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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.

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.