Lysophosphatidic Acid Stimulates Prostaglandin E2 Production in Cultured Stromal Endometrial Cells Through LPA1 Receptor

2009 ◽  
Vol 234 (8) ◽  
pp. 986-993 ◽  
Author(s):  
Izabela Woclawek-Potocka ◽  
Katarzyna Kondraciuk ◽  
Dariusz Jan Skarzynski
Keyword(s):  
Epithelial Cells ◽  
Intracellular Calcium ◽  
Mrna Expression ◽  
Lysophosphatidic Acid ◽  
Stromal Cells ◽  
Calcium Ion ◽  
Cell Types ◽  
Endometrial Cells ◽  
Intracellular Calcium Ion

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 α (TNFα; 10 ng/mL) or oxytocin (OT; 10−7 M) for 24 h. LPA treatment resulted in a dose-dependent increase of PGE2 production in stromal cells, but not in epithelial cells. LPA did not influence PGF2α 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 PGE2 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 PGE2 synthesis in stromal cells is via the influence on mRNA expression for the enzymes responsible for PGE2 synthesis— PGE 2 synthase ( PGES) and PG-endoperoxide synthase 2 ( PTGS2). We demonstrated that the stimulatory effect of LPA on PGE2 production in stromal cells is via the stimulation of PTGS2 and PGES mRNA expression in the cells. The overall results indicate that LPA stimulates PGE2 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 PGE2 production in stromal cells via the induction of PTGS2 and PGES mRNA expression.

scholarly journals Splenic Stromal Cells from Aged Mice Produce Higher Levels of IL-6 Compared to Young Mice

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Jihyun Park ◽  
Takuya Miyakawa ◽  
Aya Shiokawa ◽  
Haruyo Nakajima-Adachi ◽  
Masaru Tanokura ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Stromal Cells ◽  
The Elderly ◽  
Cell Types ◽  
Aged Mice ◽  
Chronic Inflammatory Condition ◽  
Inflammatory State ◽  
Principle Objective ◽  
Young Mice

Inflamm-aging indicates the chronic inflammatory state resulting from increased secretion of proinflammatory cytokines and mediators such as IL-6 in the elderly. Our principle objective was to identify cell types that were affected with aging concerning IL-6 secretion in the murine model. We compared IL-6 production in spleen cells from both young and aged mice and isolated several types of cells from spleen and investigated IL-6 mRNA expression and protein production. IL-6 protein productions in cultured stromal cells from aged mice spleen were significantly high compared to young mice upon LPS stimulation. IL-6 mRNA expression level of freshly isolated stromal cells from aged mice was high compared to young mice. Furthermore, stromal cells of aged mice highly expressed IL-6 mRNA after LPS injection in vivo. These results suggest that stromal cells play a role in producing IL-6 in aged mice and imply that they contribute to the chronic inflammatory condition in the elderly.

scholarly journals Regulation of Electromagnetic Perceptive Gene Using Ferromagnetic Particles for the External Control of Calcium Ion Transport

Biomolecules ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 308 ◽  
Author(s):  
Jangsun Hwang ◽  
Yonghyun Choi ◽  
Kyungwoo Lee ◽  
Vijai Krishnan ◽  
Galit Pelled ◽  
...  
Keyword(s):  
Intracellular Calcium ◽  
Cell Fate ◽  
Calcium Ion ◽  
Magnetic Particles ◽  
Unmet Need ◽  
Ion Concentration ◽  
External Control ◽  
Intracellular Calcium Ion ◽  
Ferromagnetic Particles

Developing synthetic biological devices to allow the noninvasive control of cell fate and function, in vivo can potentially revolutionize the field of regenerative medicine. To address this unmet need, we designed an artificial biological “switch” that consists of two parts: (1) the electromagnetic perceptive gene (EPG) and (2) magnetic particles. Our group has recently cloned the EPG from the Kryptopterus bicirrhis (glass catfish). The EPG gene encodes a putative membrane-associated protein that responds to electromagnetic fields (EMFs). This gene’s primary mechanism of action is to raise the intracellular calcium levels or change in flux through EMF stimulation. Here, we developed a system for the remote regulation of [Ca2+]i (i.e., intracellular calcium ion concentration) using streptavidin-coated ferromagnetic particles (FMPs) under a magnetic field. The results demonstrated that the EPG-FMPs can be used as a molecular calcium switch to express target proteins. This technology has the potential for controlled gene expression, drug delivery, and drug developments.

scholarly journals Effect of LH on prostaglandin F2α and prostaglandin E2 secretion by cultured porcine endometrial cells

Reproduction ◽  
2005 ◽  
Vol 130 (1) ◽  
pp. 105-112 ◽  
Author(s):  
Agnieszka Blitek ◽  
Adam J Ziecik
Keyword(s):  
Epithelial Cells ◽  
Prostaglandin E2 ◽  
Stromal Cells ◽  
Prostaglandin F2α ◽  
Cell Types ◽  
Oestrous Cycle ◽  
Time Dependent ◽  
Dependent Effect ◽  
Endometrial Cells ◽  
Time Dependent Effect

LH appears to be a potent stimulator of the release of endometrial prostaglandins (PGs) in the pig. The aim of the present studies was to examine the effect of LH on PGF2αand PGE2secretion by cultured porcine endometrial cells on days 10–12 and 14–16 of the oestrous cycle and to compare its action with oxytocin. A time-dependent effect of LH (10 ng/ml) on PGF2αrelease from luminal epithelial and stromal cells on days 10–12 was observed (experiment 1). The highest increase in PGF2αsecretion in response to LH was detected in stromal cells after 6 h of incubation (P< 0.001). Epithelial cells responded to LH after a longer exposure time (P< 0.01). A concentration-dependent effect of LH (0.1–100 ng/ml) on PGF2αrelease from stromal cells was examined after 6 h and from epithelial cells after 12 h (experiment 2). Effective concentrations of LH were 10 and 100 ng/ml. LH (10 ng/ml) and oxytocin (100 nmol/l) affected PGF2αand PGE2secretion from endometrial cells on days 10–12 and 14–16 of the oestrous cycle (experiment 3). LH stimulated PGF2αsecretion from both cell types and its action was more potent on days 10–12. LH induced PGE2release, especially in epithelial cells on days 14–16. A stimulatory effect of oxytocin on PGF2αwas confirmed in stromal cells, but this hormone was also shown to enhance PGE2output. These results indicated that LH, like oxytocin, a very effective stimulator of PGF2αrelease, could play an important role in the induction of luteolysis.

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

Reproduction ◽  
2001 ◽  
pp. 605-612 ◽  
Author(s):  
GT Braileanu ◽  
SM Simasko ◽  
J Hu ◽  
A Assiri ◽  
MA Mirando
Keyword(s):  
Epithelial Cells ◽  
Intracellular Calcium ◽  
Phospholipase C ◽  
Arginine Vasopressin ◽  
Stromal Cells ◽  
Calcium Concentration ◽  
Intracellular Calcium Concentration ◽  
Endometrial Cells ◽  
Lysine Vasopressin ◽  
Luminal Epithelial 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.

scholarly journals Genomic and non-genomic effects of progesterone on prostaglandin (PG) F2α and PGE2 production in the bovine endometrium

2016 ◽  
Vol 28 (10) ◽  
pp. 1588 ◽  
Author(s):  
Mariko Kuse ◽  
Ryosuke Sakumoto ◽  
Kiyoshi Okuda
Keyword(s):  
Epithelial Cells ◽  
Mrna Expression ◽  
Stromal Cells ◽  
Follicular Phase ◽  
Pge2 Production ◽  
Cell Type ◽  
Short Term ◽  
Endometrial Cells ◽  
Receptor Mrna ◽  
Short Time

Progesterone (P4) acts through different actuating pathways called genomic and non-genomic pathways. Here we investigated whether P4 regulates prostaglandin (PG) F2α (PGF) and PGE2 production in bovine endometrium through different pathways. Cultured endometrial cells were exposed to P4 for a short time (5–20 min) or bovine serum albumin (BSA)-conjugated P4 (P4-BSA) for 24 h. Progesterone treatment for 24 h stimulated PGE2 production in epithelial cells, but suppressed both PGF and PGE2 production and the expression of PG-metabolising enzymes including phospholipase A2 (PLA2) and cyclooxygenase-2 (COX2) in stromal cells. Short-term (5–20 min) P4 treatment did not affect PLA2 or COX2 transcript levels in either cell type. P4-BSA increased PGF and PGE2 production only in epithelial cells. Nuclear P4 receptor mRNA expression in endometrium was higher at the follicular phase than at the early- to mid-luteal stages, whereas membrane P4 receptor mRNA expression did not change throughout the oestrous cycle. The overall results suggest that P4 controls PG production by inhibiting enzymes via a genomic pathway and by stimulating signal transduction via a non-genomic pathway. Consequently, P4 may protect the corpus luteum by attenuating PGF production in stromal cells and by increasing PGE2 secretion from epithelial cells.

Increased mRNA expression of selected pro-inflammatory factors in inflamed bovine endometrium in vivo as well as in endometrial epithelial cells exposed to Bacillus pumilus in vitro

2016 ◽  
Vol 28 (7) ◽  
pp. 982 ◽  
Author(s):  
Martina A. Gärtner ◽  
Sarah Peter ◽  
Markus Jung ◽  
Marc Drillich ◽  
Ralf Einspanier ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Mrna Expression ◽  
Bacillus Pumilus ◽  
Cultured Cells ◽  
Inflammatory Factors ◽  
Endometrial Cells ◽  
Subclinical Endometritis ◽  
Endometrial Epithelial Cells

Endometrial epithelium plays a crucial role in the first immune response to invading bacteria by producing cytokines and chemokines. The aim of this study was to investigate the first inflammatory response of the endometrium in vivo and in vitro. Gene expression of several pro-inflammatory factors and Toll-like receptors (TLR2, -4, -6) was determined in endometrial cytobrush samples obtained from healthy cows and cows with clinical or subclinical endometritis. Endometrial epithelial cells were co-cultured with an isolated autochthonous uterine bacterial strain Bacillus pumilus. Total RNA was extracted from in vivo and in vitro samples and subjected to real-time reverse transcription polymerase chain reaction. CXC ligands (CXCL) 1/2 and CXC chemokine receptor (CXCR) 2 mRNA expression was higher in cows with subclinical endometritis and CXCL3 mRNA expression was higher in cows with clinical endometritis compared with healthy cows. B. pumilus induced cell death of epithelial cells within 24 h of co-culturing. The presence of B. pumilus resulted in significantly higher mRNA expression of interleukin 1α (IL1A), IL6, IL8, CXCL1–3 and prostaglandin–endoperoxide synthase 2 in co-cultured cells compared with untreated controls. The maximum increase was mainly detected after 2 h. These results support the hypothesis that bacterial infection of endometrial cells might induce prompt synthesis of pro-inflammatory cytokines resulting in a local inflammatory reaction.

scholarly journals Phyto- and Endogenous Estrogens Differently Activate Intracellular Calcium Ion Mobilization in Bovine Endometrial Cells

2006 ◽  
Vol 52 (6) ◽  
pp. 731-740 ◽  
Author(s):  
Izabela WOCLAWEK-POTOCKA ◽  
Krzysztof BORKOWSKI ◽  
Anna KORZEKWA ◽  
Kiyoshi OKUDA ◽  
Dariusz J. SKARZYNSKI
Keyword(s):  
Intracellular Calcium ◽  
Calcium Ion ◽  
Endometrial Cells ◽  
Intracellular Calcium Ion

Time Series Analysis of Transmesothelial Invasion by Endometrial Stromal and Epithelial Cells Using Three-dimensional Confocal Microscopy

2001 ◽  
Vol 7 (S2) ◽  
pp. 580-581
Author(s):  
CA Witz ◽  
S Cho ◽  
VE Centonze ◽  
IA Montoya-Rodriguez ◽  
RS Schenken
Keyword(s):  
Epithelial Cells ◽  
Stromal Cells ◽  
Time Course ◽  
Three Dimensional ◽  
Collagen Iv ◽  
Mesothelial Cells ◽  
Endometrial Stromal Cells ◽  
Human Collagen ◽  
Endometrial Epithelial Cells

Using human peritoneal explants, we have previously demonstrated that endometrial stromal cells (ESCs) and endometrial epithelial cells (EECs) attach to intact mesothelium. Attachment occurs within one hour and mesothelial invasion occurs within 18 hours (Figure 1). We have also demonstrated that, in vivo, the mesothelium overlies a continuous layer of collagen IV (Col IV).More recently we have used CLSM, to study the mechanism and time course of ESC and EEC attachment and invasion through mesothelial monolayers. in these studies, CellTracker® dyes were used to label cells. Mesothelial cells were labeled with chloromethylbenzoylaminotetramethylrhodamine (CellTracker Orange). Mesothelial cells were then plated on human collagen IV coated, laser etched coverslips. Mesothelial cells were cultured to subconfluence. ESCs and EECs, labeled with chloromethylfluorscein diacetate (CellTracker Green) were plated on the mesothelial monolayers. Cultures were examined at 1, 6, 12 and 24 hours with simultaneous differential interference contrast and CLSM.

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