Effects of immunization against recombinant bovine inhibin α subunit on circulating concentrations of gonadotrophins in ewes

1989 ◽  
Vol 120 (1) ◽  
pp. 59-65 ◽  
Author(s):  
J. K. Findlay ◽  
B. Doughton ◽  
D. M. Robertson ◽  
R. G. Forage

ABSTRACT Immunization of ewes against a pure recombinant preparation of the α subunit of bovine inhibin (α-bI) resulted in a three- to fourfold increase in ovulation rate, associated with antibodies in plasma recognizing pure native 31 kDa inhibin. The aim of this study was to examine the effects of this immunization on basal and GnRH-stimulated plasma concentrations of FSH and LH in ewes during the anoestrous and breeding seasons. The groups were untreated control ewes (n = 5), control ewes treated with keyhole limpet haemocyanin (KLH alone, n = 4), ewes treated with α-bI alone (n = 4) and α-bI–KLH conjugate-treated ewes (n = 3). There were no effects of immunization on basal FSH or LH in anoestrous ewes, despite the presence of antibodies recognizing 31 kDa inhibin. In the breeding season, immunization against α-bI resulted in increased basal (follicular phase, P < 0·1; luteal phase P < 0·05) and GnRH-stimulated (follicular phase only, P < 0·001) release of FSH, but not LH. The data are compatible with the hypotheses that the increase in ovulation rate in immunized ewes is due to an increase in circulating FSH concentrations and that inhibin may only have a major peripheral influence on FSH in sheep during the breeding season. Journal of Endocrinology (1989) 120, 59–65

1989 ◽  
Vol 122 (2) ◽  
pp. 509-517 ◽  
Author(s):  
R. J. E. Horton ◽  
H. Francis ◽  
I. J. Clarke

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


1990 ◽  
Vol 124 (1) ◽  
pp. 167-176 ◽  
Author(s):  
J. H. M. Wrathall ◽  
B. J. McLeod ◽  
R. G. Glencross ◽  
A. J. Beard ◽  
P. G. Knight

ABSTRACT Two experiments were conducted to explore the effectiveness of synthetic peptide-based vaccines for active and passive autoimmunization of sheep against inhibin. In the first experiment, adult Romney ewes (n = 20) were actively immunized against a synthetically produced peptide that corresponded to the N-terminus of the α-subunit of bovine inhibin (bIα(1–29)-Tyr30). This peptide was conjugated to tuberculin purified protein derivative (PPD) to increase its antigenic properties. Control groups comprised non-immunized (n = 10) and PPD-immunized (n = 10) ewes. Primary immunization (400 μg conjugate/ewe) was followed by two booster immunizations (200 μg conjugate/ewe), given 5 and 8 weeks later. Following synchronization of oestrus using progestagen sponges, ovulation rates were assessed by laparoscopy. Weekly blood samples were taken throughout the experiment. All inhibin-immunized ewes produced antibodies which bound 125I-labelled bovine inhibin (Mr 32 000), and ovulation rate in inhibin-immunized ewes (2·15 ± 0·22; mean ± s.e.m.) was significantly (P<0·01) greater than in both non-immunized (0·90 ± 0·23) and PPD-immunized (1·20 ± 0·13) control groups. Immunization against the peptide, but not against PPD alone, resulted in a modest rise in plasma FSH, with mean levels after the second boost being significantly (P<0·025) higher (22%) than those before immunization. Moreover, when blood samples were taken (2-h intervals) from randomly selected groups of control (n = 7) and inhibin-immunized (n = 7) ewes for an 84-h period following withdrawal of progestagen sponges, the mean plasma concentration of FSH during the 48 h immediately before the preovulatory LH surge was 37% greater (P< 0·025) in immunized than in control animals. However, more frequent blood sampling (every 15 min for 12 h) during follicular and mid-luteal phases of the oestrous cycle revealed no significant differences between treatment groups in mean plasma concentrations of FSH. In addition, neither mean concentrations of LH nor the frequency and amplitude of LH episodes differed between immunized and control ewes. However, the mean response of LH to a 2 μg bolus of gonadotrophin-releasing hormone, given during the luteal phase, was significantly (P<0·05) less in immunized than in control ewes. These findings indicate that active immunization of Romney ewes against a synthetic fragment of inhibin can promote a controlled increase in ovulation rate, but this response cannot be unequivocally related to an increase in plasma levels of FSH. In the second experiment, passive immunization of seasonally anoestrous ewes (mule × Suffolk crossbred; n = 6 per group) against inhibin, using an antiserum raised in sheep against a synthetic peptide corresponding to the N-terminus of the α-subunit of human inhibin promoted a rapid (<3 h), dose-dependent rise in plasma levels of FSH which remained increased (2·5-fold; P<0·001) for up to 30 h. Plasma concentrations of LH, however, were unaffected by treatment with the antiserum. It is deduced from this observation that, even in the seasonally anoestrous ewe, the ovary secretes physiologically active levels of inhibin, which exert an inhibitory action on the synthesis and secretion of FSH. Journal of Endocrinology (1990) 124, 167–176


1984 ◽  
Vol 101 (1) ◽  
pp. 75-80 ◽  
Author(s):  
P. V. Malven ◽  
D. F. B. Bossut ◽  
M. A. Diekman

ABSTRACT Mature ewes were injected intravenously with the opioid antagonist naloxone (1·1 mg/kg) during the breeding season. Ewes with luteal phase concentrations of plasma progesterone responded with a significant (P <0·05) increase in plasma LH 14–23 min after naloxone injection. In contrast, non-luteal ewes with low plasma progesterone did not respond to injection of naloxone with an LH increase. Similar treatment of castrated males (wethers) with this dosage of naloxone failed to increase plasma LH. Electroacupuncture (EA) treatment of luteal phase ewes prevented the ability of exogenous naloxone to increase plasma LH. Treatment of wethers by EA decreased significantly (P <0·01) their high basal concentrations of plasma LH, but similar EA treatment of intact ewes did not change their low basal concentrations of LH. J. Endocr. (1984) 101, 75–80


1990 ◽  
Vol 127 (2) ◽  
pp. 285-296 ◽  
Author(s):  
B. K. Campbell ◽  
D. T. Baird ◽  
A. S. McNeilly ◽  
R. J. Scaramuzzi

ABSTRACT Active immunization of sheep against androstenedione results in an increase in ovulation rate that is associated with increased plasma levels of LH and progesterone, but not FSH. Although immunized ewes have more activated follicles the secretion rate of oestradiol is not increased. An experiment was conducted to examine the effect of androstenedione immunity on the ovarian secretion and peripheral plasma concentrations of inhibin. Merino ewes in which the left ovary had been autotransplanted to a site in the neck were divided into control (n = 5) and androstenedione-immune (n = 6) groups. Ovarian and jugular venous blood was collected every 10 min at two stages of the follicular phase, 21–27 h and 38–42 h after a luteolytic dose of an analogue of prostaglandin F2α (PG), and every 15 min for 6 h on day 10 of the subsequent luteal phase. The ewes were monitored regularly for luteal function by measurement of the concentration of progesterone and preovulatory LH surges. The concentration of inhibin in jugular and ovarian venous plasma was determined by radioimmunoassay and ovarian secretion rates and peripheral concentrations are expressed as pg of 1–26 peptide fragment of the α chain. The ovarian secretion rate of inhibin tended to be greater in androstenedione-immune ewes at all stages of the oestrous cycle measured, with this difference being statistically significant (P <0·05) during the luteal phase (100±40 and 260±80 (s.e.m.) pg/min for control and immune groups respectively). The pattern of ovarian inhibin secretion exhibited pulsatile-like fluctuations which were not associated with LH pulses. Peripheral concentrations of inhibin were generally higher in immunized than in control ewes with this difference being significant (P < 0·01) from day 4 to 14 of the luteal phase (59±5 and 110±7 ng/1 for control and immune respectively). The ovarian secretion rate of immunoactive inhibin was greater (P <0·01) during the follicular phase than during the luteal phase in both groups of ewes, and peripheral concentrations of inhibin increased (P < 0·001) following injection of PG in ewes from both treatment groups. We concluded that androstenedione immunity results in an increase in ovarian inhibin secretion, an effect that can probably be attributed to the greater number of large oestrogenic follicles present in the ovaries of these ewes. Furthermore, this increase in the concentration of inhibin may override any decrease in the negative feedback effects of ovarian steroid produced by immunization and, hence, explain the paradoxical findings of normal concentrations of FSH and raised concentrations of LH in ewes which are immunized against androstenedione. Journal of Endocrinology (1990) 127, 285–296


2021 ◽  
Vol 36 (4) ◽  
pp. 662-662
Author(s):  
Ott S ◽  
Gonzalez L ◽  
Redell J ◽  
Duran J ◽  
Schatz P ◽  
...  

Abstract Objective Few studies have addressed serum levels of hypothalamic hormones, as a consequence of sports-related concussion, in concussed, female adolescent athletes. Methods Female athletes, ages 14–18, were assigned to two study groups: 1) 18 diagnosed with a sports-related concussion (SRC) and evaluated within 7 days of injury, and 2) 18 healthy participants matched on age, education, and sport. Participants self-reported menstrual cycle status, with Days 1–15 of the menstrual cycle corresponding to the Follicular phase and days &gt;15 to the Luteal phase. Blood samples were drawn for Plasma concentration measurement of follicle-stimulating hormone (FSH), luteinizing hormone (LH), thyroid-stimulating hormone (TSH), adrenocorticotropin hormone (ACTH), prolactin, growth hormone (GH) and progesterone. Using progesterone plasma concentration, &lt;=2 ng/mL corresponding to the Follicular phase and &gt; 2 ng/mL corresponding to the Luteal phase. Results Across both groups, there was agreement between self-reported and progesterone plasma concentrations menstrual cycle Follicular (71.4%) or Luteal phase (52.9%) [X2(1) = 2.33, p = 0.13, phi = 0.25] in identifying menstrual cycle phases. Within the non-concussed controls, there was 90% agreement for Follicular phase and only 33.3% agreement for Luteal phase [X2(1) = 1.55, p = 0.30, phi = 0.29], and for the concussed athletes there was 54.5% agreement for Follicular phase and 75% agreement for Luteal phase [X2(1) = 1.66, p = 0.35, phi = 0.30] Conclusions We provide preliminary evidence of disruption of the hypothalamic–pituitary-axis (HPA) following mTBI as there was a 90% agreement between self-reported follicular phase and follicular phase determined by Progesterone levels for non-concussed females as compared to 54% agreement between the same measures for concussed females.


Reproduction ◽  
2010 ◽  
Vol 139 (1) ◽  
pp. 153-161 ◽  
Author(s):  
Michiko Noguchi ◽  
Koji Yoshioka ◽  
Seigo Itoh ◽  
Chie Suzuki ◽  
Sachiko Arai ◽  
...  

We investigated changes in peripheral concentrations of inhibin A, total inhibin, steroids, and gonadotropins throughout the intact estrous cycle of the sow in relation to ovarian changes determined by daily transrectal ultrasonography. All visible follicles of 3 mm or more in diameter were classified as small (≥3 and <6 mm) or large (≥6 mm). Follicular recruitment was identified in two periods of the cycle: one from the late luteal to the follicular phase, characterized by an increase in the number of small follicles followed by the appearance of large follicles; and another during the early luteal phase, consisting only of increased numbers of small follicles. Plasma concentrations of inhibin A increased (P<0.05), coinciding with the two periods of follicle emergence. Estradiol (E2) levels increased (P<0.05) during the follicular phase, but not during the early luteal phase. An inverse relationship (P<0.01) between the patterns of inhibin and FSH concentrations was noted around the two periods of follicle emergence, but there was no relationship (P≥0.1) between the patterns of plasma E2and FSH during the early luteal phase. In conclusion, measurement of plasma inhibin A levels combined with ultrasonographic examination of the ovaries revealed two periods of synchronous follicular growth during the sow's estrous cycle. The results strongly suggest that inhibin A functions as a negative feedback regulator of FSH secretion throughout the estrous cycle, whereas E2appears to influence FSH secretion only during the follicular phase.


1996 ◽  
Vol 62 (3) ◽  
pp. 507-512 ◽  
Author(s):  
T. Sweeney ◽  
D. O'Callaghan

AbstractWhile the onset of the breeding season in ewes can be advanced up to 7 weeks following exposure to long days (44 days) before lambing, a melatonin implant from lambing and exposure to a ram from the summer solstice, this advance is reduced by the absence of a ram. The aim of this experiment was to determine if earlier exposure to the ram and/or exposure to more long days before lambing would further advance the onset of the breeding season. A further aim was to compare the ovulation rate and duration of the breeding season between ewes with a normal and advanced onset of the breeding season due to this treatment regime. Pregnant ewes lambing in mid March were allocated to treatments as follows: (1) controls, maintained on natural photoperiod; (2) 44 LD + mel, exposed to 44 long days (18L: 6D; LD) before lambing followed by a 700-mg melatonin implant after lambing; (3) 90 LD + mel, as in treatment 2, but exposed to 90 LD before lambing; (4) 90 LD + mel + ram, as in treatment 3, including continuous exposure to a vasectomized ram from 10 April. Reproductive state was assessed from serum progesterone concentrations in blood collected at 3- to 4-day intervals from 10 April and ovulation rate was determined during the luteal phase of the first five oestrous cycles in control ewes and ewes exposed to 90 LD + mel + ram. Ewes exposed to 90 LD + mel + ram commenced the breeding season 10 weeks earlier (median date 25 July, range 28 days;P< 0·001) than controls (median date 8 October, range 68 days). Overall, the onset of the breeding season was not advanced in ewes given 44 or 90 LD + mel in the absence of a ram, but a biphasic response was evident, with 40 to 50% of ewes beginning the breeding season at the same time as ewes on 90 LD + mel + ram. Ovulation rate was similar between control ewes and ewes exposed to 90 LD + mel + ram (P > 0·05). The duration of the breeding season was longer in ewes exposed to 90 LD + mel + ram (210 (s.e. 15) days;P< 0·002) than in control ewes (158 (s.e. 10) days), with no difference in date of the end of the breeding season (P > 0·05). Exposure of ewes to 90 LD before lambing in March, followed by a melatonin implant and exposure to the ram from 10 April advanced the onset and extended the duration of the breeding season without an overall effect on ovulation rate. The ram is an important component of this signal sequence.


2010 ◽  
Vol 298 (2) ◽  
pp. E222-E228 ◽  
Author(s):  
Liaqat H. Khan ◽  
Charles R. Rosenfeld ◽  
Xiao-tie Liu ◽  
Ronald R. Magness

The follicular phase of the ovine ovarian cycle demonstrates parallel increases in ovarian estrogens and uterine blood flow (UBF). Although estrogen and nitric oxide contribute to the rise in UBF, the signaling pathway remains unclear. We examined the relationship between the rise in UBF during the ovarian cycle of nonpregnant sheep and changes in the uterine vascular cGMP-dependent pathway and large-conductance Ca2+-activated K+ channels (BKCa). Nonpregnant ewes ( n = 19) were synchronized to either follicular or luteal phase using a vaginal progesterone-releasing device (CIDR), followed by intramuscular PGF2α, CIDR removal, and treatment with pregnant mare serum gonadotropin. UBF was measured with flow probes before tissue collection, and second-generation uterine artery segments were collected from nine follicular and seven luteal phase ewes. The pore-forming α- and regulatory β-subunits that constitute the BKCa, soluble guanylyl cyclase (sGC), and cGMP-dependent protein kinase G (cPKG) isoforms (cPKG1α and cPKG1β) were measured by Western analysis and cGMP levels by RIA. BKCa subunits were localized by immunohistochemistry. UBF rose >3-fold ( P < 0.04) in follicular phase ewes, paralleling a 2.3-fold rise in smooth muscle cGMP and 32% increase in cPKG1α ( P < 0.05). sGC, cPKG1β, and the BKCa α-subunit were unchanged. Notably, expression of β1- and β2-regulatory subunits rose 51 and 79% ( P ≤ 0.05), respectively. Increases in endogenous ovarian estrogens in follicular-phase ewes result in increases in UBF associated with upregulation of the cGMP- and cPKG-dependent pathway and increased vascular BKCa β/α-subunit stoichiometry, suggesting enhanced BKCa activation contributes to the follicular phase rise in UBF.


1977 ◽  
Vol 24 (3) ◽  
pp. 363-376 ◽  
Author(s):  
A. G. Wheeler ◽  
R. B. Land

SUMMARYThe patterns of seasonal variation in reproductive activity were observed over a period of 15 mo for approximately 15 females of each of three breeds: Finnish Landrace (Finn), Tasmanian Merino (Merino) and Scottish Blackface (Blackface). The incidence of oestrus was measured by teasing with vasectomized rams, and the incidence and rate of ovulation were determined frequently by laparoscopy. Luteal function was assessed from peripheral venous blood progesterone concentration on days 7 and 11 of the oestrous cycle. Nutritional status was monitored by recording body weight and plasma-free fatty acid levels throughout the study.The breeding seasons differed significantly: Finn, October to May; Merino, September to February; and Blackface, October to February. Variation in the incidence of ovulation was similar to that in the incidence of oestrus for each breed. The incidence of silent ovulation varied with the breed, being greatest in the Merino and least in the Finn. The ovulation rate varied among breeds (Finn, 2·99; Merino, 1·08 and Blackface, 1·30), and during the breeding season (e.g. Finn: November, 3·5; March, 2·6). Follicles were observed in each breed throughout the period of study.The pattern of variation in progesterone concentration was similar for each breed despite their different breeding seasons. In addition to breed differences in ovulation rate and in onset and end of the breeding season, the sensitivity to oestrogen was apparently such that, with the Finn if oestrogen secretion was high enough to stimulate ovulation it would usually also stimulate oestrus, whereas with the Merino ovulation often occurred without oestrus; this suggests that in the Merino the centres controlling ovulation are more sensitive to oestrogen than those controlling behaviour.


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