Caspase-3-mediated apoptosis and cell proliferation in the equine endometrium during the oestrous cycle

2007 ◽  
Vol 19 (8) ◽  
pp. 925 ◽  
Author(s):  
R. P. Roberto da Costa ◽  
P. M. Serrão ◽  
S. Monteiro ◽  
P. Pessa ◽  
J. Robalo Silva ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Luteal Phase ◽  
Caspase 3 ◽  
Follicular Phase ◽  
Oestrous Cycle ◽  
Nuclear Antigen ◽  
Simultaneous Increase ◽  
Endometrial Cell ◽  
Glandular Structures ◽  
Active Caspase

Cell proliferation and apoptosis are hormone-dependent physiological processes involved in endometrial growth and regression. The aims of the present study were: (1) to evaluate endometrial cell proliferation using proliferating cell nuclear antigen (PCNA) expression; (2) to evaluate the induction of endometrial cell death by the expression of active caspase-3 and the apoptotic phenotype visualised by DNA fragmentation; and (3) to relate these observations to endometrial tissue dynamics in the equine endometrium throughout the oestrous cycle. Endometria were assigned to follicular and luteal phases based on ovarian structures and plasma progesterone. Cell proliferation and active caspase-3-mediated apoptosis were expressed in both phases of the oestrous cycle. In the luteal phase, PCNA expression was higher than in the follicular phase. Highest PCNA activity was noted in the luminal and glandular structures. Active caspase-3 staining was increased in luminal epithelium and deep glandular cells during the luteal phase. However, in the follicular phase, stromal cells showed greater active caspase-3 expression. Only a few apoptotic endometrial cells were detected by terminal deoxyribonucleotidyl transferase-mediated dUTP–digoxigenin nick end-labelling (TUNEL) and these cells were mostly present in luminal and glandular structures. A simultaneous increase in DNA, cell proliferation and protein synthesis was observed in the endometrium during the mid-luteal phase. This suggests that cell hyperplasia occurs at the time the histotroph is needed for eventual embryo nourishment.

scholarly journals Apoptotic Cell Death in Ewe Endometrium during the Oestrous Cycle

2020 ◽  
Vol 71 (3) ◽  
pp. 2323
Author(s):  
S. BENBIA ◽  
Y. BELKHIRI ◽  
M. YAHIA
Keyword(s):  
Cell Death ◽  
Luteal Phase ◽  
Caspase 3 ◽  
Apoptotic Cell ◽  
Follicular Phase ◽  
Oestrous Cycle ◽  
Ki 67 ◽  
Plasma Progesterone ◽  
Endocrine Profile ◽  
Macroscopic Observation

We hypothesized that endometrial tissues from ewes undergo spatial and temporal changes. Thus, two regulatory events were investigated in this study: cell death (apoptosis) and cell proliferation. Uteri were obtained from healthy ewes at Batna abattoir (Algeria). Based on macroscopic observation of the ovaries and plasma progesterone, uteri were assigned to follicular, early and active luteal phases. Apoptosis and proliferation were assessed by detection of cleaved caspase-3 and Ki-67, respectively. Ki-67 and cleaved caspase-3 (CCP-3) were expressed in both phases of the oestrous cycle and all endometrium cells types [luminal epithelia (LE), superficial gland epithelia (SG) and deep gland epithelia (DG)]. Immunohistochemistry for cleaved caspase-3 revealed few or no apoptotic stained cells in all endometrium locations during the entire oestrous cycle. However, Ki-67 was significantly higher in the follicular phase than in the early and active luteal phase. Besides, expression of CCP-3 in LE was higher than in SG and DG at the follicular phase and early luteal phase. However, Ki -67 and CCP-3 levels in all endometrium cells types did not significantly change at active luteal phase. Therefore, it is concluded that apoptosis and proliferation were occurred in ewe endometrium in a cyclic pattern and under the influence of the endocrine profile.

Seasonal and steroid-dependent effects on the modulation of LH secretion in the ewe by intracerebroventricularly administered β-endorphin or naloxone

1989 ◽  
Vol 122 (2) ◽  
pp. 509-517 ◽  
Author(s):  
R. J. E. Horton ◽  
H. Francis ◽  
I. J. Clarke
Keyword(s):  
Breeding Season ◽  
Luteal Phase ◽  
Pulse Frequency ◽  
Opioid Peptide ◽  
Follicular Phase ◽  
Oestrous Cycle ◽  
Opioid Antagonist ◽  
Endogenous Opioid ◽  
Endogenous Opioid Peptide ◽  
Lh Secretion

ABSTRACT The natural opioid ligand, β-endorphin, and the opioid antagonist, naloxone, were administered intracerebroventricularly (i.c.v.) to evaluate effects on LH secretion in ovariectomized ewes and in ovariectomized ewes treated with oestradiol-17β plus progesterone either during the breeding season or the anoestrous season. Ovary-intact ewes were also studied during the follicular phase of the oestrous cycle. Jugular blood samples were taken at 10-min intervals for 8 h and either saline (20–50 μl), 100 μg naloxone or 10 μg β-endorphin were injected i.c.v. after 4 h. In addition, luteal phase ewes were injected i.c.v. with 25 μg β-endorphin(1–27), a purported endogenous opioid antagonist. In ovariectomized ewes, irrespective of season, saline and naloxone did not affect LH secretion, but β-endorphin decreased the plasma LH concentrations, by reducing LH pulse frequency. The effect of β-endorphin was blocked by administering naloxone 30 min beforehand. Treating ovariectomized ewes with oestradiol-17β plus progesterone during the breeding season reduced plasma LH concentrations from 6–8 μg/l to less than 1 μg/l. In these ewes, saline did not alter LH secretion, but naloxone increased LH pulse frequency and the plasma concentrations of LH within 15–20 min. During anoestrus, the combination of oestradiol-17β plus progesterone to ovariectomized ewes reduced the plasma LH concentrations from 3–5 μg/l to undetectable levels, and neither saline nor naloxone affected LH secretion. During the follicular phase of the oestrous cycle, naloxone enhanced LH pulse frequency, which resulted in increased plasma LH concentrations; saline had no effect. In these sheep, β-endorphin decreased LH pulse frequency and the mean concentrations of LH, and this effect was prevented by the previous administration of naloxone. The i.c.v. administration of β-endorphin(1–27) to luteal phase ewes did not affect LH secretion. These data demonstrate the ability of a naturally occurring opioid peptide to inhibit LH secretion in ewes during the breeding and non-breeding seasons, irrespective of the gonadal steroid background. In contrast, whilst the gonadal steroids suppress LH secretion in ovariectomized ewes during both seasons, they only appear to activate endogenous opioid peptide (EOP)-mediated inhibition of LH secretion during the breeding season. Furthermore, these data support the notion that LH secretion in ovariectomized ewes is not normally under the control of EOP, so that naloxone has no effect. Journal of Endocrinology (1989) 122, 509–517

scholarly journals Effect of ram introduction on the oestrous cycle of springbok ewes (Antidorcas marsupialis)

Reproduction ◽  
2002 ◽  
pp. 509-513 ◽  
Author(s):  
DC Skinner ◽  
SD Cilliers ◽  
JD Skinner
Keyword(s):  
Maximum Concentration ◽  
Luteal Phase ◽  
Follicular Phase ◽  
Oestrous Cycle ◽  
Arid Environments ◽  
Wild Ungulates ◽  
Plasma Progesterone ◽  
Before And After ◽  
Antidorcas Marsupialis

Springbok are aseasonally breeding wild ungulates that inhabit arid environments, and interest has been shown in domesticating them for agricultural purposes. The present study was conducted for husbandry purposes to determine the effect of introducing a vasectomized ram to an isolated herd of springbok ewes (n = 9). Blood was collected from ewes every third day, before and after introduction of a vasectomized ram. Ewes were subjected to the ram for 42 days. Plasma progesterone was measured by radioimmunoassay and was used to establish the stage of the oestrous cycle. After introduction of the ram, the variation in the timing of the follicular phase between ewes was clearly reduced, compressing the spread of oestrus in the springbok ewes from 11 to 3 days. In seven of the nine ewes, the ram was introduced during the luteal phase of the oestrous cycle, causing this cycle to be significantly longer in duration (P < 0.05) and to have a higher maximum concentration of progesterone (P < 0.001) than cycles before and after introduction of the ram. This finding implies that the mechanism of synchronization operates through a luteotrophic effect. These results indicate that rams may be used successfully to synchronize breeding in springbok.

Source of ovarian inhibin secretion during the oestrous cycle of the sheep

1989 ◽  
Vol 123 (2) ◽  
pp. 181-188 ◽  
Author(s):  
G. E. Mann ◽  
A. S. McNeilly ◽  
D. T. Baird
Keyword(s):  
Corpus Luteum ◽  
Luteal Phase ◽  
Venous Blood ◽  
Contralateral Side ◽  
Follicular Phase ◽  
Oestrous Cycle ◽  
Secretion Rate ◽  
Antral Follicles ◽  
Luteal Tissue

ABSTRACT The source of inhibin secretion by the ovary in the sheep at different stages of the oestrous cycle was investigated by in-vivo cannulation of the ovarian veins. Twenty-four Scottish Blackface ewes were allocated to four groups of six ewes, i.e. those operated on during the luteal phase (day 10), and those operated on during the follicular phase 24–30, 36 and 60 h following an injection of 125 μg cloprostenol on day 10 of the luteal phase. Samples of jugular and timed ovarian venous blood were collected under anaesthesia before and after enucleation of the corpus luteum. Ovaries were then removed and follicles dissected out. Following injection of cloprostenol, luteal regression occurred as indicated by a fall in the secretion of progesterone. The concentration of inhibin in jugular venous plasma and its ovarian secretion rate were similar at all stages of the follicular phase and during the luteal phase. In contrast, the secretion rate of oestradiol rose from 2·68 ±0·73 pmol/min during the luteal phase to 8·70± 2·24 pmol/min 24 h after injection of cloprostenol (P<0·05). Following enucleation of the corpus luteum the secretion rate of progesterone fell from 809 ± 270 pmol/min to 86 ± 30 pmol/min (P<0·001). There was also a smaller, artifactual fall in the secretion rate of oestradiol following enucleation of the corpus luteum, which was of similar size to a fall seen in the secretion rate of inhibin. This resulted in a significant (P<0·001) fall in the ratio of progesterone to inhibin, while the oestradiol to inhibin ratio remained unchanged. The secretion rate of inhibin from ovaries containing luteal tissue was similar to that from the contralateral side without luteal tissue (1·41±0·30 compared with 1·32±0·30 ng/min), while ovaries with large antral follicles secreted significantly (P< 0·001) more inhibin than those with no follicles ≥3 mm (2·28 ± 0·36 compared with 0·25 ±0·06 ng/min). From these results we conclude that, in the sheep, large antral follicles are responsible for most, if not all, the secretion of inhibin by the ovary at all stages of the oestrous cycle, and that the corpus luteum secretes little or no immunoactive or bioactive inhibin. Due to the fact that, unlike inhibin, the secretion rate of oestradiol rises during the follicular phase of the cycle, when the concentration of FSH is suppressed, it seems likely that oestradiol rather than inhibin is the major ovarian factor modulating the change in FSH secretion seen at this stage of the oestrous cycle. Journal of Endocrinology (1989) 123, 181–188

Evidence that the switch from negative to positive feedback at the level of the pituitary gland is an important timing event for the onset of the preovulatory surge in LH in the ewe

1995 ◽  
Vol 145 (2) ◽  
pp. 271-282 ◽  
Author(s):  
I J Clarke
Keyword(s):  
Pituitary Gland ◽  
Negative Feedback ◽  
Luteal Phase ◽  
Bolus Injection ◽  
Pulse Frequency ◽  
Follicular Phase ◽  
Oestrous Cycle ◽  
Lh Surge ◽  
Small Rise ◽  
Lh Secretion

Abstract Experiments were performed to test the hypothesis that there is a negative feedback 'clamp' of ovarian hormones on the hypothalamus and pituitary gland during the follicular phase of the oestrous cycle that limits the secretion of GnRH and LH. GnRH secretion was monitored by sampling the hypophysial portal blood of ewes during the luteal phase of the oestrous cycle and either 24 h or 48 h after the induction of luteolysis by the injection of cloprostenol, a prostaglandin analogue. There was an increase in GnRH pulse frequency in the transition from the luteal to the follicular phase of the cycle. A reduction in the amplitude of GnRH pulses did not occur until 48 h after cloprostenol, suggestive of negative feedback at the level of the hypothalamus that is more profound in the latter part of the follicular phase. The responsivity of the pituitary gland to GnRH was monitored in ewes during the luteal phase of the oestrous cycle and 24 h or 48 h after cloprostenol. Injections of 250 ng or 1000 ng GnRH were given (i.v.) to ewes that had been anaesthetised to suppress endogenous secretion of GnRH and LH. Using the lower dose, the responses 48 h after cloprostenol were not significantly different from those in the luteal phase. With the higher dose of GnRH, a significant (P<0·05) increase in mean responsivity was seen 48 h after cloprostenol. There was, however, a marked variation in response, with some ewes showing profound increases in LH secretion in response to GnRH and others showing responses that were similar to those obtained during the luteal phase of the cycle. These data are interpreted to mean that the secretion of LH is 'clamped' during the follicular phase of the oestrous cycle and the 'clamp' is only released near the time of the preovulatory LH surge. To test whether or not a rise in GnRH input to the pituitary gland could over-ride the 'clamp' on the pituitary secretion of LH in the late follicular phase of the cycle, sheep were treated 40 h after cloprostenol with either a bolus injection of 500 ng GnRH or four pulses of 125 ng GnRH given at 10-min intervals. These treatments caused small elevations in LH secretion but did not always cause preovulatory LH surges. In some cases, a small rise in LH secretion was induced by GnRH treatments and levels of LH in plasma returned to baseline with the preovulatory LH surge occurring a few hours later. In one clear case, a bolus injection of GnRH induced an LH surge. The overall data from the GnRH-treated groups, however, indicated a significant delay in the onset of the LH surge which may have been due to perturbation of the subcellular mechanisms in the gonadotrophs. These data were interpreted to mean that the secretion of LH from the pituitary gland is inhibited up to very soon before the onset of the LH surge. The inhibitory factor could be oestrogen but could also be some other pituitary feedback hormone such as gonadotrophin surge-attenuating factor. It is concluded that the increase in the secretion of GnRH at the time of the onset of the LH surge is closely linked to an increase in the responsivity of the gonadotrophs to GnRH. The latter is not caused by the increase in the secretion of GnRH. Journal of Endocrinology (1995) 145, 271–282

Isoliquiritigenin Suppresses Osteosarcoma U2OS Cell Proliferation and Invasion by Regulating the PI3K/Akt Signalling Pathway

Chemotherapy ◽  
2018 ◽  
Vol 63 (3) ◽  
pp. 155-161 ◽  
Author(s):  
Jing Chen ◽  
Cheng Liu ◽  
Qin-Qing Yang ◽  
Rui-Bin Ma ◽  
Ying Ke ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Protein Expression ◽  
Caspase 3 ◽  
Signalling Pathway ◽  
U2os Cell ◽  
Expression Levels ◽  
Migration Assay ◽  
Invasion And Migration ◽  
U2os Cells ◽  
Active Caspase

Aims: Isoliquiritigenin (ISL) is a flavonoid, that has been shown to have antioxidant, vasorelaxant, anti-inflammatory, and antitumor activities. This study aimed to explore the antitumor effect of ISL on human osteosarcoma U2OS cells and investigate the mechanism of this effect. Methods: The effect of ISL on osteosarcoma U2OS cell proliferation, invasion, migration, and apoptosis were determined by a CCK8 assay, a transwell invasion assay, a transwell migration assay, and fluorescence-activated cell sorting, respectively. In addition, the protein expression levels of Bcl2, Bax, active Caspase-3, Akt, mTOR, p70, and Cyclin D1 were detected by western blotting. Results: ISL suppressed cell proliferation, inhibited invasion and migration, and promoted apoptosis in U2OS cells. After treatment with ISL, the protein expression levels of Bax and active Caspase-3 increased, while the level of Bcl-2 declined significantly. Furthermore, the phosphorylation levels of Akt and mTOR declined significantly compared with that of the control. Conclusion: ISL could retard proliferation and promote apoptosis of U2OS cells possibly by suppressing the PI3K/Akt signalling pathway, indicating that it might be a potential therapeutic agent for osteosarcoma treatment.

Proliferation and apoptosis processes in the seasonal testicular development of the wild Daurian ground squirrel (Citellus dauricus Brandt, 1844)

2017 ◽  
Vol 29 (9) ◽  
pp. 1680 ◽  
Author(s):  
Yingying Han ◽  
Jinqi Zhan ◽  
Ying Xu ◽  
Fengwei Zhang ◽  
Zhengrong Yuan ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Breeding Season ◽  
Caspase 3 ◽  
Nuclear Translocation ◽  
Ground Squirrels ◽  
Nuclear Antigen ◽  
Plasma Testosterone ◽  
Testicular Development ◽  
Cyclin D2 ◽  
Proliferation And Apoptosis

The aim of the present study was to elucidate the regulatory role of cell proliferation and apoptosis in testicular development of wild Daurian ground squirrels during the breeding season (April), the non-breeding season (June) and before hibernation (September). Gross mass and hormonal analysis showed that the testis : body mass ratio and plasma testosterone concentration fluctuated seasonally, with a peak in April and lowest values in June. Similarly, spermatogenesis was fully developed in April but suppressed in June and September. Testicular decellularisation and vacuolisation was seen during the transition from the breeding to the non-breeding season. Furthermore, testicular levels of proliferating cell nuclear antigen, cyclin D2 and caspase-3 protein were significantly increased in June and September. Intriguingly, positive terminal deoxyribonucleotidyl transferase-mediated dUTP–digoxigenin nick end-labelling staining and nuclear translocation of caspase-3 in testicular germ cells appeared only during the prehibernation period, whereas accumulation of cyclin D2 in spermatocyte nuclei occurred in September. These findings demonstrate, for the first time, that both cell proliferation and apoptosis are stimulated during the prehibernation period, indicating that a hormonal-regulated balance of testicular germ cell proliferation and apoptosis may play a pivotal role in preparing for testicular recrudescence of wild Daurian ground squirrels.

Mobilization of LH secretory granules in gonadotrophs in relation to gene expression, synthesis and secretion of LH during the preovulatory phase of the sheep oestrous cycle

1995 ◽  
Vol 147 (2) ◽  
pp. 259-270 ◽  
Author(s):  
R J W Currie ◽  
A S McNeilly
Keyword(s):  
Electron Microscopy ◽  
Luteal Phase ◽  
Secretory Granules ◽  
Follicular Phase ◽  
Oestrous Cycle ◽  
Single Type ◽  
Maximum Diameter ◽  
Northern Analysis ◽  
Mrna Levels ◽  
Lh Surge

Abstract Changes in LH secretory granules in pituitary gonadotrophs throughout the sheep oestrous cycle were determined by immunogold localisation of LH at ultrastructural level by electron microscopy. Oestrous cycles in Welsh Mountain ewes were initially synchronised with progestagen sponges and studies carried out in the subsequent cycle. Animals were allocated at random to six groups each of five animals, one killed on day 12 of the luteal phase and the other groups after prostaglandin (PG)-induced luteal regression at PG plus 18 h (early follicular phase), oestrus (PG plus 33·6±1·0 h), oestrus plus 9 h just before the preovulatory LH surge, 1 h after GnRH agonist-induced LH surge at PG plus 48 h (mid-LH surge) and oestrus plus 24 h, after the preovulatory LH surge. Blood samples collected throughout confirmed the pulsatile secretion of LH before and the timing in relation to the preovulatory LH surge. Pituitaries were dissected and processed for transmission electron microscopy and frozen for later extraction of mRNA. Only a single type of LH cell was present in the sheep pituitary. In the luteal phase, LHimmunopositive secretory granules were distributed throughout the cytoplasm in 80% of cells while in 20% of cells granules were polarised to the region of the cell next to a vascular sinusoid. The percentage of polarised cells increased during the follicular phase to 45% at oestrus, 75% at oestrus plus 9 h just before the LH surge and 90% in mid-LH surge. Cell size increased in parallel with polarisation. Gonadotrophs after the LH surge were almost totally devoid of LH granules but prominent LHβ immunoreactivity was observed in the rough endoplasmic reticulum. Analysis of granule diameters revealed a single class of granules with a maximum diameter of 300 nm. Polarised cells had significantly fewer 130–150 nm granules than non-polarised cells, suggesting preferential exocytosis of LH-containing granules of this size from polarised cells. Northern analysis showed that LHβ mRNA levels decreased from luteal through the follicular phase. These results suggest that the preovulatory LH surge in sheep is not related to a change in synthesis of LH but to a progressive recruitment of gonadotrophs into a releasing state, priming, as indicated by polarisation of secretory granules to the region of the cell next to the vascular system. Journal of Endocrinology (1995) 147, 259–270

Hormone production in vivo and in vitro from follicles at different stages of the oestrous cycle in the sheep

1992 ◽  
Vol 132 (2) ◽  
pp. 225-234 ◽  
Author(s):  
G. E. Mann ◽  
A. S. McNeilly ◽  
D. T. Baird
Keyword(s):  
Luteal Phase ◽  
Follicular Phase ◽  
Oestrous Cycle ◽  
Secretion Rate ◽  
Hormone Production ◽  
Luteal Regression ◽  
Follicle Diameter ◽  
Oestradiol Production

ABSTRACT This experiment was undertaken in order to investigate the production of inhibin, oestradiol and androstenedione by ovarian follicles at different stages of the oestrous cycle in sheep. Twenty-four Scottish Blackface ewes were allocated to four groups of six ewes, i.e. those operated on during the luteal phase (day 10), and those operated on during the follicular phase 24–30, 36 and 60 h after the induction of luteal regression by an injection of 125 μg cloprostenol on day 10 of the luteal phase. Samples of jugular and ovarian venous blood were collected under anaesthesia and ovaries were then removed and all follicles larger than 3 mm diameter dissected out and incubated in medium for 2 h. After injection of cloprostenol, luteal regression occurred as indicated by a fall in the secretion rate of progesterone. The ovarian secretion rate of inhibin was similar at all stages of the follicular phase and during the luteal phase while, in contrast, the secretion rate of oestradiol was significantly (P < 0·05) elevated in the group 24 h after injection of cloprostenol. There was good correlation between the in-vivo ovarian secretion rate and production rate during incubation in vitro for both inhibin (r = 0·57) and oestradiol (r = 0·60). When follicle diameter was compared with in-vitro hormone production there was good correlation for inhibin (r = 0·72) with larger follicles producing more inhibin, while the value for oestradiol was somewhat lower (r = 0·57) owing to the presence of large atretic follicles with low oestradiol production. Androstenedione production showed a lower correlation with follicle diameter (r = 0·39). When the four time periods were compared separately, there were significantly (P < 0·05) more follicles with high in-vitro oestradiol production (> 90 fmol/min) in the group at 36 h than in the other three groups, while inhibin release in relation to follicle size was similar in the four groups. Large oestrogenic follicles were responsible for 90% of the total oestradiol production during culture while only providing 55% of the total inhibin production, with large non-oestrogenic and small follicles contributing 33% and 12% of inhibin production respectively. From the results of this study we conclude that while oestradiol is mainly produced by the large oestrogenic follicles, a considerable amount of inhibin is also produced by large non-oestrogenic and small follicles. We also found that a lack of variation in inhibin secretion rate in the intact animal was paralleled by a lack of variation in the pattern of inhibin produced from individual follicles. Journal of Endocrinology (1992) 132, 225–234

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