Mouse oocytes suppress cAMP-induced expression of LH receptor mRNA by granulosa cells in vitro

1998 ◽  
Vol 49 (3) ◽  
pp. 327-332 ◽  
Author(s):  
John J. Eppig ◽  
Frank L. Pendola ◽  
Karen Wigglesworth
Keyword(s):  
Granulosa Cells ◽  
Induced Expression ◽  
Lh Receptor ◽  
Receptor Mrna ◽  
Mouse Oocytes

scholarly journals Differential effects of prostaglandin F2alpha on in vitro luteinized bovine granulosa cells

Reproduction ◽  
2001 ◽  
pp. 245-253 ◽  
Author(s):  
SJ Tsai ◽  
MC Wiltbank
Keyword(s):  
Protein Kinase ◽  
Protein Kinase A ◽  
Granulosa Cells ◽  
Ovarian Function ◽  
Prostaglandin F2alpha ◽  
Induced Expression ◽  
Receptor Mrna ◽  
Kinase A

Prostaglandins have been implicated in various aspects of ovarian function including ovulation and luteolysis. In this study, the expression and regulation of inducible prostagland in G/H synthase (PGHS-2) and PGF(2alpha) receptors were investigated in bovine granulosa cells at various stages of differentiation. Firstly, the induction of PGF(2alpha) receptor mRNA and PGHS-2 mRNA in preovulatory granulosa cells was evaluated. Granulosa cells were collected from preovulatory follicles and cultured for 1, 4, 7 or 10 days. Cells were treated with hCG (10 iu) or with increasing doses of forskolin (0-10 micromol l(-1)) for 24 h. Forskolin increased steady-state concentrations of mRNA for PGHS-2 (> 20-fold) and PGF(2alpha) receptor (> 1000-fold) in a dose-dependent fashion. Use of selective protein kinase A inhibitor (H89) reduced both hCG- and forskolin-induced expression of PGF(2alpha) receptor mRNA and PGHS-2 mRNA. The hypothesis that luteinized granulosa cells would acquire PGF(2alpha) responsiveness similar to responses to PGF(2alpha) observed in vivo was also evaluated. Treatment with PGF(2alpha) (100 nmol l(-1)) reduced forskolin-induced expression of PGF(2alpha) receptor mRNA on days 4, 7 and 10, but not on day 1 of culture (n = 3). Treatment with PGF(2alpha) did not change forskolin-induced expression of PGHS-2 mRNA on or before day 4 of culture. In contrast, PGF(2alpha) significantly increased PGHS-2 mRNA expression in granulosa cells primed with forskolin for 7 or 10 days. In conclusion, expression of PGHS-2 and PGF(2alpha) receptor mRNA is protein kinase A-dependent in preovulatory bovine granulosa cells. Granulosa cells become PGF(2alpha)-responsive soon after expression of PGF(2alpha) receptor, whereas further differentiation is required before PGF(2alpha) induces PGHS-2 mRNA upregulation. These results demonstrate that at least two key transitions are required in PGF(2alpha)-induced luteal regression in the mid-cycle corpus luteum.

Expression of LH receptor mRNA splice variants in bovine granulosa cells: changes with follicle size and regulation by FSH in vitro

2007 ◽  
Vol 74 (6) ◽  
pp. 680-686 ◽  
Author(s):  
M.F.G. Nogueira ◽  
J. Buratini ◽  
C.A. Price ◽  
A.C.S. Castilho ◽  
M.G.L. Pinto ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Splice Variants ◽  
Lh Receptor ◽  
Mrna Splice ◽  
Receptor Mrna ◽  
Follicle Size

scholarly journals Gonadotropin-Induced Expression of Messenger Ribonucleic Acid for Cyclooxygenase-2 and Production of Prostaglandins E and F2α in Bovine Preovulatory Follicles Are Regulated by the Progesterone Receptor

Endocrinology ◽  
2006 ◽  
Vol 147 (10) ◽  
pp. 4713-4722 ◽  
Author(s):  
P. J. Bridges ◽  
C. M. Komar ◽  
J. E. Fortune
Keyword(s):  
Granulosa Cells ◽  
Medroxyprogesterone Acetate ◽  
Cyclooxygenase 2 ◽  
Induced Expression ◽  
Messenger Ribonucleic Acid ◽  
Preovulatory Follicles ◽  
Gonadotropin Surge ◽  
Induced Changes

Follicular production of prostaglandins (PGs) is essential for ovulation, but the factors mediating gonadotropin-induced secretion of PGE and PGF2α remain largely unknown. We tested the hypothesis that gonadotropin-induced changes in progesterone and its receptor (PR) mediate the increase in periovulatory PGs. Heifers were treated with PGF2α and GnRH to induce luteolysis and the LH/FSH surge (ovulation occurs ∼30 h after GnRH). Because there are two increases in intrafollicular progesterone/PR mRNA during the bovine periovulatory period, we first examined the temporal pattern of PG production by follicles collected at 0, 3.5, 6, 12, 18, and 24 h after GnRH. Although PGs did not increase in the follicular fluid until 24 h after GnRH, acute secretion of PGs by follicle wall (theca + granulosa cells) was initiated by 18 h and had increased manyfold by 24 h after GnRH. In vitro, FSH and LH induced dramatic transient increases in PG production by follicle wall and granulosa, but not theca, cells isolated from preovulatory follicles (0 h after GnRH). PG accumulation peaked on d 2 of culture, mimicking the secretion pattern after a gonadotropin surge in vivo. In cultures of follicle wall and granulosa cells, the PR antagonist mifepristone (MIFE, 1 μm) inhibited LH-induced PG secretion and the progestin medroxyprogesterone acetate (1 or 10 μm), but not the glucocorticoid dexamethasone (1 or 10 μm), overcame the effect of MIFE on PGs. Semiquantitative RT-PCR revealed that MIFE inhibited LH-induced expression of cyclooxygenase-2 mRNA in granulosa cells in vitro. Again, treatment with medroxyprogesterone acetate overcame the effect of MIFE. Together these results provide strong evidence that periovulatory increases in cyclooxygenase-2 mRNA, PGE, and PGF2α are mediated by gonadotropin-induced increases in progesterone/PR, indicating that in some species there is an important functional relationship between these pathways in the ovulatory cascade.

Cell-specific expression of aromatase and LH receptor mRNAs in rat ovary

1992 ◽  
Vol 9 (3) ◽  
pp. 309-312 ◽  
Author(s):  
P.F. Whitelaw ◽  
C.D. Smyth ◽  
C.M. Howles ◽  
S.G. Hillier
Keyword(s):  
Cytochrome P450 ◽  
Granulosa Cells ◽  
Direct Evidence ◽  
Specific Expression ◽  
Lh Receptor ◽  
Paracrine Regulation ◽  
Receptor Mrna ◽  
Rat Ovary ◽  
Per Se ◽  
Receptor Mrnas

ABSTRACT Current understanding of the endocrine and paracrine regulation of follicular oestrogen synthesis predicts that aromatase cytochrome P450 (P450arom) mRNA is inducible by FSH in granulosa cells. LH receptor mRNA is constitutively expressed in thecal/interstital cells, and is also thought to be induced in granulosa cells in response to joint stimulation by FSH and oestrogen. This study provides direct evidence that FSH induces the ovarian P450arom gene selectively, perhaps exclusively, in the granulosa cells of Graafian follicles. FSH-induction of LH receptor mRNA occurs simultaneously but is independent of oestrogen synthesis per se.

scholarly journals Inhibitory effect of retinoic acid on the development of immature porcine granulosa cells to mature cells

2000 ◽  
Vol 25 (1) ◽  
pp. 53-61 ◽  
Author(s):  
M Hattori ◽  
K Takesue ◽  
N Nishida ◽  
Y Kato ◽  
N Fujihara
Keyword(s):  
Retinoic Acid ◽  
Cell Differentiation ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Inhibitory Effect ◽  
Immature Stage ◽  
Mrna Levels ◽  
Lh Receptor ◽  
Receptor Mrna ◽  
Immature Cells

The present study investigated the effect of retinoic acid (RA) on the differentiation of granulosa cells prepared from porcine ovaries. The granulosa cells were precultured for 15 h, then cultured for 48 h with FSH and further treated for 24 h with LH in order to induce their transformation into luteal cells. After the cells had been exposed to 1 microM retinoids (RA, retinal and retinol) for 87 h, analysis of the LH receptor mRNA expression, an indicator of granulosa cell differentiation, was carried out by using semiquantitative RT-PCR. The results showed that there was a decrease in LH receptor mRNA levels, and that RA had a more potent effect on these levels than the other two retinoids. When cells were exposed to RA in the immature stage (before the addition of FSH) or the early stage of development (0-24 h after the addition of FSH), expression of LH receptor mRNA was greatly diminished. When the immature cells were cultured for 15 h with RA, then washed and cultured for 48 h with FSH and for 24 h with LH, the expression of LH receptor mRNA was not reversed. In the differentiated cells (24 h after the addition of FSH), however, RA no longer had any inhibitory effect. When the immature cells were exposed to RA, FSH-induced expression of c-fos mRNA was markedly decreased. In contrast, expression of c-jun and activating transcription factor-4 mRNAs remained constant. However, the expression of c-fos mRNA was not decreased by forskolin. The results indicate that RA is a potent inhibitor in the immature stage of porcine granulosa cell differentiation, probably through decreased expression of FSH receptor, but that RA does not inhibit differentiation in the mature stage of the cells.

scholarly journals Analysis of Ovarian Gene Expression in Follicle-Stimulating Hormone β Knockout Mice*

Endocrinology ◽  
2001 ◽  
Vol 142 (7) ◽  
pp. 2742-2751 ◽  
Author(s):  
Kathleen H. Burns ◽  
Changning Yan ◽  
T. Rajendra Kumar ◽  
Martin M. Matzuk
Keyword(s):  
Cell Cycle ◽  
Granulosa Cell ◽  
Granulosa Cells ◽  
Knockout Mice ◽  
Marker Genes ◽  
Cyclin D2 ◽  
Glycoprotein Hormone ◽  
Lh Receptor ◽  
Receptor Mrna ◽  
Deficient Mice

Abstract FSH is a heterodimeric glycoprotein hormone that is produced in the gonadotroph cells of the anterior pituitary. It acts on Sertoli cells of the testis and granulosa cells of the ovary. We previously demonstrated that FSHβ knockout female mice are infertile due to a block in folliculogenesis preceding antral stage development. To investigate aberrations of ovarian gene regulation in the absence of FSH, we analyzed the expression of several important marker genes using Northern blot and in situ hybridization techniques. Key findings are as follows: 1) Follicles of FSHβ knockout mice develop a well organized thecal layer, which is positive for P450 17α-hydroxylase and LH receptor messenger RNAs (mRNAs). This indicates that theca recruitment is completed autonomously with respect to FSH. 2) Granulosa cells in FSH-deficient mice demonstrate an increase in FSH receptor mRNA, and decreases in P450 aromatase, serum/glucocorticoid-induced kinase, and inhibin/activin subunit mRNAs. These data support studies that implicate FSH signaling cascades in the expression of these genes. 3) In contrast to the thecal layer, granulosa cell populations in FSHβ knockout mice do not accumulate LH receptor mRNA. This suggests that although the granulosa cells have a block in proliferation at the antral follicle stage in the absence of FSH, they do not initiate programs of terminal differentiation as seen in luteinizing cells of wild-type ovaries. 4) Ovaries of FSH-deficient mice demonstrate a modest decrease in cyclin D2 mRNA, without up-regulation of cell cycle inhibitor mRNAs associated with luteinization (i.e. p15, p27, and p21). Although components of the FSH null phenotype may be caused by partial cyclin D2 loss of function, these findings indicate that the mechanisms of granulosa cell cycle arrest in FSHβ knockout mice are distinct from those of cycle withdrawal at luteinization. Underscoring the usefulness of the FSH-deficient mouse model, this study clarifies aspects of gonadotropin-dependent folliculogenesis, thecal layer development, cycle control in granulosa cells, and luteinization.

scholarly journals 21-Hydroxylase-Derived Steroids in Follicles of Nonobese Women Undergoing Ovarian Stimulation for In Vitro Fertilization (IVF) Positively Correlate With Lipid Content of Luteinized Granulosa Cells (LGCs) as a Source of Cholesterol for Steroid Synthesis

2014 ◽  
Vol 99 (4) ◽  
pp. 1299-1306 ◽  
Author(s):  
Marli Amin ◽  
Ariel Simerman ◽  
Michele Cho ◽  
Prapti Singh ◽  
Christine Briton-Jones ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Mrna Expression ◽  
Granulosa Cells ◽  
Lipid Content ◽  
Mineralocorticoid Receptor ◽  
Steroid Synthesis ◽  
Vitro Fertilization ◽  
Receptor Mrna

Context: Mineralocorticoid synthesis by the nonhuman primate periovulatory follicle enhances luteinization. Whether a similar event occurs in women undergoing in vitro fertilization (IVF) is unknown. Objective: The objective of the study was to determine whether human luteinized granulosa cells (LGCs) produce mineralocorticoids derived from 21-hydroxylase activity and also express mRNA for 21-hydroxylase and the mineralocorticoid receptor. Design: This was a prospective cohort study. Setting: The study was conducted at an academic center. Patients: LGC lipid content and follicle fluid (FF) hormone analysis was performed on 27 nonobese IVF women. LGCs from six additional nonobese IVF women were used for gene expression studies. Intervention: At oocyte retrieval, FF was aspirated from the first follicle (≥16 mm in size) of each ovary and pooled LGCs were collected. Main Outcome Measures: FF steroid analysis was performed by liquid chromatography-tandem mass spectrometry. LGCs were stained with lipid fluorescent dye BODIPY FL C16 to estimate lipid content by confocal microscopy as a cholesterol source for steroidogenesis in vivo. Quantitative real-time PCR was performed using LGCs to detect 21-hydroxylase and mineralocorticoid receptor mRNA expression. Pearson correlation coefficients determined associations between FF steroid levels and LGC lipid content. Results: FF levels of the 21-hydroxylase-derived steroids, 11-deoxycorticosterone [DOC, 39.97, median (13.94–63.02) ng/mL] and 11-deoxycortisol [11DOC, 2.07 (0.69–5.01) ng/mL], along with the 21-hydroxylase precursor 17-hydroxyprogesterone [1268.21 (493.26–3558.39) ng/mL], positively correlated with LGC lipid content (84 ± 43 fluorescent units/sample) (P ≤ .05, all steroids). 21-Hydroxylase and mineralocorticoid receptor mRNA expression was detected in LGCs. Conclusions: Human LGCs likely synthesize 21-hydroxylase-derived mineralocorticoids from cholesterol-containing lipid in vivo to promote postovulatory luteinization via mineralocorticoid receptor-mediated events.

scholarly journals Ovulatory Induction of SCG2 in Human, Nonhuman Primate, and Rodent Granulosa Cells Stimulates Ovarian Angiogenesis

Endocrinology ◽  
2018 ◽  
Vol 159 (6) ◽  
pp. 2447-2458 ◽  
Author(s):  
Patrick R Hannon ◽  
Diane M Duffy ◽  
Katherine L Rosewell ◽  
Mats Brännström ◽  
James W Akin ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Messenger Rna ◽  
Growth Factor Receptor ◽  
Endothelial Cell Migration ◽  
Receptor Signaling ◽  
Signaling Mechanism ◽  
Lh Receptor ◽  
Vitro Fertilization

Abstract The luteinizing hormone (LH) surge is essential for ovulation, but the intrafollicular factors induced by LH that mediate ovulatory processes (e.g., angiogenesis) are poorly understood, especially in women. The role of secretogranin II (SCG2) and its cleaved bioactive peptide, secretoneurin (SN), were investigated as potential mediators of ovulation by testing the hypothesis that SCG2/SN is induced in granulosa cells by human chorionic gonadotropin (hCG), via a downstream LH receptor signaling mechanism, and stimulates ovarian angiogenesis. Humans, nonhuman primates, and rodents were treated with hCG in vivo resulting in a significant increase in the messenger RNA and protein levels of SCG2 in granulosa cells collected early during the periovulatory period and just prior to ovulation (humans: 12 to 34 hours; monkeys: 12 to 36 hours; rodents: 4 to 12 hours post-hCG). This induction by hCG was recapitulated in an in vitro culture system utilizing granulosa-lutein cells from in vitro fertilization patients. Using this system, inhibition of downstream LH receptor signaling pathways revealed that the initial induction of SCG2 is regulated, in part, by epidermal growth factor receptor signaling. Further, human ovarian microvascular endothelial cells were treated with SN (1 to 100 ng/mL) and subjected to angiogenesis assays. SN significantly increased endothelial cell migration and new sprout formation, suggesting induction of ovarian angiogenesis. These results establish that SCG2 is increased in granulosa cells across species during the periovulatory period and that SN may mediate ovulatory angiogenesis in the human ovary. These findings provide insight into the regulation of human ovulation and fertility.

scholarly journals Macromolecular crowded conditions strengthen contacts between mouse oocytes and companion granulosa cells during in vitro growth

2018 ◽  
Vol 64 (2) ◽  
pp. 153-160 ◽  
Author(s):  
Shizuka MIZUMACHI ◽  
Taiki ARITOMI ◽  
Kuniaki SASAKI ◽  
Kazuei MATSUBARA ◽  
Yuji HIRAO
Keyword(s):  
Granulosa Cells ◽  
Mouse Oocytes
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