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 Insulin-inducible changes in insulin receptor mRNA splice variants.

1994 ◽  
Vol 269 (49) ◽  
pp. 30769-30772
Author(s):  
S M Sell ◽  
D Reese ◽  
V M Ossowski
Keyword(s):  
Insulin Receptor ◽  
Splice Variants ◽  
Mrna Splice ◽  
Receptor Mrna

Identification of Gαs messenger ribonucleic acid splice variants in human granulosa cells

1997 ◽  
Vol 18 (1) ◽  
pp. 27-35 ◽  
Author(s):  
G N Europe-Finner ◽  
E Cartwright ◽  
J Bellinger ◽  
H J Mardon ◽  
D H Barlow ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Ovarian Function ◽  
Splice Variants ◽  
Consensus Sequence ◽  
Phosphorylation Site ◽  
Follicular Development ◽  
Protein Isoforms ◽  
Mrna Isoforms ◽  
Messenger Ribonucleic Acid

ABSTRACT Granulosa cells are essential for follicular development and corpus luteum formation and their functions are regulated by gonadotrophins through G protein-coupled receptors. The dominant second messenger pathway involves the stimulation of cyclic AMP formation by Gαs-linked receptors. In this paper we have investigated the expression of Gαs mRNA splice variants in relation to expression of Gαs protein isoforms in granulosa cells obtained from patients undergoing in vitro fertilization. We have carried out ribonuclease protection assays using cRNA riboprobes which are capable of detecting all Gαs mRNA isoforms as well as quantifying total amounts of Gαs mRNA. Granulosa cells express the message for Gαs-Large and Gαs-Small and the presence of two distinct protein products was confirmed by immunoblotting using the antibody RM/1. Moreover, the data show that a significant fraction of Gαs-Large and Gαs-Small mRNAs contain an extra CAG codon. This should generate proteins with an extra serine residue, resulting in Gαs variants with the consensus sequence of a protein kinase C phosphorylation site. These results highlight the possible interaction between different signalling pathways in the control of cAMP production and the need to investigate the relationship between Gαs variants and different adenylyl cyclase isozymes in patients with normal and abnormal ovarian function.

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.

Presence of opioid receptor-like (ORL1) receptor mRNA splice variants in peripheral sensory and sympathetic neuronal ganglia

Life Sciences ◽  
1999 ◽  
Vol 64 (22) ◽  
pp. 2029-2037 ◽  
Author(s):  
Guo-Xi Xie ◽  
Thomas Meuser ◽  
Christian Pietruck ◽  
Manohar Sharma ◽  
Pamela Pierce Palmer
Keyword(s):  
Opioid Receptor ◽  
Splice Variants ◽  
Mrna Splice ◽  
Receptor Mrna ◽  
Orl1 Receptor ◽  
Peripheral Sensory

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 Aging-related premature luteinization of granulosa cells is avoided by early oocyte retrieval

2015 ◽  
Vol 226 (3) ◽  
pp. 167-180 ◽  
Author(s):  
Yan-Guang Wu ◽  
David H Barad ◽  
Vitaly A Kushnir ◽  
Emanuela Lazzaroni ◽  
Qi Wang ◽  
...  
Keyword(s):  
Oocyte Retrieval ◽  
Pregnancy Rates ◽  
Gene Expressions ◽  
Lh Receptor ◽  
Premature Luteinization ◽  
Age Related ◽  
Follicle Size ◽  
Oocyte Donors ◽  
Infertile Patients

Why IVF pregnancy rates decline sharply after age 43 is unknown. In this study, we compared granulosa cell (GC) function in young oocyte donors (n=31, ages 21–29), middle-aged (n=64, ages 30–37) and older infertile patients (n=41, ages 43–47). Gene expressions related to gonadotropin activity, steroidogenesis, apoptosis and luteinization were examined by real-time PCR and western blot in GCs collected from follicular fluid. FSH receptor (FSHR), aromatase (CYP19A1) and 17β-hydroxysteroid dehydrogenase (HSD17B) expression were found down regulated with advancing age, while LH receptor (LHCGR), P450scc (CYP11A1) and progesterone receptor (PGR) were up regulated. Upon in vitro culture, GCs were found to exhibit lower proliferation and increased apoptosis with aging. While FSH supplementation stimulated GCs growth and prevented luteinization in vitro. These observations demonstrate age-related functional declines in GCs, consistent with premature luteinization. To avoid premature luteinization in women above age 43, we advanced oocyte retrieval by administering human chorionic gonadotropin at maximal leading follicle size of 16 mm (routine 19–21 mm). Compared to normal cycles in women of similar age, earlier retrieved patients demonstrated only a marginal increase in oocyte prematurity, yet exhibited improved embryo numbers as well as quality and respectable clinical pregnancy rates. Premature follicular luteinization appears to contribute to rapidly declining IVF pregnancy chances after age 43, and can be avoided by earlier oocyte retrieval.

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.

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