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

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

Changes in the plasma concentrations of inhibin throughout the normal sheep oestrous cycle and after the infusion of FSH

1989 ◽  
Vol 120 (2) ◽  
pp. 295-305 ◽  
Author(s):  
A. S. McNeilly ◽  
I. A. Swanston ◽  
W. Crow ◽  
C. G. Tsonis ◽  
D. T. Baird
Keyword(s):  
Follicular Fluid ◽  
Plasma Concentrations ◽  
Follicular Phase ◽  
Oestrous Cycle ◽  
Secretion Rate ◽  
Metabolic Clearance ◽  
Α Subunit ◽  
Sephadex Column ◽  
Luteal Regression ◽  
Ovarian Inhibin

ABSTRACT A radioimmunoassay for inhibin was developed using a peptide containing the 1–26 amino acid sequence of the N-terminus of the α-chain of 32 kDa porcine inhibin as immunogen, and 125I-labelled tracer. Evaluation of this assay using Sephadex column chromatography, chromatoelectrophoresis and immunoblotting confirmed that it measured all forms of inhibin present in sheep follicular fluid and was suitable for measurement of inhibin in sheep plasma. There was no evidence of the presence of free α-subunit in either sheep follicular fluid or ovarian vein plasma. The concentration of inhibin in jugular plasma throughout the follicular and luteal phases of four ewes with ovarian autotransplants was measured. The ovarian secretion of inhibin and oestradiol were also measured simultaneously throughout the follicular phase in a spontaneous cycle and after infusion of NIH-oFSH-S14 at 10 μg/h for 48 h following premature luteal regression induced by prostaglandin. The results showed: (1) no change in the peripheral concentration of inhibin throughout the cycle except an increase related to the periovulatory increase in FSH and LH. (2) Following luteal regression, the concentration of FSH fell as the secretion rate of oestradiol increased. During this time there was no significant change in the peripheral concentration of inhibin or ovarian inhibin secretion rate. (3) Following the infusion of FSH there was a marked increase in the concentration of inhibin in both ovarian and peripheral plasma and an increase in ovarian inhibin secretion rate. (4) The calculated metabolic clearance rate of inhibin, 20·3 ml/min, is similar to that of FSH. We conclude that in the ewe the ovarian inhibin secretion rate is stimulated by FSH and, although inhibin may modulate the basal secretion of FSH, a change in its secretion does not account for the fall in FSH which occurs during the follicular phase of the sheep oestrous cycle. Journal of Endocrinology (1989) 120, 295–305

STIMULATION BY HUMAN CHORIONIC GONADOTROPHIN OF OESTRADIOL PRODUCTION BY DISPERSED CELLS FROM HUMAN CORPUS LUTEUM: COMPARISON WITH PROGESTERONE PRODUCTION; UTILIZATION OF EXOGENOUS TESTOSTERONE

1981 ◽  
Vol 91 (2) ◽  
pp. 197-203 ◽  
Author(s):  
M. C. RICHARDSON ◽  
G. M. MASSON
Keyword(s):  
Luteal Phase ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Cell Suspensions ◽  
Corpora Lutea ◽  
Progesterone Production ◽  
Late Luteal Phase ◽  
Oestradiol Production ◽  
Dispersed Cells

Cell suspensions were prepared from tissue samples of human corpora lutea obtained during the mid- and late-luteal phase of the menstrual cycle. Both oestradiol and progesterone production by dispersed cells were stimulated by similar concentrations of human chorionic gonadotrophin (hCG). As the degree of stimulation of production by hCG was greater for progesterone than for oestradiol (five- to tenfold compared with two- to threefold higher than basal production), the ratio of progesterone to oestradiol produced varied according to the level of trophic stimulation. A comparison of cell suspensions prepared from mid- and late-luteal phase corpora lutea, exposed to the same concentration of hCG (10 i.u./ml) in vitro, did not reveal a shift to oestradiol production in the late-luteal phase. Provision of additional testosterone during incubation raised the level of oestradiol production by dispersed luteal cells. At an optimum concentration of testosterone (1 μmol/l), oestradiol synthesis was not raised further in the presence of hCG or N6, O2-dibutyryl cyclic AMP, suggesting a lack of induction or activation of the aromatase system by gonadotrophin in short-term cultures. Basal and stimulated levels of progesterone production were not significantly impaired in the presence of testosterone.

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 Suppression of parathyroid hormone production in vitro and in vivo by RNA interference

2009 ◽  
Vol 75 (5) ◽  
pp. 490-498 ◽  
Author(s):  
Genta Kanai ◽  
Takatoshi Kakuta ◽  
Kaichiro Sawada ◽  
Tun A. Yokoyama ◽  
Reika Tanaka ◽  
...  
Keyword(s):  
Parathyroid Hormone ◽  
Rna Interference ◽  
Hormone Production

Passively immunizing ewes against inhibin during the luteal phase of the oestrous cycle raises the plasma concentration of FSH

1989 ◽  
Vol 123 (3) ◽  
pp. 383-391 ◽  
Author(s):  
G. E. Mann ◽  
B. K. Campbell ◽  
A. S. McNeilly ◽  
D. T. Baird
Keyword(s):  
Plasma Concentration ◽  
Luteal Phase ◽  
Passive Immunization ◽  
Oestrous Cycle ◽  
Ovulation Rate ◽  
Control Group ◽  
Pituitary Cells ◽  
Marked Rise ◽  
Peripheral Plasma

ABSTRACT Passive immunization was used to investigate the importance of inhibin in the negative feedback loop regulating the production of FSH in sheep. An antiserum raised to the 1–26 peptide fragment of the N-terminus of the α-chain of porcine inhibin was first shown to neutralize the suppressive effects of inhibin on the production of FSH by dispersed ovine pituitary cells in vitro. Groups of five mature Scottish Blackface ewes on day 8 of the luteal phase of the oestrous cycle were then injected with either 10 ml plasma from normal ewes (control) or 10 ml ovine inhibin antiserum. On day 10, luteal regression was induced by an i.m. injection of cloprostenol (100 μg), and ovulation rate determined 6 days later by laparoscopy. Peripheral plasma samples were collected throughout the experimental period. Following treatment, there was no change in the peripheral plasma concentration of LH in either group. Following injection of the inhibin antiserum, the concentration of FSH rose significantly (P<0·001) compared with the control group. The concentration of FSH rose from 1·42 ± 0·06 to a maximum of 2·58 ± 0·23 (s.e.m.) μg/l by 5·6 ±0·9 h, this maximum lasting 9·0±1·1 h. By 32·8 ±6·9 h, the concentration of FSH had returned to pretreatment levels, while the titre of free antibody in the plasma of treated ewes was still high. In the treated ewes, there were one single and four double ovulations compared with three single and two double ovulations in the control group, indicating that the inhibin immunization may have resulted in an increase in ovulation rate. We conclude that the marked rise in the plasma concentration of FSH following injection of inhibin antiserum provides strong evidence that inhibin is an important factor in the regulation of FSH production by the pituitary gland at this time. Journal of Endocrinology (1989) 123, 383–391

scholarly journals Evidence for a PGF2α auto-amplification system in the endometrium in mares

Reproduction ◽  
2016 ◽  
Vol 151 (5) ◽  
pp. 517-526 ◽  
Author(s):  
Keisuke Kozai ◽  
Shota Tokuyama ◽  
Anna Z Szóstek ◽  
Yuko Toishi ◽  
Nobuo Tsunoda ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Stromal Cells ◽  
Luteal Phase ◽  
Endometrial Cell ◽  
Late Luteal Phase ◽  
Prostaglandin F ◽  
Amplification System ◽  
Prostaglandin Endoperoxide Synthase

AbstractIn mares, prostaglandin F2α(PGF2α) secreted from the endometrium is a major luteolysin. Some domestic animals have an auto-amplification system in which PGF2αcan stimulate its own production. Here, we investigated whether this is also the case in mares. In anin vivostudy, mares at the mid-luteal phase (days 6–8 of estrous cycle) were injected i.m. with cloprostenol (250 µg) and blood samples were collected at fixed intervals until 72 h after treatment. Progesterone (P4) concentrations started decreasing 45 min after the injection and continued to decrease up to 24 h (P < 0.05). In turn, 13,14-dihydro-15-keto-PGF2α(PGFM) metabolite started to increase 4h after an injection and continued to increase up to 72 h (P < 0.05). PGF receptor (PTGFR) mRNA expression in the endometrium was significantly higher in the late luteal phase than in the early and regressed luteal phases (P < 0.05).In vitro, PGF2αsignificantly stimulated (P < 0.05) PGF2αproduction by endometrial tissues and endometrial epithelial and stromal cells and significantly increased (P < 0.05) the mRNA expression of prostaglandin-endoperoxide synthase-2 (PTGS2), an enzyme involved in PGF2αsynthesis in endometrial cell. These findings strongly suggest the existence of an endometrial PGF2αauto-amplification system in mares.

In vivo effects of flavonoid EMD 21388 on thyroid hormone secretion and metabolism in rats

1991 ◽  
Vol 261 (2) ◽  
pp. E227-E232 ◽  
Author(s):  
J. P. Schroder-van der Elst ◽  
D. van der Heide ◽  
J. Kohrle
Keyword(s):  
Thyroid Hormone ◽  
Hormone Secretion ◽  
Male Rats ◽  
Intact Male ◽  
Hormone Production ◽  
Iodide Uptake ◽  
Brown Adipose ◽  
In Vivo Effects

In vitro, the synthetic flavonoid EMD 21388 appears to be a potent inhibitor of thyroxine (T4) 5'-deiodinase and diminishes binding of T4 to transthyretin. In this study, in vivo effects of long-term administration of EMD 21388 on thyroid hormone production and metabolism were investigated. Intact male rats received EMD 21388 (20 mumol.kg body wt-1.rat-1.day-1) for 14 days. [125I]T4 and 3,5,3'-[131I]triiodotyronine (T3) were infused continuously and intravenously in a double-isotope protocol for the last 10 and 7 days, respectively. EMD 21388 decreased plasma thyroid hormone concentrations, but thyrotropin levels in plasma and pituitary did not change. Plasma clearance rates for T4 and T3 increased. Thyroidal T4 secretion was diminished, but T3 secretion was elevated. Extrathyroidal T3 production by 5'-deiodination was lower. T4 concentrations were markedly lower in all tissues investigated. Total tissue T3 was lower in brown adipose tissue, brain, cerebellum, and pituitary, tissues that express the type II 5'-deiodinase isozyme due to decreased local T3 production. Most tissues showed increased tissue/plasma ratios for T4 and T3. These results indicate that this flavonoid diminished T4 and increased T3 secretion by the thyroid, probably in analogy with other natural flavonoids, by interference with one or several steps between iodide uptake, organification, and hormone synthesis.

In vitro-produced horse embryos exhibit a very narrow window of acceptable recipient mare uterine synchrony compared with in vivo-derived embryos

2019 ◽  
Vol 31 (12) ◽  
pp. 1904 ◽  
Author(s):  
Juan Cuervo-Arango ◽  
Anthony N. Claes ◽  
Tom A. E. Stout
Keyword(s):  
Pregnancy Rate ◽  
Pregnancy Loss ◽  
Oestrous Cycle ◽  
Ongoing Pregnancy ◽  
Ongoing Pregnancy Rate ◽  
Wide Range ◽  
In Pregnancy

In recent years, the number of equine invitro-produced embryos (IVP) has increased markedly; as yet, there are few reports on what constitutes an ‘ideal’ recipient for an IVP embryo. This study retrospectively investigated the effects of recipient mare oestrous cycle characteristics on the likelihood of pregnancy after transfer of IVP (n=264) and invivo-derived embryos (n=262). IVP embryos tolerated only a narrow window of recipient mare ‘synchrony’, with transfer on Day 4 after ovulation resulting in a higher likelihood of ongoing pregnancy (69%) than transfer on Days 3, 5 or 6 (53.2%, 41.3% and 23.1% respectively; P=0.02). In contrast, Day 8 invivo-derived embryos tolerated a wide range of uterine (a)synchrony, with no difference in pregnancy or pregnancy loss for recipients that ovulated between Day 4 and Day 9 before transfer. However, transferring invivo-derived embryos to recipients that had a longer oestrus preceding transfer resulted in higher Day 12 and ongoing pregnancy rate (P&lt;0.01). This effect was not significant in IVP embryos. In conclusion, Day 6–8 IVP blastocysts survive best after transfer to Day 4 recipient mares; Day 8 invivo-derived embryos survive equally well in Day 4–9 recipients, but do better in mares that have a long preceding oestrus.

RNA AND PROTEIN METABOLISM IN THE OVIDUCT AND ENDOMETRIUM OF THE EWE AT PRO-OESTRUS: REGULATION BY OESTRADIOL AND PROGESTERONE

1976 ◽  
Vol 69 (1) ◽  
pp. 57-66 ◽  
Author(s):  
B. G. MILLER
Keyword(s):  
Protein Metabolism ◽  
Hormonal Regulation ◽  
Luteal Phase ◽  
Rna Synthesis ◽  
Model System ◽  
Oestrous Cycle ◽  
Cell Content ◽  
Hormone Effects ◽  
As Effects

SUMMARY The effects on RNA and protein metabolism in the oviduct and endometrium at pro-oestrus of oestradiol and progesterone secreted during the oestrous cycle were examined, using the ovariectomized, hormone-treated ewe as a model system. Thirty ewes received hormone injections during a period of 13 days, according to schedules designed to simulate endogenous ovarian secretion of oestradiol and progesterone during the oestrous cycle. Hormone effects on RNA:DNA ratios and on rates of synthesis of protein and methylated RNA in vitro, as well as effects on oviducal and uterine weight, were examined. The results obtained suggest that endogenous ovarian hormones have the following effects in the intact ewe. The secretion of oestradiol at pro-oestrus rapidly increases rates of synthesis of protein and methylated RNA, and mean cell content of RNA in both the endometrium and oviduct. Oestradiol secreted during the previous luteal phase of the oestrous cycle markedly increases mean cell content of RNA and amounts of protein and methylated RNA synthesis occurring in both tissues at pro-oestrus. In the endometrium, progesterone secreted during the luteal phase increases the RNA: DNA ratio, and probably also the amounts of protein and methylated RNA synthesized at pro-cestrus, but there are no significant interactions between the effects of oestradiol and progesterone. Progesterone had no effect on either the amounts or rates of synthesis of protein or methylated RNA in the oviduct. The results are discussed in relation to the hormonal regulation of physiological functions of the oviduct and endometrium during the first few days after the onset of oestrus.

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