Prostaglandin F2alpha Secretion Research Articles

MANFAAT PEMBERIAN JUS NANAS DAN MADU TERHADAP PENURUNAN NYERI DESMINORE PADA REMAJA PUTRI DI SMP YP SINGOSARI DELITUA

Dysmenorrhea is abdominal pain caused by uterine contractions and occurs during menstruation. Dysmenorrhea occurs due to increased secretion of prostaglandin F2a during the luteal phase of the menstrual cycle. Increased secretion of prostaglandin F2 alpha increases the frequency of uterine contractions and as a result can cause abdominal pain in women. The effects of primary dysmenorrhea include disruption of activities, such as absence from school or work, and limitations in social life, school performance, and athletic activities. Non-pharmacological pain management includes warm compresses, drinking lots of fluids, taking regular exercise breaks, consuming nutritious food, pain relief activities such as yoga, and using herbal medicine supplements which are effective in reducing menstrual cramps. Community service was carried out at YP Singosari Delitua Middle School with a total of 20 respondents. According to the results of most students who took part in consultations at YP Singosari Deli Tua Junior High School regarding the effect of pineapple juice and honey in reducing menstrual pain in young women, the level of knowledge is still low due to lack of socialization by providing education regarding the effectiveness of pineapple juice and honey in reducing menstrual cramps. , it is hoped that students can handle this dysmenorrhoea

Roadmap to pregnancy during the period of maternal recognition in the cow: Changes within the corpus luteum associated with luteal rescue

A viable corpus luteum (CL) producing an adequate amount of progesterone is absolutely essential for establishment and maintenance of pregnancy. One function of embryonic signaling to the mother is to ensure that the CL is maintained. In ruminants, the secretion of uterine prostaglandin F2alpha (PGF2A) is the signal that initiates luteolysis. Despite many studies to determine if PGF2A secretion from the uterus is altered in early pregnancy, conflicting interpretations have led to controversy regarding the exact mechanisms by which maternal recognition of pregnancy is achieved. In addition to alteration of uterine prostaglandin metabolism, changes within the CL itself may facilitate the establishment of a successful pregnancy. These changes include alteration of luteal blood flow, prostaglandin metabolism, sensitivity to prostaglandin actions, increased steroidogenic capacity, significant intracellular molecular changes and modification of the immune cells that are within the CL. Whether these intraluteal changes are necessary to establish pregnancy is undetermined. The focus of this review will be to provide a brief historical perspective on the utero-ovarian relationships that regulate luteal lifespan and review current knowledge of the CL of pregnancy in sheep and cattle.

Regulation of embryo survival in cattle

Evidence is presented that bovine somatotrophin (bST) treatment of lactating dairy cows enhances both expression of oviductal insulin-like growth factor II (IGF-II) mRNA and endometrial insulin-like growth factor binding protein 3 (IGFBP-3) mRNA between day 3 and day 7 of the oestrous cycle. mRNA encoding growth hormone (GH) receptor in endometrial tissues increased between day 3 and day 7 of the oestrous cycle. The changes induced by bST treatment may contribute to stimulation of embryo development and increase pregnancy rates in lactating dairy cows. Additive effects of bST and rb interferon tau (rbIFN-tau) to inhibit phorbol ester induction of prostaglandin F2alpha secretion in immortalized bovine endometrial cells indicates that there is interplay between their signal transduction pathways. Non-lactating dairy cows were killed at day 17 after oestrus to evaluate the effects of pregnancy status (cyclic versus pregnant) and bST (bST versus control) treatment on endometrial gene expression. Distinctly different mRNA and protein responses were detected between cyclic and pregnant cows that were related to luteolytic-antiluteolytic drive (that is expression of progesterone receptor, oxytocin receptor, oestradiol receptor alpha and prostaglandin GH synthase 2 (PGHS-2)). The bST-induced changes in PGHS-2 protein (+), oxytocin receptor mRNA (+) and oestrogen receptor alpha protein (+) may potentially affect the mechanisms associated with maintenance of pregnancy. Two experiments were conducted to evaluate whether ovarian follicular suppression induced by biodegradable deslorelin implants would reduce either early or late embryo losses. A 450 microg deslorelin implant used to induce ovulation in a timed insemination programme decreased subsequent follicular development and tended to reduce early embryo losses, whereas a 2.1 mg deslorelin implant failed to reduce late embryonic losses when inserted on day 27 of pregnancy.

In Vivo Chronic and In Vitro Acute Effects of Di(2-Ethylhexyl) Phthalate on Pseudopregnant Rabbit Corpora Lutea: Possible Involvement of Peroxisome Proliferator-Activated Receptor Gamma1

The in vivo chronic and in vitro acute effects of di(2-ethylhexyl) phthalate (DEHP) on the reproductive function of peroxisome proliferator-activated receptor gamma (PPARG) were studied in rabbit corpora lutea (CL) at early stage (Day 4), midstage (Day 9), and late stage (Day 13) of pseudopregnancy. The rabbits were in vivo treated with DEHP for 15 days before induction of pseudopregnancy. Immunohistochemistry provided evidence for the presence of PPARG, prostaglandin endoperoxide synthase 1 (PTGS1), PTGS2, prostaglandin E2-9-ketoreductase (PGE2-9-K), and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) in all the luteal cells during pseudopregnancy. DEHP decreased progesterone plasma levels and CL production in all the luteal stages and PPARG protein and gene expressions in early and mid-CL. DEHP in vivo treatment reduced PTGS2 protein expression at the late stage and that of PGE2-9-K at all the stages, whereas PTGS1 and 3beta-HSD were not affected. In in vitro cultured CL, DEHP alone, the PPARG antagonist T0070907 alone, or DEHP plus T0070907 diminished progesterone production and 3beta-HSD activity and increased PGF2alpha and PTGS2 in early and mid-CL, whereas DEHP plus the PPARG agonist 15d-PGJ2 did not affect these hormones and enzymes. All the in vitro treatments did not affect PGE2 secretion as well as PTGS1 and PGE2-9-K enzymatic activities in all the luteal stages. These results provided evidence that DEHP favors functional luteolysis of pseudopregnant rabbit CL, with a mechanism that seems to involve PPARG expression down-regulation, an increase of PTGS2 activity and prostaglandin F2alpha secretion, 3beta-HSD down-regulation, and decrease in progesterone.

Pregnancy recognition signaling mechanisms in ruminants and pigs

Maternal recognition of pregnancy refers to the requirement for the conceptus (embryo and its associated extra-embryonic membranes) to produce a hormone that acts on the uterus and/or corpus luteum (CL) to ensure maintenance of a functional CL for production of progesterone; the hormone required for pregnancy in most mammals. The pregnancy recognition signal in primates is chorionic gonadotrophin which acts directly on the CL via luteinizing hormone receptors to ensure maintenance of functional CL during pregnancy. In ruminants, interferon tau (IFNT) is the pregnancy recognition signal. IFNT is secreted during the peri-implantation period of pregnancy and acts on uterine epithelia to silence expression of estrogen receptor alpha and oxytocin receptor which abrogates the oxytocin-dependent release of luteolytic pulses of prostaglandin F2-alpha (PGF) by uterine epithelia; therefore, the CL continues to produce progesterone required for pregnancy. Pig conceptuses secrete interferon delta and interferon gamma during the peri-implantation period of pregnancy, but there is no evidence that they are involved in pregnancy recognition signaling. Rather, pig conceptuses secrete abundant amounts of estrogens between Days 11 to 15 of pregnancy required for maternal recognition of pregnancy. Estrogen, likely in concert with prolactin, prevents secretion of PGF into the uterine venous drainage (endocrine secretion), but maintains secretion of PGF into the uterine lumen (exocrine secretion) where it is metabolized to a form that is not luteolytic. Since PGF is sequestered within the uterine lumen and unavailable to induce luteolysis, functional CL are maintained for production of progesterone. In addition to effects of chorionic gonadotrophin, IFNT and estrogens to signal pregnancy recognition, these hormones act on uterine epithelia to enhance expression of genes critical for growth and development of the conceptus.

Selective Increased Luteal Prostaglandin E2 Compared to F2 Alpha Biosynthesis and Signaling Are Critical for Luteal Protection and Resistance During Establishment of Pregnancy in Sheep.

In ruminants, pulsatile secretion of prostaglandin F2 alpha (PGF2a) by the endometrium is locally transported to corpus luteum (CL) and stimulates intraluteal PGF2a biosynthesis and induces luteal regression/luteolysis. Interestinlgy, luteolytic PGF2a fails to accomplish regression of CL of early pregnancy. This increased resistance of the CL to PGF2a may suggest existence of luteal protective mechanisms. Early physiological studies proposed prostaglandin (E2) PGE2 as luteal protective mediator because PGE2 counteracted the effects of PGF2a and protected the CL from regression. However, the molecular and cellular aspects of resistance of the CL to luteolytic PGF2a are largely unknown. The objective of this study is to determine biosynthesis, transport, and signaling of PGF2a and PGE2 in CL during the estrous cycle and early pregnancy in sheep. Suffolk cross-bred ewes were ovariohysterectomized on days 12, 14 and 16 of the estrous cycle (ECY) or pregnancy (PNY), the CLs were collected, and temporal expression of PGF2a and PGE2 biosynthetic, transport and signaling proteins were determiend by western blot. Blood samples were collected from the ovarian artery and ovarian vein and transport of PGF2a, PGFM, PGE2, and PGEM were determiend by ELISA. Results indicated that: (i) COX-2 protein was abundantly expressed on days 12-16 of ECY and PNY and not affected by the ECY or PNY status, and COX-1 protein was expressed at very low level; (ii) PGES-1 protein was abundantly expressed on days 12-14 of ECY and PNY, and then decreased (4 fold) on day 16 of the ECY compared to that of PNY, expression of PGES-2 protein was not modulated by ECY and PNY, and expression of PGES-3 protein was decreased (5 fold) on days 12-16 of ECY compared to that of PNY; (iii) expression of PGFS-AKR1C1 protein was not modulated by ECY and PNY, and PGFS-AKR1C3 protein was expressed at steady sate level on day 10-16 of ECY but decreased (several fold) on days 12-16 of PNY; (iv) expression of FP, EP1 and EP3 proteins were not modulated by ECY and PNY, but interestingly, expression of EP2 and EP4 proteins were decreased (4 fold) on day 16 of ECY compared to that of PNY; (v) PGT protein was abundantly expressed on days 12-16 of ECY and PNY and its expression was decreased on day 16 of ECY compared to that of PNY; and (vi) PGDH protein expression was increased on days 14-16 of PNY compared to that of ECY. (vi) Ovarian venous plasma concentrations of PGF2a, PGFM, PGE2 and PGEM were 1500-2000 pg/ml, 400-500 pg/ml, 1000-1250 pg/ml, and 2-5 pg/ml on days 14-16 of the estrous cycle, respectively, and 400-500 pg/ml, 2000-2500 pg/ml, 3000-3500 pg/ml, and 3-5 pg/ml on days 14-16 of pregnancy, respectively. These results together suggest that the luteal PG biosynthetic and signaling machinery is selectively directed towards PGF2a at the time of luteolysis, by contrast; towards PGE2 at the time of establishment of pregnancy. In addition, CL of pregnancy has innate capacity to catabolize PGF2 to inactive PGFM but not PGE2 to PGEM and favors intraluteal PGE2 production. Thus, increased intraluteal PGE2 production and action are the critical mechanisms that resist/protect CL of early pregnancy to luteolytic PGF2a. Selective inhibition of PGES-1 or EP2 and EP4 along with PGF2a could be the potential future therapeutic strategy to improve reproductive efficiency in ruminants. Supported by USDA award 2008-35203-19101 to JAA. (poster)

Dietary supplementation with safflower seeds differing in fatty acid composition differentially influences serum concentrations of prostaglandin F metabolite in postpartum beef cows

Synthesis and secretion of prostaglandin F2alpha (PGF2alpha) is elevated following parturition and exerts divergent effects on the re-establishment of fertile estrous cycles in cows. The objective of these experiments was to determine if oil seed supplements differing in fatty acid composition differentially influence serum concentrations of the specific PGF2alpha metabolite, PGFM. Safflower seed supplements were formulated to provide 5% of dry-matter intake as fat. In Trial 1, 24 multiparous beef cows were individually fed control (beet pulp-soybean meal) or cracked high-linoleate safflower seed (78% 18:2n-6) supplements for 80 d postpartum. Linoleate supplemented cows had greater (P < 0.001) serum concentrations of PGFM than control cows. In Trial 2, primiparous beef cows (n = 36) were individually fed control (cracked corn-soybean meal), cracked high-linoleate (76% 18:2n-6) or -oleate (72% 18:1n-9) safflower seed supplements for 92 d postpartum. As in Trial 1, serum concentrations of PGFM were greater (P < or = 0.04) in linoleate than control or oleate supplemented cows. Serum concentrations of PGFM, however, did not differ (P = 0.40) among oleate and control supplemented cows. Although potential impacts on reproductive performance remain to be proven, dietary oil supplements high in linoleate, but not oleate, increased serum concentrations of PGFM compared to control supplements.

Polyunsaturated fatty acids and bovine interferon-tau modify phorbol ester-induced secretion of prostaglandin F2 alpha and expression of prostaglandin endoperoxide synthase-2 and phospholipase-A2 in bovine endometrial cells.

Embryonic mortality in cattle may occur because of inadequate inhibition of uterine secretion of prostaglandin (PG) F2alpha mediated by bovine interferon-tau (bIFN-tau). The objectives of the present study were to determine whether polyunsaturated fatty acids inhibit secretion of PGF2alpha from bovine endometrial cells induced by stimulating protein kinase C with phorbol 12,13 dibutyrate (PDBu) and to investigate possible mechanisms of action. Confluent cells were exposed for 24 h to 100 microM of linoleic, arachidonic (AA; C20:4, n-6), linolenic (LNA; C18:3, n-3), eicosapentaenoic (EPA; C20:5, n-3), or docosahexaenoic (DHA; C22:6, n-3) acid. After incubation, cells were washed and stimulated with PDBu. The EPA, DHA, and LNA attenuated secretion of PGF2alpha in response to PDBu. The EPA and DHA were more potent inhibitors than LNA. The EPA inhibited secretion of PGF2alpha at 6.25 microM. Secretion of PGF2alpha in response to PDBu decreased with increasing incubation time with EPA. Both bIFN-tau and EPA inhibited secretion of PGF2alpha, and their inhibitory effects were additive. The bIFN-tau, but not EPA, reduced the abundance of PG endoperoxide synthase-2 (PGHS-2) mRNA. Incubation with 100 microM EPA, DHA, or AA for 24 h followed by treatment with PDBu did not affect concentrations of PGHS-2 and phospholipase A2 proteins. The EPA and DHA inhibit secretion of PGF2alpha through a mechanism different from that of bIFN-tau. The effect of EPA on PGF2alpha secretion may be caused by competition with AA for PGHS-2 activity or reduction of PGHS-2 activity. The use of EPA and DHA to inhibit uterine secretion of PGF2alpha and to improve embryonic survival in cattle warrants further investigation.

Steroidal regulation of uterine resistance to bacterial infection in livestock

Postpartum uterine infections reduce reproductive efficiency and have significant animal welfare and economic consequences. Postpartum uterine infections are classified as nonspecific, but Arcanobacterium pyogenes and Escherichia coli are usually associated with them in cattle and sheep. Pyometra is the most common type of uterine infection in dairy cattle, and it is detected almost exclusively in cows with active corpora lutea. Luteal progesterone typically down-regulates uterine immune functions and prevents the uterus from resisting infections. Progesterone also can down-regulate uterine eicosanoid synthesis. This seems to be a critical event in the onset of uterine infections, because eicosanoids can up-regulate immune cell functions in vitro. In addition, exogenous prostaglandin F2 alpha stimulates uterine secretion of prostaglandin F2 alpha and enhances immune functions in vivo. Thus, one may hypothesize that eicosanoids can override the negative effects of progesterone and that the up-regulatory effects of exogenous prostaglandin F2 alpha allow the uterus to resolve an infection, regardless of progesterone concentrations. Based on the results of studies to test that hypothesis, cows, sheep, and pigs in various physiological statuses are resistant to intrauterine infusions of Arcanobacterium pyogenes and Escherichia coli, unless progesterone concentrations are increased. In sheep and pigs, exogenous prostaglandin F2 alpha stimulates uterine production of prostaglandin F2 alpha and allows the uterus to resolve Arcanobacterium pyogenes-Escherichia coli-induced infections, even when progesterone is maintained at luteal phase concentrations before and after treatment. Prostaglandin F2 alpha is a proinflammatory molecule that stimulates the production of various proinflammatory cytokines, and it may enhance uterine production of leukotriene B4. Proinflammatory cytokines and leukotriene B4 enhance phagocytosis and lymphocyte functions. Even though there are clear associations among prostaglandin F2 alpha, leukotriene B4, proinflammatory cytokines, phagocytosis, and lymphocyte functions, the mechanism of action of exogenous prostaglandin F2 alpha in overriding the down-regulatory effects of progesterone and resolving uterine infections has not been elucidated. Defining this mechanism should yield new prevention and treatment strategies for uterine infections that do not rely on antibiotic and antimicrobial compounds.

Effects of arginine- and lysine-vasopressin on phospholipase C activity, intracellular calcium concentration and prostaglandin F2alpha secretion in pig endometrial cells.

Oxytocin and vasopressin are related peptides that have receptors in the uterus. Species from families other than Suidae produce only arginine-vasopressin; in contrast, pigs apparently express both arginine- and lysine-vasopressin. The aim of this study was to determine whether arginine- or lysine-vasopressin would activate phospholipase C, increase intracellular calcium concentration [Ca(2+)](i) and stimulate PGF(2alpha) production in enriched cultures of stromal, glandular epithelial and luminal epithelial cells from pig endometrium. Cells were obtained from gilts on day 16 after oestrus by differential enzymatic digestion and sieve separation. After 96 h in culture, the cells were treated with 0 or 100 nmol arginine- or lysine-vasopressin l(-1). The responses to 100 nmol oxytocin l(-1) and 100 nmol GnRH l(-1) were used as positive and negative controls, respectively. Consistent with previous results, oxytocin stimulated phospholipase C activity (P < 0.05), increased [Ca(2+)](i) (P < 0.05) and promoted PGF(2alpha) secretion (P < 0.05) from stromal and glandular epithelial cells. Activity of phospholipase C, [Ca(2+)](i) and PGF(2alpha) release were also increased (P < 0.05) by arginine-vasopressin in stromal cells, but the responses were less (P < 0.01) than those induced by oxytocin. An oxytocin antagonist attenuated the [Ca(2+)](i) response of stromal cells to both oxytocin and arginine-vasopressin. Sequential treatment of cells with oxytocin and arginine-vasopressin indicated that oxytocin desensitized the response to oxytocin, but arginine-vasopressin did not similarly desensitize the response to oxytocin. In glandular and luminal epithelial cells, arginine-vasopressin did not stimulate phospholipase C activity, [Ca(2+)](i) or PGF(2alpha) secretion. Neither GnRH nor lysine-vasopressin induced phospholipase C activity, increased [Ca(2+)](i) or stimulated PGF(2alpha) production in any endometrial cell type. These results indicate that oxytocin receptors can bind arginine-vasopressin more readily than they bind lysine-vasopressin. Type 1 vasopressin receptors may also exist in endometrium predominantly on cells other than stromal, glandular epithelial and luminal epithelial cells, as in previous studies both arginine-vasopressin and lysine-vasopressin stimulated phospholipase C activity in endometrial explants to a similar extent as oxytocin.

Effects of arginine- and lysine-vasopressin on phospholipase C activity, intracellular calcium concentration and prostaglandin F2alpha secretion in pig endometrial cells

Oxytocin and vasopressin are related peptides that have receptors in the uterus. Species from families other than Suidae produce only arginine-vasopressin; in contrast, pigs apparently express both arginine- and lysine-vasopressin. The aim of this study was to determine whether arginine- or lysine-vasopressin would activate phospholipase C, increase intracellular calcium concentration [Ca(2+)](i) and stimulate PGF(2alpha) production in enriched cultures of stromal, glandular epithelial and luminal epithelial cells from pig endometrium. Cells were obtained from gilts on day 16 after oestrus by differential enzymatic digestion and sieve separation. After 96 h in culture, the cells were treated with 0 or 100 nmol arginine- or lysine-vasopressin l(-1). The responses to 100 nmol oxytocin l(-1) and 100 nmol GnRH l(-1) were used as positive and negative controls, respectively. Consistent with previous results, oxytocin stimulated phospholipase C activity (P < 0.05), increased [Ca(2+)](i) (P < 0.05) and promoted PGF(2alpha) secretion (P < 0.05) from stromal and glandular epithelial cells. Activity of phospholipase C, [Ca(2+)](i) and PGF(2alpha) release were also increased (P < 0.05) by arginine-vasopressin in stromal cells, but the responses were less (P < 0.01) than those induced by oxytocin. An oxytocin antagonist attenuated the [Ca(2+)](i) response of stromal cells to both oxytocin and arginine-vasopressin. Sequential treatment of cells with oxytocin and arginine-vasopressin indicated that oxytocin desensitized the response to oxytocin, but arginine-vasopressin did not similarly desensitize the response to oxytocin. In glandular and luminal epithelial cells, arginine-vasopressin did not stimulate phospholipase C activity, [Ca(2+)](i) or PGF(2alpha) secretion. Neither GnRH nor lysine-vasopressin induced phospholipase C activity, increased [Ca(2+)](i) or stimulated PGF(2alpha) production in any endometrial cell type. These results indicate that oxytocin receptors can bind arginine-vasopressin more readily than they bind lysine-vasopressin. Type 1 vasopressin receptors may also exist in endometrium predominantly on cells other than stromal, glandular epithelial and luminal epithelial cells, as in previous studies both arginine-vasopressin and lysine-vasopressin stimulated phospholipase C activity in endometrial explants to a similar extent as oxytocin.

Interferon-tau suppresses prostaglandin F2alpha secretion independently of the mitogen-activated protein kinase and nuclear factor kappa B pathways.

Pregnancy is established in ruminants through inhibitory actions of interferon (IFN)-tau on the release of prostaglandin F2alpha (PGF), which allows the corpus luteum to survive and continue to produce progesterone. Experiments were designed to 1) delineate the signal transduction pathway coordinating the synthesis of PGF, 2) determine how rapidly recombinant bovine (rb) IFN-tau attenuated phorbol ester (PDBu)-induced secretion of PGF, and 3) establish the site at which rbIFN-tau attenuates the secretion of PGF in cultured bovine endometrial (BEND) cells. BEND cells were untreated (control) or treated for 5, 10, 60, 180, or 300 min with PDBu (100 ng/ml), rbIFN-tau (50 or 500 ng/ml), PDBu + rbIFN-tau, or PDBu + PD98059 (MEK-1 inhibitor; 50 microM). Secretion of PGF was induced (P < 0.0001) by PDBu within 180 min, but induction was inhibited 74% by the addition of rbIFN-tau (P < 0.0001) and was ablated completely by PD98059. Parallel results were obtained for cyclooxygenase (COX)-2 protein expression. PDBu induced (P < 0.05) activation of the Raf-1/MEK-1/ERK-1/2 pathway, which was obligatory for the expression of COX-2 and secretion of PGF but was not altered by cotreatment with rbIFN-tau. PDBu induced (P < 0.05) transcription of c-jun and c-fos mRNAs within 30 min; induction was inhibited (P < 0.05) by cotreatment with PD98059 but not by cotreatment with rbIFN-tau. Treatment of BEND cells with rbIFN-tau also did not attenuate PDBu-induced degradation of IkappaBalpha, suggesting that the IkappaBalpha/NFkappaB pathway is not a site of IFN-tau inhibition of PGF. However, rbIFN-tau did block transcription of the COX-2 gene induced by PDBu within 30 min. In conclusion, COX-2 expression and PGF secretion induced by PDBu is mediated through the Raf-1/MEK-1/ERK-1/2 pathway, but this pathway is not disrupted by rbIFN-tau. Because rbIFN-tau inhibits COX-2 mRNA within 30 min, we hypothesized that transcription factors activated by rbIFN-tau rapidly and directly attenuate COX-2 gene expression, thereby suppressing secretion of PGF.

Regulation of protein and prostaglandin secretion in polarized primary cultures of caprine uterine epithelial cells.

Caprine uterine epithelial (UE) cells were cultured on Matrigel-coated filters. Transmission electron microscopy revealed polarized UE cells characterized by basally located nuclei, apical microvilli, convoluted lateral membranes, and junctional complexes. Domain-specific secretion of prostaglandins and radiolabeled proteins provide further evidence of functional epithelial cell polarity. Two experiments were conducted to evaluate factors controlling prostaglandin E2 (PGE) and prostaglandin F2alpha (PGF) secretion. In experiment one, steroid-treated (estradiol, progesterone, or estradiol + progesterone) polarized UE cells were treated with interferon tau (IFNtau) and/or oxytocin (OT). Steroid treatment did not influence PGE or PGF secretion. However, analysis of variance revealed an IFNtau by OT interaction (P < .01) for both PGE and PGF This interaction was caused by a reduction in PGE and PGF secretion by cultures receiving only IFNtau and the inability of IFNtau to block OT-induced release of PGE or PGF. In experiment 2, polarized UE cells were cultured in progesterone, with or without IFNtau, and sequentially challenged with estradiol and OT. Oxytocin stimulated the release of both PGE and PGF by polarized cUE cells (P < .01) and resulted in an increased accumulation of PGE (OT*domain; P < .01) in the basal compartment. Interferon tau did not influence PGE (P < .1) secretion. However, further analysis revealed that IFNtau reduced PGF secretion and was unable to block OT-induced PGF secretion (IFNtau*OT; P < .05) by polarized UE cells. Therefore, caprine UE cells form polarized monolayers and retain responsiveness to IFNtau and OT in vitro.

Induction of human immunodeficiency virus type 1 replication in human glial cells after proinflammatory cytokines stimulation: Effect of IFN?, IL1?, and TNF? on differentiation and chemokine production in glial cells

Although evidence for human immunodeficiency virus 1 (HIV-1) presence in the central nervous system (CNS) of infected patients is well established, the intensity of viral replication within the brain is not usually known. In vitro, human embryonic microglial cells internalized HIV-1 through a CD4-dependent pathway but were not permissive to viral replication. We observed that HIV replication was induced when CNS cell cultures were stimulated for 14 days by a combination of proinflammatory cytokines including IFNgamma, IL1beta, and TNFalpha. After long-term cytokine stimulation, morphologically differentiated glial cells appeared, in which HIV-1 tat antigen was detected after infection. Thus, variations in the stage of maturation/activation of CNS cells under inflammatory conditions probably play a major role in facilitating massive production of HIV-1. We then studied the effect of prolonged cytokine stimulation on the secretion of inflammatory mediators by glial cells. An early increased secretion of prostaglandin F2alpha and chemokines (RANTES>>MIP-1alpha>>MIP-1beta) was observed, due to both microglia and astrocytes. In contrast to persistent PGF2alpha production, an extinction of RANTES and MIP-1beta but not of MIP-1alpha secretion occurred during the 14 days of stimulation and was inversely correlated with the ability of glial cells to replicate HIV-1. The study of the secretory factors produced in response to a persistent inflammation could provide a better understanding of the modulation of HIV replication in glial cells.

Oxytocin- and aluminium fluoride-induced phospholipase C activity and prostaglandin F2 alpha secretion during the ovine luteolytic period.

A series of studies was conducted to characterize changes in components of the cell signalling cascade that mediates oxytocin-induced prostaglandin F2 alpha (PGF2 alpha) synthesis at the onset of luteolysis in sheep. In the first experiment, caruncular tissue was dissected from 20 ewes on days 12-15 of the oestrous cycle, and incubated for the measurement of phospholipase C (PLC) activity or secretion of PGF2 alpha. Activation of GTP-binding proteins with aluminium fluoride stimulated both inositol phosphate accumulation and PGF2 alpha secretion on all days examined. However, oxytocin did not stimulate PLC activity or PGF2 alpha accumulation until day 13. While the ability of oxytocin to stimulate PLC activity increased after day 13, oxytocin-induced PGF2 alpha secretion declined slightly from day 13 to 15, suggesting that cell signalling components downstream from PLC modulate the response to oxytocin after day 13. Oxytocin failed to stimulate PGF2 alpha synthesis on day 14 after oestrus. Secretion of endogenous luteal oxytocin may have rendered uterine tissues collected on day 14 refractory to oxytocin in vitro. Therefore, a second study was conducted in ovariectomized, steroid replaced ewes. Ovarian steroids were administered to mimic endogenous changes in progesterone and oestradiol. The temporal patterns of PGF2 alpha synthesis in response to oxytocin and pharmacological agents were similar to uterine tissues from cyclic ewes in the first experiment; however, the magnitude of the response was less. These data suggest that oxytocin receptors are absent or are not coupled to PLC until day 13 after oestrus.

Uterine secretion of prostaglandin F2alpha stimulated by different doses of oxytocin and released spontaneously during luteolysis in cattle.

The objectives were to determine the involvement of oxytocin (OT) in the stimulation of prostaglandin F2alpha (PGF) secretion during luteolysis in cattle. On days 16-17 of the oestrous cycle, catheters were inserted into the aorta abdominalis of heifers for OT or saline infusion and into the jugular vein for blood sample collection. The following day, heifers were assigned to one of three experimental groups (Gr): Gr I - 10 IU OT (n=4); Gr II - 20 IU OT (n=4); Gr III - 50 IU OT (n=4). Blood samples were collected every 10 min during a 1-h control period before treatment and every 5-10 min for 2 h after OT treatment. In Gr IV (n=5), a catheter was inserted into the jugular vein on day 15 of the cycle and blood samples were collected every 15 min for 12 h on days 16-19. Plasma concentrations of progesterone, PGF metabolite, 13, 14-dihydro-15-keto-prostaglandin F2alpha (PGFM) and OT were determined. Within 5 min of infusion of 10 or 20 IU OT, peripheral concentrations of OT (7-12 pg/mL) increased by about 200 and 350-500 pg/mL, respectively. These doses did not affect plasma concentrations of PGFM or progesterone within 1.5 h. Fifty IU of OT increased its maximal peripheral concentration to 1500 pg/mL, which is over 20 times greater than that observed physiologically. Concentrations of plasma PGFM in Gr III increased from basal concentrations (5065 pg/mL) to 150-250 pg/mL (P < 0.01) within 10-30 min. During luteolysis, PGFM pulses ranged between 250 and 600 pg/mL on days 16-19 of the cycle (Gr IV), whereas coincident pulses of OT, and those appearing between spikes of PGFM, were never above 75 pg/mL. Only 50% of OT pulses coincided with pulses of PGFM, and 54 % of PGFM pulses coincided with a pulse of OT. Results indicate that luteolytic PGF secretion in cattle is not directly dependent upon ovarian OT.

Up-regulation of oxytocin receptors in porcine endometrium by adenosine 3',5'-monophosphate.

Porcine endometrial cells (a mixture of epithelial, stromal, and glandular cells) were examined for the presence of oxytocin (OT) receptors using a cell culture system and a 125I-labeled OT antagonist. Binding specificity was tested in displacement studies with various related peptides. Scatchard analyses revealed the presence of a binding site with a dissociation constant (Kd) = 0.9 nM and a capacity of 1.9 fmol/10(5) cells. These cells, which were obtained from prepubertal gilts and thus had not been exposed to endogenous ovarian steroids, were used as a model to evaluate the possible action of ovarian steroids and intracellular cAMP on OT receptors. Although ovarian steroids showed no effect on OT receptors, forskolin (an adenylate cyclase activator) and dibutyryl cAMP caused 1.5- to 1.6-fold increases in specific binding of OT without changing the binding affinity. When the endometrial cells were exposed to OT (0.1-1000 nM) in combination with arachidonic acid (10 microM), OT stimulated prostaglandin F2 alpha secretion in a dose-dependent manner. These results indicate the presence of functional OT receptors in prepubertal porcine endometrial cells and suggest that the concentration of OT receptors may be regulated by one or more substances that raise intracellular cAMP levels.

Changes in uterine secretion of prostaglandin F2 alpha in response to oxytocin during the estrous cycle, early pregnancy, and estrogen-induced pseudopregnancy in swine.

Thirty-one sows were used in an experiment designed to determine whether the ability of the porcine uterus to release prostaglandin (PG) F2 alpha in response to oxytocin was suppressed in pregnancy and pseudopregnancy. Sows were assigned to one of three treatment groups: nonbred (nonpregnant) controls (n = 8), pseudopregnant (5 mg estradiol benzoate, i.m., daily on Days 11-15 postestrus; n = 8), or bred (bred once daily throughout the estrous period; n = 15). Jugular venous blood samples were collected daily for quantification of progesterone. Pregnancy was determined by uterine examination at slaughter 51-72 days postmating. On the basis of progesterone and embryo recovery, bred sows were classified into three subgroups: confirmed pregnant (n = 4), suspected pregnant based on delayed luteal regression (n = 5), or bred/not pregnant (n = 6). All sows received an injection of oxytocin (30 IU, i.v.) on Days 12, 15, and 18 postestrus. Jugular venous blood samples were collected from 60 min prior to through 120 min after injection of oxytocin for quantification of 13, 14-dihydro-15-keto-PGF2 alpha (PGFM). Magnitude of response above baseline and area under the PGFM response curve (AUC) were calculated for each sow on each day and compared among treatment groups by analysis of variance. Responses in pregnant and suspected-pregnant sows were not different on any day examined; therefore the two groups were combined (n = 9) and considered pregnant for all subsequent analyses. Responses in the nonpregnant and bred/not pregnant sows were pooled and compared to the responses in the pregnant and pseudopregnant sows. Magnitudes of response were similar between these pooled groups on Day 12 (p > 0.5), but were less in pregnant and pseudopregnant sows on Days 15 and 18 (p < 0.01). When nonpregnant and bred/not pregnant sows were compared to each other, the magnitudes of the response were similar on Days 12, 15, and 18 (p > 0.3 on each day). In contrast, when pregnant and pseudopregnant sows were compared, pseudopregnant sows had a lower magnitude of response that was consistent across all 3 days (p < 0.02). Similar relationships were apparent for the AUC. These results demonstrate that uterine secretory responsiveness to oxytocin is suppressed during early pregnancy and that this effect may be mediated through estrogen secreted by conceptuses.

Differential suppression of endometrial prostaglandin F2alpha by the equine conceptus

Prostaglandin F2alpha secretion by the uterine endometrium between Days 13 and 14 postovulation causes luteal regression in mares. A mechanism involving interruption or suppression of this secretion causes pregnancy to be maintained. The present study was designed to determine the age of the conceptus when maximal suppression of PGF2alpha secretion occurs. Mares were examined daily during estrus with ultrasonography (day 0 = day of ovulation). Conceptus tissues were recovered nonsurgically on Days 9 (n = 7), 12 (n = 5), 13 (n = 5), and 16 (n = 7) and uterine biopsies on Day 14. Both uterine and conceptus tissues were washed in phosphate-buffered saline (PBS) with 100 units penicillin G/ml + 100 μg streptomycin/ml, pH 7.4. Endometrial tissue (~ 200 mg) plus conceptus tissues were incubated in 15 ml of tissue culture medium 199 (M199) + 10% fetal calf serum and 10 units penicillin G/ml and 10 μg streptomycin/ml at 37 °C under 5% CO 2: 5% O 2 : 90% N 2. Samples were taken at 4, 8, and 24 h. Two plates that contained only endometrial tissue and two additional plates with 25 mg flunixin meglumine added along with endometrial tissue were also included in the incubations. Concentrations of PGF2alpha were measured in all samples using radioimmunoassay. There was a trend toward suppression of PGF2alpha secretion by conceptus tissues, regardless of age. However, Day 12 concepti significantly suppressed PGF2alpha secretion compared with that of endometrial tissue incubated alone (P = 0.03).

Changes in peripheral concentrations of 13,14-dihydro-15-keto-prostaglandin F2 alpha induced by progesterone in swine.

This experiment was designed to examine the effects of progesterone on endogenous and oxytocin-induced secretion of prostaglandin F2 alpha (PGF2 alpha) in sows. Peripheral concentrations of 13,14-dihydro-15-keto-PGF2 alpha (PGFM) were used as an indirect measure of uterine PGF2 alpha secretion. Eight sows received twice daily injections of progesterone (160 mg/injection) on d 1 to 5 after estrus. Another eight sows received comparable injections of a corn oil injection vehicle. Each sow then received i.v. injections of oxytocin (30 IU) on d 10, 12, and 15 after estrus. Concentrations of PGFM were determined in jugular venous blood samples collected at -60, -45, -30, -15, 0, 2, 5, 10, 15, 30, 45, 60, 90, and 120 min after each oxytocin injection. The mean concentrations of PGFM in samples collected before injection of oxytocin (baseline), the magnitude of the PGFM response to oxytocin, and the area under the PGFM response curve (AUC) were calculated for the three oxytocin challenges administered to each sow. Baseline, magnitude, and AUC were low on d 10 after estrus and similar for the two treatment groups. On d 12 baseline, magnitude, and AUC remained low in the control sows; however, all three response variables increased in sows that received progesterone. By d 15, all three variables were high and similar in both treatment groups. In conclusion, progesterone, administered early in the estrous cycle, seems to promote premature secretion of PGF2 alpha as indicated by the high basal concentrations of PGFM observed before injection of oxytocin on d 12.(ABSTRACT TRUNCATED AT 250 WORDS)