scholarly journals In Vitro Exposure of Human Luteinized Mural Granulosa Cells to Dibutyl Phthalate Affects Global Gene Expression

2017 ◽  
Vol 160 (1) ◽  
pp. 180-188 ◽  
Author(s):  
Michal Adir ◽  
Mali Salmon-Divon ◽  
Catherine M. H. Combelles ◽  
Abdallah Mansur ◽  
Yoram Cohen ◽  
...  
Keyword(s):  
Gene Expression ◽  
Granulosa Cells ◽  
Dibutyl Phthalate ◽  
Global Gene Expression ◽  
In Vitro Exposure

scholarly journals Granulosa cells from human primordial and primary follicles show differential global gene expression profiles

2018 ◽  
Vol 33 (4) ◽  
pp. 666-679 ◽  
Author(s):  
E H Ernst ◽  
S Franks ◽  
K Hardy ◽  
P Villesen ◽  
K Lykke-Hartmann
Keyword(s):  
Gene Expression ◽  
Granulosa Cells ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Global Gene Expression

scholarly journals Investigating the Intrafollicular Factors Affecting Oocyte Developmental Competency

2021 ◽  
Author(s):  
◽  
Zaramasina Clark
Keyword(s):  
Gene Expression ◽  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Embryo Quality ◽  
Cumulus Cells ◽  
Significant Finding ◽  
High Quality ◽  
Final Maturation

<p>The number of cycles of assisted reproductive technologies (ART) performed increased by ~9.5 % globally between 2008 and 2010. In spite of this, the success rate in terms of delivery was only ~19.0 % (Dyer et al., 2016). This discrepancy between the demand for, and success of, these technologies necessitates the development of tools to improve ART efficiency. To facilitate this, a better understanding of how the microenvironment changes within the developing follicle to culminate in a mature, developmentally-competent oocyte is required. This study employed an in vivo and in vitro ovine model to investigate the relationship between the surrounding microenvironment and oocyte maturation, and in particular, the attainment of oocyte developmental competency and high-quality embryos.  The first objective of this PhD study was to comprehensively investigate the changing microenvironment of in vivo matured, presumptive preovulatory (PPOV) follicles from wild-type (++) and high ovulation rate (OR; I+B+) ewes. The high OR ewes were heterozygous carriers of mutations in BMP15 (I+) and BMPRIB (B+). Functional differences in follicular somatic (granulosa and cumulus) cells between these genotypes, including differential gonadotropin responsiveness of granulosa cells, composition of follicular fluid and gene expression profiles in cumulus cells were evident. These differences emerged as part of a compensatory mechanism by which oocytes from smaller follicles, containing fewer granulosa cells, achieved developmental competency in I+B+ ewes.  The second objective of this PhD study was to develop new approaches for improving current in vitro maturation (IVM) strategies. The first approach utilised in this study focused on developing biomarkers that could be used to improve prediction of developmental competency in oocytes and in vitro produced embryos. This involved interrogating the hypothesis that a combination of molecular and morphokinetic biomarkers would better predict the developmental competency of oocytes and embryos compared to using these biomarkers alone. The second approach utilised in this PhD study tested the effects of modulating IVM conditions to better mimic the follicular microenvironment of a high, compared to a low, OR species on oocyte developmental competency and embryo quality. This involved supplementing IVM media with different ratios of two oocyte-secreted growth factors, i.e. GDF9:BMP15, that were representative of low or high OR species. These approaches demonstrated significant potential and warrant further investigation.  The most significant finding of this study was that despite variances in the surrounding microenvironment during in vivo and in vitro oocyte maturation that culminated in differential gene expression patterns in cumulus cells, and divergent gonadotropin-responsiveness of granulosa cells, the gene expression signatures of developmentally-competent oocytes and the morphokinetics of high-quality embryos were unaltered. This confirms the value of developing such biomarkers for oocyte development competency and embryo quality that remain unaltered despite a changing surrounding environment. Interestingly, simulating the ratio of GDF9:BMP15 that oocytes from high OR species are exposed to during maturation improved developmental competency in oocytes as demonstrated by increased blastocyst rates. Furthermore, this study has demonstrated that combinations of molecular (cumulus cell gene expression) and morphokinetic biomarkers improved the ability to predict developmental competency in oocytes and embryos. Overall, this study revealed novel information regarding the follicular microenvironment during final maturation and identified several novel approaches to improving the efficiency of ART.</p>

scholarly journals Expression of mRNAs for Pro-and Anti-Apoptotic Factors in Granulosa Cells and Follicular Fluid of Women Undergoing in Vitro Fertilization. A Pilot Study

2021 ◽  
Author(s):  
Jozsef Bodis ◽  
Endre Sulyok ◽  
Akos Varnagy ◽  
Viktória Prémusz ◽  
Krisztina Godony ◽  
...  
Keyword(s):  
Gene Expression ◽  
In Vitro Fertilization ◽  
Mrna Expression ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Vitro Fertilization ◽  
Expression Levels ◽  
The Impact ◽  
Apoptotic Factors

Abstract BackgroundThis observational clinical study evaluated the expression levels and predictive values of some apoptosis-related genes in granulosa cells (GCs) and follicular fluid (FF) of women undergoing in vitro fertilization (IVF).Methods GCs and FF were obtained at oocyte retrieval from 31 consecutive patients with heterogeneous infertility diagnosis (age: 34.3±5.8 years, body mass index: 24.02±3.12 kg/m2, duration of infertility: 4.2±2.1 years). mRNA expression of pro-apoptotic (BAX, CASP3, CASP8) and anti-apoptotic (BCL2, AMH, AMHR, FSHR, LHR, CYP19A1) factors was determined by quantitative RT-PCR using ROCHE LightCycler 480. Results No significant difference in GC or FF mRNA expression of pro- and anti-apoptotic factors could be demonstrated between IVF patients with (9 patients) or without (22 patients) clinical pregnancy. Each transcript investigated was detected in FF, but their levels were markedly reduced and independent of those in GCs. The number of retrieved oocytes was positively associated with GC AMHR (r=0.393, p=0.029), but the day of embryo transfer was negatively associated with GC LHR (r=-0.414, p=0.020) and GC FSHR transcripts (r=-0.535, p=0.002). When pregnancy positive group was analysed separately the impact of apoptosis- related gene expressions on some selected measures of IVF success could be observed. Strong positive relationship was found between gene expression levels of pro- and anti-apoptotic factors in GCs.ConclusionOur study provides only marginal evidences for the apoptosis dependence of IVF outcome and suggests that the apoptosis process induces adaptive increases of the anti-apoptotic gene expression to attenuate apoptosis and to protect cell survival.

scholarly journals 2-Hydroxy-4-Methylselenobutanoic Acid (HMSeBA) Promote Follicle Development By Antioxidant Pathway in Gilts

2021 ◽  
Author(s):  
Yanpeng Dong ◽  
Sirun Chen ◽  
Yalei Liu ◽  
Zimei Li ◽  
Xinlin Jia ◽  
...  
Keyword(s):  
Gene Expression ◽  
Antioxidant Capacity ◽  
Granulosa Cells ◽  
Body Weight Gain ◽  
Control Diet ◽  
Negative Control ◽  
Follicle Development ◽  
Selenium Content ◽  
Total Selenium

Abstract Background Dietary 2-hydroxy-4-methylselenobutanoic acid (HMSeBA) supplementation can exert antioxidant effects in poultry, pigs and weaned pigs. However, it is unknown whether HMSeBA could improve the development of follicle by anti-oxidize effects in gilt. This study was conducted to evaluate the effects of dietary HMSeBA supplementation on the follicle development in gilt. A total of 36 gilts were randomly fed the control diet (CON, negative control), Na2SeO3 diet containing 0.3 mg Se/kg (positive control) or the HMSeBA diet containing 0.3 mg Se/kg from weaning to the 19th day after the second estrus. In another study, the effect of HMSeBA on the cells viability, proliferation, release of 17βestradiol (E2 ) and antioxidant capacity were investigated in the mouse ovarian granulosa cells in vitro. Results Results showed that HMSeBA group increased the average daily body weight gain (ADG) and decreased the ratio of feed: gain during day 120 to 176 in gilts ( P < 0.05). The selenium (HMSeBA and Na 2 SeO 3 ) increased the weight of uterine at the third estrus. There was no effect of HMSeBA on the number of large follicles (diameter >5mm), but HMSeBA decreased the gene expression of growth differentiation factor-9 ( GDF-9 ) and bone morphogenetic protein-15 ( BMP-15 ) in cumulus-oocyte complexes (COCs). HMSeBA group increased the total selenium content in serum ( P < 0.05) and liver ( P < 0.01) and tended to increase the total selenium content in ovary ( P = 0.08). HMSeBA group decreased the malondialdehyde (MDA) concentration in the serum, liver and ovary ( P < 0.05), increased the total antioxidant capacity (T-AOC) in the liver, thioredoxin reductase (TrxR) in the ovary ( P < 0.05) and increased the activity of GPx in the serum, liver and ovary ( P < 0.05). Na 2 SeO 3 supplementation decreased MDA and increased the T-AOC in liver, increased the T-SOD and TrxR in the ovary compared with control. At the transcription level, HMSeBA group increased the glutathione peroxidase 2 ( GPx2 ) and TrxR1 ( P < 0.05) expression in the liver, and increased the GPx1 expression ( P < 0.05) in the ovary of gilts compared with Na2SeO3 treatment. Besides, HMSeBA group increased the expressions of superoxide dismutase 1 ( SOD1 ) and Thioredoxin l ( Trx1 ) in the liver. In vitro experiment, HMSeBA improved granulosa cells’ proliferation and E2 secretion ( P < 0.05). HMSeBA and Na 2 SeO 3 both increased the T-AOC and decreased MDA in granulosa cells in vitro. Meanwhile, HMSeBA increased T-SOD, GPx, glutathione reductase (GR) and TrxR activity in granulosa cells in vitro. In addition, HMSeBA up-regulated SOD2 and GPx1 gene expression in the granulosa cells in vitro.Conclusion These results demonstrate directly, HMSeBA was more conducive to absorption and storage of selenium in the liver and ovary in gilt, and beneficial to exert the effect of HMSeBA on the antioxidant function in the liver and ovary of gilt. Moreover, HMSeBA has stronger antioxidant capacity in granular cells in vitro , which is more conducive to promoting follicle development. Therefore, the new type of organic selenium, HMSeBA, could be potentially useful for the control of reproductive processes in gilt.

scholarly journals Eaf1 Links the NuA4 Histone Acetyltransferase Complex to Htz1 Incorporation and Regulation of Purine Biosynthesis

2015 ◽  
Vol 14 (6) ◽  
pp. 535-544 ◽  
Author(s):  
Xue Cheng ◽  
Andréanne Auger ◽  
Mohammed Altaf ◽  
Simon Drouin ◽  
Eric Paquet ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cross Talk ◽  
Global Gene Expression ◽  
Histone Variant ◽  
Purine Biosynthesis ◽  
Expression Array ◽  
Link Type ◽  
Molecular Events ◽  
Nucleosome Disassembly

ABSTRACT Proper modulation of promoter chromatin architecture is crucial for gene regulation in order to precisely and efficiently orchestrate various cellular activities. Previous studies have identified the stimulatory effect of the histone-modifying complex NuA4 on the incorporation of the histone variant H2A.Z (Htz1) at the PHO5 promoter (A. Auger, L. Galarneau, M. Altaf, A. Nourani, Y. Doyon, R. T. Utley, D. Cronier, S. Allard, and J. Côté, Mol Cell Biol 28:2257–2270, 2008, http://dx.doi.org/10.1128/MCB.01755-07 ). In vitro studies with a reconstituted system also indicated an intriguing cross talk between NuA4 and the H2A.Z-loading complex, SWR-C (M. Altaf, A. Auger, J. Monnet-Saksouk, J. Brodeur, S. Piquet, M. Cramet, N. Bouchard, N. Lacoste, R. T. Utley, L. Gaudreau, J. Côté, J Biol Chem 285:15966–15977, 2010, http://dx.doi.org/10.1074/jbc.M110.117069 ). In this work, we investigated the role of the NuA4 scaffold subunit Eaf1 in global gene expression and genome-wide incorporation of Htz1. We found that loss of Eaf1 affects Htz1 levels mostly at the promoters that are normally highly enriched in the histone variant. Analysis of eaf1 mutant cells by expression array unveiled a relationship between NuA4 and the gene network implicated in the purine biosynthesis pathway, as EAF1 deletion cripples induction of several ADE genes. NuA4 directly interacts with Bas1 activation domain, a key transcription factor of adenine genes. Chromatin immunoprecipitation (ChIP) experiments demonstrate that nucleosomes on the inactive ADE17 promoter are acetylated already by NuA4 and enriched in Htz1. Upon derepression, these poised nucleosomes respond rapidly to activate ADE gene expression in a mechanism likely reminiscent of the PHO5 promoter, leading to nucleosome disassembly. These detailed molecular events depict a specific case of cross talk between NuA4-dependent acetylation and incorporation of histone variant Htz1, presetting the chromatin structure over ADE promoters for subsequent chromatin remodeling and activated transcription.

Growth, Drug Susceptibility, and Gene Expression Profiling of Plasmodium falciparum Cultured in Medium Supplemented with Human Serum

2007 ◽  
Vol 12 (8) ◽  
pp. 1109-1114 ◽  
Author(s):  
Kshipra Singh ◽  
Ameeta Agarwal ◽  
Shabana I. Khan ◽  
Larry A. Walker ◽  
Babu L. Tekwani
Keyword(s):  
Gene Expression ◽  
Plasmodium Falciparum ◽  
Human Serum ◽  
Global Gene Expression ◽  
Drug Susceptibility ◽  
Growth Cycle ◽  
In Vitro Cultivation ◽  
Human Malaria Parasite ◽  
In Vitro Drug Susceptibility

In vitro cultivation of Plasmodium falciparum has been extremely useful in understanding the biology of the human malaria parasite as well as research on the discovery of new antimalarial drugs and vaccines. A chemically defined serum-free medium supplemented with lipid-rich bovine serum albumin (AlbuMAX I) offers the following advantages over human serum-supplemented media for the in vitro culture of P. falciparum: 1) improved growth profile, with more than a 2-fold higher yield of the parasites at any stage of the growth cycle; 2) suitability for in vitro antimalarial screening, as the parasites grown in AlbuMAX and human serum-supplemented media show similar sensitivity to standard and novel antimalarials as well as natural product extracts in the in vitro drug susceptibility assays; and 3) DNA microarray analysis comparing the global gene expression profile of sorbitol-synchronized P. falciparum trophozoites grown in the 2 different media, indicating minimal differences. ( Journal of Biomolecular Screening 2007:1109-1114)

scholarly journals Absence of leptin expression and secretion by human luteinized granulosa cells

2003 ◽  
Vol 31 (1) ◽  
pp. 233-239 ◽  
Author(s):  
M Karamouti ◽  
P Kollia ◽  
E Karligiotou ◽  
A Kallitsaris ◽  
N Prapas ◽  
...  
Keyword(s):  
Gene Expression ◽  
Granulosa Cells ◽  
Messenger Rna ◽  
Limit Of Detection ◽  
Culture Media ◽  
Point Of View ◽  
In Vitro Fertilisation ◽  
Ob Gene ◽  
Human Granulosa Cells

Whether leptin is secreted by the human ovary is not known. The available data on leptin gene (ob gene) expression by human granulosa cells are conflicting. The aim of the present study was first to re-examine the expression of leptin messenger RNA (mRNA) by human granulosa cells and second to investigate if these cells have the ability to secrete leptin in cultures. Human luteinized granulosa cells were obtained from normal women undergoing in vitro fertilisation treatment after ovarian stimulation and follicle aspiration. The expression of ob gene was studied by Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR) both in primary granulosa cells treated immediately after oocyte recovery and in cells cultured up to 24 h under baseline and hormonally stimulated conditions (FSH: 100 ng/ml, LH: 100 ng/ml). ob mRNA transcripts were not detected in luteinized granulosa cells, while they were present in adipose tIssue cDNA. Actin gene expression was detected in all studied samples. Using a sensitive radioimmunoassay (lower limit of detection 0.05 ng/ml), leptin was undetectable in the culture media at all points during the 72 h cultures, while at the same time significant amounts of oestradiol and progesterone were produced particularly after the addition of androstendione (1 microM) to the incubation media. These results demonstrate for the first time that leptin is not secreted by human luteinized granulosa cells in cultures. From a physiological point of view, this may contribute to the development of the optimal follicular environment for oocyte maturation during the preovulatory period.

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