scholarly journals Ketoconazole Inhibits Ovulation as a Result of Arrest of Follicular Steroidogenesis in the Rat Ovary

2014 ◽  
Vol 8 ◽  
pp. CMRH.S15887 ◽  
Author(s):  
Michael Gal ◽  
Joseph Orly
Keyword(s):  
Cumulus Cell ◽  
Follicular Development ◽  
Ovarian Response ◽  
Control Group ◽  
Ovarian Steroidogenesis ◽  
Cell Complexes ◽  
Follicular Maturation ◽  
Fertility Treatments ◽  
Cell Layers ◽  
Ovulatory Follicles

Objective Ketoconazole (KCZ) is a known inhibitor of steroidogenic P450 enzymes in the adrenal cortex and the gonads. Previous studies examined the potential clinical use of KCZ for attenuation of ovarian response to gonadotropin treatments. This study aimed to use the superovuating rat model to explore the effect of KCZ on ovarian steroidogenesis, follicular function, and development toward ovulation. Methods Prepubertal rats were treated with equine chorionic gonadotropin (eCG)/human CG (hCG) resulting in multiple follicular development and ovulation. The effect of KCZ on this model was examined by administration of KCZ-gel formula and subsequent analyses of ovarian steroidogenesis, rate of ovulation, morphometric assessments of follicular parameters, and cell-specific steroidogenic maturation of the treated ovaries. Results When applied shortly before gonadotropin stimulation, KCZ markedly reduced ovarian progesterone, androstenedione, and estradiol levels down to 18.7, 36.5, and 19.0%, respectively ( P < 0.001). A single KCZ-gel administration of 6, 12, and 24 mg/rat resulted in reduction of ovulated ova/ovary down to 8.6 ± 4.9, 5.1 ± 4.3, and 2.4 ± 3.2, respectively, as compared to 13.6 ± 4.4 ova found in the oviduct of control-gel-injected animals ( P < 0.001). An alternative protocol made use of small KCZ doses injected in non-gel formula (5 mg/dose/8 hours), commenced with the eCG administration and terminated 24 hours later; this treatment readily inhibited the ovulation rates to 6.6 ± 6.6 as compared to 16.5 ± 4.1 ova/ovary in the control group ( P < 0.01). By contrast, KCZ failed to inhibit ovulation if administered 24 hours after eCG injection. Anovulation by KCZ resulted from arrest of follicular development at the stage of 800-840 μm Graafian follicles as compared to 920 μm of peri-ovulatory follicles (OFs) observed in the control group, P = 0.029. In addition, absence of CYP11A1 expression was evident in the granulosa cell layers of the growth-arrested follicles, which also lacked mucified mature cumulus cell complexes. Conclusion These results suggest that KCZ-mediated inhibition of follicular maturation probably results from impaired steroidogenesis at early phase of follicular development toward ovulation. Hence, attenuation of folliculogenesis by KCZ may be harnessed to modulate gonadotropin-ovarian stimulation in fertility treatments.

188 IMPROVEMENT OF IN VITRO CANINE OOCYTE MATURATION BY OVIDUCTAL SECRETOME

2017 ◽  
Vol 29 (1) ◽  
pp. 202 ◽  
Author(s):  
A. Lange-Consiglio ◽  
C. Perrini ◽  
P. Esposti ◽  
F. Cremonesi
Keyword(s):  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
Cumulus Cell ◽  
Follicular Development ◽  
Cumulus Cells ◽  
Chi Square ◽  
Cell Layers ◽  
Hoechst Staining

The in vitro maturation of canine oocyte is problematic because it is difficult to reproduce the oviducal microenvironment where the in vivo maturation occurs. Because cells are able to communicate with each other by paracrine action, oviducal cells could be in vitro cultivated to obtain the conditioned medium (CM) consisting of soluble factors and microvesicles (MV), which represent a carrier for nonsoluble molecules including microRNA. The aim of the present work was to investigate the effect of the addition of CM or MV, secreted by oviducal cells, to the canine in vitro maturation medium. To generate CM, cells from oviducts of 3 animals in late oestrus were cultured for 5 days at 38.5°C in a humidified atmosphere of 5% CO2. Supernatants were collected, pooled, centrifuged at 2500 × g, and stored at −80°C. Microvesicles were obtained by ultracentrifugation of CM at 100,000 × g for 1 h at 4°C and measured for concentration and size by a Nanosight instrument. Ovaries were obtained from 50 healthy domestic bitches (1–4 years old) of different breeds that underwent ovariectomy regardless of the oestrous cycle. Cumulus-oocyte complexes were released by slicing the ovarian cortex with a scalpel blade, and only Grade 1 cumulus-oocyte complexes (darkly granulated cytoplasm and surrounded by 3 or more compact cumulus cell layers) 110 to 120 µm in diameter were selected for culture. Maturation was performed at 38.5°C in a humidified atmosphere of 5% CO2 and 5% of O2 in bi-phasic systems: 24 h in SOF with 5.0 μg mL−1 of LH followed by 48 h in SOF supplemented with 10% of oestrous bitch serum and 10% CM or 50, 75, 100, or 150 × 106 MV mL−1 labelled with PKH-26. Control was the same medium without CM or MV. Oocytes were observed under a fluorescent microscope to detect metaphase II (MII), by Hoechst staining, and the incorporation of MV. Statistical analysis was performed by chi-square test. Results show that canine oviducal cells secreted MV of 234 ± 23 nm in size, underling that these MV fall within the shedding vesicles category. The incorporation of labelled MV occurred at first in cumulus cells, at 48 h of maturation, and then, at 72 h, in oocyte cytoplasm. These MV had a positive effect on maturation rate (MII) at the concentration of 75 and 100 × 106 MV mL−1 compared with CM and control (20.34 and 21.82 v. 9.09 and 3.95%, respectively). The concentration of 150 × 106 MV mL−1 provided only 9.26% of MII. To understand the role of MV, we assessed the expression of 3 microRNA (miRNA-30b, miR-375, and miR-503) that are involved in some key pathways (WNT, MAPK, ERbB, and TGFβ) regulating follicular development and meiotic resumption. The lower rate of MII with the higher concentration of MV is possibly due to the high level of miR-375, which recent literature shows to suppress the TGFβ pathway, leading to impaired oocyte maturation. In conclusion, the oviducal MV, or specific microRNA, are involved in cellular trafficking during oocyte maturation, and their possible use in vitro could facilitate the exploitation of canine reproductive biotechnologies.

116 EFFECTS OF l-GLUTAMINE ON CRYOPRESERVATION OF IMMATURE BOVINE OOCYTES

2006 ◽  
Vol 18 (2) ◽  
pp. 167
Author(s):  
C. Yamada ◽  
M. D. Goissis ◽  
H. V. A. Caetano ◽  
A. R. S. Coutinho ◽  
M. E. O. A. Assumpção ◽  
...  
Keyword(s):  
Amino Acids ◽  
In Vitro Maturation ◽  
Complex Structure ◽  
Cumulus Cell ◽  
Epifluorescence Microscopy ◽  
Control Group ◽  
Maturation Medium ◽  
Cell Layers ◽  
Bovine Oocytes

The cryopreservation of bovine oocytes remains a challenge despite significant reported progress. Immature bovine oocytes have a complex structure and the conventional cryoprotectants (penetrating cryoprotectants, sugars, and macromolecules) appear to be not sufficient to preserve them efficiently during freezing. Studies on semen and fibroblast cryopreservation indicate that amino acids, particularly l-glutamine, protect enzymes during freezing and increase the post-thaw viability. Therefore, the amino acids may optimize oocyte cryopreservation when associated with conventional cryoprotectants. This work evaluated the effect of l-glutamine on cryopreservation of immature bovine oocytes after in vitro maturation. Oocytes with homogeneous cytoplasm and several cumulus cell layers from slaughterhouse ovaries were distributed randomly in three groups: non-vitrified control, vitrified control, and vitrified with l-glutamine. Oocytes from vitrified groups were exposed for 10 min to PBS + 10% FCS + 10% ethylene glycol (EG) + 0.25 m trehalose (T), and for 30 s to PBS + 10% FCS + 25% EG + 25% dimethylsulfoxide + 0.5 m T at room temperature, adding 80 mm l-glutamine for the third group. Oocytes were loaded into OPS and plunged in liquid nitrogen. For thawing, OPS were immersed in PBS + 10% FCS + 10% EG + 1 m T for three min. Oocytes werethen placed in PBS + 10% FCS + 0.5 m T and in PBS + 10% FCS, remaining three min in each solution. For in vitro maturation, oocytes were washed three times on holding medium (TCM-HEPES + FCS + pyruvate + gentamycin), washed three times in maturation medium (TCM-bicarbonate + FCS + pyruvate + gentamycin + hCG + FSH + estradiol), and cultured in microdrops (90 μL) of maturation medium covered with mineral oil at 38.5°C under 5% CO2 in air and high humidity for 24 h. Oocytes were denuded, fixed in paraformaldehyde and triton, stained with Hoechst 33342, and evaluated under epifluorescence microscopy. Oocytes at metaphase II were considered matured. The group vitrified with l-glutamine had a significantly higher maturation rate than the group vitrified without l-glutamine; however, both had significantly lower maturation rates than the non-vitrified control group. In conclusion, l-glutamine improved the viability of vitrified oocytes. Table 1. Oocyte maturation rates of non-vitrified control, vitrified control, and vitrified with glutamine groups This work was supported by FAPESP 03/08543-1.

scholarly journals Effect of hyperandrogenism on ovarian function

Reproduction ◽  
2015 ◽  
Vol 149 (6) ◽  
pp. 577-585 ◽  
Author(s):  
Leandro M Velez ◽  
Maria F Heber ◽  
Silvana R Ferreira ◽  
Giselle A Abruzzese ◽  
Roxana M Reynoso ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Ovarian Function ◽  
Follicular Development ◽  
Female Rats ◽  
Control Group ◽  
Peroxisome Proliferator ◽  
Mrna Levels ◽  
Ovarian Steroidogenesis ◽  
Inflammatory Status

The objective of this work was to study the ovarian function when follicular development is induced during a hyperandrogenic condition. Female rats were injected with either equine chorionic gonadotropin (eCG group) to induce folliculogenesis or eCG together with DHEA to induce folliculogenesis in a hyperandrogenic condition (eCG+HA group). The control group was injected with vehicle. Ovarian mRNA levels of the peroxisome proliferator-activated receptor gamma (PPARγ) co-activator PGC1α, the PPARγ co-repressor NCoR, the main enzymes involved in the ovarian steroidogenesis (CYP17, 3β-hydroxysteroid dehydrogenase (3β-HSD), 17β-HSD, and CYP19A), and cyclooxygenase 2 (COX2) were evaluated only by real-time PCR. COX2 was evaluated by both real-time PCR and western blot. Serum steroid hormones and both the oxidative and inflammatory statuses were also quantified. We found that eCG-induced folliculogenesis induced increased mRNA levels of PGC1α and decreased those of NCoR when compared with controls. In addition, we found an increase in serum estradiol (E2) levels and enhanced mRNA expression of CYP19A. A pro-inflammatory status and a pro-oxidant status were also established. When folliculogenesis was induced in a hyperandrogenic condition, the mRNA levels of the PPARγ co-repressor NCoR remained higher than in controls and the pro-inflammatory and pro-oxidant statuses were enhanced. In addition, the enzymes involved in ovarian steroidogenesis were altered leading to the accumulation of testosterone and an unfavorable E2/testosterone ratio. These alterations led to abnormal follicular development.

Role of androgens in hCG-induced ovulation in PMSG-primed immature rats

1980 ◽  
Vol 93 (4) ◽  
pp. 505-512 ◽  
Author(s):  
J. J. Peluso ◽  
D. Stude ◽  
R. W. Steger
Keyword(s):  
Cyproterone Acetate ◽  
Cumulus Cell ◽  
Induced Ovulation ◽  
Ovulatory Follicle ◽  
Immature Rats ◽  
Cell Layers ◽  
Pre Treatment ◽  
Theca Interna ◽  
Ovulatory Follicles

Abstract. The effect of pre-treatment with the anti-androgens, cyproterone acetate (C.A.) and Flutamide (Fl) on hCG-induced ovulation in immature PMSG-primed rats was studied. The results showed that pre-treatment with either anti-androgen reduced the ovulation rate by 50%. Not all pre-ovulatory follicles (≥ 600 μm in diameter) in control and anti-androgen-treated rats ovulated in response to hCG. In control rats, hCG induced cumulus cell dispersement and resumption of meiosis in the ova of these non-ovulatory follicles; while C.A. or Fl treatment prevented these pre-ovulatory responses (P < 0.05). In a subsequent study, PMSG-primed rats were pre-treated with Fl and autopsied 8 h after hCG. In control rats, hCG reduced ovarian oestradiol content, did not alter ovarian testosterone content, increased ovarian progesterone content (P < 0.05) and increased the percentage of follicles with 3B-HSD activity within their granulosa cell layers (P < 0.05). Fl did not alter ovarian steroid content but blocked the hCG-induced appearance of 3B-HSD activity in the granulosa cells of pre-ovulatory follicles. This lack of 3B-HSD activity could result in a local decrease in progesterone within the non-ovulatory follicle. It appears that androgens, presumably synthesized in the theca interna play an important role in either ovulation or in maintaining responsiveness of the pre-ovulatory follicle to LH.

Protein sources affect follicular dynamics in ewes near the onset of the breeding season

1996 ◽  
Vol 8 (6) ◽  
pp. 1021 ◽  
Author(s):  
S Landau ◽  
JA Houghton ◽  
JR Mawhinney ◽  
EK Inskeep
Keyword(s):  
Luteal Phase ◽  
Follicular Development ◽  
Follicular Maturation ◽  
Prostaglandin F2 Alpha ◽  
Late Luteal Phase ◽  
Corn Gluten ◽  
Ovulatory Follicles ◽  
Ground Corn ◽  
Corn Grain ◽  
Follicular Dynamics

The influence of source of protein during the luteal phase before a synchronized oestrus on the dynamics of follicular development, observed daily by ultrasonography, was assessed in ewes that were beginning the sexual season. Iso-nitrogenous amounts of soybean meal (SBM) or of a corn-gluten meal-ground-corn grain mixture (CGM-GC), or an iso-energetic amount of ground-corn grain (GC), were fed from four days before to four days after treatment with prostaglandin F2 alpha (PGF2 alpha). Feeding with SBM was associated with a higher frequency of short luteal phases (P < 0.02). Dynamics of follicular population were studied in ewes that ovulated after normal cycles. More follicles > or = 2 mm in diameter were observed on the ovaries of ewes fed SBM four days before PGF2 alpha treatment (P < 0.02), but the highest number was seen in ewes fed CGM-GC at the time of injection of PGF2 alpha (P < 0.08). Ewes fed SBM had larger follicles at last detection and ovulated earlier after PGF2 alpha treatment than their counterparts fed other diets (P < 0.001). Ovulatory follicles developed over a greater range of days in ewes with twin ovulations compared with ewes with single ovulations (P < 0.08). Serum concentrations of insulin were increased after four days of feeding with CGM-GC (P < 0.01), but not with SBM or GC, and reached a peak at the time of oestrus. In summary: (1) the source of dietary protein during the late-luteal phase affected follicular maturation after PGF2 alpha treatment; (2) insulin and glucose may be involved in this response and may play a role in ovarian follicular activity; and (3) twin ovulations appeared to result from both reduced atresia and increased recruitment of follicles.

Individual and Combined Assessment of Ser680Asn FSH Receptor and FSHβ -211 G>T Gene Polymorphisms in Ovarian Response in IVF/ICSI Program

2020 ◽  
Vol 21 ◽  
Author(s):  
Elli Anagnostou ◽  
Alexia Kafkoutsou ◽  
Despina Mavrogianni ◽  
Ekaterini Domali ◽  
Evangelia Dimitroulia ◽  
...  
Keyword(s):  
Gene Polymorphism ◽  
Gene Polymorphisms ◽  
Ovarian Response ◽  
Greek Population ◽  
Control Group ◽  
Genotype Distribution ◽  
Β Subunit ◽  
Molecular Genetic Study ◽  
The Impact ◽  
First Time

Background: Molecular biology tools, such as the detection of single nucleotide polymorphisms (SNPs), have been considered to assist to the management of the ovarian stimulation protocols. Purpose: The aim of this study was to evaluate the impact of two polymorphisms, the Asn680Ser polymorphism of the FSHR gene, and the FSH β subunit (FSHβ) gene polymorphism -211 G>T, in a Greek population of women undergoing IVF/ICSI program in our center. In addition, a control group of fertile women was studied, to verify whether there are differences in the genotype distribution between fertile and infertile population for both polymorphisms, as the FSHβ gene polymorphism -211 G>T is studied for the first time in the Greek population. Results : The FSH β-211 G>T polymorphism, studied for the first time in the Greek infertile population, appears to be quite rare. When studying the two polymorphisms separately, statistically significant differences were obtained that concerned the LH levels. Discussion: According to the combination analysis of the two polymorphisms by the number of alleles, women with 2-3 polymorphic alleles needed more days of stimulation, but there were no differences in pregnancy rates. Conclusion: This molecular genetic study helps to elucidate whether the polygenic combination of the Asn680Ser and FSH β subunit -211 G>T gene polymorphisms is of additive value in the prediction of ovarian response to exogenous gonadotropins.

scholarly journals Urinary bisphenol A in women with polycystic ovary syndrome – a possible suppressive effect on steroidogenesis?

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Zora Lazúrová ◽  
Jana Figurová ◽  
Beáta Hubková ◽  
Jana Mašlanková ◽  
Ivica Lazúrová
Keyword(s):  
Polycystic Ovary Syndrome ◽  
Bisphenol A ◽  
Steroid Hormones ◽  
Polycystic Ovary ◽  
Suppressive Effect ◽  
Human Reproduction ◽  
Control Group ◽  
Ovary Syndrome ◽  
Ovarian Steroidogenesis ◽  
The Relationship

Abstract Objectives There is a growing evidence indicating an impact of endocrine distrupting chemicals such as bisphenol A (BPA) on human reproduction. Its higher levels in serum or urine have been documented in women with polycystic ovary syndrome (PCOS), however the relationship to ovarian steroidogenesis remains unclear. Aim of the study was to compare urinary BPA (U-BPA) concentrations among PCOS women and control group. Second aim was to assess the relationship of U-BPA to ovarian steroidogenesis in the group with PCOS. Methods Eighty six Caucasian women (age 28.5 ± 5.1 years) diagnosed with PCOS and 32 controls of age 24.9 ± 4.4 years were included in the study. Fasting blood samples were analyzed for biochemical parameters and steroid hormones. U-BPA was measured in the morning urine sample using high pressure liquid chromatography. Results PCOS women had significantly higher U-BPA as compared with control group (p=0.0001). Those with high levels of U-BPA (U-BPA ≥2.14 ug/g creatinine) demonstrated higher serum insulin (p=0.029) and HOMA IR (p=0.037), lower serum estrone (p=0.05), estradiol (p=0.0126), FSH (p=0.0056), and FAI (p=0.0088), as compared with low-BPA group (U- BPA <2.14 ug/g creatinine). In PCOS women, U-BPA positively correlated with age (p=0.0026; R2=0.17), negatively with estradiol (p=0.0001, R2=0.5), testosterone (p=0.0078, R2=0.15), free-testosterone (p=0.0094, R2=0.12) and FAI (p=0.0003, R2=0.32), respectively. Conclusions PCOS women have significantly higher U-BPA concentrations than healthy controls. U-BPA positively correlates with age and negatively with ovarian steroid hormones suggesting a possible suppressive effect of bisphenol A on ovarian steroidogenesis.

scholarly journals Bushen Tiaojing (II and III) Decoctions Activate MAPK14 and MAPK3/1 to Promote the Expression of Cumulus Expansion-Related Factors in Mice

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Xiao Liang ◽  
Xue Tong ◽  
Hui-lan Du ◽  
Ming He ◽  
Yu Zhang ◽  
...  
Keyword(s):  
Western Blot ◽  
Polar Body ◽  
Cumulus Cell ◽  
Serum Levels ◽  
Control Group ◽  
Distilled Water ◽  
Related Factors ◽  
Cumulus Expansion ◽  
Sequential Administration ◽  
First Polar Body

Background. Bushen Tiaojing Decoctions (BSTJ-II-D and BSTJ-III-D) are used to assist pregnancy in clinical practice. In this study, we explored the ability of sequential administration of BSTJ-II-D and BSTJ-III-D to promote cumulus cell (CC) expansion and its underlying mechanisms in controlled ovarian hyperstimulation (COH) mice. Methods. Kunming mice were randomly divided into three groups. The normal group was injected intraperitoneally with saline, and distilled water was administered orally by gavage. As the COH model, mice were injected with GnRHa, eCG, and hCG. Subsequently, the BSTJD group received BSTJ-II-D and BSTJ-III-D orally by gavage, while the control group received distilled water. We evaluated CC expansion and oocyte first polar body (PB1) extrusion under a stereomicroscope. Serum levels of follicle-stimulating hormone (FSH) were detected by radioimmunoassay. The expression of the CC expansion-related factors PTX3 and PTGS2 was detected by immunofluorescence, western blot, and quantitative real-time-polymerase chain reaction analyses (qRT-PCR). Expression of p-MAPK14, p-MAPK3/1, MAPK14, and MAPK3/1 was detected by western blot analysis. Results. Sequential administration of BSTJ-II-D and BSTJ-III-D promoted cumulus expansion and oocyte PB1 extrusion and upregulated PTX3 and PTGS2 expression at the mRNA and protein levels. Furthermore, the levels of p-MAPK14/MAPK14, p-MAPK3/1/MAPK3/1 proteins, and serum FSH in the BSTJD group were higher than those in the normal and control groups. Conclusions. Sequential administration of BSTJ-II-D and BSTJ-III-D promotes cumulus expansion and oocyte maturation in COH mice by increasing FSH expression and activating the MAPK14 and MAPK3/1 signalling pathways, thereby increasing expression of PTX3 and PTGS2.

scholarly journals Human growth hormone use in poor ovarian response – caution and opportunities

2021 ◽  
Vol 15 ◽  
pp. 263349412199942
Author(s):  
Robert J. Norman ◽  
Roger J. Hart
Keyword(s):  
Growth Hormone ◽  
Human Growth Hormone ◽  
Live Birth ◽  
Cell Function ◽  
Cumulus Cell ◽  
Human Growth ◽  
Ovarian Response ◽  
Randomised Trials ◽  
Poor Ovarian Response ◽  
Oocyte Maturity

Human growth hormone has found favour as a co-gonadotrophin in assisted reproduction particularly in the circumstances of a poor response to stimulation. Its use has been based on animal studies suggesting insulin-like growth factor-1 enhances granulosa and cumulus cell function and possibly oocyte quality. While there is limited ovarian cellular information in women, the use of human growth hormone is alleged to improve egg numbers, embryo quality, clinical pregnancies and live birth in women with a poor ovarian response. A number of cohort studies have claimed these benefits compared with prior nil treatment, but there are a limited number of quality randomised controlled studies. The few good randomised trials indicate an enhanced ovarian response in terms of oestradiol secretion and oocyte maturity with controversial improvement in ongoing pregnancy and live birth. Given the cost of the medication, the lack of convincing data on enhanced clinical outcomes and the theoretical possibility of side effects, we propose it is still too early to determine human growth hormone’s true cost-benefit for widespread use. However, a number of emerging randomised trials may tilt the equation to a positive outlook in the future. Meanwhile, the hormone should only be used after full informed consent from the patient as to its effectiveness and efficacy.

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