scholarly journals Mitochondrial Function in Modulating Human Granulosa Cell Steroidogenesis and Female Fertility

2020 ◽  
Vol 21 (10) ◽  
pp. 3592
Author(s):  
Dilip Bhargava Sreerangaraja Urs ◽  
Wen-Han Wu ◽  
Katerina Komrskova ◽  
Pavla Postlerova ◽  
Yung-Feng Lin ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Granulosa Cells ◽  
Mitochondrial Function ◽  
Oocyte Quality ◽  
Fertilization Rate ◽  
Mitochondrial Mass ◽  
Ovarian Endometrioma ◽  
The Relationship ◽  
Estradiol Levels ◽  
Follicular Fluids

Ovarian follicle steroidogenesis associated with embryo quality results in a successful pregnancy. Each follicle consists of an oocyte surrounded by granulosa cells, which secrete several steroid and peptide hormones. Follicles harvested from women who conceived after assisted reproductive therapy (ART) had significantly higher estradiol levels in follicular fluids than the follicles from women who failed to conceive after ART. The higher follicular estradiol levels correlate well with successful fertilization following ART. Mitochondria are the central sites for steroid hormone biosynthesis. The first and rate-limiting step in the biosynthesis of steroid hormones occurs in the mitochondria of granulosa cells. In the present study, we hypothesized that the mitochondria in granulosa cells are critical for maintaining oocyte quality and fertility capacity. This study aims to clarify the relationship between mitochondrial function and granulosa cell steroidogenesis, and the relationship between hormone levels and fertility capacity. Sera, follicular fluids and granulosa cells were obtained from individuals undergoing IVF-ET treatment. The oocyte numbers, oocyte quality, fertilization rate, and pregnancy rate were also recorded. The patients who provided the granulosa cells were further classified into four groups: endometriosis, ovarian endometrioma, endometriosis without ovarian endometrioma, and polycystic ovary syndrome (PCOS); patients with other female factor infertility and male factor infertility were used as controls. We measured the levels of estradiol (E2) by radioimmunoassay. Concurrently, we analyzed the mitochondrial mass and membrane potential, and apoptosis by flow cytometry using nonyl acridine orange, TMRE, Annexin V-FITC and PI. Mitochondrial morphology was visualized by transfection with pLV-mitoDsRed. In addition, we assessed the protein levels of steroidogenic enzymes, steroidogenic acute regulatory protein (StAR) and 3β-hydroxysteroid dehydrogenase (3β-HSD) by Western blot. The results showed significantly decreased serum E2 and follicular E2 levels, and decreased IVF outcomes, in the patients with endometriosis. Reduced mitochondrial mass and decreased mitochondrial membrane potential were correlated with lower E2. Furthermore, a significant decrease in StAR and 3β-HSD was found in patients with ovarian endometrioma. The enzyme levels of StAR and 3β-HSD were highly correlated with E2 levels. Finally, elevated cumulus cell apoptosis was found in the patient group with ovarian endometrioma and PCOS. In conclusion, mitochondrial dysfunction of human granulosa cells may contribute to the decline of steroidogenesis, decreased fertilization rate, oocyte maturation rate, and oocyte quality, and it can ultimately jeopardize fertility.

scholarly journals CD9 Expression by Human Granulosa Cells and Platelets as a Predictor of Fertilization Success during IVF

2010 ◽  
Vol 2010 ◽  
pp. 1-7 ◽  
Author(s):  
Carolyn R. Jaslow ◽  
Kyle S. Patterson ◽  
Shila Cholera ◽  
Lisa K. Jennings ◽  
Raymond W. Ke ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Surface Density ◽  
Venous Blood ◽  
Fertilization Rate ◽  
Fertilization Success ◽  
Vitro Fertilization ◽  
Human Granulosa Cells ◽  
Significant Relationships ◽  
The Relationship

Objective. To determine whether CD9 expression on human granulosa cells (GCs) and platelets could predict the success of conventional fertilization of human oocytes during in vitro fertilization (IVF).Methods. Thirty women undergoing IVF for nonmale factor infertility participated. Platelets from venous blood and GCs separated from retrieved oocytes were prepared for immunofluorescence. Flow cytometry quantified the percent of GCs expressing CD9, and CD9 surface density on GCs and platelets. Fertilization rate was determined for the total number of oocytes, and the number of mature oocytes per patient. Correlations tested for significant relationships (P<.05) between fertilization rates and CD9 expression.Results. CD9 surface density on human GCs is inversely correlated with fertilization rate of oocytes (P=.04), but the relationship was weak.Conclusion. More studies are needed to determine if CD9 expression on GCs would be useful for predicting conventional fertilization success during IVF.

151. LIPOTOXICITY MEDIATED ENDOPLASMIC RETICULUM STRESS, MITOCHONDRIAL DYSFUNCTION AND APOPTOSIS CONTRIBUTE IMPAIRED OOCYTE QUALITY IN RESPONSE TO OBESITY

2009 ◽  
Vol 21 (9) ◽  
pp. 69
Author(s):  
L. L. Y. Wu ◽  
X. Yang ◽  
K. R. Dunning ◽  
R. J. Norman ◽  
R. L. Robker
Keyword(s):  
Endoplasmic Reticulum ◽  
Membrane Potential ◽  
Er Stress ◽  
Mitochondrial Dysfunction ◽  
Granulosa Cells ◽  
Oocyte Quality ◽  
Developmental Competence ◽  
Tunel Staining ◽  
Marker Genes ◽  
Er Homeostasis

In obesity, accumulation of lipid in non-adipose tissues, a process termed lipotoxicity, is associated with endoplasmic reticulum (ER) stress, mitochondrial dysfunction and ultimately apoptosis . We have previously shown that diet-induced obesity in mice causes impaired oocyte developmental competence, but whether this is due to activation of lipotoxicity pathways in the ovary is not known. The present study examined the hypothesis that diet-induced lipid accumulation in the cumulus oocyte complex (COC) disrupts ER homeostasis and mitochondrial membrane potential which leads to apoptosis. COCs and mural granulosa cells were collected from ovaries of adult mice fed a high fat (HFD) or control diet for 4 weeks. ER homeostasis was assessed by measuring expression of known ER stress marker genes, GRP78, ATF4 and CHOP. COCs from mice fed HFD showed significantly increased expression of GRP78 and ATF4. There was a similar trend towards increased expression in granulosa cells. Mitochondrial function was assessed by measuring membrane potential using the dual emission probe JC-1. In COCs from mice fed HFD there were reduced numbers of active mitochondria but instead large aggregated clusters of inactive mitochondria. Apoptosis in granulosa cells was determined by DNA laddering assay which showed significantly increased DNA fragmentation in cells from mice fed HFD. Apoptosis was also assessed by TUNEL staining of paraffin embedded ovaries from identical treatment groups. Ovaries from HFD mice appeared to have increased TUNEL positivity in both granulosa and cumulus cells. Our results demonstrate that the ER stress, mitochondrial dysfunction and apoptosis are markedly increased in granulosa cells and COCs from mice fed HFD, suggesting that lipotoxicity contributes to the impaired oocyte quality and reduced fertility observed in response to obesity.

Effect of mitophagy in oocytes and granulosa cells on oocyte quality†

2020 ◽  
Author(s):  
Qiuzi Shen ◽  
Yu Liu ◽  
Honggang Li ◽  
Ling Zhang
Keyword(s):  
Cell Survival ◽  
Granulosa Cells ◽  
Mitochondrial Function ◽  
Oocyte Quality ◽  
Reproductive Technology ◽  
Developmental Competence ◽  
Mitochondrial Homeostasis ◽  
New Directions ◽  
Developmental Capacity ◽  
Assisted Reproductive Technology Treatment

Abstract Mitophagy is the process by which cells selectively remove supernumerary or damaged mitochondria through autophagy, and is crucial for mitochondrial homeostasis and cell survival. Mitochondria play vital roles in determining the developmental competence of oocytes. During the early stages of oogenesis, aberrant mitochondria can be removed by mitophagy. After oocyte formation, mitophagy is not actively initiated to clear damaged mitochondria despite the presence of mitophagy regulators in oocytes, which leads to the transmission of dysfunctional mitochondria from the oocyte to the embryo. However, granulosa cells around oocytes can improve mitochondrial function through mitophagy, thereby improving oocyte developmental capacity. Furthermore, this review discusses recent work on the substances and environmental conditions that affect mitophagy in oocytes and granulosa cells, thus providing new directions for improving oocyte quality during assisted reproductive technology treatment.

Intracellular Ca2+ and antioxidant values induced positive effect on fertilisation ratio and oocyte quality of granulosa cells in patients undergoing in vitro fertilisation

2013 ◽  
Vol 25 (5) ◽  
pp. 746 ◽  
Author(s):  
Esra Nur Tola ◽  
Muhittin Tamer Mungan ◽  
Abdülhadi Cihangir Uğuz ◽  
Mustafa Naziroğlu
Keyword(s):  
Glutathione Peroxidase ◽  
Granulosa Cells ◽  
Oocyte Quality ◽  
Poor Quality ◽  
In Vitro Fertilisation ◽  
Fertilisation Rate ◽  
Gonadotrophin Releasing Hormone ◽  
The Relationship

Oxidative stress is important for promoting oocyte maturation and ovulation within the follicle through calcium ion (Ca2+) influx. The relationship between antioxidant and cytosolic Ca2+ levels and oocyte quality and fertilisation rate in the granulosa cells of patients undergoing in vitro fertilisation was investigated. Granulosa cells were collected from 33 patients. Cytosolic free Ca2+ ([Ca2+]i) concentration, lipid peroxidation, reduced glutathione, glutathione peroxidase and oocyte quality were measured in the granulosa cells. The relationship between two drug protocols was also examined (gonadotrophin-releasing hormone antagonist and agonist protocols) and the same parameters investigated. The [Ca2+]i concentration (P < 0.001), glutathione (P < 0.05) and oocyte quality (P < 0.001) values were significantly higher in the fertilised group than in the non-fertilised group, although glutathione peroxidase activity was significantly (P < 0.05) higher in the non-fertilised group than in the fertilised group. The [Ca2+]i concentrations were also higher (P < 0.001) in the good-quality oocyte groups than in the poor-quality oocyte group. There was no correlation between the two drug protocols and investigated parameters. In conclusion, it was observed that high glutathione and cytosolic Ca2+ concentrations in granulosa cells of patients undergoing in vitro fertilisation tended to increase the fertilisation potential of oocytes.

scholarly journals Effects of metformin on mitochondrial function of leukocytes from polycystic ovary syndrome patients with insulin resistance

2015 ◽  
Vol 173 (5) ◽  
pp. 683-691 ◽  
Author(s):  
Victor M Victor ◽  
Susana Rovira-Llopis ◽  
Celia Bañuls ◽  
Noelia Diaz-Morales ◽  
Raquel Castelló ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Membrane Potential ◽  
Polycystic Ovary Syndrome ◽  
Mitochondrial Function ◽  
Polycystic Ovary ◽  
Metformin Treatment ◽  
Oxygen Species ◽  
Ovary Syndrome ◽  
Mitochondrial Mass ◽  
Beneficial Effects

ObjectiveOxidative stress and mitochondrial dysfunction are implicated in polycystic ovary syndrome (PCOS). The present study assesses the effect of metformin treatment on mitochondrial function in polymorphonuclear cells from PCOS subjects. Additionally, we evaluate endocrine parameters and levels of interleukin 6 (IL6) and tumour necrosis factor alpha (TNFα).Design and methodsOur study population was comprised of 35 women of reproductive age diagnosed with PCOS and treated with metformin for 12 weeks, and their corresponding controls (n=41), adjusted by age and BMI. We evaluated the alteration of endocrinological and anthropometrical parameters and androgen levels. Mitochondrial O2 consumption (using a Clark-type O2 electrode), membrane potential, mitochondrial mass, and levels of reactive oxygen species (ROS) and glutathione (GSH) (by means of fluorescence microscopy) were assessed in poymorphonuclear cells. H2O2 was evaluated with the Amplex RedR H2O2/Peroxidase Assay kit. IL6 and TNFα were measured using the Luminex 200 flow analyser system.ResultsMetformin had beneficial effects on patients by increasing mitochondrial O2 consumption, membrane potential, mitochondrial mass and glutathione levels, and by decreasing levels of reactive oxygen species and H2O2. In addition, metformin reduced glucose, follicle-stimulating hormone, IL6 and TNFα levels and increased dehydroepiandrosterone sulfate levels. HOMA-IR and mitochondrial function biomarkers positively correlated with ROS production (r=0.486, P=0.025), GSH content (r=0.710, P=0.049) and H2O2 (r=0.837, P=0.010), and negatively correlated with membrane potential (r=−0.829, P=0.011) at baseline. These differences disappeared after metformin treatment.ConclusionsOur results demonstrate the beneficial effects of metformin treatment on mitochondrial function in leukocytes of PCOS patients.

Metabolic Cross Talk Between Foxo3 and mTOR Is Essential for Hematopoietic Stem Cell Function

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 856-856 ◽  
Author(s):  
Pauline Rimmele ◽  
Valentina d'Escamard ◽  
Kocabas Fatih ◽  
Chengcheng Zhang ◽  
Hesham A. Sadek ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Function ◽  
Metabolic Pathways ◽  
Energy Production ◽  
Mtor Signaling ◽  
Atp Content ◽  
Mitochondrial Oxidative Phosphorylation ◽  
Hematopoietic Stem ◽  
Mitochondrial Mass

Abstract Abstract 856 Unbalanced accumulation of reactive oxygen species (ROS) compromises self-renewal of hematopoietic stem cells (HSC). Nonetheless, survival of HSC in the hypoxic niche requires mitochondrial ROS-mediated activation of hypoxic response and significant metabolic adaptation. Hypoxia favors glycolysis to mitochondrial oxidative phosphorylation for energy production. In agreement with this, increased mitochondrial function results in defective maintenance of HSC as observed in Tsc1−/− mice through overactivation of mammalian target of rapamycin (mTOR) signaling pathway. Mechanisms that regulate the strict coordination of mitochondrial function with upstream metabolic pathways that is required for HSC maintenance and balancing ROS remain largely unknown. Foxo3 Forkhead transcription factor is a strong candidate for coordinating metabolic pathways in HSC. Evidence from our laboratory and others has identified Foxo3 as a critical regulator of HSC quiescence and a key modulator of oxidative stress in HSC. To address whether Foxo3 has a more global metabolic control of HSC activity, we investigated the mitochondrial function in Foxo3 mutant HSC. To achieve this, we measured ATP content and oxygen consumption, two major defining mitochondrial parameters, using ATP Bioluminescence Assay and Oxygen Biosensor analysis respectively in freshly isolated Lin−Sca-1+cKit+(LSK) CD34−Flk2− bone marrow cells isolated from wild type and Foxo3−/− mice. We showed that loss of Foxo3 leads to significant mitochondrial defects in HSC as indicated by a strong decrease in both ATP content and oxygen consumption. In agreement with this, the glycolytic flux in Foxo3 mutant HSC, as analyzed by 13C lactate production using gas chromatography-mass spectrometry, was increased indicating a shift in the ATP production from mitochondria to the cytosolic glycolysis in Foxo3 mutant HSC. In addition, mitochondrial mass and membrane potential, that is generated during mitochondrial oxidative phosphorylation for energy production, were assessed by flow cytometry in WT and Foxo3−/− LSK using mitotracker green and JC-1 probes respectively. Loss of Foxo3 increased both mitochondrial mass and membrane potential, likely reflecting a compensatory mechanism to the defective mitochondria in Foxo3−/− HSC. These anomalies may partly contribute, in addition to the known defective ROS detoxification, to the increased ROS levels previously observed in Foxo3−/− HSC. Importantly, the mitochondrial dysfunction was not due to the abnormal increase of ROS observed in Foxo3−/− HSC since in vivo treatment of Foxo3−/− mice with ROS scavenger N-Acetyl-Cysteine (NAC) for two weeks, did not revert the increased mitochondrial membrane potential. Altogether these results strongly suggest that Foxo3 is critical for the regulation of mitochondrial function in HSC. mTOR signaling controls major cellular metabolic processes and is critical for the regulation of mitochondrial function in HSC. We have previously found that mTOR signaling is amplified by a redox-mediated mechanism in Foxo3−/− hematopoietic progenitors (Yalcin et al., 2010). Thus, we asked whether mTOR signaling is involved in the Foxo3 regulation of mitochondria function in HSC. Interestingly, in vivo treatment of Foxo3−/− mice for two weeks with rapamycin, a specific inhibitor of mTOR complex 1 (mTORC1) activity, as measured by the decrease in phosphorylation of ribosomal protein S6 in HSC, mitigates the increased mitochondrial membrane potential and normalizes ROS levels in Foxo3−/− HSC, suggesting that Foxo3 regulation of mitochondria is mediated by mTOR signaling in HSC. Notably, rapamycin treatment rescued partially Foxo3−/− HSC pool and function as measured by the number and frequency of LSK in Foxo3−/− mice, as well as their long-term repopulation ability measured by the capacity of CD48−CD150+LSK cells to repopulate the hematopoietic compartment in lethally irradiated recipient mice within 8 weeks. Taken together, our findings reveal a new function for Foxo3 in the control of mitochondria in HSC and support a model in which mitochondria is key to the maintenance of HSC. We propose that a Foxo3-mTOR signaling node partly controls mitochondrial function in HSC. These findings are likely to have an important impact on our understanding of the metabolic regulation of hematopoietic and leukemic stem cells and may be of therapeutic value. Disclosures: No relevant conflicts of interest to declare.

P–139 The role of hormones and LH receptor expression of granulosa cells collected from large and small follicles in natural/minimal stimulation cycle IVF

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
T Okubo ◽  
H Teruaki ◽  
O Noriyuki ◽  
O Kenji ◽  
S Tomoya
Keyword(s):  
Mrna Expression ◽  
Granulosa Cells ◽  
Receptor Expression ◽  
Mrna Levels ◽  
Lh Receptor ◽  
Small Follicle ◽  
Estradiol Levels ◽  
Natural Cycles ◽  
Follicular Fluids

Abstract Study question Do different follicle sizes influence gonadotropins (LH, FSH) and sex steroid (estradiol) in follicular fluids and LH receptor expression (LHCGR) in cumulus oocyte complexes (COCs)? Summary answer It was found that differences in levels of FSH, estradiol values and LHCGR mRNA expression level in COCs between small and large follicles. What is known already The maturity rate in oocytes of small follicle is significantly lower compared to that of large follicles. Study design, size, duration After obtaining written consents from 78 infertile patients, we aspirated the large (&gt;15 mm) and small (&lt;5 mm) follicles, and collected follicular fluids at oocyte retrieval. Participants/materials, setting, methods We measured levels of LH, FSH and estradiol by enzyme immunoassay from large and small follicular fluids after oocytes retrievals. All collected oocytes were distinguished from large and small follicles, we confirmed the maturity of retrieved oocytes by the presence of first polar body. Then we extracted total RNA from granulosa cells and measured mRNA expression of LHCGR, encoding the human LH receptor, by quantitative real-time PCR. Each value was normalized to ACTB mRNA levels. Main results and the role of chance LH levels were nearly equal between small and large follicles (P = 0.8356). Whereas FSH and estradiol levels were significantly lower in small follicles (P &lt; 0.0001). The expression levels of LHCGR mRNA were significantly lower in small follicles than in large follicles during natural cycles. The maturity rate in oocytes of small follicle was significantly lower compared to that of large follicles (96.0% vs. 21.7%, P &lt; 0001). Limitations, reasons for caution The main limitation of the present study was collected by 42 natural cycles and 36 mild stimulation cycles with letrozole following low-dose clomiphene. Wider implications of the findings: In spite of almost the same LH levels between two groups, the reason why the significantly lower maturation rates of oocytes collected from small follicles is poor LHCGR mRNA expression due to insufficient granulosa cells glowth because of low FSH and estradiol levels. Trial registration number Not applicable

PHYSICAL DISORDERS AFTER HYSTERECTOMY DEPEND ON MRS AND SERUM ESTRADIOL LEVELS

2016 ◽  
pp. 41-45
Author(s):  
Lam Huong Le
Keyword(s):  
Key Words ◽  
Cross Sectional Study ◽  
Sexual Disorders ◽  
Serum Estradiol ◽  
Sectional Study ◽  
Cross Sectional ◽  
Total Hysterectomy ◽  
Physical Disorders ◽  
The Relationship ◽  
Estradiol Levels

Objectives: To study the relationship between serum estradiol and physical disorders after hysterectomy. Methods: A descriptive cross sectional study on 151 women after total hysterectomy from 12/2008 to 11/2010 at Hue Cental Hospial. Results: Serious disorders in total hysterectomy group include: physical disorders (34.78%), maternal disorders (71.74%), sexual disorders (58.70%). There were significant differences these disorders between total hysterectomy group and hysterectomy with/without accessiory remove group, as well as menopause group. Conclusion: Serum estradiol levels was inversely correlated with physical disorders, martenal disorders, sexual disorders and other disorders. Key words: hysterectomy, serum estradiol

scholarly journals Constitutive Expression of TERT Enhances β-Klotho Expression and Improves Age-Related Deterioration in Early Bovine Embryos

2021 ◽  
Vol 22 (10) ◽  
pp. 5327
Author(s):  
Lianguang Xu ◽  
Muhammad Idrees ◽  
Myeong-Don Joo ◽  
Tabinda Sidrat ◽  
Yiran Wei ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Constitutive Expression ◽  
Growth Factor Receptor ◽  
Oocyte Quality ◽  
Low Fertility ◽  
Treatment Groups ◽  
Age Related ◽  
Early Embryos ◽  
Plasmid Injection ◽  
Telomere Lengthening

Age-associated decline in oocyte quality is one of the dominant factors of low fertility. Aging alters several key processes, such as telomere lengthening, cell senescence, and cellular longevity of granulosa cells surrounding oocyte. To investigate the age-dependent molecular changes, we examined the expression, localization, and correlation of telomerase reverse transcriptase (TERT) and β-Klotho (KLB) in bovine granulosa cells, oocytes, and early embryos during the aging process. Herein, cumulus-oocyte complexes (COCs) obtained from aged cows (>120 months) via ovum pick-up (OPU) showed reduced expression of β-Klotho and its co-receptor fibroblast growth factor receptor 1 (FGFR1). TERT plasmid injection into pronuclear zygotes not only markedly enhanced day-8 blastocysts’ development competence (39.1 ± 0.8%) compared to the control (31.1 ± 0.5%) and D-galactose (17.9 ± 1.0%) treatment groups but also enhanced KLB and FGFR1 expression. In addition, plasmid-injected zygotes displayed a considerable enhancement in blastocyst quality and implantation potential. Cycloastragenol (CAG), an extract of saponins, stimulates telomerase enzymes and enhances KLB expression and alleviates age-related deterioration in cultured primary bovine granulosa cells. In conclusion, telomerase activation or constitutive expression will increase KLB expression and activate the FGFR1/β-Klotho pathway in bovine granulosa cells and early embryos, inhibiting age-related malfunctioning.

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