scholarly journals Cytochrome P450-Dependent Lipid Metabolism in Preovulatory Follicles

Endocrinology ◽  
2004 ◽  
Vol 145 (11) ◽  
pp. 5097-5105 ◽  
Author(s):  
J. W. Newman ◽  
J. E. Stok ◽  
J. D. Vidal ◽  
C. J. Corbin ◽  
Q. Huang ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Lipid Metabolism ◽  
Follicular Fluid ◽  
Ovarian Tissue ◽  
Follicular Development ◽  
Epoxyeicosatrienoic Acid ◽  
Follicular Maturation ◽  
Before And After ◽  
Acid Metabolites

Abstract Estrogen biosynthesis and proteolysis are both important processes involved in ovarian follicular development, which may be influenced by cytochrome P450 (CYP)-dependent fatty acid metabolites. However, CYP-dependent lipid metabolism has not been characterized with respect to follicular maturation in vivo. Therefore, follicular fluid was collected in the hours before and after the LH surge in pigs, and concentrations of epoxy, hydroxy, and dihydroxy lipids were measured by liquid chromatography tandem mass spectrometry. Arachidonate oxidation and epoxyeicosatrienoic acid hydrolysis to dihydroxyeicosatrienoic acids (DHETs) were also assessed in thecal and granulosa tissue fractions, and the expression of CYP epoxygenases was evaluated by immunoblots using available antisera. To evaluate soluble epoxide hydrolase (sEH) expression, the porcine sEH was cloned from ovarian tissue, expressed and purified for antibody generation. The follicular fluid oxylipin concentrations ranged from 1–150 nm depending on the compound and estrous stage. The follicular fluid concentrations of CYP-dependent oxylipins increased at estrus, as did sEH expression; however, significant changes in epoxides were not observed, and the 11,12-DHET peak was delayed. The ratio of 14,15–DHET:11,12-DHET across all samples correlated with the log of follicular fluid estradiol concentrations (P < 0.01). Epoxygenase activities were similar in theca and granulosa, varying little with follicular development, whereas the decline of a single CYP2J isoform at ovulation was observed by immunoblots. The sEH activity was higher in granulosa than in theca. Finally, the dynamic changes in follicular CYP-dependent arachidonic acid metabolites and their modulatory function in vascular models suggest roles for these metabolites in follicular maturation, which may include regulation of estradiol biosynthesis and preovulatory remodeling of the follicular wall that should be fully explored in future studies.

Follicle development in cryopreserved bitch ovarian tissue grafted to immunodeficient mouse

2012 ◽  
Vol 24 (3) ◽  
pp. 461 ◽  
Author(s):  
L. Commin ◽  
S. Buff ◽  
E. Rosset ◽  
C. Galet ◽  
A. Allard ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Histological Analysis ◽  
Smooth Muscle Actin ◽  
Ovarian Tissue ◽  
Follicular Development ◽  
Slow Freezing ◽  
Angiogenic Factor ◽  
Muscle Actin ◽  
Once Daily

The present study evaluated: (1) in vivo follicular development in canine ovarian tissue after slow freezing and xenotransplantation; and (2) the use of erythropoietin (EPO) as an angiogenic factor to optimise the transplantation procedure. Frozen–thawed ovarian tissue from five bitches was grafted into severe combined immunodeficient (SCID) mice (n = 47) treated with or without EPO (500 IU kg–1, once daily for 3 days) (Groups A and B, respectively) and analysed after 0, 1, 8 or 16 weeks. Follicle grade, follicle density, follicle morphology and stromal cells density were assessed by histological analysis, whereas vascularisation of the graft was quantified by immunohistochemistry with anti-α-smooth muscle actin antibody. Despite a massive loss of follicles after grafting, secondary follicle density was higher at 8 and 16 weeks than at 1 week regardless of EPO treatment. EPO significantly improved early follicle morphology and stromal cell density after 8 weeks and blood vessel density at 16 weeks after transplantation (P < 0.05). Intact secondary follicles with more than three granulosa cells layers were observed 16 weeks after transplantation. The results suggest that canine ovarian tissue can be successfully preserved by our slow-freezing protocol because the tissue showed follicular growth after xenotransplantation. EPO treatment did not lessen the massive loss of follicles after transplantation.

Modulation of the insulin-like growth factor-binding proteins by follicle size in the human ovary

1997 ◽  
Vol 154 (1) ◽  
pp. 35-43 ◽  
Author(s):  
S C Cwyfan Hughes ◽  
H D Mason ◽  
S Franks ◽  
J M P Holly
Keyword(s):  
Molecular Weight ◽  
Conditioned Medium ◽  
Follicular Fluid ◽  
Metal Ion ◽  
Binding Proteins ◽  
Follicular Development ◽  
Lower Molecular Weight ◽  
Human Ovary ◽  
Follicular Maturation ◽  
Igfbp 3

Abstract The IGFs are believed to play an important role in the regulation of steroidogenesis and follicular maturation in the human ovary. The activities of the IGFs are regulated by a family of binding proteins (IGFBPs) which are subject to a number of potential post-translational modifications. The aim of this study was to investigate both the production and modification of the IGFBPs in follicular fluid and in medium conditioned by granulosa cells and theca from individual follicles at different stages of maturation. In follicular fluid from healthy, dominant follicles there was an increase in the amount of IGFBP-2, -3 and -4 present as lower molecular weight forms (23 kDa, 29 kDa and 16·5 kDa respectively) in comparison to that seen in atretic follicles from the same ovary. Furthermore for IGFBP-4, this fragmentation was confirmed to be attributable to the presence of a specific protease which could be inhibited not only by the addition of metal ion chelators or serine protease inhibitors, but also by the addition of other recombinant unsaturated IGFBPs, particularly IGFBP-3. IGF-I did not modulate the activity of the IGFBP-4 protease in solution but was able to prevent the inhibition seen with IGFBP-3. Analysis of granulosa cell conditioned medium from the same series of healthy and atretic follicles revealed that IGFBP-2 and -4 were the predominant IGFBPs with no fragments seen on immunoblotting. In contrast, IGFBP-3 in conditioned medium from theca of atretic follicles was always found as an intact doublet, but was found partially fragmented (30 and 32 kDa) in medium conditioned by theca from healthy, dominant follicles with the proportion of IGFBP-3 in this lower molecular weight or fragmented doublet increasing with follicular maturation. A similar situation was also found for IGFBP-4 with the progressive increase in the amount of the 15 and 16·5 kDa fragments. IGFBP-2 was always found to be intact. Finally, IGFBP production from stroma explants was also examined. This revealed a wide variation in IGFBP pattern between different ovaries, although there was a remarkable degree of consistency between different stroma explant cultures from the same ovary. Immunoblotting for IGFBP-3 revealed that, where present, it existed as both an intact and a lower molecular weight doublet and that IGFBP-2 was again always found to be intact. In conclusion we have demonstrated alterations in the proteolytic modification of the IGFBPs which differ in the various follicular compartments and are closely linked to the stage of follicular development. Journal of Endocrinology (1997) 154, 35–43

scholarly journals Placental Growth Factor Is Required for Ovulation, Luteinization, and Angiogenesis in Primate Ovulatory Follicles

Endocrinology ◽  
2017 ◽  
Vol 159 (2) ◽  
pp. 710-722 ◽  
Author(s):  
Hannah R Bender ◽  
Heidi A Trau ◽  
Diane M Duffy
Keyword(s):  
Growth Factor ◽  
Granulosa Cell ◽  
Follicular Fluid ◽  
Follicular Development ◽  
Placental Growth Factor ◽  
Vascular Endothelial ◽  
Follicle Rupture ◽  
Ovulatory Follicles ◽  
Endothelial Growth Factor

Abstract Placental growth factor (PGF) is member of the vascular endothelial growth factor (VEGF) family of angiogenesis regulators. VEGFA is an established regulator of ovulation and formation of the corpus luteum. To determine whether PGF also mediates aspects of ovulation and luteinization, macaques received gonadotropins to stimulate multiple follicular development. Ovarian biopsies and whole ovaries were collected before (0 hours) and up to 36 hours after human chorionic gonadotropin (hCG) administration to span the ovulatory interval. PGF and VEGFA were expressed by both granulosa cells and theca cells. In follicular fluid, PGF and VEGFA levels were lowest before hCG. PGF levels remained low until 36 hours after hCG administration, when PGF increased sevenfold to reach peak levels. Follicular fluid VEGFA increased threefold to reach peak levels at 12 hours after hCG, then dropped to intermediate levels. To explore the roles of PGF and VEGFA in ovulation, luteinization, and follicular angiogenesis in vivo, antibodies were injected into the follicular fluid of naturally developed monkey follicles; ovariectomy was performed 48 hours after hCG, with ovulation expected about 40 hours after hCG. Intrafollicular injection of control immunoglobulin G resulted in no retained oocytes, follicle rupture, and structural luteinization, including granulosa cell hypertrophy and capillary formation in the granulosa cell layer. PGF antibody injection resulted in oocyte retention, abnormal rupture, and incomplete luteinization, with limited and disorganized angiogenesis. Injection of a VEGFA antibody resulted in oocyte retention and very limited follicle rupture or structural luteinization. These studies demonstrate that PGF, in addition to VEGFA, is required for ovulation, luteinization, and follicular angiogenesis in primates.

scholarly journals Gonadotropin requirements for dominant follicle selection in GnRH agonist-treated cows

Reproduction ◽  
2004 ◽  
Vol 127 (6) ◽  
pp. 695-703 ◽  
Author(s):  
J H Hampton ◽  
J F Bader ◽  
W R Lamberson ◽  
M F Smith ◽  
R S Youngquist ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Gnrh Agonist ◽  
Follicular Fluid ◽  
Follicular Development ◽  
Constant Infusion ◽  
Treatment Groups ◽  
Chain Cleavage ◽  
Hybridization Intensity ◽  
Development And Differentiation ◽  
Pre Treatment

A study was conducted to examine the effects of gonadotropins on ovarian follicular development and differentiation in GnRH agonist (GnRHa)-treated cattle. Holstein cows were allotted into two pre-treatment groups: controls (n = 5) and GnRHa-treated (n = 9). Ovaries were removed from control cows on day 5 following a synchronized estrus. Treatment with GnRHa resulted in follicular arrest at <5 mm. Following follicular arrest, GnRHa-treated cows received a constant infusion of FSH for 96 h (GnRHa/FSH), with a randomly selected subset receiving hourly pulses of LH in addition to FSH during the last 48 h of infusion (GnRHa/FSH + LH). At the end of infusion, ovaries were removed, follicles were counted and measured, and follicular fluid samples were collected from large follicles (>10 mm). Differences in expression of mRNA for LH receptor, FSH receptor, cytochrome P450 side-chain cleavage, 3β-hydroxysteroid dehydrogenase, cytochrome P450 17α-hydroxylase (P450c17) and cytochrome P450 aromatase were determined in large follicles using in situ hybridization. The number of large follicles did not differ between GnRHa/FSH-treated and GnRHa/FSH + LH-treated cows (P = 0.64), but was greater than control animals (P ≤ 0.004). Follicular fluid concentrations of estradiol-17β and androstenedione were highest in GnRHa/FSH + LH-treated cows (P ≤ 0.04), intermediate in control cows, and lowest in GnRHa/FSH-treated cows. Hybridization intensity of P450c17 was greater in GnRHa/FSH + LH-treated versus control or GnRHa/FSH-treated cows (P ≤ 0.03). These results indicate that while FSH can support bovine follicular growth >10 mm, LH increases androgen production and expression of P450c17.

The role of intraovarian regulators in the aetiology of polycystic ovarian syndrome

1996 ◽  
Vol 5 (3) ◽  
pp. 151-168 ◽  
Author(s):  
Ghanim Almahbobi ◽  
Alan O Trounson
Keyword(s):  
Granulosa Cells ◽  
Follicular Development ◽  
Aromatase Activity ◽  
Lh Receptor ◽  
Follicular Maturation ◽  
Developmental Capacity ◽  
Follicle Selection

The present review demonstrates that the availability of bioactive FSH and LH in PCOS is normal and that granulosa cells of PCO are not apoptotic and instead hyperexpress functional FSH receptors and may possess intact aromatase activity. Consequently, these cells respond excessively to exogenous FSH stimulation and produce high amounts of oestradiol both in vivo and in vitro. The altered developmental capacity of follicles from PCO in vivo is most likely due to the abnormal follicular milieu of PCO and the culminating effects of intrafollicular inhibitors and stimulators. The failure of ovarian oestradiol production and follicular maturation to dominance in vivo may be due to a mechanism that interferes with the function of FSH, such as intraovarian steroids and growth factors. It has previously been shown that EGF and TGFα have inhibitory actions on follicular development, aromatization and LH receptor formation. In contrast, EGF enhances early follicular recruitment and growth. Therefore, it is hypothesized that EGF/TGFα may have a causal relationship in the mechanisms of anovulatory infertility in women with PCOS. Thus, an aberration in the regulation of follicular fluid EGF and/or TGFα may result in reduced numbers of granulosa cells, cessation of follicle selection and ultimately in the creation and maintenance of PCOS. The exact mechanism by which the hyperfunction of EGF/TGFα occurs and the trigger for this hyperactivity in the ovary remain to be determined. An experimental animal model may be required to assist such investigations in the future.

scholarly journals Analysis of the variations of follicular fluid composition during follicular growth and maturation in the mare using proton nuclear magnetic resonance (1H NMR)

Reproduction ◽  
2002 ◽  
pp. 241-248 ◽  
Author(s):  
N Gerard ◽  
S Loiseau ◽  
G Duchamp ◽  
F Seguin
Keyword(s):  
Amino Acids ◽  
Follicular Fluid ◽  
Follicular Development ◽  
Proton Nuclear Magnetic Resonance ◽  
Follicular Growth ◽  
Dominant Follicle ◽  
Follicular Maturation ◽  
H Nmr ◽  
Sugar Chains ◽  
Glucose Metabolites

Follicular development and ovulatory processes in mammals involve local biochemical changes as a result of substantial modifications in cellular metabolism, the most well known of which is steroid variation. In the present study, the intrafollicular variation of several other components was studied using proton nuclear magnetic resonance ((1)H NMR). This approach made it possible to demonstrate that the intrafollicular biochemical content changes during follicular growth and maturation. Follicular fluid was aspirated by ovarian puncture of the dominant follicle at various physiological stages of its development: early dominant, late dominant and preovulatory. Serum samples were collected during each puncture session. (1)H NMR was used to evaluate intrafollicular and circulating glycoconjugates (sugar chains and N-acetyl groups), lipoproteins (CH(3) and CH(2) groups), glucose metabolites (trimethylamines, acetate and lactate), amino acids (glutamine/glutamate and alanine), creatine/creatinine and polyamines. Follicular fluids were assayed by radioimmunoassay for oestradiol and progesterone contents. The intrafollicular contents of alanine and lipoproteins (CH(3) groups) decreased in the dominant follicle during growth, whereas concentrations of progesterone and oestradiol increased significantly. After injection of gonadotrophin to induce ovulation, follicular maturation was characterized by a decrease in glycoconjugates (sugar chains), trimethylamines and acetate, a decrease in oestradiol concentration, and a further increase in CH(3) groups of lipoproteins and progesterone. The results from the present study showed a clear correlation between the intrafollicular content of alanine and that of oestradiol. A correlation between progesterone and glycoconjugates (sugar chains) was also observed. Therefore, (1)H NMR was shown to be effective for studying specific changes in the biochemical composition of the follicular fluid that occur during follicular development. For the first time, the variation of several compounds (glycoconjugates, lipoproteins, glucose metabolites, amino acids and polyamines) in relation to growth and maturation was demonstrated. Some of these changes could be of crucial importance for follicular maturation and ovulation as well as for oocyte maturation and further fertilization.

scholarly journals 170 FOLLICULAR FLUID CONCENTRATION AND OOCYTE QUALITY FROM TOGGENBURG GOATS FED WITH UREA

2009 ◽  
Vol 21 (1) ◽  
pp. 184
Author(s):  
E. A. M. Amorim ◽  
L. S. Amorim ◽  
C. A. A. Torres ◽  
J. D. Guimãres ◽  
J. F. Fonseca ◽  
...  
Keyword(s):  
Follicular Fluid ◽  
Follicular Development ◽  
Cumulus Cells ◽  
Oocyte Quality ◽  
Urea Concentration ◽  
Follicular Aspiration ◽  
Randomized Design ◽  
Fluid Concentration

Protein and urea concentrations impair oocyte and embryo development in vivo and in vitro through an unclear mechanism. A possible way to understand this process is to determine the concentration of hormones and metabolites in follicular fluid associated with normal development. The objective of this study was to determine the effect of dietary urea levels on follicular fluid concentration of hormones and metabolites and oocyte quality. A trial was conducted with 9 nonpregnant and nonlactating Saanen goats, which had been distributed in a randomized design and fed with diets with 0 (n = 4) and 2.4% of urea in the total dry matter (DM) of the diet (n = 5). Before follicle aspiration by laparotomy, the goats were synchronized by inserting intravaginal sponges containing 60 mg of acetate medroxyprogesterone (Progespon®, Sintex) for 10 days followed by 125 μg of cloprostenol (Ciosin® Coopers) 48 h before the removal of the sponge. The sponge was removed immediately before the follicular aspiration. The follicular development was stimulated with 70 mg of NIH-FSH-P1 (Folltropin V® Vetrepharm) i.m., and 300 IU of eCG i.m., (Novormon® Sintex) given 36 h before the follicular aspiration. Fluid from the 2 lartest follicles of each ovary were analyzed to determine the concentration of estradiol, progesterone, and testosterone by quimioluminesence, and glucose and urea concentrations were measured by enzymatic kit. The other follicles in each ovary were aspired with new needles and syringes and the oocyte quality was recorded. Oocytes were classified according to cytoplasma aspect and number of granulosa cells: Class A (dark cytoplasm and uniform aspect) with 3 (AMG) and <3 layers of cumulus cells (AmG); class B (cytoplasm with color alterations, desuniform aspect and vacuoles) with 3 (BMG) and <3 layers of cumulus cells (BmG); number of partially denuded oocytes (PD) and number of denuded oocytes (DO). Data were analyzed by ANOVA and treatment difference separated by SNK test. Follicular fluid estradiol concentration was lower in goats fed with urea (4.02 ± 0.16; 4.97 ± 0.18 ng mL–1; P < 0.05), progesterone concentration did not differ between treatments (2.48 ± 0.58; 3.37 ± 0.52 ng mL–1; P > 0.05), testosterone concentration was lower in the control animals (1.17 ± 0.48; 3.20 ± 0.43 ng mL–1; P < 0.05). The glucose (91.44 ± 3.60; 84.78 ± 5.58 mg dL–1) and urea concentration (23.04 ± 1.06; 18.00 ± 2.35) were greater in the animals treated with 2.4% compared with 0% of urea (P < 0.05), respectively. The number of oocytes in the different categories was not affected by treatment (P > 0.05): AMG 1.20 ± 1.09 v. 0.50 ± 0.57, AmG 4.20 ± 2.16 v. 3.50 ± 3.10, BMG 0.40 ± 0.54 v. 0.25 ± 0.50, BmG 1.40 ± 0.54 v. 1.75 ± 1.25, DO 10.20 ± 3.76 v. 11.50 ± 5.44, in the 0 and 2.4% of urea groups respectively. Only the number of PD (1.60 ± 0.54 v. 3.50 ± 1.91) recovered from animals treated with 2.4% was greater than in controls (P < 0.05). The hormone and metabolites concentration in follicular fluid as well as the oocyte quality was affected by the urea concentration of the diet. Supported by grant from: CNPq, FAPEMIG, Shering Plough®, Tecnopec®, Carbogel®.

Metabolomic Analysis of SCD During Goose Follicular Development: Implications for Lipid Metabolism

2020 ◽  
Author(s):  
Xin Yuan ◽  
Shenqiang Hu ◽  
Liang Li ◽  
Hehe Liu ◽  
Hua He ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Expression Patterns ◽  
Follicular Development ◽  
Pcr Analysis ◽  
Qrt Pcr ◽  
Preovulatory Follicles ◽  
Liquid Chromatography Tandem Mass

Abstract Background Stearoyl-CoA desaturase (SCD) is known to be an important rate-limiting enzyme in the production of MUFA. The role of this enzyme in goose follicular development is poorly understood. To investigate the metabolic mechanism of SCD during goose follicular development, we observed SCD expression patterns during follicular development in vivo and in vitro using quantitative reverse-transcription (qRT)-PCR. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to determine a cellular model of SCD function in granulosa cells (GCs) via SCD overexpression and knockdown.Results qRT-PCR analysis showed that SCD was abundantly expressed in the GC layer, and was upregulated in preovulatory follicles. Peak expression was found in F1 and prehierarchal follicles with diameters of 4–6 mm and 8–10 mm, respectively. We further found the mRNA expression and corresponding enzyme activity to occur in a time-dependent oscillation in vitro, beginning on the first day of GC culture. By using LC-MS/MS, we identified numerous changes in metabolite activation and developed an overview of multiple metabolic pathways, ten of which were associated with lipid metabolism and enriched in both the overexpressed and the knockdown groups.ConclusionsWe confirmed cholesterol and pantothenol or pantothenate as potential metabolite biomarkers for studying SCD-related lipid metabolism in goose GCs.

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