dinoprost and lysophosphatidic-acid

Topics: Animals; Buffaloes; Cyclooxygenase 2; Dinoprost; Dinoprostone; Estrous Cycle; Female; Lysophospholipids; Nitric Oxide; Receptors, Lysophosphatidic Acid; Signal Transduction; Uterus

In previous work, an EP2 prostanoid receptor (EP2R) agonist in vivo increased mRNA expression of luteal LH receptors (LHR), unoccupied and occupied luteal; LHR, and circulating progesterone, while an EP3R or FPR agonist decreased; mRNA expression of luteal LHR, unoccupied and occupied luteal LHR, and; circulating progesterone. An EP4R and lysophosphatidic acid (LPA) LPA2R and LPA3R agonists were reported to inhibit luteal function and sirtuins have been proposed to increase prostaglandin synthesis. The objectives were to determine; whether an EP4R, LPA2R, or LPA3R agonist affect ovine luteal function in vivo or; in vitro. In addition, whether sirtuin (SIRT)-1, 2, or 3; LPA2R or LPA3R; or EP1R, EP2R, EP3R, or EP4R agonists affect caruncular endometrial PGF

Topics: Alprostadil; Animals; Corpus Luteum; Dinoprost; Endometrium; Female; Lysophospholipids; Progesterone; Prostaglandins E; Receptors, Lysophosphatidic Acid; Receptors, Prostaglandin E; Receptors, Prostaglandin E, EP4 Subtype; RNA, Messenger; Sheep; Sirtuins

We examined whether lysophosphatidic acid affects prostaglandin biosynthesis, transport, and signalling in bovine steroidogenic luteal cells. The aim of the present study was to determine the influence of LPA on PGE2 and PGF2α synthesis and on the expression of enzymes involved in PG biosynthesis (PTGS2, mPGES-1, cPGES, mPGES-2, PGFS and 9-KPR), prostaglandin transporter (PGT), and prostaglandin receptors (EP1, EP2, EP3, EP4 and FP) in bovine steroidogenic luteal cells. We found that LPA inhibited PGF2α synthesis in steroidogenic luteal cells. Moreover, LPA increased mPGES1 and cPGES and decreased PGFS expression in cultured bovine steroidogenic luteal cells. Additionally, LPA stimulated EP2 and EP4 receptor and PGT expression. This study suggests that LPA activity in the bovine CL directs the physiological intraluteal balance between the two main prostanoids towards luteotropic PGE2.

Topics: Abattoirs; Animals; Biological Transport; Cattle; Cells, Cultured; Dairying; Dinoprost; Dinoprostone; Estrous Cycle; Female; Gene Expression Regulation; Hydroxyprostaglandin Dehydrogenases; Intramolecular Oxidoreductases; Isoenzymes; Luteal Cells; Luteinizing Hormone; Lysophospholipids; Organic Anion Transporters; Prostaglandin-E Synthases; Receptors, Prostaglandin E, EP2 Subtype; Receptors, Prostaglandin E, EP4 Subtype; Signal Transduction

Lysophosphatidic acid (LPA) together with its active G protein-coupled receptors are present in the corpus luteum (CL) of the cow. Under in vivo conditions, LPA stimulated P4 and PGE2 secretion during the luteal phase of the estrous cycle in heifers. Furthermore, LPA maintained P4 synthesis and actions in the bovine CL in vitro. However, the effect of this phospholipid on nitric oxide (NO)-induced functional and structural luteolysis has not been investigated. The aim of the present work was to determine the effects of LPA on 1) NO-induced functional luteolysis, 2) NO-dependent PG synthesis, and 3) NO-induced structural luteolysis in cultured steroidogenic luteal cells. We documented that LPA reversed the inhibitory effect of NONOate, an NO donor, on P4 synthesis and PGE2/PGF2alpha ratio in cultured steroidogenic luteal cells. Additionally, LPA inhibited NO-induced apoptosis in cultured steroidogenic luteal cells via abrogation of the NO-dependent stimulatory influence on proapoptotic TNFalpha/TNFR1 and Fas/FasL expression, Caspase 3 activity, and the Bax/Bcl2 ratio during luteal regression in the bovine CL. In conclusion, this study proves that in the presence of LPA, NO cannot induce luteolytic capacity acquisition, leading to functional and structural luteolysis of bovine luteal cells.

Topics: Animals; Apoptosis; Cattle; Cells, Cultured; Dinoprost; Estrous Cycle; Female; Gonadal Steroid Hormones; Luteal Cells; Luteolysis; Lysophospholipids; Nitric Oxide; Nitric Oxide Donors

The bovine corpus luteum (CL) is a transient gland with a life span of only 18 days in the cyclic cow. Mechanisms controlling CL development and secretory function may involve factors produced both within and outside this gland. Although luteinizing hormone (LH) surge is the main trigger of ovulation and granulosa cells luteinization, many locally produced agents such as arachidonic acid (AA) metabolites, growth factors and cytokines were shown to complement gonadotropins action in the process of CL development. Bovine CL is a highly vascular gland, where the very rapid angiogenesis rate (until Day 5 of the cycle) results in the development of a capillary network, endowing this gland with one of the highest blood flow rate per unit mass in the body. Angiogenesis in the developing CL is later followed by either controlled regression of the microvascular tree in the non-fertile cycle or maintenance and stabilization of the blood vessels, as seen during pregnancy. Different luteal cell types (both steroidogenic and accessory luteal cells: immune cells, endothelial cells, pericytes and fibroblasts) are involved in the pro- and/or anti-angiogenic responses. The balance between pro- and anti-angiogenic responses to the main luteolysin - prostaglandin F2α (PGF2α) could be decisive in whether or not PGF2α induces CL regression. Fibroblast growth factor-2 (FGF2) may be one of the factors that modulate the angiogenic response to PGF2α. Manipulation of local production and action of FGF2 will provide new tools for reproductive management of dairy cattle. Luteolysis is characterized by a rapid decrease in progesterone production, followed by structural regression. Factors like endothelin-1, cytokines (tumour necrosis factorα, interferons) and nitric oxide were all shown to play critical roles in functional and structural regression of the CL by inhibiting steroidogenesis and inducting apoptosis.

Topics: Animals; Cattle; Corpus Luteum; Cytokines; Dinoprost; Endothelin-2; Female; Gonadal Steroid Hormones; Leukotrienes; Luteinizing Hormone; Luteolysis; Lysophospholipids; Neovascularization, Physiologic; Prostaglandins; Vascular Endothelial Growth Factor A

Lysophosphatidic acid (LPA) is becoming a new player in regulation of the reproductive processes of domestic animals. In the present study, we examined whether LPA modulates prostaglandin (PG) synthesis in the bovine endometrium at the time of the early maternal pregnancy recognition compared with the respective days of the estrous cycle and the enzymatic mechanism of this action. Bovine epithelial and stromal endometrial cells isolated from the uteri on days 8-10 of the estrous cycle and pregnancy were cultured with LPA for 24 h. LPA increased PGE(2) production in stromal cells during the estrous cycle and early pregnancy. On days 8-10 of pregnancy, LPA inhibited PGF(2alpha) production in epithelial cells. LPA stimulated PGES mRNA expression in stromal cells during both examined phases and inhibited PGFS mRNA expression in epithelial cells on days 8-10 of pregnancy. The overall results indicate that LPA may serve as a luteotropic factor during the luteal phase of the estrous cycle and early pregnancy stimulating PGE(2) synthesis and mRNA expression for PGES in stromal cells. Moreover, during early pregnancy, LPA might protect bovine CL and early embryo development by decreasing PGF(2alpha) synthesis and mRNA expression for PGFS in the epithelial cells of the bovine endometrium.

Topics: Animals; Cattle; Dinoprost; Dinoprostone; Endometrium; Estrous Cycle; Female; Gene Expression Regulation; Lysophospholipids; Models, Biological; Pregnancy; Pregnancy, Animal; RNA, Messenger; Time Factors; Uterus

Lysophosphatidic acid (LPA) has been shown to be a potent modulator of prostaglandin (PG) secretion during the luteal phase of the estrous cycle in the bovine endometrium in vivo. The aims of the present study were to determine the cell types of the bovine endometrium (epithelial or stromal cells) responsible for the secretion of PGs in response to LPA, the cellular, receptor, intracellular, and enzymatic mechanisms of LPA action. Cultured bovine epithelial and stromal cells were exposed to LPA (10(-5)-10(-9) M), tumor necrosis factor alpha (TNFalpha; 10 ng/mL) or oxytocin (OT; 10(-7) M) for 24 h. LPA treatment resulted in a dose-dependent increase of PGE(2) production in stromal cells, but not in epithelial cells. LPA did not influence PGF(2alpha) production in stromal or epithelial cells. To examine which type of LPA G-protein-coupled receptor (LP-GPCR; LPA1, LPA2, or LPA3) is responsible for LPA action, stromal cells were preincubated with three selected blockers of LPA receptors: NAEPA, DGPP, and Ki16425 for 0.5 h, and then stimulated with LPA. Only Ki16425 inhibited the stimulatory effect of LPA on PGE(2) production and cell proliferation in the stromal cells. LPA-induced intracellular calcium ion mobilization was also inhibited only by Ki16425. Finally, we examined whether LPA-induced PGE(2) synthesis in stromal cells is via the influence on mRNA expression for the enzymes responsible for PGE(2) synthesis-PGE(2) synthase (PGES) and PG-endoperoxide synthase 2 (PTGS2). We demonstrated that the stimulatory effect of LPA on PGE(2) production in stromal cells is via the stimulation of PTGS2 and PGES mRNA expression in the cells. The overall results indicate that LPA stimulates PGE(2) production, cell viability, and intracellular calcium ion mobilization in cultured stromal endometrial cells via Ki16425-sensitive LPA1 receptors. Moreover, LPA exerts a stimulatory effect on PGE(2) production in stromal cells via the induction of PTGS2 and PGES mRNA expression.

Topics: Animals; Calcium Signaling; Cattle; Cell Survival; Cells, Cultured; Cyclooxygenase 2; Dinoprost; Dinoprostone; Diphosphates; Endometrium; Epithelial Cells; Female; Glycerol; Intracellular Space; Intramolecular Oxidoreductases; Isoxazoles; Lysophospholipids; Propionates; Prostaglandin-E Synthases; Receptors, Lysophosphatidic Acid; RNA, Messenger; Stromal Cells

Recent data suggest that G protein-coupled receptors (GPCRs), including those for PTH and prostaglandins (PGs), contribute to the proliferation and differentiation of osteoblasts in vivo. To understand how these signals are transduced, we studied activation of the ERK1/2 MAPK cascade in cultures of differentiating TMOb murine osteoblasts. In TMOb cells, stimulation of endogenous Gs/Gq-coupled PTH receptors, Gq-coupled PGF2 alpha receptors, and Gi/Gq-coupled lysophosphatidic acid receptors, but not Gs-coupled PGE2 receptors, caused a rapid 5- to 10-fold increase in ERK1/2 phosphorylation. GPCR-stimulated ERK1/2 activation coincided with increased tyrosine phosphorylation of epidermal growth factor (EGF) receptors and was blocked by the EGF receptor inhibitor, tyrphostin AG1478, and the metalloprotease inhibitor, batimastat, suggesting that the response involved transactivation of EGF receptors through the proteolytic release of an EGF receptor ligand. To further examine the mechanism of PTH-stimulated EGF receptor transactivation, we employed COS-7 cells expressing the rat PTH receptor. Here, stimulation with PTH(1-34) caused proteolysis of hemagglutinin epitope-tagged heparin binding-EGF, increased tyrosine autophosphorylation of EGF receptors, and AG1478-sensitive ERK1/2 activation. When PTH receptor-expressing COS-7 cells were placed in a mixed culture with cells lacking the PTH receptor but expressing a green fluorescent protein-tagged ERK2, stimulation with PTH(1-34) induced phosphorylation of green fluorescent protein-ERK2 that was abolished by either batimastat or tyrphostin AG1478. These data suggest that autocrine/paracrine cross-talk between EGF receptors and Gi- or Gq/11-coupled GPCRs represents the predominant mechanism of GPCR-mediated activation of ERK1/2 in cultured TMOb osteoblasts.

Topics: Animals; Cell Differentiation; Cells, Cultured; Cyclic AMP-Dependent Protein Kinases; Dinoprost; Enzyme Activation; Epidermal Growth Factor; ErbB Receptors; Lysophospholipids; Mice; Mice, Transgenic; Mitogen-Activated Protein Kinase 1; Mitogen-Activated Protein Kinase 3; Mitogen-Activated Protein Kinases; Osteoblasts; Parathyroid Hormone; Phosphorylation; Protein Kinase C; Quinazolines; Rats; Receptors, G-Protein-Coupled; Receptors, Lysophosphatidic Acid; Receptors, Prostaglandin; Signal Transduction; Transcriptional Activation; Tyrphostins