Cloning, functional expression, and characterization of the human prostaglandin E2 receptor EP2 subtype.

Bastien, Lison; Sawyer, Nicole; Grygorczyk, Ryszard; Metters, Kathleen M.; Adam, Mohammed

doi:10.1016/s0021-9258(17)32654-6

Cited by 221 publications

(20 citation statements)

References 20 publications

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“…Using EP 2 -specific primers, we amplified an EP 2 receptor fragment from the ovine fetal DA that had 90, 87, and 85% homology with the corresponding se-quences from the human, mouse, and rat EP 2 receptors, respectively (4,21,35). Using EP 4 specific primers, we also amplified an EP 4 fragment, which was virtually identical to the reported ovine EP 4 receptor sequence (GenBank accession AF035418) and had 94% homology with the corresponding sequence of the human EP 4 receptor (29).…”

Section: Competitive Displacement Of [ 3 H]pgementioning

confidence: 99%

Characterization of PGE₂receptors in fetal and newborn lamb ductus arteriosus

Bouayad

Kajino

Waleh

et al. 2001

American Journal of Physiology-Heart and Circulatory Physiology

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Although the role of PGE2 in maintaining ductus arteriosus (DA) patency is well established, the specific PGE2 receptor subtype(s) (EP) involved have not been clearly identified. We used late gestation fetal and neonatal lambs to study developmental regulation of EP receptors. In the fetal DA, radioligand binding and RT-PCR assays virtually failed to detect EP1 but detected EP2, EP3D, and EP4 receptors in equivalent proportions. In the newborn lamb, DA total density was one-third of that found in the fetus and only EP2 was detected. Stimulation of EP2 and EP4 increased cAMP formation and was associated with DA relaxation. Though stimulation of EP3 inhibited cAMP formation, it surprisingly relaxed the fetal DA both in vitro and in vivo. This EP3-induced relaxation was specifically diminished by the ATP-sensitive K(+) (K(ATP)) channel blocker glibenclamide. In conclusion, PGE2 dilates the late gestation fetal DA through pathways that involve either cAMP (EP2 and EP4) or K(ATP) channels (EP3). The loss of EP3 and EP4 receptors in the newborn DA is consistent with its decreased responsiveness to PGE2.

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Section: Competitive Displacement Of [ 3 H]pgementioning

confidence: 99%

Characterization of PGE₂receptors in fetal and newborn lamb ductus arteriosus

Bouayad

Kajino

Waleh

et al. 2001

American Journal of Physiology-Heart and Circulatory Physiology

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“…After extraction of total RNA from HGF, cDNAs were synthesized from 2 gg of total RNA with RAV2 reversetranscriptase and oligo(dT)primers (Takara Co., Shiga, Japan), as previously described (Kimmel and Berger, 1987). The specific primer pairs for human EP1, EP2, EP3, EP4, and B-actin were selected, according to the cDNA sequences reported by Funk et al (1993), Regan et al (1994), Adam et al (1994), Bastien et al (1994), and Ponte et al (1984), respectively. The primers are: EP1 sense primer, 5 '-TCTACCTCCCTGCAGCGGCCACTG-3', antisense primer, 5'-GAAGTGGCTGAGGCCGCTGTGC-CGGGA-3'; EP2 sense primer, 5'-TTCATCCGGCACGGGC-GGACCGC-3', antisense primer, 5 '-GTCAGCCTGTTTACTG-GCATCTG-3'; EP3 sense primer, 5'-GAGCACTGCAAGACA-CACACGGAG-3', antisense primer, 5'-GATCTCCCATGGG-TATTACTGACAA-3'; EP4 sense primer, 5'-CCTCCTGAGAA-AGACAGTGTC-3', antisense primer, 5'-AGGACTCAGA-GAGTGTCTT-3'; and B-actin sense primer, 5'-GTGGGCA-TGGGTCATCAGAAGGAT-3 ', antisense primer, 5'-CTCCT-TAATGTCACGCACGATTTC-3'.…”

Section: Cell Stimulationmentioning

confidence: 99%

Cyclooxygenase-2-dependent Prostaglandin E2 Down-regulates Intercellular Adhesion Molecule-1 Expression via EP2/EP4 Receptors in Interleukin-1ß-stimulated Human Gingival Fibroblasts

et al. 2000

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Prostaglandin E2 (PGE2), which exerts its actions via EP receptors (EP1, EP2, EP3, and EP4), is a bioactive metabolite of arachidonic acid produced by cyclooxygenase (COX)-1 and/or COX-2. We have previously demonstrated that PGE2 down-regulates intercellular adhesion molecule-1 (ICAM-1) expression in interleukin-1beta (IL-1beta)-stimulated human gingival fibroblasts (HGF). In the present study, we investigated which COX was involved in down-regulation of ICAM-1 expression by PGE2 in IL-1beta-stimulated HGF and which subtypes of EP receptors modulated the ICAM-1 expression. NS-398, a specific COX-2 inhibitor, completely inhibited PGE2 production by IL-1beta-stimulated HGF, as did indomethacin, a COX-1/COX-2 inhibitor. Northern blot analysis and immunocytochemical staining showed that mRNA and protein of COX-2 were expressed in IL-1beta-challenged HGF, but not in unstimulated HGF, and that the expression of mRNA and protein of COX-1 was similar both in unstimulated and in stimulated cells. NS-398 and indomethacin enhanced ICAM-1 expression in IL-1beta-challenged HGF. EP1, EP2, and EP4 receptor mRNA was expressed in HGF according to reverse-transcription/polymerase chain-reaction. PGE2, 11-deoxy-PGE1 (a selective EP2/EP4 agonist), and Butaprost (a selective EP2 agonist) attenuated IL-1beta-elicited ICAM-1 expression, although Butaprost was less potent than PGE2 and 11-deoxy-PGE1. AH-23848B, an EP4 antagonist, antagonized the inhibitory effect of IL-1beta-elicited ICAM-1 expression by PGE2. Sulprostone, an EP1/EP3 agonist, had no effect on IL-1beta-elicited ICAM-1 expression. Analysis of these data suggests that COX-2-derived PGE2 down-regulates ICAM-1 expression via EP2/EP4 receptors in IL-1beta-stimulated HGF.

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“…All EP receptors are heterotrimeric GTP-binding protein (G-protein)-coupled rhodopsin-type receptors (Watabe et al, 1993;Sugimoto et al, 2000). The primary signaling pathways of the four EP receptors are as follows: EP1 couples to elevation of intracellular calcium levels [Ca2+]i (Funk et al, 1993), EP2 and EP4 couple to an increase in intracellular cAMP accumulation (Regan et al, 1994;Bastien et al, 1994), and EP3 couples to a decrease in intracellular cAMP concentrations (Boie et al, 1997).…”

Section: Introductionmentioning

confidence: 99%

Regional Distribution of the Prostaglandin E<SUB>2</SUB> Receptor EP1 in the Rat Brain: Accumulation in Purkinje Cells of the Cerebellum

Candelario‐Jalil

Slawik

Ridelis

et al. 2005

JMN

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Prostaglandin E2 (PGE2), is a major prostanoid produced by the activity of cyclooxygenases (COX) in response to various physiological and pathological stimuli. PGE2 exerts its effects by activating four specific E-type prostanoid receptors (EP1, EP2, EP3, and EP4). In the present study, we analyzed the expression of the PGE2 receptor EP1 (mRNA and protein) in different regions of the adult rat brain (hippocampus, hypothalamus, striatum, prefrontal cerebral cortex, parietal cortex, brain stem, and cerebellum) using reverse transcription- polymerase chain reaction, Western blotting, and immunohistochemical methods. On a regional basis, levels of EP1 mRNA were the highest in parietal cortex and cerebellum. At the protein level, we found very strong expression of EP1 in cerebellum, as revealed by Western blotting experiments. Furthermore, the present study provides for the first time evidence that the EP1 receptor is highly expressed in the cerebellum, where the Purkinje cells displayed very high immunolabeling of their perikaryon and dendrites, as observed in the immunohistochemical analysis. Results from the present study indicate that the EP1 prostanoid receptor is expressed in specific neuronal populations, which possibly determine the region-specific response to PGE2.

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Cloning, functional expression, and characterization of the human prostaglandin E2 receptor EP2 subtype.

Cited by 221 publications

References 20 publications

Characterization of PGE₂receptors in fetal and newborn lamb ductus arteriosus

Characterization of PGE₂receptors in fetal and newborn lamb ductus arteriosus

Cyclooxygenase-2-dependent Prostaglandin E2 Down-regulates Intercellular Adhesion Molecule-1 Expression via EP2/EP4 Receptors in Interleukin-1ß-stimulated Human Gingival Fibroblasts

Regional Distribution of the Prostaglandin E<SUB>2</SUB> Receptor EP1 in the Rat Brain: Accumulation in Purkinje Cells of the Cerebellum

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Cloning, functional expression, and characterization of the human prostaglandin E2 receptor EP2 subtype.

Cited by 221 publications

References 20 publications

Characterization of PGE2receptors in fetal and newborn lamb ductus arteriosus

Characterization of PGE2receptors in fetal and newborn lamb ductus arteriosus

Cyclooxygenase-2-dependent Prostaglandin E2 Down-regulates Intercellular Adhesion Molecule-1 Expression via EP2/EP4 Receptors in Interleukin-1ß-stimulated Human Gingival Fibroblasts

Regional Distribution of the Prostaglandin E<SUB>2</SUB> Receptor EP1 in the Rat Brain: Accumulation in Purkinje Cells of the Cerebellum

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Characterization of PGE₂receptors in fetal and newborn lamb ductus arteriosus

Characterization of PGE₂receptors in fetal and newborn lamb ductus arteriosus