Effects of testosterone on pedicle formation and its transformation to antler in castrated male, freemartin and normal female red deer (Cervus elaphus).

Pedicles and antlers are male deer secondary sexual characters. As such, development of these structures is under the control of androgen hormones. Pedicle growth is caused by increasing and elevated plasma testosterone (T) levels, whereas first antler transformation from a fully formed pedicle occurs when the T levels are decreasing. Castration prior to pedicle initiation abrogates future pedicle and antler formation. Female deer also have the potential to develop pedicles and antlers, but they do not normally express this phenotype due to lack of sufficient androgen stimulation. Previous studies have shown that female white-tailed deer could be readily induced to grow pedicles as well as antlers by singular administration of exogenous androgens (EA), but in red deer (Cervus elaphus) singular or irregular EA treatment could only stimulate castrated male, normal or ovariectomised females to grow pedicles, but not antlers. The present study was set out to test whether these EA-induced pedicles in red deer failed to give rise to antlers was because they were constitutively incapable of doing so, or because the plasma T profile naturally exhibited in intact stags was not achieved by the androgen treatment used in these previous studies. Eight castrated red deer stag calves, 3 freemartins (females which were born co-twin to males), and 3 normal female red deer were used in the present study and treated with EA, either as biweekly injections for the castrates or as implants for freemartin and females until the late stage of pedicle growth. Blood sampling was carried out biweekly for the analyses of plasma T and IGF1 concentration. The results showed that the natural plasma T profile in the experimental deer was successfully mimicked through regular EA treatment and subsequent withdrawal at late pedicle growth stage. All castrated males, 2 out of 3 freemartin, and 1 out of 3 normal female red deer formed not only pedicles, but also antlers. Based on these results, we conclude that EA-induced pedicles at least in red deer of the genus Cervus, like those in the genus Odocoileus, are constitutively capable of giving rise to antlers, if they are of sufficient height.

Effects of photoperiod on the cessation of growth during autumn in male red deer and growth hormone and insulin-like growth factor-I secretion.

Male red deer undergo seasonal cycles of food intake and growth rate, which are high during spring and low during winter, despite high quality food ad libitum. Hormonal profiles during the cessation of growth in autumn and the potential role of photoperiod in the timing of the observed changes have been investigated. Whether this seasonal decrease in growth affected the response of GH and IGF-I to fasting was also examined. Two groups of six male 1-year-old red deer were exposed to different photoperiods after the summer solstice. One group (C) was given a simulated natural photoperiod while the other group (SS) was maintained on a summer solstice photoperiod (16L:8D). GH was measured in blood collected continuously and divided into pools every 5 min for 24 h in the fed state and after a 48-h fast on two occasions; the first was in November before photoperiod manipulation began and the second was in April approximately 16 weeks after initiating treatments. IGF-I, prolactin, and testosterone were measured in weekly samples. Individual live weight and group food intake were also measured each week. The normal growth pattern seen in the C group was delayed in the SS group. Thus, from 7 March until the second GH sampling on 11 April the live weight of deer in group C fell; in contrast, deer in group SS continued to grow (-43 vs 186 g/day s.e.d. = 65.5, P < 0. 01). Food intake changes reflected the pattern of growth in both groups. Mean GH (P < 0.05), GH pulse amplitude (P < 0.01), and IGF-I (P < 0.001) declined in both groups from November to April. This decline was more marked in group C and in April these parameters were all lower in group C than in group SS (GH, P < 0.05; IGF-1, P < 0.01). Prolactin levels in April were also lower in group C than in group SS (P < 0.01); testosterone was not affected by treatment. Fasting increased mean GH and GH pulse amplitude in both groups in November (P < 0.05). In April, the fasting response differed between the groups. In group C, mean GH, pulse amplitude, and pulse frequency were all greater in the fasted state than in the fed state (P < 0.05), while in group SS there were no significant differences (P > 0.05). IGF-I was lower in the fasted state than in the fed state at both sampling dates (P < 0.001). The seasonal decline in food intake and growth is associated with decreased GH, IGF-I, and prolactin concentrations, and increased testosterone and the GH response associated with fasting. All these changes except those of testosterone were delayed or reduced by continued exposure to a summer solstice photoperiod in autumn. The decreased photoperiod in autumn may thus influence the normal timing of the seasonal growth cycle.

Effects of season, protein and nutritional state on glucose tolerance during an annual cycle of growth in young red deer stags.

Two hypotheses were tested in gonad-intact, young (aged 6-18 months), growing red deer stags during an annual growth cycle. First, that glucose clearance rate is faster during summer than during winter. Secondly, that increased dietary protein availability will enhance winter growth. Stags were randomly assigned into one of two groups: group 1 (n = 5) had 16% while group 2 (n = 6) had 48% of dietary protein naturally protected against fermentative degradation in the rumen. Total crude protein and energy remained similar for each diet (12 and 14% respectively for protein and 11 MJ metabolisable energy/kg dry matter). Stags were kept indoors in individual pens for 12 months and given monthly intravenous glucose tolerance tests (IVGTT), at a dose of 200 mg/kg, in the fed and fasted (48 h) states to determine both growth and steady-state tissue requirements. Protein level had no effect on food intake, weight gain, insulin kinetics, or glucose clearance rate. In the fed state, insulin peak (highest level' after IVGTT) increased (P < 0.01) from October (139 pmol/l) to December (247 pmol/l) (S.E.D. = 42) and remained elevated during the summer, before declining (P < 0.01) from February (223 pmol/l) to April (130 pmol/l) (S.E.D. = 25). Glucose clearance rate was faster (P < 0.05) in December (1.69 litres/min) than June (0.61 litres/min) in the fed state (S.E.D. = 0.30), and decreased (P < 0.05) from February (1.75 litres/min) to April (0.92 litres/min) (S.E.D. = 0.39). During fasting, the pattern of glucose clearance was similar to that observed in the fed state, but the amplitude was lower, while the pattern for insulin peak was similar to that of the fed state. We concluded first, that additional protected protein does not benefit growth during winter. Secondly, we concluded from the fasted, steady-state data that stags are insulin resistant during summer. Thirdly, despite insulin resistance, data on the fed state demonstrated that stags have higher tissue energy requirements during summer growth.

Prolactin does not enhance glucose-stimulated insulin release in red deer stags.

Red deer stags have a seasonal pattern of insulin secretion that is characterized by both elevated basal and glucose-stimulated insulin release in summer compared with winter. Since the seasonal timing of this pattern is similar to that of prolactin and growth rate, the objectives of this study were: first, to determine whether prolactin is associated with the enhanced secretion of insulin during the summer growth period, and second, to determine whether a chronic reduction in plasma prolactin levels would alter body composition. Prolactin was suppressed in plasma using a long-acting form of the dopamine agonist bromocriptine (parlodel LA), which was administered at one of four doses (0-0.3 mg/kg) to each of four groups of castrate stags. Bromocriptine was administered during two 6-wk periods; the first in winter and the second in summer. During the sixth wk of each period, each animal was given three IVGTT at the following glucose doses (10 mg/kg, 70 mg/kg, and 200 mg/kg). Two d later, ovine prolactin was administered to each animal (0.08 mg/kg) and a single IVGTT (70 mg/kg) was given 2 hr later. Body composition was determined by the tritriated water dilution method at the beginning and end of each 6-wk treatment. Chronic suppression of prolactin during winter or summer did not significantly alter the amount of insulin released after each IVGTT, nor did it significantly alter body composition. Furthermore, acute administration of prolactin did not significantly enhance the release of insulin following an IVGTT, during winter or summer treatment periods. It is concluded that elevated levels of prolactin in summer do not enhance the release of insulin to glucose in red deer. Furthermore, a reduction in growth rate following a reduction in plasma prolactin is not associated with a change in body composition.

Effects of glucose or insulin infusions on growth hormone secretion in male red deer.

The growth hormone (GH) secretory pattern in male red deer is associated with the seasonal growth cycle. During this cycle metabolic state changes from weight gain in spring to weight loss in winter. However, short-term metabolic changes due to feeding could also alter the GH pattern. To investigate the effect of such changes on GH secretion, the acute feedback of blood glucose level on the GH secretory pattern was examined. Six yearling male red deer were infused iv with glucose (G; 150 mg/kg/hr) or insulin (I; 30 mU/kg/hr) for a 12-hr period, 1 week apart. GH was measured in jugular venous blood every 10 min, for 12 hr before, during, and 6 hr after the infusions. Glucose, insulin, IGF-1, and haematocrit were also measured. There was no difference (P > 0.05) in glucose levels between G and I prior to infusions (5.8 vs 6.0 mmol/liter, SED = 0.42). Glucose levels rose to 8.7 mmol/liter during G and fell to 3.4 mmol/liter (SED = 0.72, P <0.001) during I, then returned towards normal postinfusion. Insulin levels increased during G and I (P < 0.01) with no difference (P > 0.05) between G and I during preinfusion (163 +/- 7.6 pmol/liter) or infusion (259 vs 264 +/- 16. 5 pmol/liter) periods. There were no differences (P > 0.05) in GH secretory characteristics, mean IGF-1, or haematocrit between G and I. However, there were significant effects of infusion within the treatments. Mean GH declined (P < 0.05) from 1.8 ng/ml (both treatments) preinfusion to 1.13 and 1.31 ng/ml during G and I infusion, respectively. GH pulse amplitude was lower during I infusion (5.6 ng/ml vs 8.2 ng/ml preinfusion, P < 0.05, SED = 1.0) and the change in amplitude from preinfusion to infusion differed (P > 0.05) with an increase in G and a decrease in I (+0.6 and -2.6, SED = 1.1). IGF-1 levels were stable and averaged 555 and 520 ng/ml (SED = 34.9) for G and I, respectively. Haematocrit declined from 34.3 +/- 1.85% over the first 4 hr of sampling to 25.7 +/- 0.97% for the remainder of the sampling period. The finding that there were no major alterations in GH secretory patterns during 12 hr of hypoglycemia and hyperglycemia suggests that GH secretion in the male red deer is relatively insensitive to short-term changes in metabolic state.

Effects of season and nutrition on growth hormone and insulin-like growth factor-I in male red deer.

GH and insulin-like growth factor (IGF)-I are important components of the growth axis. We undertook to determine how plasma levels of these hormones altered with different seasonal and nutritional states in young male red deer to provide an insight into how the growth axis changes under these conditions. Growth rate alters dramatically with season in male red deer, providing an opportunity to sample the same animals at two different growth rates within a short period of time. GH was measured every 15 min for 24 h in the fed state and after a 48-h fast, during slow growth in winter (23 June to 16 July), and during rapid growth in spring (8 September to 2 October). At the end of each sampling period, the animals were treated with N-methyl-D, L-aspartic acid (NMDA) (5 mg/kg live weight) and sampled for a further 1 h, 45 min. Glucose and IGF-I were measured hourly during each sampling period. Live weight was measured at weekly intervals. GH was secreted in a characteristic pattern in which pulses tended to occur in rapid succession, termed a volley, that was separated from the subsequent volley by a period of baseline GH levels, termed a latent period. There were more GH pulses/24 h in the fasted state than in the fed state in winter (12.4 vs. 7.8, standard error of the difference [SED] = 1.07, P < 0.001) and in spring (11.5 vs. 8.8, SED = 1.04, P < 0.05). The increased number of GH pulses in the fasted state could be attributed to a higher number of pulses per volley (winter = 3.7 vs. 2.5, SED = 0.16, P < 0.001; spring = 3.1 vs. 2.8, SED = 0.19). Consequently, the volleys were wider in the fasted state than the fed state (winter = 197 min vs. 122 min, SED = 25, P < 0.05; spring = 173 min vs. 154 min, SED = 24, P > 0.05), and the latent periods between volleys were shorter in the fasted state than the fed state (winter = 175 min vs. 280 min, SED = 14, P < 0.001; spring = 183 min vs. 262 min, SED = 11, P < 0.001). The main differences between seasons in the fed state were larger amplitude pulses (12.4 vs. 8.3 ng/ml, SED = 1.57, P < 0.05) and higher mean GH concentrations (4.1 vs. 2.3 ng/ml, SED = 0.44, P < 0.01) in spring than in winter. The number of volleys and the intravolley pulse interval did not change significantly with nutritional state or season. NMDA administration was followed by an increase in GH with higher GH levels found in the fed state than in the fasted state in both seasons. Fed animals also had a larger initial increase in GH (until 60 min post NMDA) than fasted animals in spring (P < 0.01). Plasma IGF-I was higher in the fed state than the fasted state in both winter (315 vs. 221 ng/ml, SED = 21.0, P < 0.001) and spring (651 vs. 494 ng/ml, SED = 37.5 P < 0.001) and in the fed state was higher in spring than in winter (SED = 29.1, P < 0.001). Blood glucose was higher in the fed state than fasted state in winter (6.1 vs. 5.5 mmol/l, SED = 0.07, P < 0.001) and there was a strong trend toward this same effect in spring although it did not reach statistical significance (6.0 vs. 5.7 mmol/l, SED = 0.26, P > 0.05). Growth rate in winter at 117 g/day was less than that in spring when 220 g/day was recorded (SED = 36.8, P < 0.05). These results demonstrate that the secretory pattern of GH and plasma IGF-I levels alter in response to changes in season and nutrition. The alterations in response to a 48-h fast show that the control of GH and IGF-I secretion may be rapid and is probably a response to maintain energy balance, whereas alterations with season reflect long term control that underlies the seasonal growth pattern of the animal.

Histogenesis of antlerogenic tissues cultivated in diffusion chambers in vivo in red deer (Cervus elaphus).

In a previous study we showed that formation of deer pedicle and first antler proceeded through four ossification pattern change stages: intramembranous, transition, pedicle endochondral, and antler endochondral. In the present study antlerogenic tissues (antlerogenic periosteum, apical periosteum/perichondrium, and apical perichondrial of pedicle and antler) taken from four developmental stages were cultivated in diffusion chambers in vivo as autografts for 42-68 days. The results showed that all the cultivated tissues without exception formed trabecular bone de novo, irrespective of whether they were forming osseous, osseocartilaginous, or cartilaginous tissue at the time of initial implant surgery; in two cases in the apical perichondria from antler group, avascularized cartilage also formed. Therefore, the antlerogenic cells, like the progenitor cells of somatic secondary type cartilage, have a tendency to differentiate into osteoblasts and then form trabecular bone. Consequently, the differentiation pathway whereby antlerogenic cells change from forming osteoblasts to forming chondroblasts during pedicle formation is caused by extrinsic factors. Both oxygen tension and mechanical pressure are postulated to be the factors that cause this alteration of the differentiation pathway.

Role of steroids in antler growth of red deer stags.

A series of six studies were carried out in red deer stags to test hypotheses concerning the importance of steroid control of velvet antler growth and to investigate mechanisms by which these hormones exert their effects. Medroxyprogesterone acetate (MPA) an LH inhibitor administered to stags during hard antler caused premature antler casting, reduced subsequent antler weight and caused a reduction in the LH and testosterone responses to GnRH. In two separate studies blockade of testosterone receptors with cyproterone acetate (CPA) administered to stags, either during early velvet antler growth or during the hard antler stage, significantly reduced LH and testosterone responses to GnRH. In both studies antler length, but not weight, was increased by CPA treatment. In another study testosterone implants were used to prevent the gradual decline in plasma testosterone levels normally observed during winter. Implants were removed 3 weeks before the anticipated date of antler casting. The implants significantly increased plasma testosterone levels and subsequent antler growth (expressed as a proportional increase compared with the previous year) compared with untreated controls. To determine whether the annual cycle of plasma testosterone response following GnRH stimulation was due simply to a lack of LH stimulation, ovine LH was injected on six occasions at defined stages of the antler cycle to red deer stags and the testosterone response measured. The testosterone responses were low at antler casting and during velvet antler growth compared with antler cleaning and peak rut. It appears low testosterone levels are due, in part, to a loss of responsiveness by the testes to LH as well as a low level of secretion of LH during the antler growing season. Finally synthetic ACTH was injected at the same defined stages of antler growth as in the previous study to determine whether cortisol and adrenal androgen production altered with the stage of the antler cycle. No significant differences were found in the dehydroepiandrosterone (DHEA) response, but cortisol responses were higher from late velvet antler growth to peak rut, compared with the times of antler casting and early velvet growth. Overall it was concluded that velvet antler growth can occur without testosterone stimulation during the period of velvet growth, but the data reinforce the concept that the timing of antler growth is linked to the annual cycle of testosterone.

Effects of unilateral cranial sympathectomy either alone or with sensory nerve sectioning on pedicle growth in red deer (Cervus elaphus).

In a previous study (Li et al. [1993], J. Exp. Zool., 267:188-197) sensory nerve sectioning had no effect on the timing of pedicle growth. The aim of the present study was to determine whether sensory nerve sectioning in conjunction with sympathectomy would influence pedicle growth. Twelve intact male red deer calves were allocated to treatment before any pedicle growth as follows: 1) unilateral sensory nerve removal (USX, n = 5), 2) unilateral superior cervical ganglionectomy (SGX, n = 4), or 3) both USX and SGX (SG/USX, n = 3). The calves were observed weekly. In all cases the untreated side was the control. Pedicle initiation was measured with a pedicle detector and after initiation, growth was measured with a ruler. When the treated pedicles reached a length of 60 mm the calves were killed and tissues from the pedicle were examined immunohistochemically for nerves. No large bundles of nerves were observed in the treated pedicle although a few fine fibres were present. All calves grew pedicles. There were no significant differences in the timing of pedicle initiation either within treatment or between treatments. All denervated pedicles grew faster than controls and were consequently higher at examination. The fact that pedicle growth took place despite reduced innervation indicates that a continuous neural connection is not a pre-requisite for normal pedicle growth.

Pedicle and antler development following sectioning of the sensory nerves to the antlerogenic region of red deer (Cervus elaphus).

Sensory nerves supplying the deer antlerogenic region were sectioned about 60 days prior to pedicle initiation to determine the extent of neural influence on pedicle and first antler growth. Our results from a combination of histological examination and immunohistochemical localization showed that all 12 antlerogenic regions were successfully deprived of sensory nerve supply, but in 10 of 12 cases there was partial regeneration during the experimental period. In the two cases where no sensory reinnervation occurred, pedicle growth did not show any differences compared with partially sensory reinnervated or intact pedicles. With or without reduced sensory nerve supply, first antlers were initiated, grown, cleaned of velvet, cast, and regenerated in the normal way, but they were smaller than controls. Consequently, we conclude that a sensory nerve supply is not necessary for normal pedicle formation and for the first antler cycle, but plays a role in determining antler size.

Seasonal pattern of luteinizing hormone and testosterone pulsatile secretion in young adult red deer stags (Cervus elaphus) and its association with the antler cycle.

Blood from stages aged 15 months (n = 6) was sampled at monthly intervals every 30 min for 24 h for 12 months, at 45 degrees S in New Zealand. Three extra samplings each for 24 h were carried out at about the anticipated time of antler casting. All samples were analysed for luteinizing hormone (LH) and testosterone and the resulting data further analysed by the Pulsar pulse detection routine. The animals were kept indoors under natural daylength and were fed ad libitum. All animals were weighed, antler status and size recorded and testes diameter was measured on each sampling day. Mean LH and testosterone pulsatily and plasma concentration varied seasonally. LH pulse frequency was low during autumn (2.5 pulses in 24 h), winter (1.0-1.5 pulses in 24 h) and early spring (1 pulse in 24 h) and lowest in late spring (0.2 pulse in 24 h) before rising in summer (1.0-4.0 pulses in 24 h). LH pulse amplitude and mean plasma concentration were low (< 1 ng ml-1) from March to November (autumn-spring); both rose to a peak in January (summer) of 3.4 and 1.6 ng ml-1, respectively. Testosterone pulse frequency was generally similar to LH except that slightly more pulses of testosterone than of LH were detected from March to November and more pulses of LH from November to February (summer). Testosterone pulse amplitude fell from March to November (5.3 ng ml-1 to undetectable) although there was a conspicuous peak in July (midwinter) of almost 5 ng ml-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Behavioural and heart rate responses to velvet antler removal in red deer.

Heart rate and behaviour during and following velvet antler removal were monitored in yearling red deer stags to determine the extent to which this procedure was perceived by the deer to be aversive. Nine stags normally kept at pasture were habituated over 5 weeks to the following daily handling procedure. Each deer was fitted with a harness containing a heart rate monitor. It was then allowed to run through a fixed course in a deer yard, restrained for 40 s in a mechanical deer crush, and then confined for 3.5 h with the remainder of the group of stags in an indoor pen containing food and water. In Week 6, the deer were subjected to either restraint for 6 minutes (the control treatment) or removal of one velvet antler under local anaesthesia. Each velvet antler was removed on separate occasions, either on Days 1 and 2 (five deer) or Days 3 and 4 (four deer). The control treatment was applied to all deer when velvet antler was not being removed, and on Day 5. Heart rate and behaviour (time taken to enter the treatment area, and number of struggles made during restraint) were measured before and during treatment, and post-treatment activities were recorded at 0, 1 and 3 h (indoors), and at 6 and 9 h (at pasture). Heart rate was higher during the second velvet antler removal treatment than during the first, but lower during the second control treatment than the first (P<0.05). During velvet antler removal, stags struggled more, and after the treatment flicked their ears, shook their heads, and groomed themselves more than control stags (P<0.05). Stags whose velvet antler had been removed spent less time eating than control stags, and spent progressively more time sitting during the 3.5 h of confinement (P<0.05). However, during the paddock observation at 9 h post-treatment, stags which had had their velvet antler removed grazed more than control stags (P<0.05). The increase in heart rate over the two velvet antler removal treatments and the greater amount of struggling during velvet antler removal indicated that it was more aversive than the control treatment. Post-treatment differences in behaviour may have been due to pain following velvet antler removal.

Temporal changes in LH and testosterone and their relationship with the first antler in red deer (Cervus elaphus) stags from 3 to 15 months of age.

Blood samples were taken from six tame red deer stags at 3-15 months of age once a month from a jugular catheter every 30 min for 24 h to investigate hormonal secretion during puberty and during growth of the pedicle and first antler. All plasma samples were analysed for LH and testosterone concentrations and the resultant data were analysed using the PULSAR pulse detection routine. In addition each stag was injected wih gonadotrophin-releasing hormone (GnRH; 20 ng/kg body weight) after the above samples had been taken and the bleeding regimen was continued for a further 2 h. Body weight, antler size and status (i.e. whether the stags had a pedicle or antler) were also recorded. The pulsatile secretion of LH could be considered to have occurred in three phases. The first of these was one of development, with the LH pulse frequency increasing to 8 pulses/24 h, the second a phase of regression, with a decrease in LH pulse frequency to 2 pulses/24 h, and finally a second phase of development characterized by increased LH pulse frequency to 12 pulses/24 h. Testosterone secretion generally followed the same pattern. During the period before the permanent bony pedicles grew, there were less than five LH pulses/24 h. When the pedicles were growing, LH and testosterone pulsatile secretion increased but the pulse frequency of both hormones fell during velvet antler growth.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of melatonin implants on reproductive seasonality of male red deer (Cervus elaphus).

Red deer stags were treated with melatonin implants in 2 experiments designed to examine the control of reproductive seasonality. In Exp. 1, stags (n = 24) were allocated to 4 treatment groups: 2 groups were treated with 3 implants per stag each month from 8 November to 5 February (EM) or 9 December to 5 February (LM), 1 untreated group of control stags remained with the melatonin-treated stags (CC) and the other untreated control group remained isolated (IC). Melatonin treatment advanced the seasonal changes in scrotal circumference, liveweight, antler state and coat type compared with control stags. The extent of advancement was greater in EM than LM stags. In EM and LM stags, size of testes regressed rapidly and antlers were cast shortly after melatonin implants became exhausted in March. This was followed by an additional antler cycle and reproductive development and decline from June to November. EM and LM stags became synchronized with control stags 14-15 months after melatonin treatment began. The extra cycle of seasonal changes was more pronounced in EM than in LM stags. In Exp. 2, stags (n = 30) were allocated to 6 treatment groups: 4 groups were treated with 3 implants per stag at monthly intervals for 6 months from 22 June (J), 4 August (A), 16 September (S) and 23 October (O), a further group of stags was treated in the same manner for 12 months from 22 June (Y), and the remaining group was untreated (C). Compared with control stags, testicular regression and antler casting was delayed in Groups J, A and Y. These events occurred at the same time as in control stags in Groups S and O. Subsequent reproductive development was advanced in Groups S and O and delayed in Groups J, A and Y. The results demonstrated that treatment with melatonin implants in November or December advanced reproductive development. However, when stags were treated with melatonin implants from June to August, reproductive development was delayed, indicating a change in response to melatonin treatment during the year. The change in response to melatonin treatment between late winter and early spring was interpreted as a resetting of an endogenous circannual rhythm caused by a photoperiodic cue responsible for initiating the final stages of reproductive regression.

Influence of food intake but independence of body weight on puberty in female sheep.

The effect of maintaining female sheep at a body weight intermediate between the normal weight for puberty (30-35 kg) and 20 kg (puberty suppressed) on the onset of oestrous cycles was studied. In addition, the influence of ad-libitum food intake or insulin infusion was studied in animals previously maintained at 20 kg. Coopworth ewe lambs (10 weeks old) were allocated to one of 6 treatments: (A) ad-libitum fed (n = 6), (B) ad-libitum fed to 28 kg then maintained at that weight (n = 6), (C) ad-libitum fed to 24 kg then maintained at that weight (n = 6), (D) maintained at 20 kg until Week 29 and then fed ad libitum (n = 6), (E) maintained at 20 kg and infused with 0.1 U insulin/kg/24 h for 2 weeks from 29-31 weeks of age (n = 5), (F) maintained at 20 kg (n = 6). The lambs were penned indoors under natural photo-period, which was decreasing virtually throughout the study, and fed a pelleted concentrate diet which was recorded daily. They were blood sampled twice a week, and plasma was analysed for progesterone. Puberty was defined as the date when plasma concentrations of progesterone first exceeded 1 ng/ml. In addition, ewes in Groups D, E and F were blood sampled every 10 min for 8 h on Days 0 and +12 of the insulin infusion or access to ad-libitum feeding and the plasma was analysed for luteinizing hormone (LH).(ABSTRACT TRUNCATED AT 250 WORDS)

Pulsatile growth hormone, insulin-like growth factors and antler development in red deer (Cervus elaphus scoticus) stags.

Plasma samples taken every 30 min over a 26-h period each month from six 4- to 15-month-old red deer stags were analysed for GH. In addition, two samples taken at 10.00 and 22.00 h were analysed for insulin-like growth factor-I (IGF-I) and insulin-like growth factor-II (IGF-II). A concentrate diet was available ad libitum. Food intake, body weight and antler status were recorded. Concentrations of GH were analysed using the PULSAR peak detection routine. Secretion of GH was pulsatile in every month of sampling, but the pattern of pulsatility differed seasonally. During the autumn and early winter (April-June in the Southern hemisphere) GH pulses were frequent and of low amplitude. In contrast, GH pulses in spring (August-September) were of high amplitude and high frequency resulting in a high mean level of GH circulating in the plasma. In early summer (November) the GH pulse amplitude was much lower and pulse frequency fell. There was a rise in GH pulse frequency not accompanied by an increase in GH pulse amplitude in summer (December-January). GH pulse amplitude seemed to be the main determinant of mean GH plasma level. Secretion of IGF-I was raised 1 month after peak monthly mean GH secretion. There was little consistent relationship between concentrations of IGF-II and mean daily GH. Concentrations of GH correlated positively and significantly with liveweight gain and antler growth rate with a delay of 1 month. Significantly positive correlations between concentrations of IGF-I, liveweight gain and antler growth rate were observed. It is considered that the spring and summer (September-December) seasonal acceleration of liveweight gain and antler development in stags could be a consequence of high winter/early spring (August-September) GH pulse frequency and amplitude resulting in increased concentrations of IGF-I, particularly in October.

LH and testosterone responses to GnRH in red deer (Cervus elaphus) stags kept in a manipulated photoperiod.

Six red deer stags from age 4 months were kept in a light-proof room under an artificial photoperiod consisting of 5.5 cycles of alternate 2-month periods of 16 h light and 8 h dark (16L:8D) and 8L:16D. At 2 or 3 weekly intervals from 10 months of age through 4 cycles, the stags were anaesthetized with xylazine and challenged i.v. with 10 micrograms GnRH. Blood samples were withdrawn immediately before and 10 and 60 min after injection. LH and testosterone concentrations were measured in all samples by RIA. Antler status was recorded daily. Peak LH values on each sampling day occurred in the sample taken 10 min after GnRH stimulation while peak testosterone occurred in the sample taken at 60 min. There were 4 cycles of LH and testosterone secretion accompanied by 4 antler cycles in the stags. The highest LH responses were detected during short days (8L:16D), and the highest testosterone responses were detected around the time of the change from short to long days. The responses of both hormones were lowest at the end of periods of long days or the beginning of short days. The increased pituitary LH response to GnRH was evident 4 weeks after the change to short days which are stimulatory for gonadal development. Antler casting occurred at the end of long days and cleaning at the end of short days. It is considered that antler cycles were due to the ability of the stags to vary release of LH and testosterone in response to changes in the artificial photoperiod.

Elevated plasma IGF 1 levels in stags prevented from growing antlers.

We have previously shown that plasma IGF 1 concentration is positively correlated with the rate of antler growth and have proposed that IGF 1 is stimulatory for antler growth in the red deer stag. Therefore to partly resolve the question of whether the IGF 1 was of circulating or local origin in relation to its effect on antler growth, we surgically prevented stags from growing antlers. We recorded significantly elevated plasma levels of IGF 1 in the non-antlered stags compared with normal antlered stags during the antler growth periods. This result is consistent with a hypothesis that the antler is a target organ for IGF 1 and that prevention of antler growth removed a population of IGF 1 receptors. IGF 1 is already known to stimulate body growth but this work points strongly to the possibility that plasma IGF 1 may stimulate individual organ growth in an endocrine manner.

Plasma LH and testosterone responses to gonadotrophin-releasing hormone in adult red deer (Cervus elaphus) stags during the annual antler cycle.

Eight adult red deer stags were given an i.v. injection of synthetic gonadotrophin-releasing hormone (GnRH) on seven occasions at various stages of the antler cycle, namely hard antler in late winter, casting, mid-velvet growth, full velvet growth, antler cleaning and hard antler both during the rut and in mid-winter. The stags were allocated at random on each occasion to one of four doses, i.e. 1, 3, 10 or 95 micrograms GnRH. Blood samples were taken before GnRH injection and for up to 2 h after injection. Pituitary and testicular responses were recorded in terms of plasma LH and testosterone concentrations. There was an increase in plasma concentration of LH after the GnRH injection in all stags at all stages of the antler cycle. Dose-dependent responses of LH to GnRH in terms of area under the curve were apparent at all stages of the antler cycle. The lowest responses were recorded at casting, during velvet antler growth and at the rut sampling. The pattern of testosterone response reflected the inter-relationship of the antler and sexual cycles with very low testosterone responses occurring at casting and during velvet antler growth. The responses were higher at antler cleaning and then increased to a maximum at the rut before declining to reach their nadir at casting. The results are consistent with a hypothesis that the antler cycle, as a male secondary sexual characteristic, is closely linked to the sexual cycle and its timing is controlled by reproductive hormones. Low plasma concentrations of testosterone, even after LH stimulation, are consistent with the hypothesis that testosterone is unnecessary as an antler growth stimulant during growth.

Carotid artery exteriorisation in the red deer.

A technique for the surgical relocation of the carotid artery, to permit repeated percutaneous puncture, in red deer, is described. An incision was made through the skin distal to the ramus of the jaw parallel to and dorsal to the superficial jugular vein. The brachiocephalicus muscle was divided by blunt dissection to reveal the carotid artery. The carotid artery was dissected free of connective tissue and the vagus nerve and enclosed in a polythene prosthesis. The brachiocephalicus muscle was sutured dorsal to the now enclosed artery. The wound was closed taking care that the line of sutures did not overlie the prosthesis. Repeated percutaneous puncture of the artery was possible for periods of up to 18 months.