Biomedical subjects

R J Scaramuzzi

Publications and source records attributed to R J Scaramuzzi.

At least 37 records · Page 2Linked to original sources

PubMed1999

Effect of progesterone on the GnRH-induced secretion of oestradiol and androstenedione from the autotransplanted ovary of the anoestrous ewe.

Two experiments were conducted during the anoestrous period in Border Leicester x Merino ewes with ovarian autotransplants to study the effects of a single injection of 20 mg progesterone on follicular steroid secretion. The aim of these experiments was to determine whether pretreatment with a 20 mg intramuscular injection of progesterone could reduce GnRH-induced ovarian steroid secretion in anoestrous ewes. In both experiments, an injection of 150 ng GnRH induced an LH pulse in all ewes with a maximum concentration 10 min (the first post-injection sample) after injection. Oestradiol and androstenedione secretion increased progressively after the GnRH-induced LH pulse and reached maximum rates of secretion between 60 and 90 min before decreasing slowly to pre-injection rates at 150 min. There were no differences in the pattern of secretion of oestradiol (measured in both experiments) or androstenedione (measured only in Expt 2). In Expt 1, the injection of progesterone 72 h before the challenge with GnRH had no effect on the maximum rate of oestradiol secretion from the autotransplanted ovary. However, in Expt 2, when progesterone was given either 36 or 60 h before GnRH, there was a significant suppression in the maximum rate of secretion of both oestradiol and androstenedione between 60 and 90 min after GnRH injection. These data show that pretreatment of anoestrous sheep with progesterone can suppress LH-stimulated steroid secretion from the ovary and indicate that progesterone may have a direct effect on oestrogenic follicles in sheep.

Analysis of Variance↗

PubMed1999

Examination of the relative role of FSH and LH in the mechanism of ovulatory follicle selection in sheep.

The GnRH-antagonist suppression-ovarian autotransplant model (n = 18) was used to examine the relative roles of temporal changes in FSH and LH stimulation on follicle development and selection. Follicle development was stimulated by infusion with oFSH for 3 days and treatments applied for 60 h after progestagen sponge withdrawal and before delivery of an ovulatory stimulus. In Expt 1, there was continuous infusion of FSH with or without small amplitude high frequency LH pulses, or withdrawal of FSH with or without pulsatile LH. In Expt 2, there was acute or gradual withdrawal of FSH at sponge withdrawal with pulsatile LH. The patterns of follicle development and basal and pulsatile ovarian hormone secretion were determined. The maintenance of FSH throughout the artificial follicular phase resulted in multiple follicle development and ovulation (3.3 +/- 0.3). Pulsatile LH stimulated steroid secretion (P < 0.001) but had little effect on ovulation rates (3.8 +/- 0.8) when FSH was maintained. However, withdrawal of FSH in the absence of LH resulted in atresia of the ovulatory follicles and anovulation whereas, when FSH was withdrawn in the presence of LH, preovulatory follicle development was maintained in some animals (3/6 and 5/9 in Expts 1 and 2, respectively) and these ewes had lower (P < 0.05) ovulation rates (1-2 ovulations per ewe). When FSH was withdrawn gradually in the presence of pulsatile LH, 9/9 animals ovulated with ovulation rates in the normal range. These results indicate that ovulatory follicles can transfer their gonadotrophic dependence from FSH to LH. It is hypothesized that the ability of a follicle to respond to this switch in gonadotrophic support is central to the mechanism of follicle selection.

Analysis of Variance↗

PubMed1999

The effect of a direct arterial infusion of insulin and glucose on the ovarian secretion rates of androstenedione and oestradiol in ewes with an autotransplanted ovary.

Improving ewe nutrition even for short periods will increase ovulation rate. The increased nutrients must in some way affect the number of follicles that develop to the pre-ovulatory stage. One possible mechanism is that a nutrient or a metabolic hormone that responds to nutrition might act directly on the ovary to influence follicle development and/or follicle selection. In the study described here, insulin and glucose, alone or together, were infused directly into the ovarian artery of ewes with an autotransplanted ovary, for 13.5 h on day 11 of the oestrous cycle. The pattern of androstenedione and oestradiol secretion in response to a GnRH-stimulated LH pulse was measured 2.5 h before and 12.5 h and 24.5 h after the start of the infusion. Glucose or insulin infused alone had no effect on the secretion of androstenedione and oestradiol. However, when infused together, they decreased significantly the secretion of androstenedione and, to a lesser extent, oestradiol. We suggest that the sudden availability of additional glucose and insulin increases insulin-stimulated glucose uptake by the follicle. This leads to an inhibition of LH-stimulated steroidogenesis by the ovarian follicle which occurs in the absence of any detectable changes in circulating plasma concentrations of FSH. These results show that insulin and glucose act together to influence ovarian function directly and suggest that the effects of short-term nutrition on ovulation rate may be mediated by a direct ovarian action of insulin and glucose.

Analysis of Variance↗

PubMed1998

Ovarian function in ewes made hypogonadal with GnRH antagonist and stimulated with FSH in the presence or absence of low amplitude LH pulses.

This study examined the effect of LH pulses, of similar amplitude and frequency to those found in the luteal phase, on the pattern of hormone secretion and follicle development in GnRH antagonist-suppressed ewes stimulated with exogenous FSH. This experiment was conducted on ewes with ovarian autotransplants in a continuous study. Follicle development was suppressed in 18 ewes by 3 weeks of GnRH antagonist treatment (50 micrograms/kg per 4 days s.c.), and was then stimulated by infusion of ovine (o)FSH (5 micrograms NIADDK-oFSH-16/h i.v.) for 3 days. In addition to FSH, 10 animals received pulses of LH (2.5 micrograms NIADDK-oLH-26 i.v.) every 4 h for the entire period of the FSH infusion. The follicle population was determined by daily ultrasound. Samples of ovarian and jugular venous blood were collected at 4-h intervals over the period of the FSH infusion and there were three periods of intensive blood sampling (15-min intervals for 2.5 h at 24, 48 and 72 h after the start of the FSH infusion) when the steroidogenic capacity of the follicles in all 18 ewes was tested around an LH challenge (2.5 micrograms i.v.). GnRH antagonist treatment resulted in a 57% decrease in FSH concentrations and prevented ovarian follicle development beyond 3 mm in diameter. Infusion of FSH resulted in a 60% increase in FSH concentrations and stimulated the development of large antral follicles and a coincident increase in ovarian androstenedione, inhibin and oestradiol secretion in both experimental groups. In the absence of 4-hourly LH pulses basal steroid secretion was negligible (< 1 ng/min; P < 0.001). Daily LH challenges, however, revealed no difference in the steroidogenic capacity of the follicle population in either experimental group. Similarly, LH pulses had no effect on the growth rate and number of antral follicles stimulated by FSH infusion, or the pattern of ovarian inhibin secretion. In conclusion, these results show that while FSH alone can stimulate the development of ovulatory sized follicles in ewes made hypogonadal with GnRH antagonist, physiological patterns of LH stimulation have no deleterious effects on FSH-stimulated follicle development and are essential for normal steroidogenesis.

Androstenedione↗

PubMed1997

The effect of the infusion of insulin during the luteal phase of the estrous cycle on the ovulation rate and on plasma concentrations of LH, FSH and glucose in ewes.

The role of insulin in mediating pituitary responses to nutrition was investigated in 30 mature Border Leicester X Merino ewes. The ewes were infused with saline (n = 15) or bovine insulin at 0.4 IU/kg/d (n = 15) for 72 h during the luteal phase of the estrous cycle The ewes were housed in individual pens and were fed, ad libitum, a diet of low quality straw. Their estrous cycles were synchronized with prostaglandin (PG), with infusions given over Days 9 to 11 of the estrous cycle. A further injection of PG was given at the end of the infusion, and the subsequent ovulation rate was determined by endoscopy 12 d later. Blood samples were collected every 4 h from Day 8 until 52 h after the final PG injection for the determination of plasma FSH, insulin and glucose concentrations. On Day 11 blood samples were also taken every 20 min for 24 h for the determination of LH pulse characteristics. During the infusion of insulin, its concentration rose 4-fold and remained elevated until the end of infusion, when it fell to pretreatment concentrations. Glucose concentrations were significantly reduced during the insulin infusion and rose to pretreatment concentrations after infusion. In control ewes glucose and insulin concentrations did not change. Ovulation rate of treated ewes was not affected by the insulin (1.9 +/- 0.07) compared with that of control ewes (2.0 +/- 0.10). Neither were FSH concentrations affected by treatment with insulin, although a significant interaction of treatment with time was observed in the 36 h after infusion. The pre-ovulatory decline in FSH concentrations was delayed by about 8 h in the insulin treated ewes. The mean (+/- SEM) LH pulse frequency (4.3 +/- 0.4 vs 1.8 +/- 0.3 pulses per 24 h) and the mean (+/- SEM) concentration of LH (0.48 +/- 0.04 vs 0.32 +/- 0.03 ng/ml) were both significantly reduced by insulin. These results indicate that insulin-induced hypoglycaemia inhibits LH secretion in cyclic ewes and implicates insulin as a mediator of normal hypothalamo-pituitary function.

Journal Article↗

PubMed1997

Ovulation rate and the concentrations of LH, FSH, GH, prolactin and insulin in ewes infused with tryptophan, tyrosine or tyrosine plus phenylalanine during the luteal phase of the oestrous cycle.

Dietary amino acid precursors for cathecholamineric and serotonergic neurotransmitters may be important in the mechanism of nutritional effects on ovulation rate. This paper reports the results of three experiments that examined the effect of such amino acids on ovulation rate and the concentrations of FSH and LH in sheep. In three separate experiments, groups of ewes were infused, over Days 9 to 13 of the oestrous cycle, with either tryptophan (n = 11), tyrosine (n = 11) or a mixture of tyrosine and phenylalanine (n = 11). Control ewes (n = 12 in each experiment) were infused with a vehicle over the same period. None of the amino acids infused effected ovulation rate or plasma concentrations of LH, FSH, GH or prolactin. The infusion of a mixture of tyrosine and phenylalanine increased insulin concentrations. The infusion of these amino acids was not associated with changes in gonadotrophin concentrations and therefore the effect of nutrition on ovulation rate in ewes does not seem to involve an increase in the availability of tryptophan, tyrosine or phenylalanine. Increasing the uptake of other amino acids that compete with tryptophan, tyrosine or phenylalanine for the large neutral amino acid transporter may cause a decrease in the availability of tryptophan, tyrosine or phenylalanine thereby eliciting the effects of nutrition on ovulation rate. However, this hypothesis remains to be tested.

Animals↗

PubMed1997

The circulating concentrations of FSH, LH and prolactin in the oestradiol-implanted ovariectomized ewe treated with caffeine.

Caffeine, a trimethylxanthine alkaloid, is a psycho-active drug that effects a wide range of physiological systems, including the reproductive system. Reports of infants with intra-uterine growth retardation and lowered birth weight as a result of in utero exposure to caffeine, are increasing. The drug is also known to alter steroidogenesis but it is not certain whether this is a direct and/or an indirect effect with the involvement of the central nervous system. Thus, an experiment was designed to determine the effect of acute caffeine administration on the circulating concentrations of gonadotrophins and prolactin in the ovariectomized oestradiol-implanted ewe. A single intravenous dose of caffeine (20 mg kg-1 bodyweight) did not affect circulating gonadotrophin concentrations with the parameters for the pulsatile secretion of luteinizing hormone (LH) and the mean concentration of follicle stimulating hormone (FSH) being similar in both experimental and control groups. Circulating prolactin levels, on the other hand, were significantly (P < 0.01) elevated following intravenous treatment with caffeine. The effect was immediate following caffeine administration with elevated concentrations being maintained over the next 3 h before their return to pre-treatment concentrations. The response was bi-phasic with peaks of prolactin concentrations at 1 and 3 h. The results of this experiment show that acute caffeine exposure does not affect the secretion of gonadotrophins from the anterior pituitary gland. Furthermore, they show that acute administration of caffeine stimulates prolactin secretion via an action that is independent of oestradiol feedback and which we suggest, may involve the ACTH/adrenal axis.

Animals↗

PubMed1997

Use of a GnRH antagonist in conjunction with low amplitude, high frequency LH pulses to induce follicular growth without an LH surge and ovulation in ewes.

The present study was undertaken to develop an experimental sheep model which could be used to investigate the abnormal follicle growth that is associated with the absence of the LH surge. On Day 10 of the oestrous cycle, 16 ewes were treated with an analogue of prostaglandin (cloprostenol; PG) and blood sampled every 4 h thereafter to determine the normal timing of the preovulatory LH surge. Three oestrous cycles later, all ewes were simultaneously treated with PG and a gonadotrophin-releasing hormone (GnRH) antagonist ([Ac-DNal1, D4C1Phe2, DTrp3, DArg6, DAla10] GnRH.HOAc; 50 micrograms kg-1 subcutaneously). Group 1 ewes (n = 6) received no further treatment. Group 2 ewes (n = 5) were additionally treated for a total of 7 days, starting at the time of PG injection, with purified ovine luteinising hormone (LH; preparation P 3 R3-5; equivalent to 1.25 micrograms NIH-oLH-S26) administered i.v. over a 2 min period. For the first 24 h, LH was given at 3 h intervals for 12 h, then every 2 h for 12 h, and thereafter hourly for 6 days. Group 3 ewes (n = 5) were treated as Group 2 but at 72 h received an additional antagonist injection (50 micrograms kg-1 subcutaneously). Mean values of LH from 24 to 96 h were significantly lower in untreated controls and Group 1 than in the other two groups (0.58 +/- 0.2 ng ml-1 and 0.55 +/- 0.2 ng ml-1 vs. 1.63 +/- 0.5 and 1.68 +/- 0.6 ng ml-1, respectively; P < 0.01). After treatment with PG alone in the untreated control group, the preovulatory LH surge began in all ewes at 59.9 +/- 2.8 h after PG. All Group 1 ewes also had an LH surge but the period from PG injection to the onset of the surge was 124 +/- 17.3 h (range 96-152 h). Only two of the Group 2 ewes had an LH surge (at 160 and 168 h, respectively) and no surge was detected in Group 3 ewes. In Group 1, mean values of follicle-stimulating hormone (FSH; 0.78 +/- 0.07 ng ml-1) were not affected by treatment with antagonist alone; however, in the two groups receiving exogenous LH pulses there was a marked decrease in FSH concentrations during the period 24-96 h. Progesterone concentrations increased 9 days after PG treatment in five out of six ewes in Group 1. In Group 2, there was evidence of a variable luteinisation response, but in Group 3 progesterone remained less than 0.08 ng ml-1 throughout. Endoscopy 112-115 h after PG confirmed that none of the 16 antagonist-treated ewes had ovulated; an event normally expected approximately 80 h after PG in sheep. The experimental protocol of Group 3 provides the basis for a model which will enable examination of the long-term functional capacity of ovarian follicles which have not been exposed to an LH surge.

Analysis of Variance↗

PubMed1997

The distribution of ovulations from the ovaries of merino and Border Leicester x merino ewes and its effect on the survival of their embryos.

The distribution of ovulation between the right and the left ovary was recorded using endoscopy, in 2806 ewes over a 5-year period. Fifteen separate tests were conducted as part of the development programme for a commercial twinning vaccine. There were significantly more ovulations on the right ovary (53.4%) compared to the left ovary (46.6%; P < 0.001). The distribution of ovulation between the ovaries was not influenced by either the breed of sheep or prior immunisation against the steroid hormones androstenedione or testosterone. These findings suggest that the hormonal control of folliculogenesis and ovulation rate is modulated by unknown local factors within the ovary and its vasculature. The site of ovulation had no effect on embryo survival, and embryos from unilateral ovulations were just as likely to survive as were embryos from bilateral ovulations. However, embryo survival was influenced by ovulation rate, and ewes with ovulation rates of four or more had reduced litter sizes and lower embryo survival.

Animals↗

PubMed1997

An evaluation of transovarian uptake of metabolites using arterio-venous difference methods in dairy cattle.

Arterio-venous (A-V) difference techniques were used in cattle to examine ovarian energy metabolism, cholesterol uptake and steroid hormone outputs. Catheters were inserted into the ovarian vein and facial artery, and Transonic flow transducers were placed around the ovarian A-V plexus. Further, in some cows, the effects of a challenge with GnRH were examined. Glucose uptake and lactate output were significant in most individual cows. Nonesterified fatty acids (NEFA) uptake were not significant in any cow in dioestrus. Ovarian uptake of beta-Hydroxy-butyrate (3-OHB) was significant in 4 cows in dioestrus. Cholesterol uptake was significant in only 1 cow. Oxygen uptake was significant in all cows at all stages of the oestrous cycle. All cows had significant output of progesterone and oestradiol-17 beta. These data show that the bovine ovary utilises significant amounts of glucose, and Respiratory quotient (RQ) estimates demonstrated that glucose was the primary fuel used by the ovary. The significant output of lactate suggested that anaerobic pathways were mainly used for glucose oxidation. The observed uptakes of 3-OHB indicated that the ovary utilises 3-OHB as a source of energy. Cholesterol uptake was not a rate-limiting factor for steroid hormone production in the ovary. Despite the high metabolic rate in the luteal ovary, the small difference in PO2 between arterial and ovarian venous blood indicated that the ovary consumes only a small proportion of available oxygen. GnRH had no significant effect on the uptake of metabolites and energy metabolism, but it increased OBF and the output of progesterone and oestradiol-17 beta. The use of A-V methods to determine the metabolic needs of the ovary is useful in understanding the means by which nutrition can influence fertility.

Analysis of Variance↗

PubMed1997

Studies on the role of LH in the maturation of the pre-ovulatory follicle in a sheep using a GnRH-antagonist.

A model designed to mimic the action of LH in preovulatory follicle development was evaluated. The model involved treatment of ewes with GnRHa (50 micrograms kg-1 24 h-1; GnRHa) at the time of luteal regression followed by exogenous LH administered as a series of bolus injections of varying frequency and amplitude. Ovarian responses to this regimen were compared to data derived from the same animals during a normal follicular phase. In experiment 1 four ewes with an autotransplanted ovary were treated with GnRHa followed by hourly LH (2.5 micrograms NIH-oLH-S25 i.v.) for 60 h at which time a further bolus of LH (25 micrograms i.v.) was given to mimic the LH surge. Plasma FSH profiles and the pattern and rate of secretion of oestradiol, inhibin, androstenedione and testosterone by the ovary in normal and GnRHa treated ewes were similar, but ovulation did not occur following the induced follicular phase. Experiment 2 examined the ability of a revised LH regimen to induce ovulation in normal intact ewes. Ewes (n = 11) were treated with GnRHa at the time of luteal regression followed by LH injections (1.25 micrograms i.v.) administered every 3 h for 12 h, every 2 h for the next 12 h and every hour for the following 36 h (3,2,1 regimen). After 60 h of LH injections the animals received an 8 h infusion of oLH (25 micrograms min-1) designed to mimic the pre-ovulatory LH surge. This treatment resulted in normal follicular phase patterns of LH and FSH and had no effect on the incidence (100%) or number (1.6 +/- 0.1) of ovulations when compared to a normal cycle (100% and 1.8 +/- 0.2). Experiment 3 examined the pattern of ovarian follicular development and hormone secretion stimulated by the 3,2,1 LH regimen in ewes with autotransplanted ovaries (n = 8). The LH regimen stimulated normal pre-ovulatory follicular development (normal 1.4 +/- 0.2; induced 1.6 +/- 0.2) and a normal pattern of ovarian oestradiol and androstenedione secretion. Pulse analysis showed, however, that baseline steroid secretion was lower in induced animals during the late follicular phase reflecting lower baseline LH at this stage. We conclude that administration of a GnRHa at the time of luteal regression in conjunction with frequent low amplitude pulses of exogenous LH induce follicular and hormonal patterns that closely mimic those observed following normal luteal regression and that this regimen represents a useful model to study the role of LH in the control of follicular selection.

Androstenedione↗

PubMed1997

Endocrine activity of induced persistent follicles in sheep.

This experiment was designed to examine gonadotropin requirements for the induction and maintenance of persistent ovarian follicles in sheep. At the time of prostaglandin (PG) treatment on the tenth day of an induced estrous cycle, 8 ewes (with one ovary autotransplanted to the neck) received an injection of a GnRH antagonist ([Ac-d-Nal1, d-4-C-1-Phe2, d-Trp3, d-Arg6, d-Ala10] GnRH.HOAc; 50 microg/kg s.c.), and continuous hourly injections of exogenous ovine LH (equivalent to 1.25 microg NIH-oLH-S26) began simultaneously with this first antagonist injection (time zero). Antagonist was given three times at 3-day intervals. On Day 6, LH injections were stopped in 4 ewes (group 2) but continued in 4 other ewes (group 1) until the end of the 10-day experiment. Ovarian vein blood was sampled daily every 15 min for a 2-h period around two injections of exogenous LH (this sampling included group 2 after Day 6). Additional jugular and ovarian vein blood samples were collected every 8 h throughout the experiment. Daily ultrasound examination revealed the presence of at least one large follicle (range 4- to 7.5-mm diameter) from Day 3 to Day 10 in all ewes, but no new growing follicles (> 2 mm) were detected for at least 6 days. After Day 2, secretion of estradiol was positively correlated with that of inhibin (r = 0.83, p < 0.001), whereas FSH concentrations were inversely related to inhibin (r = -0.71, p < 0.001) and estradiol (r = -0.81, p < 0.001). In the absence of an LH surge, estradiol and androstenedione secretion (range 5-20 ng steroid/min) was maintained from Day 1 to Day 8 in group 1; but in group 2, secretion decreased abruptly when the LH injections stopped. Thus, continued low-amplitude, high-frequency LH pulses were required to maintain estradiol secretion when concentrations of FSH were < 0.5 ng/ml. However, estradiol and androstenedione secretion decreased (and FSH concentrations increased) between Days 8 and 10 in the ewes that received continued LH injections (group 1), showing that atresia in estrogenic follicles was not due to a lack of gonadotropin availability but to changes within the follicle. For the first 3 days after administration of PG, androstenedione secretion was greater than that of estradiol (p < 0.05), but from Day 4 to 6 the secretion rates were similar (p < 0.1), suggesting that aromatase may be limiting in the first 3 days whereas provision of androstenedione precursors was altered as the follicle persisted. In group 2 on Days 7 and 8 when hourly LH injections had stopped, neither androstenedione nor estradiol secretion increased after one test injection of LH; in contrast, androstenedione but not estradiol secretion increased after a second LH test injection 1 h later, suggesting that secretion of androstenedione is controlled by repeated exposure to LH. In conclusion, persistent estrogenic follicles were produced in the follicular phase in sheep by treatment with a combination of GnRH antagonist and hourly pulses of LH. Secretion of estradiol was dependent on continued hourly LH pulses of approximately 1 ng/ml and the follicles remained estrogenic for 8 days, after which time the ability to secrete estradiol and androstenedione declined even with continued LH injections.

Androstenedione↗

PubMed1996

The ovarian secretion of androstenedione and oestradiol during late pregnancy and the early postpartum period in sheep with an autotransplanted ovary.

During late pregnancy in the ewe, ovarian function is suppressed by placental steroids and following parturition ovarian function is restored. This experiment determined the ovarian secretion of oestradiol and androstenedione during late pregnancy and the early postpartum period in ewes. Six ewes with ovarian autotransplants were transplanted with three day 6 embryos and three gave birth on day 147. Ovarian and jugular blood sampled were collected on three different occasions. On each occasion a 4 h period of sampling was followed by a 6 or 8 h period during which the ewes were challenged with 150 ng of gonadotropin-releasing hormone (GnRH). Basal secretion of oestradiol and androstenedione was 0.3 +/- 0.1 and 10.5 +/- 3.0 ng min-1, respectively, on day 120 of pregnancy. Oestradiol secretion remained low on days 7 and 21 postpartum (0.4 +/- 0.3 and 0.3 +/- 0.1 ng min-1, respectively). Androstenedione secretion (ng min-1) on days 7 and 21 postpartum was 2.5 +/- 0.5 and 4.1 +/- 1.8, respectively. The injection of GnRH on day 121 of pregnancy produced luteinizing hormone (LH) release with a peak concentration of 0.6 +/- 0.1 ng mL-1, that did not stimulate steroid secretion. On day 8 postpartum GnRH injection induced LH release with a peak concentration of 3.9 +/- 1.1 ng mL-1 that stimulated secretion of oestradiol (0.2 +/- 0.1 to 2.1 +/- 0.9 ng min-1; P < 0.01) and androstenedione (2.3 +/- 0.6 to 17.1 +/- 6.9 ng min-1; P < 0.001). Similar effects were seen on day 22 postpartum; GnRH injection induced LH release with a peak concentration of 4.7 +/- 1.4 ng mL-1 that stimulated secretion of oestradiol (0.2 +/- 0.1 to 3.7 +/- 1.1 ng min-1; P < 0.001) and androstenedione (4.2 +/- 2.6 to 29.5 +/- 9.2 ng min-1; P < 0.01). These results suggest that the suppression of ovarian function during late pregnancy in the ewe is reversed by 7 days postpartum.

Androstenedione↗

PubMed1996

Exogenous interferon delays luteal regression in red deer hinds (Cervus elaphus) by suppressing steroid-induced endometrial oxytocin sensitivity.

Three groups of intact hinds (n = 10-18) and one group of ovariectomized hinds were treated with progesterone by mean, of Controlled Internal Drug Releasing (CIDR) devices for 13 days (device removal = Day 0). Group 1 served as controls; group 2 received injections of 4 mg recombinant bovine interferon-alpha,1 twice daily from Days 13 to 21; group 3 was run with a stag from Days 0 to 3, and all hinds were subsequently diagnosed pregnant; group 4 (ovariectomized) was treated with CIDR devices and estradiol to mimic steroid secretion during the estrous cycle. Progesterone profiles were determined from thrice-weekly plasma samples from Days -13 to 28. Rectal temperature was measured in a subset of groups 1 and 2 from Days 9 to 21. Oxytocin-induced prostaglandin F2 alpha release was measured in a subset of groups 1, 2, and 4 on Days 2, 4, 10, 16, and 18. Data are presented as means +/- SEM. Exogenous interferon delayed luteolysis (> or = 28 vs. 21.2 +/- 0.55 days, P < 0.0005) and induced transient pyrexia after the first injection (39.89 +/- 0.11 vs. 38.88 +/- 0.19 degrees C, p < 0.0005). Incidence of oxytocin-induced PGF2 alpha release in control hinds was greater on Days 2 and 18 than on Days 4 and 10 (8/8 and 7/8 vs. 3/8 and 0/8, respectively; p < 0.05) and was greater in control than in interferon-treated hinds on Days 16 and 18 (5/8 and 7/8 vs. 1/8 and 1/8, respectively; p < 0.05). Profiles of plasma progesterone concentration and oxytocin sensitivity in steroid-treated ovariectomized hinds did not differ from those in control hinds. These results suggest that steroid-controlled uterine oxytocin sensitivity is important in luteolysis and is suppressed by the administration of interferon, the putative embryonic pregnancy recognition signal in red deer.

Administration, Intravaginal↗

PubMed1996

Induction and maintenance of oestradiol and immunoreactive inhibin production with FSH by ovine granulosa cells cultured in serum-free media.

A serum-free ovine granulosa cell culture system is described that allows the induction of FSH-responsive oestradiol production by undifferentiated cells from small (< 3.5 mm) follicles (P < 0.001) and the maintenance of oestradiol production by differentiated cells from large (> or = 3.5 mm) follicles. Physiological doses of FSH stimulated (P < 0.01) proliferation of cultured granulosa cells from both small and large follicles. The synthesis of immunoreactive inhibin and progesterone by granulosa cells from small and large follicles increased (P < 0.01) with time of culture, and was not dependent on FSH. Inhibin secretion expressed on a per cell basis was not FSH responsive. Insulin and insulin-like growth factor I (IGF-I), in the presence of FSH, stimulated (P < 0.001) cell proliferation and oestradiol and inhibin production by granulosa cells from small and large follicles. There was a significant (P < 0.001) interaction between insulin and IGF-I in the stimulation of granulosa cell proliferation and differentiation. Both epidermal growth factor (EGF) and transforming growth factor alpha (TGF-alpha) in the presence of FSH stimulated cellular proliferation (P < 0.001) in a dose-responsive manner and concomitantly inhibited (P < 0.001) oestradiol and inhibin secretion. The development of this granulosa cell culture system will make it possible to study, in vitro, the cascade of events that controls granulosa cell differentiation and ultimately follicle selection in sheep.

Animals↗

PubMed1996

The effects of N-methyl-D,L-aspartic acid and aspartic acid on the plasma concentration of gonadotrophins, GH and prolactin in the ewe.

Aspartic acid is a neurotransmitter in the central nervous system that acts via the glutamate receptor and the analogue, N-methyl-D,L-aspartic acid (NMA) is an agonist that stimulates GnRH secretion. Under normal dietary conditions, the plasma concentration of aspartic acid in ewes is low and if increased by improved nutrition may increase the brain concentration of aspartic acid leading to increased gonadotrophin secretion. In two experiments we investigated the effects of NMA on pituitary hormone concentrations and the effects of aspartic acid on ovulation rate and pituitary hormone concentrations. The intravenous injection of NMA into cycling ewes resulted in an immediate (within 15 min) release of a pulse of LH and of GH and a prolonged (up to 1 h) suppression of prolactin secretion. There were marked differences in responsiveness to NMA between individual ewes. The intravenous infusion of aspartic acid for 5 days in the late luteal phase of the oestrous cycle did not affect ovulation rate but reduced the mean LH (P < 0.05) and FSH (P < 0.05) concentrations in plasma. The frequency of LH pulses also tended to be lower (P < 0.1) in ewes infused with aspartic acid. It is suggested that the decrease in gonadotrophin secretion in ewes infused with aspartic acid is due to effects on the hypothalamus or the anterior pituitary gland which are not related to increased levels of ovarian feedback. These changes are likely to involve decreased GnRH secretion.

Animals↗

PubMed1996

The effect of ovarian arterial infusion of human recombinant inhibin and bovine follicular fluid on ovarian hormone secretion by ewes with an autotransplanted ovary.

Recombinant human inhibin A (rhInh) or steroid-free bovine follicular fluid (bFF) were infused into the ovarian artery of anoestrous ewes with ovarian autotransplants induced to ovulate with a pulsatile regimen of GnRH applied after a 10-day pretreatment with progestagen sponges. In the period 12-24 h after sponge withdrawal ewes received ovarian arterial infusions of saline (n = 6), 0.3 micrograms rhInh/h (n = 5), 1.6 micrograms rhInh/h (n = 5) or 25 microliters bFF/h (n = 4). Controls had a normal follicular phase with an LH surge 43 +/- 3 h after sponge withdrawal which resulted in ovulation (six out of six). Both doses of rhInh increased ovarian venous inhibin concentrations in a dose-related fashion (P < 0.05) but resulted in depressions (P < 0.05) in FSH concentrations of similar magnitude. Both doses of rhInh acutely inhibited ovarian oestradiol and androstenedione secretion (P < 0.01) but at the end of rhInh infusion oestradiol secretion was quickly re-established without a corresponding increase in FSH. LH surges were detected in five out of five and three out of five ewes infused with low and high doses of rhInh respectively, and progesterone concentrations during the subsequent luteal phase were depressed (P < 0.05). Infusion of bFF had no effect on inhibin or FSH concentrations but resulted in acute inhibition (P < 0.01) of ovarian oestradiol, androstenedione and inhibin secretion, a delay (P < 0.05) in the time to the LH surge and a depression (P < 0.05) in luteal-phase progesterone concentrations. In conclusion, while the depression in FSH induced by rhInh cannot be excluded as a cause for the inhibitory effects of rhInh treatment on ovarian function, such a mechanism cannot fully explain the ovarian responses obtained to rhInh infusion. These results therefore support a direct ovarian role for inhibin in the modulation of ovarian function in addition to its indirect role in controlling FSH. This conclusion is supported by the demonstration that bFF can induce similar inhibitory effects on ovarian function without changing FSH.

Androstenedione↗

PubMed1995

Ovulation rate and the concentrations of gonadotrophic and metabolic hormones in ewes fed lupin grain.

An experiment in which a lupin grain supplement was fed to ewes (n = 11) over days 2-13 of the oestrous cycle was carried out. A group of 12 ewes was used as a control and not fed the supplement. Plasma concentrations of LH and GH (20 min intervals) and FSH, insulin and prolactin (hourly intervals) were determined in plasma samples collected every 20 min over 24 h on day 11 of the oestrous cycle. The changes were related to increases in ovulation rate. Ovulation rate was increased (2.5 +/- 0.2 versus 1.9 +/- 0.2 for lupin-supplemented and control groups, respectively; P = 0.073) in the group that received the lupin supplement, but this increase was not associated with significant changes in either LH or FSH concentrations on day 11 of the oestrous cycle. Lupin supplementation had significant effects on the plasma concentrations of prolactin, GH and insulin. There was a transient increase (P < 0.05) in the concentration of prolactin 4-8 h after feeding, whereas insulin concentrations were increased immediately after feeding (P < 0.02) and were still high 24 h later (P < 0.02). Growth hormone concentrations were reduced in ewes fed with lupin grain (P < 0.001). These metabolic responses initiated by feeding a high energy and protein supplement such as lupin grain may be related to changes in ovulation rate. In particular, the sustained increases in insulin concentrations suggest that an increased supply of glucose to the follicle mediates nutritionally stimulated increases in ovulation rate.

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