scholarly journals OVARIAN GRANULOSA CELLS FROM WOMEN WITH PCOS EXPRESS LOW LEVELS OF SARS-COV-2 RECEPTORS AND CO-FACTORS

2021 ◽  
Author(s):  
Aalaap Anand Naigaonkar ◽  
Krutika Madhukar Patil ◽  
Shaini Joseph ◽  
Indira Hinduja ◽  
Srabani Mukherjee
Keyword(s):  
Granulosa Cells ◽  
Reproductive Tract ◽  
Polycystic Ovary ◽  
Ovarian Follicle ◽  
Cell Types ◽  
Female Reproductive Tract ◽  
Vitro Fertilization ◽  
Ovarian Granulosa Cells ◽  
Lower Expression

Purpose: Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection is global pandemic with more than 3 million deaths so far. Female reproductive tract organs express coronavirus-associated receptors and factors (SCARFs); suggesting they may be susceptible to SARS-CoV-2 infection however the susceptibility of ovary/follicle/oocyte to the same is still elusive. Co-morbidities like obesity, type-2 diabetes mellitus, cardiovascular disease etc. increase the risk of SARS-CoV-2 infection. These features are common in women with polycystic ovary syndrome (PCOS), warranting further scope to study SCARFs expression in ovary of these women. Materials and methods: SCARFs expression in ovary and ovarian tissues of women with PCOS and healthy women was explored by analyzing publically available microarray datasets. Transcript expression of SCARFs were investigated in mural and cumulus granulosa cells (MGCs and CGCs) from control and PCOS women undergoing in vitro fertilization (IVF). Results: Microarray data revealed that ovary expresses all genes necessary for SARS-CoV-2 infection. PCOS women mostly showed down-regulated/unchanged levels of SCARFs. MGCs and CGCs from PCOS women showed lower expression of receptors ACE2, BSG and DPP4 and protease CTSB than in controls. MGCs showed lower expression of protease CTSL in PCOS than in controls. Expression of TMPRSS2 was not detected in both cell types. Conclusions: Human ovarian follicle may be susceptible to SARS-CoV-2 infection. Lower expression of SCARFs in PCOS indicate that the risk of SARS-CoV-2 infection to the ovary may be lesser in these women than controls. This knowledge may help in safe practices at IVF settings in the current pandemic. Keywords: SARS-CoV-2, COVID-19, Ovarian granulosa cells, Oocyte, PCOS, IVF

scholarly journals LRH-1 High Expression in the Ovarian Granulosa Cells of PCOS Patients

2021 ◽  
Author(s):  
Xiao Yang ◽  
Qiumin Wang ◽  
Ying Wang ◽  
Tian Song ◽  
Yanjun Zheng ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Polycystic Ovary ◽  
Pearson Correlation ◽  
Statistical Significance ◽  
Control Group ◽  
Vitro Fertilization ◽  
Real Time Quantitative Pcr ◽  
Ovarian Granulosa Cells

Abstract Purpose: Polycystic ovary syndrome (PCOS) is considered as one of the most common endocrine disorder with heterogeneity. There are also reports that liver receptor homolog 1 [LRH-1 or nuclear receptor subfamily 5 group A member 2 (NR5A2)] plays an important role in the reproductive system. But up to now, there are no reports related to the link with PCOS and LRH-1. In this study, we aimed to detect the LRH-1 expression in the ovarian granulosa cell of PCOS patients and explore the potential relationship between LRH-1 and PCOS. Methods: 146 follicular fluid sample were collected in this study, including 72 from PCOS patients and 74 from control patients who underwent intracytoplasmic sperm injection (ICSI) or in vitro fertilization-embryo transfer (IVF-ET). The ovarian granulosa cells were extracted from the patient's follicular fluid by magnetic-activated cell sorting (MACS) method, and the real-time quantitative PCR (qRT-PCR) was used to measure the expression of LRH-1 in ovarian granulosa cells. Then we analyzed the correlation between the expression level of LRH-1 and the clinical characteristics of patient by using Pearson Correlation analysis. Results: The expression of LRH-1 was significantly higher in PCOS patients ovarian granulosa cells than that in the control patients [vs(1.38±0.47)vs(1.03±0.32), t=5.327, p<0.0001], and it was positively correlated with antral follicles counting (AFC) (r=0.3607, p<0.0001)and the serum AMH(r=0.2662, p=0.0012)\LH(r=0.2518, p=0.0022)\T(r=0.2516, p=0.0022) in all patients. No statistical significance between LRH-1 and BMI, FSH, HOMA-IR, DHE-S, progesterone. Conclusions: Compared with the control group, we found that LRH-1 was highly expressed in the ovarian granulosa cells of PCOS patients. Our study has revealed the relationship between the LRH-1 expression and PCOS, which suggested that LRH-1 may play an important role in ovulation disorders. While this finding provided new ideas for the study of the pathogenesis, it also provided a theoretical basis for the clinical diagnosis and treatment for PCOS.

scholarly journals ANGPTL4 expression in ovarian granulosa cells is associated with polycystic ovary syndrome

2021 ◽  
Author(s):  
Qi Jiang ◽  
Yanjun Zheng ◽  
Ping Li ◽  
Yuehong Bian ◽  
Wenqi Wang ◽  
...  
Keyword(s):  
Polycystic Ovary Syndrome ◽  
Granulosa Cells ◽  
Polycystic Ovary ◽  
Clinical Information ◽  
Positive Association ◽  
Control Group ◽  
Ovary Syndrome ◽  
Vitro Fertilization ◽  
Ovarian Granulosa Cells

ABSTRACT Objectives:To characterize the expression of ANGPTL4 in ovarian granulosa cells (GCs) and its association with polycystic ovary syndrome. Design: A retrospective study. Setting: University-based center for reproductive medicine. Participants:This study included 104 PCOS patients and 112 control women undergoing in vitro fertilization-embryo transfer (IVF-ET) from the reproductive hospital affiliated with Shandong University between 2019 and 2021. Methods: The mRNA expression of ANGPTL4 in GCs were assessed by reverse transcription and real-time quantitative (RT-q)PCR, then clinical information for these patients were reviewed and analyzed. Main outcome measures: ANGPTL4 expression in GCs in participants, correlation between ANGPTL4 expression level and metabolic characteristics of patients and predictive value of ANGPTL4 expression for PCOS. Results:The RT-qPCR results showed that ANGPTL4 expression in the control group is significantly lower than that in the PCOS group(P=0.000). It indicated positive association with AMH(r=0.211), HOMA-IR(r=0.174), LDL/HDL(r=0.176), ApoB/ApoAI(r=0.155) and TC/HDL(r=0.189). Additionally, the ANGPTL4 expression in the ovarian granulosa cells might be a independent factor in PCOS(OR:3.345, 95%CI:1.951–5.734) and served as a good predictor for PCOS (AUC0.704, 95%CI 0.633-0.774,P＜0.001). Conclusions:For the first time our study revealed on the higher ANGPTL4 expression in ovarian GCs with PCOS, and its association with glucose and lipid metabolism showed that ANGPTL4 might be a predictor for PCOS and play an important role in metabolism and pathogenesis of PCOS. Funding: National Key R&D Program of China (2018YFC1003202, 2016YFC1000604) and Taishan scholar project special funds (No. ts201712103). Key words: polycystic ovary syndrome, angiopoietin-like protein 4, mRNA, ovarian granulosa cell, glycolipid metabolism

Development of parthenogenetic and fertilized mouse embryos in the uterus and in extra-uterine sites

Development ◽  
1975 ◽  
Vol 34 (2) ◽  
pp. 387-405
Author(s):  
S. A. Iles ◽  
M. W. McBurney ◽  
S. R. Bramwell ◽  
Z. A. Deussen ◽  
C. F. Graham
Keyword(s):  
Reproductive Tract ◽  
Neural Tissue ◽  
Cell Types ◽  
Mouse Embryos ◽  
Female Reproductive Tract ◽  
Embryonic Cells ◽  
Haploid Embryos ◽  
Keratinized Epithelium ◽  
Mouse Eggs

Mouse eggs were activated with hyaluronidase in vitro and subsequently transferred to the oviduct. In the female reproductive tract they formed morulae and blastocysts which died soon after implantation. Haploid blastocysts were transferred beneath the kidney capsule and here some formed disorganized egg-cylinder structures in a week. Morulae and blastocysts from haploid and diploid parthenogenones were also transferred beneath the testis capsule. Two to four months later the growths which had formed were sectioned. They contained neural tissue, pigment, keratinized epithelium, glandular epithelium, ciliated epithelium, cartilage, bone, muscle, adipose tissue, and haemopoietic tissue. The range of cell types was similar to that produced by fertilized control blastocysts except that the parthenogenones did not form identifiable yolk-sac carcinoma or embryonal carcinomacells. The growths from haploid and diploid parthenogenones in the testis were stained with Feulgen and their DNA content measured. Growths from diploid embryos contained the normal diploid amount of DNA while growths from haploid embryos contained less than this amount. Cell cultures were prepared from the growths. The cells which were investigated contained no Y chromosome, suggesting that they were derived from the embryonic cells rather than the cells of the male host. These cells contained a near diploid chromosome number, although some of them were originally derived from haploid embryos.

scholarly journals CatSperζ regulates the structural continuity of sperm Ca2+ signaling domains and is required for normal fertility

eLife ◽  
2017 ◽  
Vol 6 ◽  
Author(s):  
Jean-Ju Chung ◽  
Kiyoshi Miki ◽  
Doory Kim ◽  
Sang-Hee Shim ◽  
Huanan F Shi ◽  
...  
Keyword(s):  
Reproductive Tract ◽  
Female Reproductive Tract ◽  
Targeted Disruption ◽  
Vitro Fertilization ◽  
Male Subfertility ◽  
Epididymal Spermatozoa ◽  
Mammalian Female ◽  
Signaling Domains

We report that the Gm7068 (CatSpere) and Tex40 (CatSperz) genes encode novel subunits of a 9-subunit CatSper ion channel complex. Targeted disruption of CatSperz reduces CatSper current and sperm rheotactic efficiency in mice, resulting in severe male subfertility. Normally distributed in linear quadrilateral nanodomains along the flagellum, the complex lacking CatSperζ is disrupted at ~0.8 μm intervals along the flagellum. This disruption renders the proximal flagellum inflexible and alters the 3D flagellar envelope, thus preventing sperm from reorienting against fluid flow in vitro and efficiently migrating in vivo. Ejaculated CatSperz-null sperm cells retrieved from the mated female uterus partially rescue in vitro fertilization (IVF) that failed with epididymal spermatozoa alone. Human CatSperε is quadrilaterally arranged along the flagella, similar to the CatSper complex in mouse sperm. We speculate that the newly identified CatSperζ subunit is a late evolutionary adaptation to maximize fertilization inside the mammalian female reproductive tract.

scholarly journals Gene Expression in Granulosa Cells From Small Antral Follicles From Women With or Without Polycystic Ovaries

2019 ◽  
Vol 104 (12) ◽  
pp. 6182-6192 ◽  
Author(s):  
Lisa Ann Owens ◽  
Stine Gry Kristensen ◽  
Avi Lerner ◽  
Georgios Christopoulos ◽  
Stuart Lavery ◽  
...  
Keyword(s):  
Gene Expression ◽  
Granulosa Cells ◽  
Polycystic Ovary ◽  
Ovarian Tissue ◽  
Polycystic Ovaries ◽  
Vitro Fertilization ◽  
Antral Follicles ◽  
Oocyte Aspiration ◽  
And Function

Abstract Context Polycystic ovary syndrome (PCOS) is the most common cause of anovulation. A key feature of PCOS is arrest of follicles at the small- to medium-sized antral stage. Objective and Design To provide further insight into the mechanism of follicle arrest in PCOS, we profiled (i) gonadotropin receptors; (ii) characteristics of aberrant steroidogenesis; and (iii) expression of anti-Müllerian hormone (AMH) and its receptor in granulosa cells (GCs) from unstimulated, human small antral follicles (hSAFs) and from granulosa lutein cells (GLCs). Setting GCs from hSAFs were collected at the time of cryopreservation of ovarian tissue for fertility preservation and GLCs collected during oocyte aspiration before in vitro fertilization/intracytoplasmic sperm injection. Participants We collected hSAF GCs from 31 women (98 follicles): 10 with polycystic ovaries (PCO) and 21 without. GLCs were collected from 6 women with PCOS and 6 controls undergoing IVF. Main Outcome Measures Expression of the following genes: LHCGR, FSHR, AR, INSR, HSD3B2, CYP11A1, CYP19, STAR, AMH, AMHR2, FST, INHBA, INHBB in GCs and GLCs were compared between women with PCO and controls. Results GCs in hSAFs from women with PCO showed higher expression of LHCGR in a subset (20%) of follicles. Expression of FSHR (P < 0.05), AR (P < 0.05), and CYP11A1 (P < 0.05) was lower, and expression of CYP19A1 (P < 0.05), STAR (P < 0.05), HSD3B2 (P = NS), and INHBA (P < 0.05) was higher in PCO GCs. Gene expression in GL cells differed between women with and without PCOS but also differed from that in GCs. Conclusions Follicle arrest in PCO is characterized in GCs by differential regulation of key genes involved in follicle growth and function.

Capacitation mechanisms, and the role of capacitation as seen in eutherian mammals

1996 ◽  
Vol 8 (4) ◽  
pp. 581 ◽  
Author(s):  
RA Harrison
Keyword(s):  
Cell Death ◽  
Reproductive Tract ◽  
Physiological State ◽  
Hormonal Control ◽  
Female Reproductive Tract ◽  
Functional Heterogeneity ◽  
Vitro Fertilization ◽  
Range Of Functions

Capacitation, the process whereby spermatozoa are rendered capable of interacting with and fertilizing the egg, was discovered more than 40 years ago. However, our understanding of it is still far from satisfactory. Several factors conspire to obfuscate studies of capacitation mechanisms: the inherent functional heterogeneity of sperm populations, the range of functions used as parameters of capacitation (whence the endpoint of the process has become conceptually uncertain), and the several profound differences between model in vitro fertilization (IVF) systems and the situation in vivo in the female reproductive tract. Recent investigations in the author's laboratory have shown that bicarbonate/CO2, an essential component for successful IVF, causes rapid changes in lipid architecture of the sperm plasma membrane and slower changes in surface coating. These changes are accompanied by membrane destabilization and cell death. Evidence suggests that bicarbonate's actions are mediated through cyclic nucleotide signalling. Of particular note is the heterogeneity in rate of response to bicarbonate shown by individual cells in the sperm populations. Taken together with other observations, the findings suggest that capacitation is a series of positive destabilizing events that eventually lead to cell death. The 'capacitated' state would then be a window of destabilization within which spermatozoa can undergo a zona-induced acrosome reaction and display hyperactivated motility. Further along the destabilization pathway, spontaneous acrosome reactions would occur before total membrane degeneration. In vivo, capacitation would be a conflict between destabilization and sperm survival. Concentrations of bicarbonate are maintained low in the cauda epididymidis, where sperm survive for long periods, and one may speculate that hormonal control of local bicarbonate/CO2 in oviducal 'storage' sites in the female tract could allow 'safe' sequestering of live spermatozoa until around the time of ovulation; the environment may then change to produce a 'capacitating' effect, whence, due to the inherent functional heterogeneity of the sequestered population, small numbers of capacitated spermatozoa are released sequentially. In this way, a succession of spermatozoa in the correct physiological state may be provided for the freshly ovulated egg.

scholarly journals The Cultivation of Human Granulosa Cells

2008 ◽  
Vol 51 (3) ◽  
pp. 165-172 ◽  
Author(s):  
Lenka Brůčková ◽  
Tomáš Soukup ◽  
Jiří Moos ◽  
Martina Moosová ◽  
Jana Pavelková ◽  
...  
Keyword(s):  
Growth Factors ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Doubling Time ◽  
Ovarian Follicle ◽  
Vitro Fertilization ◽  
Thecal Cells ◽  
Cell Doubling Time

The major functions of granulosa cells (GCs) include the production of steroids, as well as a myriad of growth factors to interact with the oocyte during its development within the ovarian follicle. Also FSH stimulates GCs to convert androgens (coming from the thecal cells) to estradiol by aromatase. However, after ovulation the GCs produce progesterone that may maintain a potential pregnancy. Experiments with human GCs are mainly focused on the purification of GCs from ovarian follicular fluid followed by FACS analysis or short-term cultivation. The aim of our study was to cultivate GCs for a long period, to characterize their morphology and phenotype. Moreover, we have cultivated GCs under gonadotropin stimulation in order to simulate different pathological mechanisms during folliculogenesis (e.g. ovarian hyperstimulation syndrome). GCs were harvested from women undergoing in vitro fertilization. Complex oocyte-cumulus oophorus was dissociated by hyaluronidase. The best condition for transport of GCs was optimized as short transport in follicular fluid at 37 °C. GCs expansion medium consisted of DMEM/F12, 2 % FCS, ascorbic acid, dexamethasone, L-glutamine, gentamycine, penicillin, streptomycin and growth factors (EGF, bFGF). GCs transported in follicular fluid and cultivated in 2 % FCS containing DMEM/F12 medium supplemented with follicular fluid presented increased adhesion, proliferation, viability and decreased doubling time. Cell viability was 92 % and mean cell doubling time was 52 hrs. We have optimized transport and cultivation protocols for long-term cultivation of GCs.

scholarly journals sRAGE Downregulates the VEGF Expression in PCOS Ovarian Granulosa Cells

2020 ◽  
Author(s):  
Jingyan Wang ◽  
Yichun Guan ◽  
Yi Liu ◽  
Liang Wang ◽  
Zhan Zhang ◽  
...  
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Granulosa Cells ◽  
Ovarian Tissue ◽  
Rt Pcr ◽  
Vegf Expression ◽  
Vitro Fertilization ◽  
Protein Levels ◽  
Ovarian Granulosa Cells

Abstract Objective High expression of VEGF in ovarian tissue, serum and follicular fluid of PCOS women is involved in the physiological and pathogenesis processes of PCOS. Our objective was to investigate the effect of sRAGE on VEGF expression and EGF-like growth factor in PCOS ovarian granulosa cells.Methods We collected ovarian granulosa cells of PCOS patients who underwent in vitro fertilization (IVF). Then treatment ovarian granulosa cells with different concentrations of sRAGE. Levels of VEGF, AREG, BTC and EREG mRNA were examined by quantitative RT-PCR. The protein levels of VEGF, AREG, BTC and EREG were measured by ELISA.Results Treatment with sRAGE decrease the production of VEGF, and the effects were dependent on the concentrations of sRAGE (P < 0.05). Simultaneously, the expression of the EGF-like growth factors AREG, BTC and EREG were decreased, and the expression were dependent on the concentrations of sRAGE (P < 0.05).Conclusions sRAGE may downregulate VEGF expression in PCOS ovarian granulosa cells,and EGF-like growth factor pathway may be involved in this process.

Simvastatin inhibits Wnt/β-catenin pathway in uterine leiomyoma

Endocrinology ◽  
2021 ◽  
Author(s):  
Malak El Sabeh ◽  
Subbroto Kumar Saha ◽  
Sadia Afrin ◽  
Mostafa A Borahay
Keyword(s):  
Uterine Leiomyoma ◽  
Reproductive Tract ◽  
Controlled Trial ◽  
Active Form ◽  
Cell Types ◽  
Female Reproductive Tract ◽  
Benign Tumors ◽  
Therapeutic Effects

Abstract The Wnt/β-catenin pathway is upregulated in uterine leiomyomas, the most common benign tumors in the female reproductive tract. Simvastatin is an anti-hyperlipidemic drug, and previous in vitro and in vivo reports showed it may have therapeutic effects in treating leiomyomas. The objective of this study is to examine the effects of simvastatin on the Wnt/β-catenin signaling pathway in leiomyoma. We treated primary and immortalized human leiomyoma cells with simvastatin and examined its effects using RT-qPCR, Western Blotting, and immunocytochemistry. We also examined the effects using human leiomyoma tissues from an ongoing, randomized controlled trial where women with symptomatic leiomyoma received simvastatin (40mg) or placebo for 3 months prior to their surgery. The results of this study reveal that simvastatin significantly reduced the expression of Wnt4 and its co-receptor LRP5. After simvastatin treatment, levels of total β-catenin and its active form, non-phosphorylated β-catenin, were reduced in both cell types. Additionally, simvastatin reduced the expression of Wnt4 and total β-catenin, as well as non-phosphorylated β-catenin protein expression in response to estrogen and progesterone. Simvastatin also inhibited the expression of c-Myc, a downstream target of the Wnt/β-catenin pathway. The effect of simvastatin on non-phosphorylated-β-catenin, the key regulator of the Wnt/β-catenin pathway, was recapitulated in human leiomyoma tissue. These results suggest that simvastatin may have a beneficial effect on uterine leiomyoma through suppressing the overactive Wnt/β-catenin pathway.

scholarly journals Integration of Sperm Motility and Chemotaxis Screening with a Microchannel-Based Device

2010 ◽  
Vol 56 (8) ◽  
pp. 1270-1278 ◽  
Author(s):  
Lan Xie ◽  
Rui Ma ◽  
Chao Han ◽  
Kai Su ◽  
Qiufang Zhang ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Reproductive Tract ◽  
Cumulus Cells ◽  
Female Reproductive Tract ◽  
Straight Channel ◽  
Vitro Fertilization ◽  
Mammalian Sperm ◽  
Reaction Capability ◽  
Mammalian Female

BACKGROUND Sperm screening is an essential step in in vitro fertilization (IVF) procedures. The swim-up method, an assay for sperm motility, is used clinically to select the ideal sperm for subsequent manipulation. However, additional parameters, including acrosome reaction capability, chemotaxis, and thermotaxis, are also important indicators of mammalian sperm health. To monitor both sperm motility and chemotaxis simultaneously during sperm screening, we designed and constructed a microdevice comprising a straight channel connected with a bibranch channel that mimics the mammalian female reproductive tract. METHODS The width and length of the straight channel were optimized to select the motile sperms. We selectively cultured cumulus cells in the bibranch channel to generate a chemoattractant-forming chemical gradient. Sperm chemotaxis was represented by the ratio of the sperm swimming toward different branches. RESULTS The percentage of motile sperms improved from 58.5% (3.8%) to 82.6% (2.9%) by a straight channel 7 mm in length and 1 mm in width. About 10% of sperms were found to be chemotactically responsive in our experiment, which is consistent with previous studies. CONCLUSIONS For the first time, we achieved the combined evaluation of both sperm motility and chemotaxis. The motile and chemotactically responsive sperms can easily be enriched on a lab-on-a-chip device to improve IVF outcome.

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