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Effect of GnRHa ovulation trigger dose on follicular fluid characteristics and granulosa cell gene expression profiles.

ABSTRACT: A recent dose-finding study showed no significant differences in number of mature oocytes, embryos and top-quality embryos when triptorelin doses of 0.2, 0.3 or 0.4 mg were used to trigger final oocyte maturation in oocyte donors co-treated with a gonadotropin-releasing hormone (GnRH) antagonist. This analysis investigated whether triptorelin dosing for triggering final oocyte maturation in oocyte donors induced differences in follicular fluid (FF) hormone levels and granulosa cell gene expression.This single-centre, randomised, parallel, investigator-blinded trial was conducted in oocyte donors undergoing a single stimulation cycle at IVFMD, My Duc Hospital, Ho Chi Minh City, Vietnam, from August 2014 to March 2015. A total of 165 women aged 18-35 years with body mass index <28 kg/m2, anti-Müllerian hormone >1.25 ng/mL, and antral follicle count ?6 were randomised to three different triptorelin doses for trigger. The main outcome was concentration of steroid hormones in FF collected from the first punctured follicle on each side. Moreover, luteinising hormone receptor (LHR), 3?-hydroxy-steroid-dehydrogenase (3ßHSD) and inhibin-Ba (INHB-A) gene expression in cumulus and mural granulosa cells were investigated in a subset of women from each group.Progesterone and oestradiol levels in FF did not differ significantly by trigger doses; findings were similar for 3?HSD, LHR and INHB-A gene expression in both cumulus and mural granulosa cells.In women co-treated with a GnRH antagonist, no significant differences in FF steroid levels and granulosa cell gene expression were seen when different triptorelin doses were used to trigger final oocyte maturation.

SUBMITTER: Vuong TNL

PROVIDER: S-EPMC5401703 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature

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Effect of GnRHa ovulation trigger dose on follicular fluid characteristics and granulosa cell gene expression profiles.

Vuong Thi Ngoc Lan TNL Ho M T MT Ha T Q TQ Jensen M Brehm MB Andersen C Yding CY Humaidan P P

Journal of assisted reproduction and genetics 20170214 4

<h4>Purpose</h4>A recent dose-finding study showed no significant differences in number of mature oocytes, embryos and top-quality embryos when triptorelin doses of 0.2, 0.3 or 0.4 mg were used to trigger final oocyte maturation in oocyte donors co-treated with a gonadotropin-releasing hormone (GnRH) antagonist. This analysis investigated whether triptorelin dosing for triggering final oocyte maturation in oocyte donors induced differences in follicular fluid (FF) hormone levels and granulosa ce ...[more]

PMID: 28197932

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Project description:BackgroundSuperovulation treatment had some adverse effects on maturity and development of oocytes. Can superovulation dose of gonadotropins (Gns) affect the transcriptome of granulosa cells and follicular fluid (FF) hormone levels?MethodsOne leading pre-ovulatory follicle per subject was used from three natural-cycle and four Gn-stimulated patients. Granulosa cells and FF samples were collected from the same leading follicle of each patient. RNA was extracted from granulosa cells and subjected to deep sequencing and analysis. Follicle-stimulating hormone (FSH), estradiol (E2), androstenedione (AND), testosterone (T), luteinizing hormone (LH), and progesterone (P4) levels in FF were measured by immunoassays. Student's t test was used for statistical analysis.ResultsA total of 715 genes were up-regulated, and 287 genes were down-regulated, in the Gn-stimulated group relative to the control group. Gene Ontology analysis revealed that the down-regulated genes were enriched in cell cycle and meiosis pathways, primarily those associated with follicle or oocyte maturation and quality. On the other hand, the up-regulated genes were enriched in functions related to immunity and cytokine-cytokine receptor interactions. Compared to the follicles of natural cycle, the E2 and LH concentrations were significantly reduced (P < 0.001), the P4 concentration was significantly increased (P = 0.003), and the concentrations of FSH, T and AND had no difference in the follicles of Gn-stimulated cycle.ConclusionsCell cycle- and meiosis-associated genes were down-regulated by Gns stimulation, whereas immune- and cytokine-associated genes were up-regulated. Hormone levels were also affected by Gns stimulation. Compared with natural-cycle follicles,putative markers associated with oocyte quality and follicle maturation were significantly different from those in Gn-stimulated follicles. Hormone levels in follicles were compatible with the steroidogenic patterns of granulosa cell, which reflects the follicle maturation and oocyte quality.

Project description:INTRODUCTION: Defensins are cytotoxic peptides and have a well-defined role in host defense. Human alpha defensins 1-3 (HNP1-3) are primarily produced by peripheral neutrophils and constitute about 50% of the azurphil granule protein. Studies have suggested that peripheral neutrophils and the resident neutrophils in the ovary enhance the release of IL-8 and TNF-alpha that play a role in ovulation and influence fertilisation rate and IVF outcome. The production of HNP1-3 by follicular fluid and its role in ovulation has never studied. The aim of this study was to demonstrate the presence of HNP1-3 in follicular fluid and to ascertain its correlation with fertilisation rate and IVF outcome. METHOD: Women attending the Reproductive Medicine Unit at Liverpool Women's Hospital UK, for IVF treatment were invited to participate in the study. Sixty-three patients were recruited for the study and underwent controlled ovarian stimulation and oocyte retrieval according to the unit's protocol. Fluid from the first follicle only was collected to minimise blood contamination of the sample and HNP1-3 was estimated using ELISA technique. RESULTS: HNP1-3 was detected in follicular fluid samples. The concentration did not correlate with the fertilisation rate (r=0.01). The concentrations were also not significantly different in the women who did or did not become pregnant following treatment. Subgroup analysis showed that women with endometriosis were not more likely to have higher levels of the HNP1-3 when compared with controls (male factor infertility group). CONCLUSION: This is the first study to show the expression of HNP1-3 in follicular fluid. HNP1-3 concentrations did not correlate with fertilisation rate or IVF outcome. It did not show an increased expression of HNP1-3 in fluid collected from women with endometriosis suggesting that inflammatory processes associated with endometriosis do not influence HNP1-3 concentration in the follicular fluid. Further studies to evaluate the correlation between HNP1-3 and IL-8 and TNF-alpha may clarify the role of defensins in ovulation.

Project description:BackgroundNerve growth factor (β-NGF) from llama seminal plasma has been described as a potent ovulatory and luteotrophic molecule after intramuscular or intrauterine infusion in llamas and alpacas. We tested the hypothesis that systemic administration of purified β-Nerve Growth Factor (β-NGF) during the preovulatory stage will up-regulate steroidogenic enzymes and Vascular Endothelial Growth Factor (VEGF) gene expression in granulosa cells inducing a change in the progesterone/estradiol ratio in the follicular fluid in llamas.MethodsExperiment I: Female llamas (n = 64) were randomly assigned to receive an intramuscular administration of: a) 50 μg gonadorelin acetate (GnRH, Ovalyse, Pfizer Chile SA, Santiago, Chile, n = 16), b) 1.0 mg of purified llama β-NGF (n = 16), or c) 1 ml phosphate buffered saline (PBS, negative control group, n = 16). An additional group of llamas (n = 16) were mated with a fertile male. Follicular fluid and granulosa cells were collected from the preovulatory follicle at 10 or 20 h after treatment (Time 0 = administration of treatment, n = 8/treatment/time point) to determine progesterone/estradiol concentration and steroidogenic enzymes and VEGF gene expression at both time points. Experiment II: Granulosa cells were collected from preovulatory follicles from llamas (n = 24) using ultrasound-guided transvaginal follicle aspiration for in vitro culture to determine mRNA relative expression of Steroidogenic Acute Regulatory Protein (StAR) and VEGF at 10 or 20 h (n = 4 replicates) and progesterone secretion at 48 h (n = 4 replicates) after LH or β-NGF treatment.ResultsExperiment I: There was a significant increase in the progesterone/estradiol ratio in mated llamas or treated with GnRH or purified β-NGF. There was a significant downregulation in the mRNA expression of Aromatase (CYP19A1/P450 Arom) for both time points in llamas mated or treated with GnRH or llama purified β-NGF with respect to the control group. All treatments except β-NGF (20 h) significantly up regulated the mRNA expression of 3-beta-hydroxysteroid dehydrogenase (HSD3B) whereas the expression of StAR and Side-Chain cleavage enzyme (CYP11A1/P450scc) where significantly up regulated only by mating (20 h), or β-NGF at 10 or 20 h after treatment. VEGF was up regulated only in those llamas submitted to mating (10 h) or treated with purified β-NGF (10 and 20 h). Experiment II: Only β-NGF treatment induced an increase of mRNA abundance of StAR from llama granulosa cells at 20 h of in vitro culture. There was a significant increase on mRNA abundance of VEGF at 10 and 20 h of in vitro culture from granulosa cells treated with β-NGF whereas LH treatment increases VEGF mRNA abundance only at 20 h of in vitro culture. In addition, there was a significant increase on progesterone secretion from llama granulosa cells 48 h after LH or β-NGF treatment.ConclusionsSystemic administration of purified β-NGF from llama seminal fluid induced a rapid shift from estradiol to progesterone production in the preovulatory follicle. Differences in gene expression patterns of steroidogenic enzymes between GnRH and mated or β-NGF-treated llamas suggest local effects of seminal components on the preovulatory follicle.

Dataset Information

Effect of GnRHa ovulation trigger dose on follicular fluid characteristics and granulosa cell gene expression profiles.

Publications

Effect of GnRHa ovulation trigger dose on follicular fluid characteristics and granulosa cell gene expression profiles.

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