scholarly journals Presence of LH receptor mRNA in granulosa cells as a potential marker of oocyte developmental competence and characterization of the bovine splicing isoforms

Reproduction ◽  
2003 ◽  
Vol 125 (3) ◽  
pp. 437-446
Author(s):  
C Robert
Keyword(s):  
Granulosa Cells ◽  
Developmental Competence ◽  
Lh Receptor ◽  
Receptor Mrna ◽  
Oocyte Developmental Competence ◽  
Potential Marker ◽  
Splicing Isoforms

scholarly journals Presence of LH receptor mRNA in granulosa cells as a potential marker of oocyte developmental competence and characterization of the bovine splicing isoforms

Reproduction ◽  
2003 ◽  
pp. 437-446 ◽  
Author(s):  
C Robert ◽  
D Gagne ◽  
JG Lussier ◽  
D Bousquet ◽  
FL Barnes ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Culture Media ◽  
Protein Product ◽  
Blastocyst Stage ◽  
Culture Conditions ◽  
Developmental Competence ◽  
Lh Receptor ◽  
Potential Marker ◽  
A Cell

As the expression of the LH receptor (LH-R) in granulosa cells is thought to be associated with later stages of folliculogenesis, this study was undertaken to evaluate the presence of LH-R mRNA as a suitable marker for developmental competence of oocytes. Granulosa cells and cumulus-oocyte complexes (COCs) were recovered from cows that had received ovarian stimulation. The COCs were subjected to embryo production procedures in vitro to assess the embryonic potential of the oocyte, and the corresponding granulosa cells were used to evaluate the presence of LH-R mRNA by RT-PCR. The presence of LH-R transcripts in granulosa cells is not a key characteristic of a follicle bearing a competent oocyte, although a higher proportion of oocytes reach the blastocyst stage when LH-R mRNA is detected in the granulosa cells. Different LH-R isoforms were cloned and sequence discrepancies among six of the isoforms enabled the design of specific oligonucleotides to study the presence of the isoforms in different follicular cells. All LH-R transcripts studied and the 80 kDa protein product corresponding to the full length receptor were found in granulosa cells of small (< 4 mm) and large (> 5 mm) follicles. When the granulosa cells were cultured, the transcripts were downregulated by the culture conditions; downregulation was more acute in granulosa cells from small follicles. The addition of LH to the culture media enhanced LH-R mRNA downregulation. The presence of several LH-R transcript isoforms was tissue specific and in the theca cells LH-R mRNA was restricted mainly to cells from larger follicles. This finding indicates that the expression and the splicing of LH-R mRNA are regulated in a cell-specific and follicular size-specific manner.

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.

Relationship between donor animal age, follicular fluid steroid content and oocyte developmental competence in the pig

2003 ◽  
Vol 15 (2) ◽  
pp. 81 ◽  
Author(s):  
Christopher G. Grupen ◽  
Stephen M. McIlfatrick ◽  
Rodney J. Ashman ◽  
Andrew C. Boquest ◽  
David T. Armstrong ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Developmental Competence ◽  
Molar Ratios ◽  
Oocyte Developmental Competence ◽  
Trophectoderm Cells ◽  
Maturation Medium

The developmental competence of oocytes recovered from the ovaries of slaughtered prepubertal and adult pigs was evaluated after in vitro maturation, parthenogenetic activation and culture in vitro. In addition, the effect of prepubertal and adult follicular fluid (FF) on the developmental competence of prepubertal and adult oocytes was investigated. When matured in adult FF, the rates of cleavage (92 v. 73%; P < 0.01) and blastocyst formation (57 v. 38%; P < 0.05) were greater for adult oocytes than for prepubertal oocytes. Blastocysts derived from adult oocytes had more trophectoderm cells (43 v. 30; P < 0.05) and total cells (51 v. 36; P < 0.05) than blastocysts derived from prepubertal oocytes. The developmental competence of prepubertal oocytes was not affected by the FF donor age, whereas the developmental competence of adult oocytes was. Blastocysts derived from adult oocytes matured in adult FF had more trophectoderm cells (38 v. 24; P < 0.005), inner cell mass cells (7 v. 3; P < 0.01) and total cells (45 v. 27; P < 0.001) than blastocysts derived from adult oocytes matured in prepubertal FF. Characterization of the steroid content of the FF used to supplement the maturation medium revealed that adult FF contained more progesterone (42 v. 23 ng mL−1; P < 0.005) and androstenedione (70 v. 16 ng mL−1; P < 0.05) than prepubertal FF. In addition, the molar ratios of progesterone to androstenedione, androstenedione to 17β-oestradiol and androstenedione to testosterone differed (P < 0.05) between prepubertal and adult FF. The results support the hypothesis that a greater proportion of adult oocytes than of prepubertal oocytes has completed ‘oocyte capacitation’. The differences in FF steroid content are indicative of the different follicular environments from which the prepubertal and adult oocytes were isolated, and may be attributed to the observed effects on oocyte developmental competence.

The Effect of Granulosa Cells Apoptosis on the Cumulus Expansion and the Developmental Competence of Bovine Oocytes

2014 ◽  
Vol 997 ◽  
pp. 251-254 ◽  
Author(s):  
Wei Guo Feng ◽  
Zhi Fang Pan
Keyword(s):  
Granulosa Cells ◽  
Culture System ◽  
Developmental Competence ◽  
Bovine Oocyte ◽  
Cumulus Expansion ◽  
Oocyte Developmental Competence ◽  
Bovine Oocytes

In this study, we used the well in the well (WIW) culture system to study the effect of granulosa cells apoptosis on the cumulus expansion and the developmental competence of bovine oocytes. Results: The apoptosis of granulosa cells affect the cumulus expansion of bovine oocytes significantly. Especially when the percentage of granulosa cells apoptosis exceed 40%, the cumulus expansion was worse. The cumulus expansion affect the oocyte developmental competence of bovine oocytes significantly. The developmental competence of bovine oocyte increases with the increasing of cumulus expansion.

The Effects of Floating Granulosa Cells Density on the Developmental Competence of Bovine Oocytes: A Study Using the Well-in-Well Culture System

2014 ◽  
Vol 884-885 ◽  
pp. 544-547
Author(s):  
Wei Guo Feng ◽  
Chun E Ren ◽  
Zhi Fang Pan
Keyword(s):  
Granulosa Cells ◽  
Culture System ◽  
Developmental Competence ◽  
Group System ◽  
Apoptosis Rate ◽  
Oocyte Developmental Competence ◽  
Bovine Oocytes ◽  
Hatching Rates

In this study, we succeed to establish the well in the well (WIW) culture system. Using this system we studied the effect of floating granulosa cells density on oocyte developmental competence. Results: Apoptosis rate of the floating cells were high (73.576.3%) and did not differ (P > 0.05) among follicles of different sizes. Hatching rates of blastocysts did not differ (P > 0.05) between oocytes cultured in the WIW system (13.3%) and those cultured in the group system (16.3%). In 4-8mm follicles, oocyte developmental competence is higher (P < 0.05) in follicles with 1030×105/mL floating cells than in follicles with < 10 or >30%×105/mL floating cells.

scholarly journals Luteinizing Hormone Receptor mRNA Down-Regulation Is Mediated through ERK-Dependent Induction of RNA Binding Protein

2011 ◽  
Vol 25 (2) ◽  
pp. 282-290 ◽  
Author(s):  
Bindu Menon ◽  
Megan Franzo-Romain ◽  
Shadi Damanpour ◽  
K. M. J. Menon
Keyword(s):  
Granulosa Cells ◽  
Rna Binding ◽  
Binding Protein ◽  
Primary Cultures ◽  
Binding Activity ◽  
Luteinizing Hormone Receptor ◽  
Down Regulation ◽  
Lh Receptor ◽  
Receptor Mrna ◽  
Mrna Binding

Abstract The ligand-induced down-regulation of LH receptor (LHR) expression in the ovaries, at least in part, is regulated by a posttranscriptional process mediated by a specific LH receptor mRNA binding protein (LRBP). The LH-mediated signaling pathways involved in this process were examined in primary cultures of human granulosa cells. Treatment with 10 IU human chorionic gonadotropin (hCG) for 12 h resulted in the down-regulation of LHR mRNA expression while producing an increase in LHR mRNA binding to LRBP as well as a 2-fold increase in LRBP levels. The activation of ERK1/2 pathway in LH-mediated LHR mRNA down-regulation was also established by demonstrating the translocation of ERK1/2 from the cytosol to the nucleus using confocal microcopy. Inhibition of protein kinase A using H-89 or ERK1/2 by U0126 abolished the LH-induced LHR mRNA down-regulation. These treatments also abrogated both the increases in LRBP levels as well as the LHR mRNA binding activity. The abolishment of the hCG-induced increase in LRBP levels and LHR mRNA binding activity was further confirmed by transfecting granulosa cells with ERK1/2 specific small interfering RNA. This treatment also reversed the hCG-induced down-regulation of LHR mRNA. These data show that LH-regulated ERK1/2 signaling is required for the LRBP-mediated down-regulation of LHR mRNA.

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