OTR mRNA (P < 0.

Androgens and estradiol-17beta production by porcine uterine cells: In vitro study.

Porcine (Sus scrofa domestica) uterine slices harvested during both early pregnancy and luteolysis produce steroid hormones. The aim of the present study was to determine (1) which porcine separated uterine cells secrete androgens: androstenedione (A(4)) and testosterone (T), and estradiol-17beta (E(2)) in culture; (2) if the production of A(4), T and E(2) in the uterine cells is regulated by P4 and OT; (3) if uterine tissues expressed cytochrome P450arom gene (CYP19). Uteri were collected on Days 14 to 16 of early pregnancy and the estrous cycle.

The effect of oxytocin on progesterone secretion, phosphoinositide hydrolysis and intracellular mobilisation of Ca2+ in porcine luteal cells.

Oxytocin (OT) is involved in the regulation of steroid secretion by the corpus luteum (CL) in pigs, but OT signal transduction in the porcine CL has not been identified. In this study, the effects of OT on in vitro progesterone (P4) secretion, phosphoinositide (PI) hydrolysis and intracellular mobilisation of Ca2+ ([Ca2+]i) were investigated in porcine luteal cells during the early (days 3-5), mid(days 8-10) and late luteal phases (days 12-14) of the oestrous cycle. Basal concentrations of P4 and accumulation of inositol phosphates (IPs) were higher (P < 0.