ABSTRACT

Uterine weight, RNA, DNA, protein content, in-vitro rate of protein synthesis, cytosol oestrogen and progesterone receptors were examined after administration of oestradiol to ovariectomized animals and on days 0, 5, 9 and 13 of the non-pregnant cycle and day 13 of pregnancy. In ovariectomized animals, oestradiol increased uterine weight, RNA: DNA and protein: DNA ratios and the concentration of cytosol receptors for oestradiol and progesterone. During the oestrous cycle there was a linear increase in uterine weight and a significant effect of the corpus luteum on the weight of the ipsilateral uterus. Changes in RNA, DNA and protein content between days 0 and 5 were not observed, but between days 5 and 13 RNA: DNA and protein: DNA ratios increased and the DNA: tissue weight ratio decreased. Thus, cellular hypertrophy and/or increased metabolic activity rather than hyperplasia occur over this period, which is coincident with the known rise in plasma progesterone levels. The rate of in-vitro protein synthesis (per unit tissue protein) during the non-pregnant cycle was greatest at day 0. These changes in uterine metabolic activity were associated with alterations in cytosol receptor concentrations for both steroids. Cytosol progesterone receptor concentrations were highest at day 0 after which they declined to a minimum at day 13. Cytosol oestradiol receptor concentrations, however, rose between days 0 and 5 and then declined. Although lutectomy on day 8 of the cycle does not interfere with the development of a histologically normal luteal phase, high peripheral progesterone levels which occur after day 8 in intact animals are associated with major increases in uterine metabolic activity.

The unilateral effect of the corpus luteum on uterine weight was associated with a decrease in DNA: g tissue ratio and an increase in rate of in-vitro protein synthesis indicating hypertrophy and/or extracellular accumulation of secreted material as well as enhanced metabolic activity. There was a significant effect of pregnancy on uterine weight at day 13 and this was associated with an increase in DNA content of both uteri. There was a unilateral effect of pregnancy on RNA: DNA ratio and in-vitro rate of protein synthesis, but not on uterine weight.

J. Endocr. (1986) 108, 201–210

ABSTRACT

Radioimmunoassays were established for the measurement of total androgens and the specific measurement of testosterone and 5α-dihydrotestosterone in peripheral plasma of the brush-tail possum. Androgen concentrations were measured in blood collected from indwelling jugular cannulae (i) to determine whether the normal pattern of androgen secretion in this species was episodic and (ii) to attempt to relate total androgen and the pattern of testosterone secretion to the changes previously reported in prostatic, but not epididymal, weight in the breeding season. Blood was collected from restrained animals at varying time-intervals during daylight hours and darkness. Despite an apparent good adaptation to the sampling procedure there was generally a progressive decline in plasma androgen level during the collection period. This was true for animals bled during or out of the breeding season. There was no significant seasonal effect on the androgen concentration in the initial blood sample. When less restraint was used, two of three animals showed fluctuations in androgen levels over the 7-h sampling period. Testosterone levels in blood obtained by cardiac puncture were four- to nine-fold higher than those of 5α-dihydrotestosterone but levels of these androgens in samples obtained during the breeding season were not significantly different from those obtained out of season. The results do not argue for a pulsatile release of testosterone in the possum but do demonstrate a marked capacity for changes in the peripheral androgen concentration. There was a poor correlation between testosterone and 5α-dihydrotestosterone levels and prostatic weight.

J. Endocr. (1985) 105, 63–70

Abstract

Pituitary adenylate cyclase-activating polypeptide (PACAP) and vasoactive intestinal polypeptide (VIP) are known to stimulate adenylate cyclase activity in rat pituitary cells but no direct effects have been reported on sheep pituitary cells. In this study we determined whether either peptide could stimulate intracellular cAMP accumulation in dispersed sheep pituitary cells in primary culture. Time course studies with PACAP showed that tachyphylaxis developed rapidly and so a short incubation time (5 min) was used to define the dose–response relationship. PACAP dose-dependently stimulated intracellular cAMP levels with a half-maximum response at 2·9 ± 0·2 nmol/l (n=4). In contrast, VIP only caused a small increase in intracellular cAMP levels at the highest dose tested (1 μmol/l). The VIP antagonist [4C1-d-Phe⁶,Leu¹⁷]VIP had no effect on the cAMP response to either PACAP or VIP while the peptide PACAP(6–38), a putative PACAP antagonist, blocked the cAMP response to PACAP. The desensitisation to PACAP was further investigated by pretreating cells with PACAP for 30 min. After a further 15 min in culture medium alone, these cells showed no cAMP response to subsequent treatment with PACAP but could respond to forskolin. When a longer incubation period of 240 min was used between the first and second treatment with PACAP, a partial return in responsiveness to PACAP was observed. In summary, these results show that PACAP activates adenylate cyclase in sheep pituitary cells but that there is rapid development of tachyphylaxis. Experiments with the antagonists suggest that the response to PACAP is via the PACAP type I receptor. In contrast, physiological doses of VIP do not stimulate cAMP accumulation in sheep pituitary cells.

Journal of Endocrinology (1996) 148, 545–552

ABSTRACT

Experiments were conducted to investigate whether prolactin suppresses the corpus luteum during lactational quiescence in the Bennett's wallaby. In the first experiment, pouch young were removed from lactating wallabies (day 0) which were then treated daily for 7 days with either saline, or 8 mg domperidone or 2 mg ovine prolactin. In the saline-injected animals there was a transient peak in progesterone concentrations on day 4 and birth on day 28. The transient progesterone peak and births were significantly (P <0·01) delayed by 5 and 8 days in animals treated with domperidone and ovine prolactin respectively. In the second experiment, four groups of lactating wallabies were treated on day 0 with either 60 mg bromocriptine (groups C and D) or the vehicle (groups A and B). On days 0–6, groups B and D were injected daily with 2 mg ovine prolactin while groups A and C received the vehicle. In group C, three pouch young died 14–29 days after administration of bromocriptine, and there was a transient rise in progesterone on day 4 in all animals, indicating that bromocriptine resulted in immediate reactivation of the quiescent corpus luteum. New births occurred in two animals on day 28. In group D, which received bromocriptine followed by ovine prolactin for 7 days, all the original pouch young remained alive at the end of the experiment. Four of the animals from this group showed a transient progesterone peak on day 11, with births in two animals on days 35 and 36 indicating that the effects of bromocriptine were prevented whilst ovine prolactin was being administered. In one animal given ovine prolactin alone (group B), there was a transient progesterone peak and birth on days 12 and 35 respectively, suggesting that removal of exogenous prolactin may also act to terminate reproductive quiescence in some animals. In summary, these results support the hypothesis that prolactin suppresses the corpus luteum during lactational quiescence, and that the effects of bromocriptine are due to suppression of endogenous prolactin.

J. Endocr. (1988) 119, 405–411

ABSTRACT

It is well known that prolactin secretion is inhibited by dopamine acting via the pituitary dopamine D₂ receptor. Dopamine D₁ receptor analogues also affect prolactin levels although the mechanisms and physiological significance are poorly understood. The present study of the ewe was undertaken to characterize the effects of the D₁ receptor agonist SKF 38393 and antagonist SCH 23390 on prolactin in this species and to determine whether the prolactin response to both drugs requires an intact hypothalamo-pituitary axis. Ovariectomized ewes were injected intravenously with vehicle, 0·2, 2 or 20 mg SKF 38393 (D₁ agonist) or SCH 23390 (D₁ antagonist). At the 20 mg dose, plasma prolactin concentrations were significantly (P <0·01) increased by each drug and returned within an hour to control levels. When injected directly into the lateral ventricles of the brain (intracerebroventricular (i.c.v.) injection), a 100-fold lower dose of SKF 38393 (0·2 mg; P <0·05) was sufficient to stimulate prolactin secretion. In contrast, i.c.v injection of SCH 23390 (0·02 and 0·2 mg) had no effect on prolactin levels and at no dose was there evidence for suppression of prolactin levels. These results are in accord with earlier studies in the rat which suggested that the D₁ agonist stimulated prolactin secretion via a direct effect on central dopamine D₁ receptors whereas the

D₁ antagonist interacted with the pituitary dopamine D₂ receptor to increase prolactin secretion. In a further experiment this hypothesis was tested in hypothalamo-pituitary disconnected ewes which were infused with dopamine (0·5 μg/kg per min) for 3 h. After 2 h of the dopamine infusion, animals were challenged with intravenous injections of the vehicle, 20 mg SKF 38393, 20 mg SCH 23390 or 2 mg domperidone (dopamine D₂ antagonist). Infusion of dopamine was followed by a significant (P <0·05) decline in prolactin concentrations so that after 2 h prolactin levels were 40% of the preinfusion value. Following injection of the vehicle, SCH 23390 or SKF 38393, prolactin levels continued to decline for the remainder of the experiment. As expected, injection of domperidone was followed by a significant (P <0·05) increase in prolactin to reach peak levels after 30 min. These results demonstrate that peripheral injections of the dopamine D₁ agonist SKF 38393 or antagonist SCH 23390 increase prolactin secretion in the ewe. The prolactin response to either drug requires an intact hypothalamo-pituitary axis indicating that SKF 38393 and SCH 23390 act at some central site(s) which is linked with hypothalamic secretion of prolactin-releasing or -inhibiting factors.

Journal of Endocrinology (1993) 137, 457–464

ABSTRACT

The quantitatively major steroid hormones in ovarian and adrenal venous plasma of the female brush-tail possum were identified by gas chromatography–mass spectrometry. The ovarian vein plasma samples all contained oestradiol and its concentration was highest during the pro-oestrous phase of the reproductive cycle. During this phase the concentration of progesterone was below the limit of detection but at day 13 of the oestrous cycle and pregnancy, the concentration of progesterone exceeded that of oestradiol. Cortisol and corticosterone were the major steroid hormones found in all adrenal vein samples with cortisol predominant. Androgens with a 3-oxo structure, if present, were below the limits of detection in all plasma samples. Radioimmunoassays for the measurement of progesterone and oestradiol in peripheral plasma were used to follow changes in the concentrations of these steroids during the reproductive cycle.

Progesterone in serial blood samples was low at oestrus, rose gradually until day 7 and then increased more rapidly to reach a peak level of 21–29 nmol/l at around day 13. Any differences between the pregnant and non-pregnant cycles were minor. Oestradiol was only detected around oestrus when levels were variable (53·3±20·92 (s.e.m.) pmol/l; n = 4). The results indicate that the reproductive cycle of the brush-tail possum is characterized by a single peak of oestradiol at around pro-oestrus followed by gradually increasing levels of progesterone. Pregnancy appears to have no influence on the circulating concentrations of oestradiol or progesterone.

J. Endocr. (1985) 105, 53–62

ABSTRACT

Non-domesticated seasonally breeding ungulates exhibit marked seasonal changes in metabolic rate, voluntary food intake (VFI), pelage growth and moult and hormone secretion. It is not known whether these seasonal rhythms are regulated by the same central processes which control the onset and termination of the breeding season. Here we compare two closely related deer species which have significantly different mating and calving seasons. Seasonal changes in VFI, liveweight, coat growth, plasma prolactin and tri-iodothyronine (T₃), and the timing of the breeding season were examined over a 15-month period in six adult post-pubertal red and Père David's deer from January to April the following year. The timing of the seasonal changes in prolactin, T₃, VFI and coat growth were all significantly advanced by 56, 23, 60 and 54 days respectively in the Père David's deer. The times of onset and termination of the breeding season of Père David's deer were also significantly advanced by 90 days, but in both species, the breeding season was of similar duration (160 ± 5 (s.e.m.) days). Changes in liveweight of adult red deer could be explained by changes in VFI rather than efficiency of utilization. This was not the case in Père David's deer and may indicate seasonal changes in the efficiency of energy utilization. In order to establish whether these species differences develop with age, we undertook a second study in which seasonal changes in VFI, growth, plasma prolactin concentrations and the timing of the onset of the breeding season were recorded for ten red deer and six Père David's deer from 6 to 18 months of age. Both species exhibited a similar decline in VFI in the first autumn of life. Subsequently, the Père David's deer exhibited an advance in the timing of the seasonal peak in VFI and prolactin (21 and 66 days respectively); puberty occurred 3 months earlier than in red deer. The earlier breeding season of the Père David's deer was associated with a significant advance in a range of seasonal endocrine and physiological parameters. These species differences may develop with age. Our data indicate that seasonal patterns of metabolism and growth may be closely linked to those mechanisms which also regulate the onset and termination of the breeding season.

Journal of Endocrinology (1989) 122, 733–745

ABSTRACT

Seventeen red deer hinds were housed in individual pens and from 28 February until 11 November were injected each week with vehicle (group A; n = 6) or 5 (group B; n = 6) or 12·5 mg (group C; n = 5) of a long-acting formulation of bromocriptine. Liveweight and voluntary food intake (VFI) were recorded for each hind, and blood was collected for determination of progesterone, prolactin, tri-iodothyronine (T₃) and cortisol concentrations by radioimmunoassay. Treatment with the high dose of bromocriptine was associated with a significant (P <0·05) reduction in VFI, with the effect being greatest between March and July. There was no treatment effect on liveweight, but there was a significant (P <0·01) interaction between time and treatment due to the faster rate of weight gain in control animals at the beginning of the experiment. Changes in liveweight could be explained by changes in VFI rather than by changes in the efficiency of utilization of intake. Termination of the breeding season was significantly (P <0·01) delayed by 54 days in group C hinds. Growth of the summer coat and subsequent winter coats was delayed by 1 and 3 months respectively in group C hinds, and in groups B and C the duration that animals were in summer coat was increased by about 1 month. The seasonal increase in prolactin concentrations was seen in all groups, but levels were significantly (P <0·05) lower in group C hinds. Concentrations of T₃ and cortisol were not affected by bromocriptine.

J. Endocr. (1988) 119, 413–420

ABSTRACT

Pituitary glands and corpora lutea collected at various stages of the reproductive cycle of the tammar wallaby (Macropus eugenii), were extracted and fractionated by high-performance liquid chromatography, and specific radioimmunoassays were used to measure mesotocin ([Ile⁸]-oxytocin) and oxytocin. Mesotocin, but not oxytocin, was identified in extracts of pituitary; the mean concentration of mesotocin in this tissue was 0·75 nmol/g wet weight. Neither mesotocin nor oxytocin was detected in extracts of corpus luteum. In female Bennett's wallabies passively immunized against mesotocin during seasonal reproductive quiescence, there was no significant effect on peripheral progesterone concentrations and there were no births, matings or changes in vaginal smears in the 2 months following treatment. Thus mesotocin is unlikely to act as a systemic luteostatic agent during seasonal quiescence.

J. Endocr. (1988) 117, 367–372

ABSTRACT

Prolactin concentration was measured in plasma collected each week for 13 months from lactating and non-lactating Bennett's wallabies (Macropus rufogriseus rufogriseus). In non-lactating animals, prolactin concentrations decreased towards the end of the study but such changes did not appear to fit a seasonal pattern. Prolactin concentrations were low during early lactation and at a similar level to non-lactating animals, increased significantly during late pouch life (February–May), and then returned to non-lactating levels at a time coincident with permanent exit of the joey from the pouch. Temporary removal of joeys from their mothers in April was followed by a rapid decline in prolactin concentrations which remained low for 24 h until the joey was returned to its mother, whereupon prolactin concentrations increased significantly within 2 h.

The effect of a single injection of bromocriptine (5 mg/kg) on lactation, embryonic diapause and plasma prolactin concentrations was examined at two stages of lactation. In November (lactational diapause), bromocriptine had no effect on prolactin concentrations but two out of four suckling joeys died on days 13 and 14 after treatment, and three out of four females gave birth on days 27, 27 and 28. Bromocriptine treatment in April (seasonal diapause) was followed by a significant reduction in prolactin concentrations and reduced growth rate of joeys belonging to treated females. New births were not observed.

In view of the effect of bromocriptine on plasma prolactin concentrations in late lactation and the demonstration that domperidone (a dopamine antagonist) significantly increases plasma prolactin concentrations, it would seem that dopamine can act as a prolactin inhibitory hormone in this as in other mammalian species.

J. Endocr. (1986) 110, 59–66