258. Characterisation of ovarian follicular growth in the brushtail possum

2005 ◽  
Vol 17 (9) ◽  
pp. 104
Author(s):  
B. Mester ◽  
B. P. Thomson ◽  
D. C. Eckery
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Follicular Phase ◽  
Brushtail Possum ◽  
Follicular Growth ◽  
Ovulatory Follicle ◽  
Follicle Selection ◽  
Ovarian Follicular Growth ◽  
Very High ◽  
Selection Of

The number and size of follicles selected for ovulation differ between species. The aim of this study was to characterise antral follicular growth and determine the size when selection of the ovulatory follicle occurs in the monovular brushtail possum. For this study, antral follicles ≥ 1 mm were dissected from the ovaries of 31 adult female possums at different reproductive states and follicular fluid and granulosa cells were harvested from each individual follicle. Selection of the ovulatory follicle in the brushtail possum occurred when follicles reached between 2.5 and 2.8 mm in diameter. Based on the analysis of steroids in follicular fluid, before selection, most follicles produced varying amounts of oestradiol (E2), but very few if any produced progesterone (P4). After selection, the selected follicle continued to produce increasing amounts of E2 and P4, whereas most other follicles were steroidogenically inactive. Near the time of ovulation, presumably after the LH surge, P4 became the predominant steroid produced by the selected follicle and most other follicles once again produced varying amounts of E2. The number of granulosa cells per follicle was highly variable, but tended to increase with increasing diameter. Cell viability was very high, averaging about 95%. Interestingly, the morphology of granulosa cells changed markedly after selection becoming larger and granular in appearance. The weights of the vaginal cul-de-sac and uteri correlated well with the presence of a selected follicle. In ovaries from pregnant animals (n = 3), follicles grew up to 3.5 mm, and although they reached the size of a selected follicle during the follicular phase, E2 production by the other follicles was not suppressed and the weights of the cul-de-sac were less than those from non-pregnant animals with similar sized follicles. During anoestrus (n = 4), follicles did not grow beyond 2 mm and produced very little steroids.

scholarly journals The role of IGFs in the regulation of ovarian follicular growth in the brushtail possum (Trichosurus vulpecula)

Reproduction ◽  
2010 ◽  
Vol 140 (2) ◽  
pp. 295-303 ◽  
Author(s):  
Jennifer L Juengel ◽  
Lisa J Haydon ◽  
Brigitta Mester ◽  
Brian P Thomson ◽  
Michael Beaumont ◽  
...  
Keyword(s):  
Granulosa Cell ◽  
Granulosa Cells ◽  
Cell Function ◽  
Antral Follicle ◽  
Brushtail Possum ◽  
Follicular Growth ◽  
Theca Cells ◽  
Ovarian Follicular Growth

IGFs are known to be key regulators of ovarian follicular growth in eutherian mammals, but little is known regarding their role in marsupials. To better understand the potential role of IGFs in the regulation of follicular growth in marsupials, expression of mRNAs encoding IGF1, IGF2, IGF1R, IGF-binding protein 2 (IGFBP2), IGFBP4 and IGFBP5 was localized by in situ hybridization in developing ovarian follicles of the brushtail possum. In addition, the effects of IGF1 and IGF2 on granulosa cell function were tested in vitro. Both granulosa and theca cells synthesize IGF mRNAs, with the theca expressing IGF1 mRNA and granulosa cell expressing IGF2 mRNA. Oocytes and granulosa cells express IGF1R. Granulosa and theca cells expressed IGFBP mRNAs, although the pattern of expression differed between the BPs. IGFBP5 mRNA was differentially expressed as the follicles developed with granulosa cells of antral follicles no longer expressing IGFBP5 mRNA, suggesting an increased IGF bioavailability in the antral follicle. The IGFBP protease, PAPPA mRNA, was also expressed in granulosa cells of growing follicles. Both IGF1 and IGF2 stimulated thymidine incorporation but had no effect on progesterone production. Thus, IGF may be an important regulator of ovarian follicular development in marsupials as has been shown in eutherian mammals.

scholarly journals Involvement of miRNAs in equine follicle development

Reproduction ◽  
2013 ◽  
Vol 146 (3) ◽  
pp. 273-282 ◽  
Author(s):  
S N Schauer ◽  
S D Sontakke ◽  
E D Watson ◽  
C L Esteves ◽  
F X Donadeu
Keyword(s):  
Cell Survival ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Ovarian Follicle ◽  
Follicle Development ◽  
Previous Evidence ◽  
Ovarian Follicle Development ◽  
Fluid Levels ◽  
Follicle Selection

Previous evidence fromin vitrostudies suggests specific roles for a subset of miRNAs, including miR-21, miR-23a, miR-145, miR-503, miR-224, miR-383, miR-378, miR-132, and miR-212, in regulating ovarian follicle development. The objective of this study was to determine changes in the levels of these miRNAs in relation to follicle selection, maturation, and ovulation in the monovular equine ovary. In Experiment 1, follicular fluid was aspirated during ovulatory cycles from the dominant (DO) and largest subordinate (S) follicles of an ovulatory wave and the dominant (DA) follicle of a mid-cycle anovulatory wave (n=6 mares). Follicular fluid levels of progesterone and estradiol were lower (P<0.01) in S follicles than in DO follicles, whereas mean levels of IGF1 were lower (P<0.01) in S and DA follicles than in DO follicles. Relative to DO and DA follicles, S follicles had higher (P≤0.01) follicular fluid levels of miR-145 and miR-378. In Experiment 2, follicular fluid and granulosa cells were aspirated from dominant follicles before (DO) and 24 h after (L) administration of an ovulatory dose of hCG (n=5 mares/group). Relative to DO follicles, L follicles had higher follicular fluid levels of progesterone (P=0.05) and lower granulosa cell levels ofCYP19A1andLHCGR(P<0.005). Levels of miR-21, miR-132, miR-212, and miR-224 were increased (P<0.05) in L follicles; this was associated with reduced expression of the putative miRNA targets,PTEN,RASA1, andSMAD4. These novel results may indicate a physiological involvement of miR-21, miR-145, miR-224, miR-378, miR-132, and miR-212 in the regulation of cell survival, steroidogenesis, and differentiation during follicle selection and ovulation in the monovular ovary.

Expression of follicle-stimulating hormone and luteinising hormone binding sites in the bitch ovary during the follicular phase

2008 ◽  
Vol 20 (8) ◽  
pp. 925 ◽  
Author(s):  
Marie Saint-Dizier ◽  
Nina Jaffré ◽  
Karine Reynaud ◽  
Benoît Remy ◽  
Sandra Thoumire ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Binding Sites ◽  
Specific Binding ◽  
Follicular Phase ◽  
Follicular Growth ◽  
Physiological Control ◽  
Lh Surge ◽  
Specific Binding Sites ◽  
Follicle Diameter ◽  
High Concentrations

In the female dog, in contrast with most mammals, the growing follicle starts to luteinise several days before ovulation. Little is known about the physiological control of the final follicular growth in this species. In order to better understand the pituitary regulation of follicular growth, specific binding sites for FSH and LH were localised and quantified by autoradiography using [125I]-porcine (p) gonadotrophins on ovarian sections (7 μm) from adult Beagle bitches during the follicular phase. Follicles were analysed either before the LH surge (n = 4 bitches; n = 117 follicles) or after the LH surge and before ovulation (n = 5 bitches; n = 110 follicles). FSH binding sites were specifically and homogeneously expressed at high levels on granulosa cells of all healthy follicles from the preantral stage onwards. In contrast, LH binding sites were detected homogeneously and at high levels only on granulosa cells of follicles larger than 1 mm in diameter, including luteinised follicles. Theca binding of LH (but not FSH) was also observed, but only when using high concentrations of [125I]-pLH. The overall incidence of atresia was 45.8% and was dependent upon follicular diameter. Quantitative analysis of labelling showed that atretic follicles had reduced levels of both FSH and LH binding sites compared with healthy follicles. In healthy follicles, levels of both FSH and LH binding sites changed with follicle diameter. Compared with other mammals, the acquisition of LH binding on canine granulosa cells occurs in smaller sized follicles relative to the size of ovulation.

scholarly journals Peptide Regulators in the Ovarian Follicle

1981 ◽  
Vol 34 (6) ◽  
pp. 491 ◽  
Author(s):  
James M Hammond
Keyword(s):  
Luteinizing Hormone ◽  
Oocyte Maturation ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Ovarian Follicle ◽  
Follicle Stimulating Hormone ◽  
Current Evidence ◽  
Follicular Growth ◽  
Maturation Inhibitor

Data generated within the last several years have shown that follicular fluid contains substances, presumably peptide in nature, which exert potentially important effects on granulosa cells and the oocyte. This review briefly summarizes the current evidence concerning the nature and importance of these putative regulators including the luteinization inhibitor, oocyte maturation inhibitor, inhibitors of the binding of luteinizing hormone and follicle stimulating hormone, stimulators of ornithine decarboxylase and intrafollicular peptides with gonadotrophin-releasing activity. Although the existence of such activities has been clearly demonstrated, the evidence for a regulatory role of these agents in the control of ovarian physiology is not compelling. However, their occurrence must now be taken into account in our attempts to understand the mechanism of follicular growth, differentiation and atresia.

scholarly journals ACTIVITY AND CONTENT OF SUPEROXIDEDISMUTASE IZOZYMES IN GRANULOSE CELLS FROM COW OVARY FOLLICLES

2020 ◽  
Vol 21 (2) ◽  
pp. 33-39
Author(s):  
Yu. V. Bodnar ◽  
N. V. Kuzmina ◽  
D. D. Ostapiv ◽  
S. W. Kawa ◽  
O. I. Chajkovska ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Corpus Luteum ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Physiological State ◽  
Ovarian Follicles ◽  
Extracellular Proteins ◽  
Cytotoxic Action ◽  
Follicular Growth ◽  
Sod Activity

The activity and content of superoxide dismutase isoforms (SOD) in granulosa cells from cow ovarian follicles were studied for research after slaughter of cows ovaries were selected, which according to the physiological state were divided into groups: with "fresh" ovulation, at the site of the ovulated follicle there is a hole, no corpus luteum or diameter up to 5 mm, red color (n = 14); with "early corpus luteum", diameter 10-20 mm, color red or brown (n = 41); with “late corpus luteum", diameter 5–15 mm, color yellow (n = 32); "follicular growth", without the corpus luteum (n = 84). The ovaries of cows with small (<4 mm), medium (4 - 7 mm) and large (> 7 mm) follicles were used. Antral fluid was obtained from the follicles, from which granulosa cells were isolated. Cells were suspended according to the volume of follicular fluid in the medium Dulbeccos modified Eagle medium (DME) with the addition of estrus serum of cows, follicular fluid, insulin and heparin. In cell culture, protein concentration, activity, and superoxide dismutase isozymes were determined. It was found that granulosa cells are characterized by SOD activity - 12.4 ± 0.74 IU / mg protein (6.8 ± 1.72 - 19.8 ± 3.75 IU / mg protein). The activity of SOD in the culture of granulosa cells had 5–6 isoforms of the enzyme. It was found that isoforms at the site of localization are divided into cytosolic, mitochondrial and extracellular proteins of SOD. The cytosolic isoform were represented by 3 - 4, and mitochondrial and extracellular have one active protein of the enzyme. he activity of the enzyme and the content of isoforms depended on the size of the follicles from which the cells are removed and the physiological state of the ovaries. The studied indicators characterize the intensity of oxidative metabolism as a whole in cells and in their individual parts and organelles. For cultivation, it is advisable to select granulosa cells from ovarian follicles of "follicular growth" and "late corpus luteum" because they are characterized by consistently high activity of SOD, which protects intracellular components from the cytotoxic action of superoxide anion.

scholarly journals Relationships between the antral follicle count, steroidogenesis, and secretion of follicle-stimulating hormone and anti-Müllerian hormone during follicular growth in cattle

2019 ◽  
Vol 17 (1) ◽  
Author(s):  
Kenichiro Sakaguchi ◽  
Yojiro Yanagawa ◽  
Koji Yoshioka ◽  
Tomoko Suda ◽  
Seiji Katagiri ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Follicle Stimulating Hormone ◽  
Growth Dynamics ◽  
Antral Follicle ◽  
Antral Follicle Count ◽  
Blood Collection ◽  
Follicular Growth ◽  
Stimulating Hormone

Abstract Background The antral follicle count (AFC) in mammalian ovaries positively correlates with female fertility. To clarify the causes of differences in fertility between low and high AFC cows, we investigated follicular growth dynamics and hormone concentrations in plasma, follicular fluid, and in vitro growth (IVG) media at different stages of follicular growth. Methods Seven cows were divided into high AFC (n = 4, > 30 follicles) and low AFC (n = 3, < 30 follicles) groups based on the peak AFC detected by ultrasonography. These cows were subjected to estrous synchronization, daily ovarian ultrasonography, and blood collection. Their follicular fluid was collected from dominant follicles at different stages (selection, luteal, and ovulatory phases). In another experiment, we cultured oocyte-cumulus-granulosa cell complexes collected from early antral follicles (< 1 mm) for 12 days. Estradiol-17β (E2), testosterone (T), progesterone (P4), and anti-Müllerian hormone (AMH) concentrations in follicular fluids and plasma were measured. Plasma follicle-stimulating hormone (FSH) concentrations were examined. E2, P4, and AMH concentrations were also measured in IVG media. Results The numbers of small (< 4 mm) and intermediate (4–8 mm) follicles were larger in the high AFC group than in the low AFC group (P < 0.05). The number of intermediate follicles was stable in the low AFC group, indicating consistent development. However, the number of these follicles fluctuated in the high AFC group. Plasma FSH concentrations were higher, whereas E2 and T concentrations were lower in the low AFC group (P < 0.05). E2 concentrations and the E2/P4 ratio in ovulatory follicles and IVG media on day 8 were higher in the high AFC group (P < 0.05). AMH concentrations in plasma and IVG media (P < 0.01) were higher in the high AFC group. Conclusions The weaker response to FSH of granulosa cells caused low E2 production in the low AFC group, resulting in high FSH concentrations and the consistent development of intermediate follicles. Conversely, higher E2 concentrations suppressed FSH secretion in the high AFC group. Granulosa cells in the high AFC group had the ability to produce more AMH than those in the low AFC group throughout IVG culture.

scholarly journals CYTOMORPHOLOGICAL RESEARCH OTSYT-CUMULUS COMPLEXES RABBIT FROM WITH OVARIAN AT DIFFERENT STAGES OF THE ESTROUS CYCLE

2017 ◽  
Vol 53 ◽  
pp. 279-284
Author(s):  
A. B. Zyuzyun
Keyword(s):  
Estrous Cycle ◽  
Luteal Phase ◽  
Follicular Phase ◽  
The Body ◽  
Follicular Growth ◽  
Research Results ◽  
Significant Difference ◽  
Further Development ◽  
Ovarian Follicular Growth

The analysis of the research results revealed that the largest number (86.4%) of oocytes suitable for further development outside the body can be obtained with ovarian follicular phase of growth. It should be noted that statistically significant difference was observed between the groups OCC rabbit derived from ovaries at different phases of the estrous cycle by the number oocytes unsuitable for further cultivation. Thus, the phase of the ovarian follicular growth of gametes was obtained only 13.6% of ovarian and with signs of ovulation and the luteal phase, 35.4% and 31.4% respectively. When comparing the results of the analysis of cytogenetic preparations oocytes from ovaries removed rabbit at different stages of the estrous cycle, found that regardless of the phase of the estrous cycle Yachnik mostly larger number of oocytes were under dyploteny. The largest number of gametes with diffuse chromatin at the stage dyploteny (37.3%) received from the stage ovarian follicular growth. At the stage of fibrillar dyploteny increasing number of gametes was removed from ovarian luteal phase of the estrous cycle. In step dyploteny visible bivalent were more likely gametes obtained from stage ovarian follicular growth (18,1%, p <0,05). The highest percentage of oocytes degeneration chromatin was observed in the group removed from the ovaries to the rabbit lyutealniy phase (21.6%).

scholarly journals Variation in antral follicle development during the follicular phase of the oestrous cycle in red deer (Cervus elaphus) hinds

Reproduction ◽  
2001 ◽  
pp. 697-705 ◽  
Author(s):  
BJ McLeod ◽  
LM Meikle ◽  
MW Fisher ◽  
TR Manley ◽  
DA Heath ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Follicular Fluid ◽  
Red Deer ◽  
Antral Follicle ◽  
Follicular Phase ◽  
Oestrous Cycle ◽  
Aromatase Activity ◽  
Follicle Development ◽  
Follicle Diameter ◽  
Follicle Size

The aim of this study was to quantify antral follicle populations in cyclic red deer hinds and to monitor follicle development leading to ovulation. Oestrus was synchronized with exogenous progesterone and ovaries were recovered approximately 0, 12, 24 or 36 h (follicular phase) or 10 days (luteal phase) after progesterone withdrawal (n = 5 per group). All follicles > or = 2 mm in diameter were dissected out, health status was assessed, follicular fluid oestradiol content was measured, granulosa cells were harvested and their capacity for oestradiol and cAMP production was determined. The time of oestrus and the preovulatory LH surge were monitored in five control hinds. Deer ovaries contained 26.6 +/- 3.45 (mean +/- SEM) follicles > or = 2 mm in diameter (range 4-81), with at least one large antral follicle (diameter: 8.3 +/- 0.38 mm) per hind. There was a strong correlation between follicle size and granulosa cell population (r(2) = 0.676). Approximately half (50.7%) of the follicles were classified as healthy, with the percentage classified as atretic decreasing with increasing follicle size. Neither the total number of antral follicles nor their size distribution differed significantly among groups. There were significantly more (P < 0.05) healthy follicles at 24 h after progesterone withdrawal than at 0 h, when large oestrogenic follicles had fewer granulosa cells, lower follicular fluid oestradiol concentrations and lower aromatase activity (P < 0.05) than did those from other groups. In summary, antral follicle development in red deer is similar to that in other monovulatory ruminants, and at least one large follicle is present at all stages of the oestrous cycle.

scholarly journals Gonadotrophin responsiveness, aromatase activity and insulin-like growth factor binding protein content of bovine ovarian follicles during the first follicular wave

Reproduction ◽  
2001 ◽  
pp. 561-569 ◽  
Author(s):  
FM Rhodes ◽  
AJ Peterson ◽  
PD Jolly
Keyword(s):  
Growth Factor ◽  
Granulosa Cells ◽  
Follicular Fluid ◽  
Ovarian Follicles ◽  
Aromatase Activity ◽  
Insulin Like Growth Factor ◽  
Dominant Follicle ◽  
Factor Binding ◽  
Follicular Wave ◽  
Selection Of

The aim of this study was to examine the function of granulosa cells and hormone concentrations in follicular fluid in bovine ovarian follicles during selection of the first dominant follicle. Ovaries were obtained from beef heifers on days 1-5 after ovulation: follicles > 4 mm in diameter were dissected and follicular fluid and granulosa cells were collected from individual follicles. Oestradiol production by granulosa cells after culture with testosterone was used to determine aromatase activity and responsiveness to gonadotrophins was determined by cAMP production after culture with FSH or LH. Concentrations of oestradiol, progesterone and insulin-like growth factor binding proteins (IGFBPs)-4 and -5 were measured in follicular fluid. Follicles were classified as largest or smaller (days 1 and 2), or dominant or subordinate (days 3-5). Aromatase activity was greater in granulosa cells from the largest follicle than in granulosa cells from smaller follicles on days 1, 3, 4 and 5 (P < 0.05). Responsiveness to LH was not detected in granulosa cells on day 1, but from day 2 to day 5 cells from the largest follicle were significantly more responsive than cells from smaller follicles (P < 0.05). Responsiveness to FSH was detected in granulosa cells from all follicles from day 1 onwards and did not differ between cells from the largest follicle or smaller follicles on any day. Follicular fluid concentrations of oestradiol and the ratio of oestradiol:progesterone were greater and concentrations of IGFBP-4 and -5 were lower in the largest follicle than in smaller follicles from day 2 to day 5 (P < 0.05). In conclusion, selection of the dominant follicle is associated with increased granulosa cell aromatase activity followed by increased cAMP response to LH and follicular fluid oestradiol concentrations, and decreased follicular fluid concentrations of IGFBP-4 and -5 within 2 days after ovulation.

HORMONAL REGULATION OF THE NOTCH PATHWAY GENES IN THE GRANULOSA CELLS DURING GONADOTROPIN INDUCED OVARIAN FOLLICULAR GROWTH

2007 ◽  
Vol 77 (Suppl_1) ◽  
pp. 119-119
Author(s):  
Raj Duggavathi ◽  
Charles Thomas ◽  
Johan Auwerx ◽  
Kristina Schoonjans ◽  
Bruce Murphy
Keyword(s):  
Granulosa Cells ◽  
Hormonal Regulation ◽  
Notch Pathway ◽  
Follicular Growth ◽  
Ovarian Follicular Growth
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