Search

Even in the absence of stressors, temperament is associated with changes in the concentration of stress-responsive hormones and, possibly because of such changes, temperament can affect metabolism. We tested whether, in sheep bred for temperament for 14 generations, "nervous" females have greater concentrations of stress-responsive hormones in the absence of stressors than "calm" females, and whether these differences are associated with changes in the concentrations of metabolic hormones. In resting "calm" (n = 8) and "nervous" (n = 8) sheep, concentrations of cortisol, prolactin, leptin, and insulin were measured in blood plasma sampled via jugular catheter every 20 min for 24 h. The animals were individually penned, habituated to their housing and human handling over 7 wk, and fed before sampling began. Diurnal variation was evident for all hormones, but a 24-h cortisol pattern was detected in only 7 individuals. There was no effect of temperament on any aspect of concentrations of cortisol or prolactin, but "calm" animals had greater concentrations of insulin in the early afternoon than "nervous" animals (14.5 ± 1.1 vs 10.0 ± 1.6 μU/mL; P = 0.038), and a similar tendency was seen for leptin ( P = 0.092). We conclude that selection for temperament affects the concentration of metabolic hormones in the absence of stressors, but this effect is independent of stress-responsive hormones.

Cattle farmers in Ghana want larger animals with better milk yield and have therefore focused on improving the indigenous tropical Sanga breed by crossing with the temperate Friesian breed. However, in the Friesian–Sanga crossbred, a major limitation to reproductive performance is extended postpartum anoestrus, perhaps exacerbated by under-nutrition under field conditions. We therefore tested whether the breeds vary in the circulating concentrations of metabolic hormones that mediate the effects of nutrition on the reproductive axis during the postpartum period. Two herds comprising 16 Sanga, and 16 Friesian–Sanga cows, were grazed separately on natural pastures of similar nutritive value with no feed supplements. Every week during weeks 1 to 10 postpartum, the cows were weighed, body condition score (BCS) was assessed, and blood was sampled. Resumption of ovarian activity, detected from progesterone profiles, was used to classify cows as ‘early-cycling’ (≤45 days), ‘late-cycling’ (46–70 days) or ‘non-cycling’ (after 70 days). Resumption of ovarian cyclicity, body weight and body condition score did not differ between breeds, but daily milk yield was lower for Sanga (1.01±0.02 kg/day) than for Friesian–Sanga crossbreds (1.57±0.02 kg/day; P

We tested the effect of dose of GnRH superagonist on pituitary and testicular function in a study with four groups of four male dogs. The Controls received blank implants and the other three groups received implants containing 3, 6 or 12 mg deslorelin (D-Trp6-Pro9-des-Gly10-GnRH ethylamide). In all deslorelin-treated groups, there was initially an acute increase in plasma concentrations of LH and testosterone, followed by declines such that both hormones became undetectable after approximately 12 days. There was a dose-response in some of these early aspects of the hormone profiles. With respect to long-term effects of treatment, the 12-mg dose had significantly greater effects than the smaller doses for the duration of minimum testicular volume [366 +/- 77, mean +/- SEM (3 mg), 472 +/- 74 (6 mg), and 634 +/- 59 (12 mg) days], absence of ejaculate [416 +/- 88 (3 mg), 476 +/- 83 (6 mg), and 644 +/- 67 (12 mg) days], undetectable plasma concentrations of LH and testosterone [367 +/- 64 (3 mg), 419 +/- 72 (6 mg), and 607 +/- 69 (12 mg) days], the delay until complete recovery of LH and testosterone secretion [394 +/- 65 (3 mg), 484 +/- 72 (6 mg) and 668 +/- 47 (12 mg) days], and the delay until testes had regrown to normal volume [408 +/- 77 (3 mg), 514 +/- 74 (6 mg), 676 +/- 59 (12 mg) days]. The time taken to restore full ejaculates was also longest for the 12-mg dose: 716 +/- 67 (12 mg) days vs 440 +/- 66 (3 mg) and 538 +/- 83 (6 mg) days after implantation. There was no correlation between delay to recovery of normal ejaculate quality and body mass. We conclude that the dose-response relationship with deslorelin implants is not expressed with respect to the degree of suppression of reproduction, but on the maximum duration of suppression and thus to delay until recovery.

Penguins show varying degrees of brood reduction behavior, from obligate brood reducers to brood maximizers, and we hypothesize that this is associated with differences in prolactin secretion. To address this hypothesis, we determined the breeding season prolactin profile of the yellow-eyed penguin (Megadyptes antipodes) for comparison with those of other penguin species found in the literature. We also measured sex steroid plasma concentrations to better characterize the reproductive cycle of the species. Plasma concentrations of prolactin increased from early in the season, reaching a peak during late incubation, and remained elevated up to the guard period. This general pattern was similar to that of other penguins for which we have corresponding data. However, we found that throughout the laying period, prolactin titers in yellow-eyed penguins remained elevated while they fell to basal levels after the laying of the first egg in macaroni penguins, which corresponds to differences in incubation behavior during this time. We conclude, therefore, that differences in the brood reduction behavior in penguins, may be reflected in the pattern of PRL concentrations around the time of egg laying.

In mature Merino rams, nutrition is one of the external cues that most strongly affects the reproductive centres of the preoptic-hypothalamic continuum. The signalling pathways that link dietary status and the activity of the neurones that produce gonadotrophin-releasing hormone signals are thought to be partly hormonal in nature to reflect the amount of body reserves. Among the hormones thought to be involved are insulin and leptin. This study tested whether recombinant bovine leptin infused (0.4 microg/h) into the third cerebral ventricle would stimulate pulsatile luteinizing hormone (LH) secretion in mature Merino rams when their energy status was low or decreasing, during both chronic (fasting) and acute reductions of energy balance. Leptin may interact with other hormones that depend on energy availability, so we also monitored changes in circulating concentrations of insulin, thyroid hormones, growth hormone, prolactin and adrenocorticotrophin. Overall, our data do not support this hypothesis. The dietary regimes induced clear responses in the metabolic profiles of the animals but there was no clear effect of central leptin administration on LH pulse frequency. The relationships between the hormonal systems measured in the present study add weight to the contention that leptin plays only a permissive role in the nutritional control of the reproductive axis and that other hormonal signals (particularly insulin) or pathways are acting in concert with leptin to stimulate the reproductive axis.

Orexins exert their effects through two specific receptors (OX1R and OX2R) that have been found mainly in the brain and also in peripheral tissues of rats and humans. Here, we demonstrate expression of mRNA encoding for ovine OX1R and OX2R in central and peripheral tissues of sheep. Gene expression for orexin receptors in the hypothalamus and the preoptic area was localised by in situ hybridisation. OX1R was detected in arcuate nuclei (ARC), median eminence (ME), the lateral hypothalamic nuclei and preoptic area (POA) and it was scattered along the third ventricle from the paraventricular (PVN) to the ventromedial hypothalamic nuclei (VMH). OX2R was localised in the PVN, ARC, ME, ventral VMH and a small region of the ventral POA. Gene expression for OX1R and OX2R in central and peripheral tissues was analysed using quantitative real time RT-PCR. Both orexin receptor genes were expressed in the hypothalamus, POA, hippocampus, amygdala, olfactory bulb, pineal gland and recess and pituitary gland, whereas only OX1R mRNA was detected in the testis, kidney and adrenal gland. The expression of the genes for orexin receptors in this range of ovine tissues suggests roles for orexins in multiple physiological functions, with actions at both central and peripheral levels.

反芻動物での血中レプチン濃度の調節機構には未解明な点が多い．そこで短期絶食後に再給飼およびVFA類似の糖源性溶液の経口投与を行い，これらの処置中の血中レプチン濃度の変化を調べ，グルコース・インシュリン・インシュリン様成長因子（IGF-1）の濃度変化との関係について調べた．雄ヒツジに対し48時間の無給餌をした後に給飼を再開し，同時に水または糖源性溶液を12時間間隔で2日間経口投与した．処置中は12時間間隔で採血し，レプチン濃度を牛レプチン用ラジオイムノアッセイにより測定した．絶食によってグルコースと全ホルモンの血中濃度が低下したが，給飼再開後に水群ではレプチンのみが有意に増加した．糖源性溶液投与によってグルコースとインシュリン濃度が高値で推移し，再給飼によるレプチン濃度増加効果を増幅させた．したがってレプチン濃度はエネルギバランスの変動に応じて短期間に変化する．レプチン濃度調節機構へのグルコース・インシュリンなどの関与については更に研究が必要である．

Veterinary practitioners frequently encounter disorders of the reproductive system in avian patients. Management of these disorders relies on manipulating reproduction by modifying the environment, diet, and social interactions, and by the use of pharmacologic agents and surgery, with varying levels of success and side effects. An alternative is to use the gonadotropin- releasing hormone (GnRH) agonist deslorelin to suppress the pituitary-gonadal axis. To determine the efficacy of deslorelin in domestic pigeons (Columba livia), male (n ¼ 10) and female (n ¼ 10) birds each were implanted intramuscularly with a single long-acting implant containing 4.7 mg deslorelin. Untreated males (n ¼ 11) and females (n ¼ 10) were used as controls. The baseline serum concentration of luteinizing hormone (LH) was assayed at 7, 28, 56, and 84 days after treatment, and egg production was recorded weekly. In females, deslorelin administration significantly reduced serum LH concentrations compared to pretreatment levels at 7, 28, 56, and 84 days (P , .05). In males, deslorelin significantly reduced LH concentrations at 7, 28, and 56 days (P , .05). Female birds treated with deslorelin laid significantly fewer eggs over the course of the study (mean ¼ 1.46, SEM ¼ 0.84) compared with controls (mean ¼ 5.54, SEM ¼ 0.88). Deslorelin treatment had no discernible effect on body weight. Deslorelin is effective for controlling egg laying in female pigeons for at least 49 days, but further research is required to determine the effects on male fertility and the duration of action in both sexes.

African wild dogs are one of the most endangered canid species, with free-living populations declining as a consequence of habitat loss, disease and human conflict. Captive breeding is considered an important conservation strategy, but is hampered by a poor overall understanding of the reproductive biology of the species. To improve our basic knowledge, we studied hormone patterns in 15 female wild dogs using non-invasive faecal collections. By comparing longitudinal hormone profiles with behavioural and anatomical changes, females could be allocated among three reproductive classes: pregnant (n=1), pseudopregnant (n=9) and acyclic (n=4). We also monitored a single female in which contraception was induced with a deslorelin implant. Comparison of pseudopregnant and acyclic females showed that, in both classes, faecal oestradiol concentrations increased from anoestrus to pro-oestrus then declined into the oestrous and dioestrous phases. Progestagen concentrations rose steadily from anoestrus to the dioestrous phase in both pseudopregnant and acyclic females and, pseudopregnant females had significantly higher concentrations of progestagens than acyclic females in all phases of the oestrous cycle. Most females classed as pseudopregnant were found in female-only groups, suggesting that wild dogs are spontaneous ovulators. Furthermore, only one adult female did not ovulate, so suppression of reproduction in subordinates is likely to be behavioural rather than physiological.

The stimulatory effect of nutritional supplementation on ovarian activity in sheep has been linked to an increase in glucose availability that, with insulin, directly decreases follicular steroidogenesis. Glucose uptake occurs by glucose transporters, but it is not known which glucose transporters are present in the sheep ovary or whether they are affected by nutritional stimulation. The aim of this study was to determine whether widely distributed glucose transporter 1 (GLUT1) or insulin-responsive GLUT4 are present in the granulosa or theca cells of sheep ovarian follicles, and whether their concentrations are affected by nutritional stimulation. Merino ewes (n = 49-51 per group) were stimulated nutritionally for 5 days before luteolysis with lupin grain or with one of two regimens of a glucogenic mixture, administered orally, which increases blood glucose concentrations towards the upper end of the normal range. Water was used as a control. Ovaries (n = 3 per group) were dissected and the granulosa cells and thecal shell from individual follicles were examined for glucose transporters using western blotting. GLUT1 concentration was 7-18 times higher in the granulosa than in the theca cells. GLUT4 was detected at a similar concentration in both types of cell. Nutritional treatment had no effect on the concentration of GLUT1 or GLUT4 in either tissue, and did not increase ovulation rate, despite increased concentrations of glucose and insulin. Concentrations of glucose transporters were not correlated with follicular concentrations of oestradiol or androstenedione. The presence of GLUT1 and GLUT4 in the granulosa and theca of sheep follicles indicates that the transporters have a role within the ovary in the modulation of follicular function.

The objective of this study was to establish, for a short-day breeding bird, the male emu, whether the breeding season is principally controlled by changes in photoperiod, and to investigate the endocrine mechanisms involved. Two groups of adult males were subjected to three alternating periods of 150-185 days of 14 h light/day (LD) and 10 h light/day (SD) terminating in a 360-day period of LD or SD. Transfer from LD to SD led to increases in plasma concentrations of luteinizing hormone (LH) and testosterone, after 82 +/- 8 and 73 +/- 3 (SEM) days, and an increase in prolactin concentrations after 115 +/- 12 days. Concentrations of LH and testosterone began to decrease before transfer back to LD, at a time when prolactin concentrations were approaching peak values. Transfer from LD to 360 days of SD resulted in increases in LH and testosterone concentrations, and these terminated after an increase in prolactin concentrations. After transfer from SD to 360 days of LD, plasma concentrations of LH and testosterone began to increase, after delays of 222 +/- 24 and 225 +/- 13 days, and were high at the end of the study, while prolactin values remained depressed throughout. These observations clearly show that seasonal breeding in the emu is directly controlled by changes in photoperiod. The dynamics of the hormonal responses to change of photoperiod suggest that, despite being short-day breeders, the photoregulation of breeding in emus involves mechanisms that are currently accepted for birds, rather than mechanisms that have been proposed for short-day breeding mammals. The initiation of breeding in emus is due to dissipation of photorefractoriness by short days which leads to an increase in the secretion of gonadotrophins to levels that are sufficient to support full reproductive condition. The termination of breeding, while days are still short, is due to the antigonadotrophic action of prolactin which, unusually for birds, increases while the days are still short. In conclusion, breeding activity in male emus is strongly controlled by photoperiod. Emus are short-day breeders, but the central mechanisms that regulate the secretion of reproductive hormones seem to be similar to those previously proposed for long-day breeding birds. The pattern of prolactin secretion in emus suggests an important role for this hormone in the termination of the breeding cycle.

Changes in food intake affect the reproductive axis in both sexes, and the nutritional signals involved and the sites that receive those signals are now beginning to be unravelled. Our studies have focussed on the mature male sheep, a model in which high food intake stimulates GnRH-LH pulse frequency for only 10-20 days but continues to promote testicular growth over several months. Different signals and different target organs seem to be responsible for these short- and long-term responses. Short-term dietary treatments lead to changes in blood concentrations of glucose, fatty acids, insulin and leptin, and concentrations of glucose, insulin, leptin and some amino acids in cerebrospinal fluid. It seems unlikely that amino acids affect GnRH-LH secretion directly in sheep. Intracerebroventricular infusions of insulin specifically increase LH pulse frequency, but intravenous, intra-abomasal or intracerebroventricular infusions of glucose have no effect, despite their effects on cerebrospinal fluid insulin concentrations. The addition of fatty acids to the diet also increases LH pulse frequency, but does not affect the concentrations of insulin or leptin in the cerebrospinal fluid. It appears that acute responses to changes in nutrition involve a range of alternative pathways, possibly including interactions among insulin, leptin and energy substrates. Effects of long-term dietary treatments on testicular size are only partly dependent on the GnRH-LH system (that is, on brain control) and so must also depend on other, as yet unknown, pathways. Concepts of 'metabolic sensing and integration' are being developed from the basis of existing knowledge of the central control of appetite and reproduction.

In mature male sheep, the level of nutrition acutely influences the secretion of reproductive hormones. The mechanism involved is not fully understood but findings in humans and laboratory rodents would suggest a major role for leptin that is secreted from adipose tissue and then travels via the circulation to the central nervous system. Before we can begin to test this hypothesis, we need to be able to measure leptin concentrations in blood plasma and cerebrospinal fluid. We have therefore developed a radioimmunoassay using antibodies raised against biologically active recombinant bovine-ovine leptin. Using this assay, we found that plasma concentrations of leptin were highly correlated to back-fat thickness and to the ratio of back-fat thickness to liveweight, in female and castrated male sheep. Plasma concentrations of leptin were higher in female sheep than in castrated or intact male sheep. Serial samples (every 5 min) suggested that the secretion of leptin in male sheep is episodic but it does not appear to show clear pulsatility, increases post-prandially, or a diurnal rhythm. Leptin concentrations in both plasma and cerebrospinal fluid increased within 5 days in male sheep fed a diet with a high content of energy and protein that also stimulates the secretion of LH pulses. These data suggest that in sheep, as in other species, leptin production is correlated with the mass of adipose tissue and that the hormone passes from the circulation to the cerebrospinal fluid and then to hypothalamic sites. There, it may affect appetite and perhaps GnRH secretion. The role of leptin in the link between nutrition and reproduction needs further investigation.

The male reproductive cycles of three species of vespertilionid bat (Nyctophilus geoffroyi, N. major, andVespadelus regulus) are described. Descriptions are based on the degree of testicular and cauda epididymaldevelopment estimated from field captures, and for N. geoffroyi, on histological examination of fixedspecimens and assays of their testicular testosterone concentrations. All species were found to undergo adissociated pattern of reproduction; the activity of the testes and accessory glands of reproduction wereasynchronous. Spermatogenesis occurred in summer/early autumn (December–March) and testes generallyregressed before mating began in autumn (March–May). Spermatozoa were stored in the caudaepididymides during the mating period. Leydig cells of N. geoffroyi involuted prior to mating but theprostate gland remained enlarged and secretory during winter (June–August), regressing in late spring(November). Testicular testosterone concentrations were greatest when Leydig cells were maximallyenlarged, and concentrations were low during the mating period. The reproductive cycle of all speciesapparently involved asynchrony between primary and secondary sexual function, which is typical ofvespertilionid bats inhabiting temperate latitudes. As mating occurs when sperm stores cannot bereplenished, selection for prudent sperm allocation may be expected.Key words: vespertilionid bat, reproduction, testes, spermatogenesis, sperm storage

The effects of nutrition on the testis were investigated in groups of five mature Merino rams that were fed either a sub-maintenance (low) diet or a supra-maintenance (high) diet for 69 days. Testosterone, oestradiol and inhibin were measured in blood plasma sampled simultaneously from jugular and testicular veins after an i.v. injection of 200 ng ovine LH kg-1. Plasma concentrations of testosterone, inhibin and oestradiol were higher in testicular than in jugular vein plasma for both diets (P < 0.01). After the LH injection, jugular plasma testosterone increased more rapidly (P < 0.01) in rams fed the high diet than in rams fed the low diet. This was not seen in the testicular vein. Oestradiol concentrations were higher in rams on the high diet than in those on the low diet, in both the testicular (P < 0.0001) and the jugular vein (P < 0.02). Diet did not affect inhibin concentrations. Testes were surgically removed and processed for light microscopy. Testicular mass and seminiferous tubule length and diameter were higher with the high diet than the low diet (P < 0.01). The number of Sertoli cell nuclei per testis was also affected (high diet: 120 +/- 6 x 10(8); low diet: 77 +/- 7 x 10(8); P < 0.001), whereas the proportion of testis occupied by Sertoli cell nuclei was not affected. The number of Leydig cells per testis was not affected by diet, but Leydig cells occupied a greater volume of testis in rams on the high diet than in those on the low diet (P < 0.001). The effects of nutrition on Leydig and Sertoli cells are consistent with changes in the endocrine and exocrine functions of the testis. The finding that Sertoli cell population was altered in adult rams may be explained by the GnRH-independent effects of nutrition.

Australian cashmere goat bucks exhibit marked seasonal changes in gonadotrophin secretion and growth. We investigated the mechanisms underlying these changes by testing the effects of castration and gonadal steroid replacement in grazing bucks at 29 °S, 153 °E over a period of 2 years. Three year old bucks were either left intact ( n = 6), castrated ( n = 6), or castrated and treated with testosterone propionate ( n = 5) or oestradiol-17β ( n = 5). Oestradiol- 17β (E 2 ) was provided by 2 subcutaneous Silastic ® implants while testosterone propionate (TP) was provided by implanting four 23.5 mg pellets subcutaneously every 12 weeks. Intact bucks exhibited marked circannual cycles of plasma LH, FSH, prolactin and testosterone concentration, body mass, paired testicular mass and male odour. Castration resulted in long term increases in LH and FSH concentration and abolition of the circannual pattern of change in these hormones without affecting prolactin concentrations. It also reduced the magnitude of seasonal changes in body mass. Treatment of castrates with TP, which produced peripheral concentrations of testosterone in the lower physiological range (mean 1.2 ± 0.08 μg L −1 ), had no effect on plasma concentrations of LH, FSH or prolactin. On the other hand treatment with E 2 , which produced E 2 concentrations in the mid physiological range (mean 12.3 ± 1.08 ng L −1 ), re-imposed a circannual pattern in plasma LH and FSH, albeit significantly different from that of intact bucks. It had no effect on prolactin concentrations. Treatment with both steroids resulted in growth cycles of intermediate magnitude between those of intact bucks and castrates. We conclude that the circannual cycle of gonadotrophin concentrations in goat bucks is dependent upon the presence of the testis, and suggest that circulating metabolites of testosterone, including oestradiol, are more important than testosterone itself in the maintenance of this cycle. We also suggest that the seasonal growth cycle observed in these bucks is driven by factors other than prevailing nutrition, probably photoperiod, and is mediated by both steroid-dependent and independent mechanisms.

The frequency of multiple ovulations in mature, cyclic ewes is strongly influenced by the level of nutrition, but it is difficult to demonstrate concurrent changes in plasma concentrations of gonadotropins. The failure to do so might be a consequence of rapid compensation by the homeostatic feedback mechanism linking secretion by the hypothalamus/pituitary gland and ovarian hormones. Most experimental models have examined the components of the homeostatic feedback system after steady state relationships had been established. We hypothesised that the effects of nutrition might be observed more readily if the system were disrupted and then examined while equilibrium was being re-established. This hypothesis was tested in three experiments in Merino ewes by allowing gonadotropin secretion to escape feedback for 5 days after ovariectomy and then replacing ovarian hormones and examining effects of feeding regimen on the return of plasma concentrations of FSH to baseline values. In all three experiments, oestrogen replacement caused plasma concentrations of FSH to decline more rapidly (P < 0.05) in ewes fed at 0.5x maintenance, than in ewes fed at 1.4x maintenance, with groups fed at maintenance being intermediate. No effect of diet was observed on plasma FSH concentrations in the absence of oestradiol, and neither progesterone nor charcoal-treated bovine follicular fluid influenced the effect of nutrition. Plasma concentrations of oestradiol were 9.8% lower on average (NS) in ewes fed above maintenance than in the sheep fed below maintenance over the three experiments, suggesting that there may have been a reduced clearance of oestradiol which contributed to the result. We conclude that feeding regimen affects the secretion or clearance of gonadotropins in mature ewes, as in the mature ram, and that this is one mechanism by which ovulation rate may be affected.

In mature Merino rams, changes in diet to below or above the requirements for maintenance of body weight lead to changes in gonadotrophin secretion and testicular growth. However, the effects on testicular growth persist for much longer than those on LH and FSH secretion so that the gonadal and gonadotrophin responses are poorly correlated over time. This suggests that the gonadal effects may be partly independent of changes in the hypothalamic secretion of GnRH, an hypothesis tested in this study. In a short-term experiment (November, late spring, non-breeding season), we tested whether a high frequency of exogenous GnRH pulses could override the endogenous system and mimic the change in gonadotrophins seen in rams fed a high plane of nutrition. Mature Merino rams (scrotal circumference (mean ± s.e.m.) 33·6 ±0·5 cm, body weight (mean ± s.e.m.) 59·0 ± 0·9 kg) were fed 900 g chaff+1·6 kg lupin grain (High diet) or 360 g chaff+60 g lupin grain (Low diet) and infused with 8 pulses of GnRH or saline daily for 5 weeks (n=5/group). Blood was sampled every 20 min for 12 h on days – 1 and 14 relative to the start of treatments. Relative to pre-treatment levels, LH pulse frequency and FSH concentrations were decreased on day 14 in saline-infused rams fed the Low diet and increased in saline-infused rams fed the High diet (P This model was used for a more complete endocrine analysis in a longer experiment designed to test the hypothesis that the effect of nutrition on testicular growth is partly independent of changes in the secretion of GnRH. The same treatments were imposed for 35 days on a different group of similar rams in March (autumn, mid-breeding season). Body weight and scrotal circumference were measured weekly and blood was sampled on days – 1 and 14. On days – 1 and 35, testosterone secretion in response to LH was tested by injecting exogenous ovine LH (NIADDK-oLH-25; 200 ng/kg body weight) to all rams. Body weight increased in rams fed the High diet and decreased in those fed the Low diet (PPPP In conclusion, we have shown that in mature rams pulsatile exogenous GnRH cannot fully reproduce the effect that feeding a high diet has on testicular growth, suggesting that the effect of nutrition on testicular growth is partly independent of changes in the secretion of GnRH. Our results also show that (i) testicular growth induced by nutrition is not associated with changes in plasma concentrations of inhibin, or peripheral concentrations of testosterone after a pulse of LH, suggesting a dissociation of the endocrine and spermatogenic functions of the testis; (ii) changes in diet alter the secretion of gonadotrophins primarily by changing GnRH pulse frequency; and (iii) an exogenous GnRH pulsatile regimen can override endogenous secretion of a similar pulse frequency. Journal of Endocrinology (1995) 147, 75–85

Erratum to “Relationships between plasma concentrations of leptin and other metabolic hormones in GH-transgenic sheep infused with glucose” [Domest Anim Endocrinol 24 (2003) 219–229] H. Kadokawa a,b,c,∗, J.R. Briegel a, M.A. Blackberry b, D. Blache b, G.B. Martin b, N.R. Adams a a CSIRO Division of Livestock Industries, Private Bag #5, PO Wembley, WA 6913, Australia b School of Animal Biology, Faculty of Natural & Agriculture Sciences, University of Western Australia, Crawley, 6009 WA, Australia c Department of Animal Production and Grassland, National Agriculture Research Center for Hokkaido Region, Hitsujigaoka 1, Toyohira, Sapporo 062-8555, Japan

The effects of dietary zinc deficiency on testicular development in young Merino rams (initial live mass, 22 kg) were tested. Four groups of five rams were fed ad libitum with diets containing 4, 10, 17 or 27 micrograms Zn g-1. To control the effects of loss of appetite caused by zinc deficiency, a fifth group (pair-fed control) was fed the diet containing 27 micrograms Zn g-1, but the amount of feed offered was restricted to that eaten voluntarily by the zinc deficient (4 micrograms Zn g-1) rams they were paired with. After 96 days on the diets, epididymal and testicular masses did not differ significantly between the animals fed 10, 17 or 27 micrograms Zn g-1 ad libitum, but were significantly lower in pair-fed controls, and lowest in the zinc-deficient animals. Testicular responsiveness to LH, as measured by testosterone production, increased substantially in most rams as the experiment progressed, the only exception being the zinc-deficient group, in which the response to LH was lower than in any of the other groups. Testicular concentrations of zinc and testosterone were lower in the zinc-deficient animals than in all the other groups. Plasma inhibin concentrations fell as the experiment progressed in rams fed 17 and 27 micrograms Zn g-1 ad libitum, but not in the other groups. The pair-fed control rams had smaller seminiferous tubules and less lumen development than did the controls fed ad libitum (27 micrograms Zn g-1), which were similar to the animals fed 10 or 17 micrograms Zn g-1. In zinc-deficient rams, the tubule development was further retarded and the interstitial regions were more extensive than in the other groups. We conclude that the overall effect of zinc deficiency on testicular development is due to a combination of a non-specific effect (low gonadotrophin concentrations caused by the low feed intake) and a specific effect due to the lack of zinc. The zinc-specific effect is localized within the testis where it reduces the development of the capacity to produce testosterone, leading to low intratesticular concentrations of testosterone, a critical factor for the growth, development and function of the seminiferous tubules.

The effects of nutrition on the hypothalamo-pituitary-gonadal axis were studied in three groups of six mature Merino rams that were fed for 56 days with a ration that maintained their initial live mass (intermediate diet: 675 g chaff plus 175 g lupins), the same ration with a lupin supplement (high diet: 675 g chaff plus 825 g lupins), or about half of the intermediate ration (low diet: 475 g chaff plus 125 g lupins). Lupin seed provides a highly (95%) digestible source of energy and protein. Plasma concentrations of LH, FSH, testosterone and inhibin were measured in blood samples collected over 24 h on the day before dietary treatments began (day-1), then on days 0, 1, 5, 14, 28 and 56. Compared with the intermediate diet, the high diet significantly increased live mass within 14 days and testicular size within 28 days, and these differences increased steadily throughout the experiment. Plasma FSH concentrations and LH pulse frequency increased within 5 days, but these effects were maintained for only 14 days. Decreasing the nutritional status reduced live mass and testicular size within 7 days, led to a low LH pulse frequency that persisted throughout the experiment, but did not affect FSH concentrations. Significantly less testosterone was secreted over 24 h in the low dietary group than in the intermediate or high group until day 28. The high group tended to secrete more than the intermediate group, but only at the beginning of the experiment when LH pulse frequencies differed between these groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Temperament influences the expression of maternal behaviour in sheep under experimental field conditions. We investigated whether maternal behaviour between ewes selected for a calm or nervous temperament is independent from environmental conditions. In addition, the level of maternal behaviour expressed by mothers is correlated with the concentration of hormones during the peripartum period. Therefore, we investigated whether the selection for temperament had resulted in hormonal differences between the two lines with regard to the hormones that could be involved in the onset of maternal behaviour. Oestradiol, progesterone and cortisol concentrations from 4 days before parturition to 24 h after parturition were determined from blood samples collected from 10 calm and 12 nervous ewes. Behavioural interactions between ewe and lamb were also recorded for 2 h starting at parturition. Mothers of both temperament lines showed adequate maternal behaviour under the controlled conditions of the study. Therefore, the results of the study do not suggest that selection for a calm or nervous temperament has profoundly affected the intrinsic ability of mothers to display adequate maternal behaviour. Hormonal differences between the two temperament lines were generally small and their possible influence on the display of maternal behaviour in the two temperament lines would have to be demonstrated.

We have developed an experimental model in which groups of ewes are simultaneously experiencing the first ovarian follicular wave of their oestrous cycle. We used this ‘first-wave model’ in a 2×2 factorial experiment (ten ewes per group) to study the effect of body condition (BC) and a short-term supplement on follicular dynamics and ovulation rate. The ‘first-wave’ was established by giving ewes three injections of prostaglandin (PG), 7 days apart. The 6-day supplement (lupin grain) began 2 days after the second PG injection and continued until the third. Follicles were studied by ultrasound, and blood was sampled to measure glucose and hormones. The supplement increased (PPP=0.06). The supplement tended to increase the number of 3 mm follicles (P=0.06). Compared with low-BC ewes, high-BC ewes had more follicular waves (PPP=0.09). Leptin and insulin concentrations remained high until the end of supplementation in high-BC ewes, whereas they decreased after the third day of supplementation in low-BC ewes. In conclusion, high concentrations of metabolic hormones in fat ewes are associated with the development of more follicular waves. When a supplement is superimposed on this situation, changes in glucose and metabolic hormones allow more follicles to be selected to ovulate.

The present study tested whether exogenous gonadotrophin-releasing hormone (GnRH) and luteinising hormone (LH) can stimulate LH and testosterone secretion in dogs chronically treated with a GnRH superagonist. Twenty male adult dogs were assigned to a completely randomised design comprising five groups of four animals. Each dog in the control group received a blank implant (placebo) and each dog in the other four groups received a 6-mg implant containing a slow-release formulation of deslorelin (d-Trp6-Pro9-des-Gly10–LH-releasing hormone ethylamide). The same four control dogs were used for all hormonal challenges, whereas a different deslorelin-implanted group was used for each challenge. Native GnRH (5 μg kg–1 bodyweight, i.v.) was injected on Days 15, 25, 40 and 100 after implantation, whereas bovine LH (0.5 μg kg–1 bodyweight, i.v.) was injected on Days 16, 26, 41 and 101. On all occasions after Day 25–26 postimplantation, exogenous GnRH and LH elicited higher plasma concentrations of LH and testosterone in control than deslorelin-treated animals (P < 0.05). It was concluded that, in male dogs, implantation of a GnRH superagonist desensitised the pituitary gonadotrophs to GnRH and also led to a desensitisation of the Leydig cells to LH. This explains, at least in part, the profound reduction in the production of androgen and spermatozoa in deslorelin-treated male dogs.

The effects of social stimuli on avian reproductive behaviors such as breeding schedules and courtship behaviors are well known due to numerous field studies. However, studies that have simultaneously examined the effects of social stimuli on reproductive behavior and the mediating endocrine mechanisms have been largely restricted to captive populations, which may not be representative of free-living populations. This study, conducted over two breeding seasons, aimed to simultaneously measure the effects of experimentally increasing auditory stimuli on the breeding schedule and endocrinology (levels of total androgen, estradiol, progesterone and prolactin) on free-living yellow-eyed penguins (Megadyptes antipodes). The yellow-eyed penguin is the least colonial of all penguins, nesting far apart from each other under dense vegetation, and, therefore, is presumed to experience much lower levels of social stimuli than other penguins. Egg laying was significantly more synchronous and tended to be earlier when birds were exposed to playbacks of the calls of conspecifics in 1 year of the study. We also found that levels of total androgen and estradiol of males in 1 year, and prolactin in another year, were proportionally higher among treated birds compared control birds that received no artificial auditory stimuli. These results show that even among supposedly solitary nesters, social stimuli could still play a role in influencing reproductive behavior and physiology. For the first time in free-living seabirds, we have demonstrated that behavioral responses to increased social stimuli are associated with hormonal changes.

The transgenic sheep used in this study expressed an additional copy of the gene for ovine growth hormone (GH), so they had continuously high plasma concentrations of GH. They were used to test whether the GH transgene affected plasma concentrations of the metabolic hormones leptin, insulin-like growth factor 1 (IGF-1) and insulin, and whether these effects were associated with changes in conception, pregnancy or parturition following artificial insemination. Compared with control animals, the GH-transgenic sheep had higher bodyweight, lower body condition score and less subcutaneous fat (Pthinsp

In mediterranean environments, pastures are very poor during the autumn and consequently small ruminants, such as sheep, would have been losing body mass for many months so, during mating, gamete production would be depressed in both sexes. Effectively, the nutritive requirements for a photo-period-driven, annual reproductive cycle are out of phase with seasonal changes in food availability. The problem could be overcome through more flexible timing of reproduction, perhaps explaining variations in seasonality between breeds that originate from differing latitudes. To study these concepts and the mechanisms involved, the endogenous rhythms and responses to photoperiod were compared in rams of ‘mediterranean origin’ (Merino) and ‘temperate origin’ (Suffolk). Groups of 16 rams of each breed were given a constant food supply and subjected to 16 months of constant equinoctial photoperiod (12L : 12D) or simulated ‘mediterranean’ changes in daylength (from 14L : 10D to 14D : 10L). With nutritional and photoperiodic inputs held constant, Merino and Suffolk rams showed similar endogenous rhythms in reproductive activity. Under constant nutritional inputs and a mediterranean photoperiodic cycle, the endogenous rhythms were modified differently in the two breeds, with the Merinos starting and finishing their seasons about 2 months earlier than the Suffolks. These observations partially explain the patterns observed in rams kept under field conditions. It is now necessary to test whether the rhythms of reproduction in these breeds are also modified by changes in nutrition and social cues.

Interactions between testosterone, estradiol, and inhibin in the control of gonadotrophin secretion in males are poorly understood. Castrated rams were treated with steroid-free bovine follicular fluid (bFF), testosterone, or estradiol and for 7 d (2 x 2 x 2 factorial design). Given independently, none of the exogenous hormones affected follicle-stimulating hormone (FSH) concentrations, but the combination of one or both steroids with bFF reduced FSH secretion. Testosterone and estradiol reduced luteinizing hormone (LH) pulse frequency (there was no synergism), and bFF had no effect. Plasma prolactin concentrations were not affected by any treatment. To locate the central sites of steroid action, castrated rams were bilaterally implanted in the preoptic area (POA), ventromedial nucleus (VMH), or arcuate nucleus (ARC). These implants did not affect FSH or prolactin concentrations, or LH pulse amplitude. The frequency of the LH pulses was not affected by testosterone in any site. Estradiol located in the ARC, but not the POA or VMH, decreased LH pulse frequency. In summary, FSH secretion is controlled by synergistic interactions between inhibin and estradiol or testosterone, whereas GnRH/LH pulse frequency is controlled by testicular steroids. Estradiol acts partly, at least, in the ARC, but the central site of action, testosterone remains unknown.

Concentrations of circulating progesterone and oestradiol were measured in 96 free-ranging, female Australian sea lions Neophoca cinerea from Kangaroo Island, South Australia. There was a marked increase in the concentrations of both hormones (progesterone from approximately 12 ng ml-1 to approximately 24 ng ml-1; oestradiol from approximately 1.5 pg ml-1 to approximately 14 pg ml-1) about 3.5 months after the probable date of mating, reaching peak values in the 5 months after parturition. Progesterone concentrations remained at peak concentrations for about 2 months, decreasing at approximately 8 months to concentrations approximating those of the first 3 months after parturition. Oestradiol concentrations decreased, after reaching a peak, to 3-4 pg ml-1 at about 8 months after parturition. The timing of the increase in the concentrations of circulating progesterone and oestradiol provides evidence that the blastocyst reactivates and implants between 3.5 and 5 months of pregnancy in Australian sea lions, indicating an embryonic diapause of similar duration to that of other pinnipeds. This would suggest a prolonged postimplantation period of up to 14 months (to fit with the gestation period of 18 months reported for this species) the longest postimplantation period recorded for pregnancy in any pinniped.

The equilibrium of the brain-pituitary-testicular axis is controlled by negative feedback exerted primarily through changes in the circulating concentrations of gonadal steroids. This is usually studied in gonadectomised animals treated with single large doses or constant low levels of exogenous steroid. However, the feedback system probably also contains dynamic components, perhaps expressed as delays to changes in GnRH secretion following a change in steroid concentration. These delays must be measured without interference from surgical procedures, including anaesthesia, bias associated with changes in pituitary responsiveness (which affect the efficiency of pulse detection), and chronic side-effects of gonadectomy. We used a GnRH antagonist ['Antarelix': Ac-D-Nal, D-Cpa, D-Pal, Ser, Tyr, D-Hci, Leu, Lys-(iPr), Pro, D-Ala-NH2] to transiently block LH and steroid secretion (in effect, inducing and reversing castration) in mature male sheep, and measured GnRH secretion into cerebrospinal fluid (CSF) in the third cerebral ventricle. The CSF was withdrawn with a peristaltic pump at a rate of 2 ml/h and pooled every 20 min. Jugular plasma was sampled every 20 min and analysed for testosterone and LH pulses. The antagonist (500 microg i.v.) was injected after 6 h of baseline sampling and the study continued for a further 24 h. The pulses of LH and testosterone disappeared shortly after antagonist injection, with delays of 20 +/- 12 min for LH and 80 +/- 29 min for testosterone. This led to an increase in GnRH pulse frequency, starting 300 +/- 54 min after antagonist injection. Secretion of LH and testosterone pulses resumed at 553 +/- 38 and 530 +/- 30 min (after antagonist injection), and GnRH pulse frequency returned to baseline values after 170 +/- 42 min (relative to LH) and 117 +/- 35 min (relative to testosterone). The consistent nature of these responses across the group of animals suggests that this can be used to test the effects of exteroceptive factors on the dynamics of negative feedback.

This study was designed to assess the effect of feeding level and dietary energy source on luteal function, systemic progesterone concentration and embryo survival in gilts during early gestation. At Day 0 of pregnancy, 104 gilts were allocated to one of four experimental diets (LStarch: 1.2 × maintenance requirement (M) Starch diet (43.3% starch), n = 31; HStarch: 2.4 × M Starch diet (43.3% starch), n = 21; HFat: 2.4 × M Fat diet (13.5% fat), n = 23; and HFibre: 2.4 × M Fibre diet (7.2% fibre), n = 23). On Day 5 of gestation, no significant difference in circulating concentration of systemic progesterone was seen among the treatments. However, on Day 15 of pregnancy, gilts on the HStarch diet had a significantly lower concentration of systemic progesterone than did gilts on both the LStarch and HFat diets (P < 0.05; 24.8 ± 2.4 v. 32.7 ± 2.4 and 36.1 ± 2.1 ng/mL, respectively). At Day 35 of gestation, there was also a tendency for gilts on the HStarch and HFat diets to have a higher total luteal weight than for gilts on the LStarch diets (7.2 ± 0.2 and 7.1 ± 0.2 v. 6.7 ± 0.2 g (P < 0.05)). No difference in embryo survival was seen among the treatments. From the present study, we can conclude that altering feeding level and dietary energy source did not affect embryo survival, despite the fact that systemic progesterone concentrations were affected on Day 15 of gestation. Also, luteal weight was greater for those gilts on the high feeding level than for those on the low feeding level when fed the same energy source.

P. Celi1*, G.B. Martin2, D. Blache2, P.E. Vercoe2, M. Blackberry2 and R.L. Tellam3 1University of Basilicata, Department of Animal Production Science, Potenza; 2T he University of Western Australia, Animal Science, Nedlands, WA, Australia; 3CSIRO, T ropical Agriculture Australia *Correspondence: Pietro Celi, University of Basilicata, Department of Animal Production Science, V ia N. Sauro, 85, 85100 Potenza, Italia E-mail: celi@unibas.it

An enzyme immunoassay with an anti-bovine-LH antibody (518B7) was applied to female western grey kanagaroos (Macropus fuliginosus ocydromus) and black-flanked rock wallabies (Petrogale lateralis lateralis). Validation showed parallelism to the assay standard curve, and significant increases in plasma LH concentrations after challenging animals with intramuscular GnRH.

Methods for the measurement of marsupial LH invariably rely upon the similarity of the LH molecule between different species and usually use anti-ovine or anti-bovine LH antibody and an ovine or bovine labelled LH preparation. Initial attempts to measure plasma LH in the Western Grey Kangaroo with assays using antibodies to 4 different isoforms of ovine LH raised in 7 different rabbits were unsuccessful. An enzymeimmunoassay (EIA) developed for the Asian elephant (Zoo Biology 23:45–63) was then applied to the Western Grey Kangaroo and the Black-flanked Rock Wallaby. This EIA has an anti-bovine-LH monoclonal antibody (518B7 provided by Dr Jan Roser, University of California, Davis, USA), biotinylated ovine LH label and bovine LH standard (NIADDK-oLH-26 and NIH-bLH-B10, both provided by Dr Janine Brown and Nicole Abbondanza, Smithsonian Institute, Front Royal, Virginia USA). Technical validation showed that serial dilution down to 1:8 of plasma from 7 individuals of each species showed parallelism to the assay standard curve, and control samples (1.24–5.30 ng/mL) had between-assay coefficients of variation 0.0005) were seen after GnRH for both the Western Grey Kangaroo (from 5.0 ± 0.8 ng/mL to 9.4 ± 1.2 ng/mL; n = 19) and the Black-flanked Rock Wallaby (from 6.0 ± 0.7 ng/mL to 10.6 ± 0.6 ng/mL; n = 28). In conclusion, this assay can be successfully used to measure LH in these two species.

Grazing saltbush reduces productivity in sheep mostly because the high salt intake decreases feed intake and challenges the metabolism of the animal. However, little is known of the effect of salt load on the endocrine control systems that regulate voluntary feed intake and metabolism. Plasma concentrations of leptin, insulin and cortisol and blood glucose were monitored in wethers fed for 2 weeks with either a control diet (adequate salt) fed ad libitum, a high salt diet (20% of dry matter) fed ad libitum or a group fed the control diet with an intake restricted to that of the high salt ad libitum group (control pair-fed). High salt intakes reduced voluntary feed intake within 1 day and circulating concentrations of insulin and glucose within 2 weeks. Liveweight and leptin concentrations were not specifically affected by the high intake of salt but decreased in response to the decrease in intake. Cortisol secretion was not affected. Although salt intake had a specific effect on insulin and glucose (over and above the effect of reduced feed intake alone), the reduction in insulin would be expected to increase rather than decrease appetite and feed intake. Therefore, insulin, leptin and cortisol do not appear to play major roles in the control of feed intake in sheep consuming high levels of salt.

Merino sheep developed in Mediterranean regions so are well adapted to acute changes in food availability. However, restricted intake during pregnancy, especially in animals that are pregnant over the dry summer, could limit the positive effects of a winter rainy season on fetal development. In this study, we tested whether the level of nutrition during pregnancy and during pre-pubertal development affected blood concentrations of insulin and leptin, scrotal circumference and age of puberty in male Merino lambs fed with pasture. During dry weather, pregnant sheep were supplemented ad libitum with hay and lupin grain (Lupinus angustifolius) to compensate for decreases in pasture supply. Puberty was detected using a standardised behavioural test with oestrous ewes. Lambs were considered pubertal if they displayed mounting in two successive weekly tests. There were no differences in plasma concentrations of insulin or leptin. The values for both hormones simply displayed the same pattern, with a rise after feeding and a fall during non-feeding periods. There was no difference among treatments in either scrotal growth or age to puberty (Table 1). This might be because the dietary treatments, being administered by food restriction under field conditions, would not have the same effects as severe undernutrition that has been used in laboratory studies. Alternatively, Merino sheep may have a greater capacity to cope with mild nutritional stress.

In Merino rams, an increase in the plane of nutrition increases the frequency of luteinising hormone (LH) pulses within a few days and this response is correlated with changes in the circulating concentrations of metabolic hormones. To analyse early dynamic aspects of these responses, we studied mature rams fed with diets that contained either low or high amounts of energy and protein. Jugular blood was sampled every 20 min for 96 h, including a control period of 24 h before the change of diet in the high-diet group. In the high-diet group, a significant increase in LH pulse frequency was first detected 6 h after the increase in nutrition on Day 1 and the frequency remained significantly elevated throughout the 72-h treatment period, except for a 12-h period on Day 2. Following the change of diet, insulin concentrations increased within 3 h and leptin concentrations increased within 7 h, after which time the concentrations of both hormones remained high. Dietary treatment did not affect the concentrations of thyroxine or insulin-like growth factor-I, but the high diet increased the concentrations of tri-iodothyronine. These observations are consistent with insulin and leptin playing a role in the early activation of the gonadotrophin-releasing hormone–LH axis by nutritional inputs.

In the present study, we tested the effect of treatment with a slow-release implant containing the gonadotrophin-releasing hormone agonist DeslorelinTM (Peptech Animal Health Australia, North Ryde, NSW, Australia) on pituitary and testicular function in mature male dogs. Four dogs were treated with Deslorelin (6-mg implant) and four were used as controls (blank implant). In control dogs, there were no significant changes over the 12 months of the study in plasma concentrations of luteinising hormone (LH) or testosterone, or in testicular volume, semen output or semen quality. In Deslorelin-treated dogs, plasma concentrations of LH and testosterone were undetectable after 21 and 27 days, testicular volume fell to 35% of pretreatment values after 14 weeks and no ejaculates could be obtained after 6 weeks. Concentrations returned to the detectable range for testosterone after 44 weeks and for LH after 51 weeks and both were within the normal range after 52 weeks. Semen characteristics had recovered completely by 60 weeks after implantation. At this time, the testes and prostate glands were similar histologically to those of control dogs. We conclude that a single slow-release implant containing 6 mg Deslorelin has potential as a long-term, reversible antifertility agent for male dogs.

Rams of a 'Mediterranean breed' (Merino) and a 'temperate breed' (Suffolk) were compared to determine how much of the differences between their reproductive seasons is owing to variation in their responses to photoperiodic and nutritional cues. In a previous study, both nutritional and photoperiodic inputs were held constant, and it was found that the two breeds show similar endogenous rhythms and, when the animals are challenged with a Mediterranean photoperiodic cycle, these endogenous rhythms are similarly modified. The present study tested whether an annual cycle in the supply of forage might modify the patterns that are generated by the interaction between photoperiod and endogenous rhythms. Both breeds were subjected to a simulated `Mediterranean' annual cycle in photoperiod (10L : 14D to 14D : 10L) and provided with either constant food supply or a simulated `Mediterranean' annual cycle in food supply. In Merino rams, testicular growth responded to photoperiod, but nutrition dominated those responses. In Suffolk rams, changes in testicular size can be completely out of phase with changes in body mass because they are driven primarily by photoperiod, with only subtle responses to changes in diet. The cycle of testicular growth in the Suffolk was driven by changes in the secretion of gonadotrophins (follicle-stimulating hormone concentrations and luteinizing hormone pulse frequency). By contrast, in the Merino, the nutritionally driven seasonal cycle of testicular growth was associated primarily with changes in body mass and this relationship could not always be explained by changes in gonadotrophin secretion. Melatonin secretion was not affected by food supply. Thus, the `Mediterranean' and `temperate' genotypes have similar endogenous rhythms that are similarly modified by photoperiod but, with respect to seasonal changes in nutrition, they differ in both the nature of their reproductive response and the physiological mechanisms that mediate those responses.

The aim of the present study was to determine whether leptin might play a role in the gonadotrophic response of mature merino rams to changes in the level of nutrition in rams fed ad libitum. Recombinant bovine leptin was infused intracerebroventricularly and voluntary food intake (VFI) and luteinizing hormone (LH) pulse frequency were measured. In Experiment 1, rams (n = 5) were infused for 24 h per day for 5 days with vehicle or with leptin (0.04, 0.4 and 4.0g h –1 ). All doses decreased both VFI and LH pulse frequency. In Experiment 2, rams were infused for 24 h per day for 5 days with vehicle (n = 10) or leptin (4 g h –1; n= 5); a sub-group of 5 controls was pair-fed to the leptin-infused group to control for effects of changes in feed intake. LH pulse frequency was reduced equally in both the leptin-infused and pair-fed groups. Leptin did not affect other systems controlled by the hypothalamic–pituitary axis. Thus, rather than stimulate LH secretion, intracerebral leptin specifically inhibits it by reducing food intake, so it is unlikely that effects of nutrition on the reproductive axis in mature rams involves leptin as a single blood-borne signal. A range of nutritional or metabolic inputs may be needed, and perhaps interconnections between neural centres that control appetite and reproduction.

Both testosterone and its aromatized metabolite, oestradiol-17beta, are known to act centrally on the secretion of GnRH, but the major site of aromatization is not clear as aromatase activities are found in numerous tissues including brain and testis. Here, we tested the importance of central aromatization of testosterone using a non-steroidal aromatase inhibitor, fadrozole. To distinguish between testicular and non-testicular sites, five intact and five testosterone-infused castrated rams (600 microg kg(-1) per 24 h for 3 days) were given four injections of fadrozole (i.m.; 500 microg kg(-1)) at 48, 52, 64 and 68 h relative to the start of testosterone infusion. Control rams (n = 5) received vehicle only. Fadrozole treatment decreased plasma oestradiol-17beta concentrations and increased the LH pulse frequency in both intact rams and testosterone-treated castrates, suggesting that non-testicular sites of aromatization are important in the control of pulsatile LH secretion. To test the importance of central aromatization, intact rams (n = 5) were infused into the third ventricle with vehicle (artificial cerebrospinal fluid) or with fadrozole (20 and 200 microg kg(-1) per day). After two weeks, the same two doses of fadrozole were infused intravenously instead of intracerebrally. Central infusion of fadrozole did not affect plasma oestradiol concentrations but increased LH pulse frequency. Only the highest dose increased LH pulse frequency when infused intravenously. In conclusion, central aromatization is involved in the control of pulsatile LH secretion in male sheep.

This study tested whether the effects of nutrition on gonadotrophin secretion and testicular growth in mature rams are due to increases in the supply of glucose, fatty acids (FA) or amino acids. Responses to protein (casein) and glucose, alone or in combination, were compared with responses to lupin grain and responses to a combination of protein, glucose and FA (acetate, propionate and vegetable oil). Glucose and casein were infused intra-abomasally whereas lupins and FA were added to the diet. Lupin feeding decreased blood growth hormone (GH) concentrations, but increased pulsatile luteinizing hormone (LH) secretion and increased the concentrations of follicle-stimulating hormone (FSH), prolactin, glucose and insulin. These effects were associated with testicular growth. Glucose or casein increased insulin concentrations and decreased GH concentrations, but did not affect gonadotrophins or testicular growth. There was no synergism between casein and glucose. Responses elicited by adding FA to the glucose+casein treatment were similar to those observed with lupins. In conclusion, the reproductive axis does not seem to be closely linked with dietary intakes of amino acids or with circulating concentrations of glucose, insulin or GH. However, the energetic components of the diet, particularly the fatty acids, appear to play a key role in the reproductive responses to changes in nutrition.

In small ruminants, reproductive wastage due to early embryo mortality is a major industry issue because it reduces reproductive efficiency and limits productivity. In sheep, early embryo mortality appears to be caused by reductions in progesterone concentrations when animals are over-fed, but this concept has not been studied in goats. Therefore we tested whether a supplement of lupin grain affects circulating progesterone concentrations in Cashmere goats during non- breeding season. We allocated 23 females into two groups: Controls were fed to ensure maintenance of body mass (85% chaff, 15% lupins head daily); Supplemented goats were fed twice their daily requirements for maintenance. All animals were anovulatory and treated with CIDRs to supply exogenous progesterone at a relatively constant rate. Nutritional treatments lasted for 18 days, and coincided with the presence of CIDRs. Leptin and insulin concentrations were increased (p < 0.05) by supplementation, but progesterone concentrations did not significant differ between groups at any time during the experiment. We conclude that a dietary supplement that elicits major changes in energy homeostasis does not reduce progesterone concentrations in goats and is thus unlikely to affect embryo mortality.

Growth hormone (GH), thyroxine (T4) and tri-iodothyronine (T3) are known to be involved in the regulation of growth and development in a variety of avian species. It has been suggested that an absence of GH and thyroid hormones in ostriches is the cause of their neoteny, a phenomenon in which juvenile characteristics are retained into adulthood. Neoteny is typical of all ratites, the single group of flightless birds that includes the ostrich, but similar endocrine studies have not been performed for other members of the group, such as the emu. To test the neoteny hypothesis further, in the present study we measured the plasma concentrations of T4, T3 and GH in emus during embryonic development and from hatching to 1 year of age. Concentrations of T4 and GH increased during the last weeks of incubation, whereas concentrations of T3 were highly variable. After hatching, the concentrations of both thyroid hormones were high during the first 3 days of life and then fell to a constant low level. Plasma concentrations of GH were high at the time of hatching and decreased gradually over the first 22 weeks of age; thereafter, the concentrations of GH were highly variable. No correlation was observed between hormone concentrations and live weight at any time. These results support the hypothesis that thyroid function is abnormally low in ratites, whereas patterns of GH secretion are similar to those observed in other birds. Dysfunction of the thyroid axis could explain, in part, the neotenous physical aspect of adult emus.

Metabolic status is a powerful regulator of reproductive activity, but the metabolic mediators involved and the relationships between fat reserves, food intake and the systems that control reproduction are not fully understood. In this study with mature male Merino sheep, we tested whether the effect of an acute nutritional stimulus on pulsatile LH secretion depended on body condition. Two groups of rams ("Fat" and "Lean") were fed differentially for 4 months to achieve high or low levels of body mass and body condition score. Half of each group was then assigned to be fed either their maintenance requirement or twice their maintenance requirement and, 7 days later, plasma samples were collected every 20 min for 24 h. All samples were used for the analysis of LH pulses and pooled samples were used for the measurement of metabolic hormone concentrations. In the rams that were fed the maintenance diet, the frequency of LH pulses was similar for the Fat and Lean groups, but plasma concentrations of leptin and insulin were significantly higher in the Fat group than in the Lean group. Following an acute increase in food intake, plasma concentrations of insulin were significantly increased in both Fat and Lean rams, but plasma leptin concentrations were increased only in Fat rams and LH pulse frequency was increased only in Lean rams. We concluded that the secretion of LH and leptin, but not insulin, is differentially influenced by nutritional status and body condition and that the role of leptin in the central regulation of the GnRH-LH system is probably permissive.