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General Comment |
Glutathione peroxidase (EC 1.11.1.9 ) catalyzes the reduction of hydrogen peroxide, organic hydroperoxide, and lipid
peroxides by reduced glutathione and functions in the protection of cells against oxidative damage. This enzyme, found
mainly in the cytosol of mammalian cells, is unusual in its content of a selenocysteine residue in its active site that is
encoded by a TGA opal codon .
This gene is FSH induced. Identification of differential gene expression in in vitro FSH treated pig granulosa cells using suppression subtractive hybridization. Bonnet A et al. ABSTRACT: FSH, which binds to specific receptors on granulosa cells in mammals, plays a key role in folliculogenesis. Its biological activity involves stimulation of intercellular communication and upregulation of steroidogenesis, but the entire spectrum of the genes regulated by FSH has yet to be fully characterized. In order to find new regulated transcripts, however rare, we have used a Suppression Subtractive Hybridization approach (SSH) on pig granulosa cells in primary culture treated or not with FSH. Two SSH libraries were generated and 76 clones were sequenced after selection by differential screening. Sixty four different sequences were identified, including 3 novel sequences. Experiments demonstrated the presence of 25 regulated transcripts. A gene ontology analysis of these 25 genes revealed (1) catalytic; (2) transport; (3) signal transducer; (4) binding; (5) anti-oxidant and (6) structural activities. These findings may deepen our understanding of FSH's effects. Particularly, they suggest that FSH is involved in the modulation of peroxidase activity and remodelling of chromatin.
NCBI Summary:
Glutathione peroxidase catalyzes the reduction of hydrogen peroxide, organic hydroperoxide, and lipid peroxides by reduced glutathione and functions in the protection of cells against oxidative damage. Human plasma glutathione peroxidase has been shown to be a selenium-containing enzyme. GPX3 expression appears to be tissue-specific.
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Comment |
RUNX2, GPX3 and PTX3 gene expression profiling in cumulus cells are reflective oocyte/embryo competence and potentially reliable predictors of embryo developmental competence in PCOS patients. Huang X 2013 et al.
BACKGROUND
Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder in women. The developmental competence of oocytes and embryos in PCOS patients is reduced to a certain extent (comparing to non-PCOS patients, the high quality embryo rate was decreased by 16% from the data of our centre) during the in vitro fertilization (IVF) process. Cross-talk between the oocyte and cumulus cells is critical for oocyte maturation and embryo competence. In this study, we have evaluated the transcription of specific genes in cumulus cells harvested from pre-ovulatory follicles of PCOS patients before IVF, according to individual oocyte nuclear maturity and developmental competence. Seven genes (RUNX2, PSAT1, ADAMTS9, CXCL1, CXCL2, CXCL3, and ITGB5) were targeted from our previous cDNA microarray data which isolated genes related to oocyte nuclear maturation in PCOS patients. Two additional genes which had been found to be associated with oocyte maturation or embryo quality in non-PCOS patients (GPX3 and PTX3) were also studied.
METHODS
The mRNA expression levels of cumulus cells were detected by qRT- PCR.
RESULTS
Consistent with our previous cDNA microarray data, with the exception of GPX3 and PTX3, the selected 7 genes were related to oocyte nuclear maturation in PCOS patients. Noticeably, the expression level of RUNX2 was lower in cumulus cells derived from oocytes that could develop into blastocysts than the level of expression from oocytes that could not. The PTX3 expression level was significantly lower in cumulus cells from oocytes with two normal pronuclei than that from oocytes that formed >2 pronuclei (MPN) after fertilization. GPX3 mRNA levels were decreased in cumulus cells isolated from oocytes that developed into blastocysts with high potential development competence.
CONCLUSIONS
Several cumulus cell genes were associated with oocyte maturation, fertilization and embryo quality in PCOS patients. RUNX2 and GPX3 are candidate genetic markers in the monitoring of embryo quality for PCOS patients, whereas PTX3 mainly played a role in fertilization process. Together with morphological evaluation, cumulus cells genes may serve as biomarkers of oocyte and embryo selection during the IVF process for PCOS patients and may advance our understanding of PCOS.
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Comment |
Transcriptome Comparisons Identify New Cell Markers for Theca Interna and Granulosa Cells from Small and Large Antral Ovarian Follicles. Hatzirodos N et al. (2015) In studies using isolated ovarian granulosa and thecal cells it is important to assess the degree of cross contamination. Marker genes commonly used for granulosa cells include FSHR, CYP19A1 and AMH while CYP17A1 and INSL3 are used for thecal cells. To increase the number of marker genes available we compared expression microarray data from isolated theca interna with that from granulosa cells of bovine small (n = 10 for both theca and granulosa cells; 3-5 mm) and large (n = 4 for both theca and granulosa cells, > 9 mm) antral follicles. Validation was conducted by qRT-PCR analyses. Known markers such as CYP19A1, FSHR and NR5A2 and another 11 genes (LOC404103, MGARP, GLDC, CHST8, CSN2, GPX3, SLC35G1, CA8, CLGN, FAM78A, SLC16A3) were common to the lists of the 50 most up regulated genes in granulosa cells from both follicle sizes. The expression in theca interna was more consistent than in granulosa cells between the two follicle sizes. Many genes up regulated in theca interna were common to both sizes of follicles (MGP, DCN, ASPN, ALDH1A1, COL1A2, FN1, COL3A1, OGN, APOD, COL5A2, IGF2, NID1, LHFP, ACTA2, DUSP12, ACTG2, SPARCL1, FILIP1L, EGFLAM, ADAMDEC1, HPGD, COL12A1, FBLN5, RAMP2, COL15A1, PLK2, COL6A3, LOXL1, RARRES1, FLI1, LAMA2). Many of these were stromal extracellular matrix genes. MGARP, GLDC, CHST8, GPX3 were identified as new potential markers for granulosa cells, while FBLN5, OGN, RAMP2 were significantly elevated in the theca interna.//////////////////
Endocrine society meeting 2002
Identification and Characterization of Glutathione Peroxidase 3 as an In Vivo Marker of Estrogen Receptor Beta Activity in Mouse Ovarian Granulosa Cells.
Susan L Fitzpatrick, Dana M Banas, Georgius deHaan, Donald E Frail, Robert A Mastroeni Women's Hlth Res Inst, Wyeth Pharmaceuticals, Collegeville, PA
The ovary is the primary site for estrogen synthesis in sexually mature animals and humans and it is also a target for estrogen action. For example, in mice and rats, granulosa cell numbers increase following in vivo treatments with estradiol (E2) and the ovary has been demonstrated to contain both estrogen receptor alpha (ER- ) and beta (ER- ). In order to identify specific genes that were regulated by estradiol, immature female mice (day 21-22) were injected with 500 g (40-60 mg/kg) E2 daily for 3 days. An Affymetrix gene chip analysis was performed using mRNA from the granulosa cells and genes regulated at least 3 fold up or down were further evaluated. Glutathione peroxidase 3 (GPx3, also called plasma GPx) mRNA expression increased 3-4 fold in the presence of estradiol. Additional experiments demonstrated an increase of 5-10 fold following E2 using quantitative RT-PCR. GPx3 mRNA expression also increased following treatment (50 mg/kg) with DES and moxestrol but not with 2-hydroxy-estrone. In addition, GPx3 expression increased in granulosa cells treated with E2 for 2 days in vitro although to a lesser degree than seen in vivo (2-4 fold vs. 5-10 fold). The in vivo induction of GPx3 appeared to be species- dependent as only a 1.5 fold change was observed in immature rats treated with E2 (1.5 mg/day, 30-50 mg/kg). To determine whether ER- was involved in the regulation of GPx3, ER- knockout mice were treated similarly with E2. Unlike wild-type mice, ER- KO mice did not demonstrate any increase in GPx3 mRNA expression in granulosa cells. In wild-type mice, propyl pyrazole triol (PPT), an ER- selective ligand, at 50 mg/kg increased uterine wet weight but did not stimulate GPx3 mRNA expression. In contrast, diarylpropionitrile (DPN), an ER- selective ligand, at 50 mg/kg stimulated GPx3 mRNA expression nearly 3-fold and had little uterotropic activity. This suggests that in mouse granulosa cells, induction of GPx3 mRNA expression by estradiol requires the presence of ER- .
Differential gene expression in cumulus cells as a prognostic indicator of embryo viability: a microarray analysis. van Montfoort AP et al. Besides the established selection criteria based on embryo morphology and blastomere number, new parameters for embryo viability are needed to improve the clinical outcome of IVF and more particular of elective single embryo transfer (eSET). Genome-wide gene expression in cumulus cells was studied, since these cells surround the oocyte inside the follicle and therefore possibly reflect oocyte developmental potential. Early cleavage (EC) was chosen as a parameter for embryo viability. Gene expression in cumulus cells from eight oocytes resulting in an EC embryo (EC-CC; n=8) and from eight oocytes resulting in a non-EC (NEC) embryo (NEC-CC; n=8) was analysed using microarrays (n=16). A total of 611 genes were differentially expressed (P < 0.01), mainly involved in cell cycle, angiogenesis, apoptosis, epidermal growth factor, fibroblast growth factor and platelet-derived growth factor signalling, general vesicle transport and chemokine and cytokine signalling. Of the 25 selected differentially expressed genes analysed by quantitative real-time PCR (qRT-PCR) 15 (60%) genes could be validated in the original samples. Of these 8 (53%) could also be validated in 24 (12-EC-CC and 12 NEC-CC) extra independent samples. The most differentially expressed genes among these were CCND2, CXCR4 , GPX3 , CTNND1 DHCR7 , DVL3 , HSPB1 and TRIM28 , which probably point to hypoxic conditions or a delayed oocyte maturation in NEC-CC samples. This opens up perspectives for new molecular embryo or oocyte selection parameters which might also be useful in countries where the selection has to be made at the oocyte stage before fertilization instead of at the embryonic stage.
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Comment |
Differential genome-wide gene expression profiling of bovine largest and second-largest follicles: identification of genes associated with growth of dominant follicles. Hayashi KG et al. ABSTRACT: BACKGROUND: Bovine follicular development is regulated by numerous molecular mechanisms and biological pathways. In this study, we tried to identify differentially expressed genes between largest (F1) and second-largest follicles (F2), and classifythem by global gene expression profiling using a combination of microarray and quantitative real-time PCR (QPCR) analysis. The follicular status of F1 and F2 were further evaluated in terms of healthy and atretic conditions by investigating mRNA localization of identified genes. METHODS: Global gene expression profiles of F1 (10.7 +/- 0.7 mm) and F2 (7.8 +/- 0.2 mm) were analyzed by hierarchical cluster analysis and expression profiles of 16 representative genes were confirmed by QPCR analysis. In addition, localization of six identified transcripts was investigated in healthy and atretic follicles using in situ hybridization. The healthy or atretic condition of examined follicles was classified by progesterone and estradiol concentrations in follicular fluid. RESULTS: Hierarchical cluster analysis of microarray data classified the follicles into two clusters. Cluster A was composed of only F2 and was characterized by high expression of 31 genes including IGFBP5, whereas cluster B contained only F1 and predominantly expressed 45 genes including CYP19 and FSHR. QPCR analysis confirmed AMH, CYP19, FSHR, GPX3, PlGF, PLA2G1B, SCD and TRB2 were greater in F1 than F2, while CCL2, GADD45A, IGFBP5, PLAUR, SELP, SPP1, TIMP1 and TSP2 were greater in F2 than in F1. In situ hybridization showed that AMH and CYP19 were detected in granulosa cells (GC) of healthy as well as atretic follicles. PlGF was localized in GC and in the theca layer (TL) of healthy follicles. IGFBP5 was detected in both GC and TL of atretic follicles. GADD45A and TSP2 were localized in both GC and TL of atretic follicles, whereas healthy follicles expressed them only in GC. CONCLUSION: We demonstrated that global gene expression profiling of F1 and F2 clearly reflected a difference in their follicular status. Expression of stage-specific genes in follicles may be closely associated with their growth or atresia. Several genes identified in this study will provide intriguing candidates for the determination of follicular growth.
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