Primate Granulosa Cell Response via Prostaglandin E2 Receptors Increases Late in the Periovulatory Interval1

Markosyan, Nune; Dozier, Brandy L.; Lattanzio, Frank A.; Duffy, Diane M.

doi:10.1095/biolreprod.106.053769

Cited by 22 publications

(36 citation statements)

References 48 publications

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“…hCG treatment in vitro increased progesterone at all times as expected; the addition of fluprostenol with hCG did not alter progesterone when compared to hCG only. Moreover, fluprostenol did not alter media cAMP (not shown) or intracellular calcium when determined as previously described ((Markosyan et al 2006), not shown). Overall, PTGFRs in monkey granulosa cells were not localized to any specific region of the cell, and a selective PTGFR agonist did not alter progesterone production by granulosa cells obtained after the ovulatory stimulus.…”

Section: Resultssupporting

confidence: 65%

“…The cells were cultured on fibronectin-coated surfaces in chemically-defined, serum-free DMEM-Ham’s F12 medium containing insulin, transferrin, selenium, aprotinin, and human low-density lipoprotein as previously described (Markosyan et al 2006). The PTGFR agonist fluprostenol (1 μM; Cayman Chemical, Ann Arbor, MI), hCG (20 IU/ml, Sigma), the general cyclooxygenase inhibitor indomethacin (0.1 μM; Sigma), the phospholipase C (PLC) inhibitor U73122 (100 μM; Cayman), the protein kinase C (PKC) inhibitor Ro31-3220 (1 μM; CalBiochem, Billerica, MA), the mammalian target of rapamycin (mTOR) inhibitor rapamycin (1 μM; Invitrogen, Rockville, MD), the 3β-hydroxysteroid dehydrogenase inhibitor trilostane (250 ng/ml, Stegram Pharmaceuticals, Sussex, UK), progesterone (0.1 μM; Sigma), the aromatase inhibitor letrozole (4,4-[1,2,3-triazol-lyl-methylene]-bis-benzonitrite; 0.5 μM; Norvartis Pharm AG, Basel, Switzerland), estradiol benzoate (0.1 μM; Sigma), and the estrogen receptor antagonist ICI182,780 (1 μM; Tocris Biosciences, Bristol, UK) were added to cultures as indicated.…”

Section: Methodsmentioning

confidence: 99%

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Estrogen promotes luteolysis by redistributing prostaglandin F2α receptors within primate luteal cells

Kim¹,

Markosyan²,

Pepe³

et al. 2015

Reproduction

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Prostaglandin F2α (PGF2α) has been proposed as a functional luteolysin in primates. However, administration of PGF2α or prostaglandin synthesis inhibitors in vivo both initiate luteolysis. These contradictory findings may reflect changes in PGF2α receptors (PTGFR) or responsiveness to PGF2α at a critical point during the life span of the corpus luteum. The current study addressed this question using ovarian cells and tissues from female cynomolgus monkeys and luteinizing granulosa cells from healthy women undergoing follicle aspiration. PTGFRs were present in the cytoplasm of monkey granulosa cells, while PTGFRs were localized to the perinuclear region of large, granulosa-derived monkey luteal cells by mid-late luteal phase. A PTGFR agonist decreased progesterone production by luteal cells obtained at mid-late and late luteal phases but did not decrease progesterone production by granulosa or luteal cells from younger corpora lutea. These findings are consistent with a role for perinuclear PTGFRs in functional luteolysis. This concept was explored using human luteinizing granulosa cells maintained in vitro as a model for luteal cell differentiation. In these cells, PTGFRs relocated from the cytoplasm to the perinuclear area in an estrogen- and estrogen receptor-dependent manner. Similar to our findings with monkey luteal cells, human luteinizing granulosa cells with perinuclear PTGFRs responded to a PTGFR agonist with decreased progesterone production. These data support the concept that PTGFR stimulation promotes functional luteolysis only when PTGFRs are located in the perinuclear region. Estrogen receptor-mediated relocation of PTGFRs within luteal cells may be a necessary step in the initiation of luteolysis in primates.

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Section: Resultssupporting

confidence: 65%

Section: Methodsmentioning

confidence: 99%

Estrogen promotes luteolysis by redistributing prostaglandin F2α receptors within primate luteal cells

Kim¹,

Markosyan²,

Pepe³

et al. 2015

Reproduction

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“…Briefly, total RNA obtained from granulosa cells was reverse transcribed using an oligo dT-anchor primer. cDNA was amplified by PCR with an PCR anchor and a gene specific forward primer for the EP3 common region (5'-TTATCAGGAGCGGAGAC-3') previously designed in our laboratory (Markosyan et al 2006). To obtain the full-length cDNA sequence of the EP3 common region, conventional PCR was performed using a forward primer 5'-GTAAACGCCGACCTCC-3' and a reverse primer 5'- GCAAAACTTTCGAAGAAGG-3'.…”

Section: Methodsmentioning

confidence: 99%

“…EP3 receptors were not detected in mock-transfected or empty vector-transfected CHO cells by immunocytochemistry using an EP3 antibody, following methods previously described by this laboratory (Markosyan et al 2006). Mock-transfected and empty vector-transfected CHO cells did not respond to PGE2 or sulprostone with alterations in intracellular calcium or cAMP.…”

Section: Methodsmentioning

confidence: 99%

“…EP3 receptors are expressed in mural and cumulus granulosa cells of ovarian follicles, with increased expression after the ovulatory gonadotropin surge (Tsai et al 1996, Calder et al 2001, Markosyan et al 2006, Bridges & Fortune 2007, Harris et al 2011). High EP3 expression in bovine cumulus cells correlates with improved quality of the oocyte and the surrounding cumulus (Calder et al 2001).…”

Section: Introductionmentioning

confidence: 99%

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EP3 receptor isoforms are differentially expressed in subpopulations of primate granulosa cells and couple to unique G-proteins

Kim¹,

Dozier

Kerry

et al. 2013

Reproduction

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Prostaglandin E2 produced within the ovarian follicle is necessary for ovulation. Prostaglandin E2 is recognized by four distinct G-protein coupled receptors. Among them, PTGER3 (also known as EP3) is unique in that mRNA splicing generates multiple isoforms. Each isoform has a distinct amino acid composition in the C-terminal region, which is involved in G-protein coupling. To determine if monkey EP3 isoforms couple to different G-proteins, each EP3 isoform was expressed in Chinese hamster ovary (CHO) cells, and intracellular signals were examined after stimulation with the EP3 agonist sulprostone. Stimulation of EP3 isoform 5 (EP3-5) reduced cyclic adenosine monophosphate (cAMP) in a pertussis toxin-sensitive manner, indicating involvement of Gαi. Stimulation of EP3-9 increased cAMP, which was reduced by the general G-protein inhibitor GDP-β-S, and also increased intracellular calcium, which was reduced by pertussis toxin and GDP-β-S. So, EP3-9 likely couples to both Gαs and a pertussis toxin-sensitive G-protein to regulate intracellular signals. Stimulation of EP3-14 increased cAMP, which was further increased by pertussis toxin, so EP3-14 likely regulates cAMP via multiple G-proteins. Granulosa cell expression of all EP3 isoforms increased in response to an ovulatory dose of hCG. Two EP3 isoforms were differentially expressed in functional subpopulations of granulosa cells. EP3-5 was low in granulosa cells at the follicle apex while EP3-9 was high in cumulus granulosa cells. Differential expression of EP3 isoforms may yield different intracellular responses to prostaglandin E2 in granulosa cell subpopulations, contributing to the different roles played by granulosa cell subpopulations in the process of ovulation.

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Gonadotropin‐dependent regulation of the prostaglandin E2 receptor in equine preovulatory follicles during the ovulatory process in mares

Sayasith

Bouchard

Doré

et al. 2008

Molecular Reproduction Devel

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The objectives of the study were to clone the primary structure of the prostaglandin E2 receptor subtype 2 (PTGER2) cDNA and to characterize its regulation in equine follicles during gonadotropin-induced ovulation. Results from DNA isolation indicated that the equine PTGER2 cDNA encodes a predicted 353-amino acid protein, which is highly similar (76-85%) to known mammalian homologues. The regulation of PTGER2 was studied by semi-quantitative RT-PCR/Southern blot using preparations of theca interna and mural granulosa cells isolated from equine follicles 0-39 hr post-treatment with human chorionic gonadotropin (hCG). Results indicated that a significant increase of PTGER2 mRNA occurred at 24 and 39 hr post-hCG in granulosa cells, and 30 and 33 hr post-hCG in theca cells (P < 0.05). Immunohistochemical staining and immunoblotting performed on equine follicular samples showed a corresponding increase of PTGER2 protein in both cell types after treatment with hCG. Levels of PTGER2 mRNA were also high in uterus, thymus and spleen, but moderate to low in other tested tissues. In the ovary, the expression of PTGER4 mRNA was observed and predominantly occurred in granulosa cells, with highest abundance of transcripts observed at 12 and 39 hr post-hCG. Thus, this study reports for the first time in mares that the ovulatory process is accompanied by the gonadotropin-dependent up-regulation of PTGER2 and PTGER4, which may in turn regulate PGE2-mediated preovulatory effects.

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Primate Granulosa Cell Response via Prostaglandin E2 Receptors Increases Late in the Periovulatory Interval1

Cited by 22 publications

References 48 publications

Estrogen promotes luteolysis by redistributing prostaglandin F2α receptors within primate luteal cells

Estrogen promotes luteolysis by redistributing prostaglandin F2α receptors within primate luteal cells

EP3 receptor isoforms are differentially expressed in subpopulations of primate granulosa cells and couple to unique G-proteins

Gonadotropin‐dependent regulation of the prostaglandin E2 receptor in equine preovulatory follicles during the ovulatory process in mares

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