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

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.

Reproduction ◽  
2000 ◽  
pp. 311-323 ◽  
Author(s):  
JL Hilton ◽  
GE Sarty ◽  
GP Adams ◽  
RA Pierson

The magnetic resonance images and maps of bovine ovaries acquired at defined phases of follicular development and regression were studied to determine whether magnetic resonance image attributes of the follicular antrum reflect the physiological status of dominant and subordinate ovarian follicles. Ovariectomies were performed at day 3 of wave one, day 6 of wave one, day 1 of wave two and at >/= day 17 after ovulation. The timings of ovariectomies were selected to acquire growing, early static, late static and regressing follicles of the first wave and preovulatory follicles of the ovulatory wave. Pre-selection and subordinate follicles were also available for analysis. Serum samples were taken on the day of ovariectomy and follicular fluid samples were taken after imaging. Numerical pixel value and pixel heterogeneity in a spot representing approximately 95% of the follicular antrum were quantified in T(1)- and T(2)-weighted images. T(1) and T(2) relaxation rates (T(1) and T(2)), proton density, apparent diffusion coefficients and their heterogeneities were determined from the computed magnetic resonance maps. The antra of early atretic dominant follicles showed higher T(2)-weighted mean pixel value (P < 0.008) and heterogeneity (P < 0. 01) and lower T(2) heterogeneity (P < 0.008) than growing follicles. Subordinate follicles in the presence of a preovulatory dominant follicle had higher T(1), T(1) heterogeneity, proton density, proton density heterogeneity, and lower mean pixel value in T(1)-weighted images than subordinate follicles of the anovulatory wave (P < 0.04). T(1) relaxation rate heterogeneity and proton density heterogeneity were positively correlated with follicular fluid oestradiol concentration (r = 0.4 and 0.3; P < 0.04). T(2) relaxation rate heterogeneity was positively correlated with follicular fluid progesterone concentration (r = 0.4; P < 0.008). Quantitative differences in magnetic resonance image attributes of the antrum observed among phases of follicular development and regression coincided with changes in the ability of the dominant follicle to produce steroid hormones and ovulate, and thus were indicative of physiological status and follicular health.


2021 ◽  
Vol 10 ◽  
Author(s):  
Catherine Deborde ◽  
Blandine Madji Hounoum ◽  
Annick Moing ◽  
Mickaël Maucourt ◽  
Daniel Jacob ◽  
...  

Abstract The long-term effect of a plant (P)-based diet was assessed by proton nuclear magnetic resonance (1H-NMR) metabolomics in rainbow trout fed a marine fish meal (FM)–fish oil (FO) diet (M), a P-based diet and a control commercial-like diet (C) starting with the first feeding. Growth performances were not heavily altered by long-term feeding on the P-based diet. An 1H-NMR metabolomic analysis of the feed revealed significantly different soluble chemical compound profiles between the diets. A set of soluble chemical compounds was found to be specific either to the P-based diet or to the M diet. Pterin, a biomarker of plant feedstuffs, was identified both in the P-based diet and in the plasma of fish fed the P-based diet. 1H-NMR metabolomic analysis on fish plasma and liver and muscle tissues at 6 and 48 h post feeding revealed significantly different profiles between the P-based diet and the M diet, while the C diet showed intermediate results. A higher amino acid content was found in the plasma of fish fed the P-based diet compared with the M diet after 48 h, suggesting either a delayed delivery of the amino acids or a lower amino acid utilisation in the P-based diet. This was associated with an accumulation of essential amino acids and the depletion of glutamine in the muscle, together with an accumulation of choline in the liver. Combined with an anticipated absorption of methionine and lysine supplemented in free form, the present results suggest an imbalanced essential amino acid supply for protein metabolism in the muscle and for specific functions of the liver.


2015 ◽  
Vol 27 (1) ◽  
pp. 178
Author(s):  
K. M. Lemes ◽  
L. A. Silva ◽  
E. C. C. Celeghini ◽  
M. A. Alonso ◽  
G. Pugliesi ◽  
...  

The postpartum period is characterised by the rapid uterine involution process and return of ovarian activity (foal heat), resulting in a fertile oestrus in most of the mares. However, the follicular development and selection processes during this period are not completely known in horses. We aimed to study the characteristics of follicular growth and vascular perfusion in the ovary during the early postpartum period in mares that demonstrated oestrous behaviour and had early (<10 days) or late (≥10 days) ovulation. Ten mares were scanned daily from the first day postpartum (Day 1) until the day of the first postpartum ovulation (Day 0). The animals were split in the early (n = 3) and late (n = 7) ovulation groups (averaged interval between parturition and ovulation: 8.0 ± 0.0 and 14.7 ± 1.2 days, respectively). For ultrasound exams a Duplex B-mode and colour Doppler instrument (M5VET®, Mindray, Shenzhen, China) was used with a multifrequency linear probe. Data were analysed for the main effects of group, day, and their interaction using the PROC MIXED procedure of SAS software (version 9.3, SAS Institute Inc., Cary, NC, USA). For the follicular growth, no difference (P > 0.05) was detected between the groups when the data were analysed for the days relative to ovulation (from Day 7 to Day 1). However, the dominant follicle was larger (P < 0.05) in the early-ovulated group (37.2 ± 1.6 v. 21.9 ± 1.1) in all days during early postpartum (Day 1 to Day 7). The number of follicles with >25 mm diameter was also greater (P < 0.05) in the early-ovulated group (1.1 ± 0.1 v. 0.1 ± 0.1) during the first 3 days postpartum. In addition, the late-ovulated mares showed greater number of follicles with 20–25 mm during Day 4 to Day 7 (2.0 ± 0.2 v. 0.7 ± 0.1). For the blood flow characteristics, no difference (P > 0.05) was detected in the coloured signals of blood flows in the follicular wall of the dominant follicle or in the ovarian pedicle ipsilateral to the largest follicle. Therefore, the characteristics of the follicle growth on the preceding days of ovulation were similar between the early- and late-ovulated mares and consistent with the follicular dynamics expected in non-pregnant and non-lactating mares. However, when the data were analysed for the days relative to parturition, a greater follicle development was present in mares that ovulate earlier during the postpartum period (<10 days). In conclusion, the results suggest that important events may occur previous to the parturition, resulting in an early follicle development, mainly in those mares that show heat signs and ovulate within 10 days postpartum. Research was supported by FAPESP process number 2010/10692-9 and CNPq process number 135954/2011-8.


1997 ◽  
Vol 154 (1) ◽  
pp. 35-43 ◽  
Author(s):  
S C Cwyfan Hughes ◽  
H D Mason ◽  
S Franks ◽  
J M P Holly

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


2001 ◽  
Vol 72 (3) ◽  
pp. 547-554 ◽  
Author(s):  
A.C.O. Evans ◽  
P. Duffy ◽  
K. M. Quinn ◽  
P. G. Knight ◽  
M. P. Boland

AbstractThe aim was to examine statistically the changes among days in the numbers of follicles relative to the growth of large follicles to test the hypothesis that follicular development occurs in a wave-like fashion in anoestrous ewes. The relationships among the patterns of circulating concentrations of FSH, oestradiol and inhibin-A and the pattern of follicular growth as well as relationships among follicular fluid steroid and inhibin-A concentrations were also studied. The ovaries of 11 ewes were examined daily using transrectal ultrasonography for 14 days and blood samples were collected every 8 h for 9 days. Five ewes were ovariectomized on the last day of ultrasound scanning. One to three identified follicles (a cohort) emerged every 2 to 5 days (mean 2·9 (s.e. 0·2) days) in individual ewes. The numbers of 4 and 5 mm follicles were fewest (P < 0·05) before and greatest (P < 0·05) 1 day after cohort emergence. This change in the numbers of follicles indicates a wave-like pattern of follicular growth. FSH concentrations were greatest (P < 0·05) on the day before wave emergence and lowest (P < 0·05) on the day of wave emergence. Peripheral concentrations of oestradiol and inhibin-A did not fluctuate (P > 0·05) in association with the emergence of follicular waves. The follicles that were collected at ovariectomy originated from one of three different waves. Oestradiol and inhibin-A concentrations in follicular fluid and the oestradiol-to-progesterone ratio were not different among the largest follicles of successive waves, when follicles were collected at the same time, indicating that new waves of follicles developed before the demise of old waves.In conclusion, waves of follicles emerged about every 3 days in anoestrous ewes (defined as significant changes in numbers of follicles) and were associated with fluctuations in FSH concentrations but not peripheral oestradiol or inhibin-A concentrations. New follicular waves also emerged in the presence of steroidogenically active (positive oestradiol-to-progesterone ratio), inhibin-A producing follicles from a previous wave suggesting that follicles do not exert functional dominance during the non-breeding season.


1976 ◽  
Vol 68 (3) ◽  
pp. 461-468 ◽  
Author(s):  
J. TH. J. UILENBROEK ◽  
E. ARENDSEN DE WOLFF-EXALTO ◽  
M. A. BLANKENSTEIN

SUMMARY Follicular development and serum gonadotrophin levels were studied in female rats after neonatal androgen administration. After injection of 1250 μg testosterone propionate (TP) on day 5 after birth the composition of the follicular population was altered: at nearly all ages the number of pre-antral follicles (follicular volume 2–20 × 105 μm3) was lower than in oil-treated rats, in some cases the number of small antral follicles (21–249 × 105 μm3) was also lower. Furthermore levels of serum follicle-stimulating hormone and luteinizing hormone were decreased from day 7 to day 20 suggesting that the high gonadotrophin levels before day 20 are of importance for normal follicular development. In contrast, final follicular maturation in TP-treated rats was enhanced; at day 35 more large antral follicles (follicular volume ≥ 500 × 105μm3) were present in TP-treated rats than in oil-treated rats. The presence of more large antral follicles was accompanied by higher plasma oestradiol concentrations, higher uterine weights and advanced vaginal opening. These results demonstrate an inhibition of normal follicular growth and an acceleration of ovarian maturation after neonatal androgen administration.


2016 ◽  
Vol 5 (1) ◽  
pp. 43-48
Author(s):  
Manoj Bhattarai

Background Ultrasonography is the first line imaging modality for evaluation of ovaries, monitoring ovarian follicular development and detecting ovulation in infertile women; thus plays a significant role in infertility management. This study was undertaken to evaluate the pattern of ovarian follicular growth and to predict and detect ovulation in infertile women by transvaginal sonography in eastern region of Nepal.Material and Methods Hospital based prospective cross-sectional study on 100 infertile patients referred for ultrasonographic monitoring of ovarian follicle was conducted over duration of 26 months. Serial transvaginal sonography of the patients was performed using standard procedure daily from day 10 of menstrual cycle till detection of ovulation. Identification of ovarian dominant follicle, monitoring of dominant follicle development and detection of ovulation was assessed in relation to the day of menstrual cycle.Results Increase in mean diameter of the dominant follicle was seen in serial ultrasound scan till ovulation, which occurred in all cases by day 16 of menstrual cycle. The average daily follicular growth rate ± SD from day 10 of menstrual cycle till detection of ovulation was 2.2 ± 0.2 mm per day and the mean diameter ± SD of dominant follicle on the day prior to ovulation was 21.4 ± 2.8 mm (range: 17.2 – 26.3 mm).Conclusion Transvaginal sonography is an excellent method for monitoring of ovarian follicular development and shows a linear increase in mean diameter of dominant follicle from day 10 of menstrual cycle till detection of ovulation.Journal of Nobel Medical CollegeVolume 5, Number 1, Issue 8, January-July 2016, 43-48


Endocrinology ◽  
2004 ◽  
Vol 145 (11) ◽  
pp. 5097-5105 ◽  
Author(s):  
J. W. Newman ◽  
J. E. Stok ◽  
J. D. Vidal ◽  
C. J. Corbin ◽  
Q. Huang ◽  
...  

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 &lt; 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.


Reproduction ◽  
2013 ◽  
Vol 146 (3) ◽  
pp. 221-231 ◽  
Author(s):  
Michael J Bertoldo ◽  
Lydie Nadal-Desbarats ◽  
Nadine Gérard ◽  
Alexis Dubois ◽  
Patricia K Holyoake ◽  
...  

The microenvironment of the developing follicle is critical to the acquisition of oocyte developmental competence, which is influenced by several factors including follicle size and season. The aim of this study was to characterise the metabolomic signatures of porcine follicular fluid (FF) collected from good and poor follicular environments, using high-resolution proton nuclear magnetic resonance (1H-NMR) spectroscopy. Sow ovaries were collected at slaughter, 4 days after weaning, in summer and winter. The contents of small (3–4 mm) and large (5–8 mm) diameter follicles were aspirated and pooled separately for each ovary pair. Groups classified as summer-small (n=8), summer-large (n=15), winter-small (n=9) and winter-large (n=15) were analysed by1H-NMR spectroscopy. The concentrations of 11 metabolites differed due to follicle size alone (P<0.05), including glucose, lactate, hypoxanthine and five amino acids. The concentrations of all these metabolites, except for glucose, were lower in large FF compared with small FF. Significant interaction effects of follicle size and season were found for the concentrations of glutamate, glycine,N-acetyl groups and uridine. Succinate was the only metabolite that differed in concentration due to season alone (P<0.05). The FF levels of progesterone, androstenedione and oestradiol were correlated with the concentrations of most of the metabolites examined. The results indicate that there is a distinct shift in follicular glucose metabolism as follicles increase in diameter and suggest that follicular cells may be more vulnerable to oxidative stress during the summer months. Our findings demonstrate the power of1H-NMR spectroscopy to expand our understanding of the dynamic and complex microenvironment of the developing follicle.


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