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ABSTRACT

In the mediobasal hypothalamus (MBH) of prooestrous rats or acutely ovariectomized oestrogentreated adults a marked but short-lived increase in adrenergic activity occurs at 16.00 h, 2 h before the oestrogen-dependent surge of gonadotrophins at 18.00 h. In this study oestrogen-stimulated (noon on day 1) 22-day-old female rats were used which are known to produce surge levels of prolactin at 18.00 h on day 2 and surges of both prolactin and LH at 18.00 h on day 3; although similar treatment of 18-day-old animals or oil-treated 22-day-old rats failed to produce these effects. Radioenzymatic assays of adrenaline concentrations and of the activity of its synthesizing enzyme (phenylethanolamine-N-methyl transferase; PNMT, EC 2.1.1.28) in the MBH of oestrogen-treated 22-day-old rats showed significant (P< 0·05–0·01) increases in both parameters at 16.00 h (i.e. 2 h before surge levels of gonadotrophins) on days 2 and 3 when compared with other times of day. Such effects were not seen in oil-treated 22-day-old animals or in oestrogen-treated 16-day-old rats. Noradrenaline and dopamine concentrations in the MBH of oestrogen-treated 22-day-old rats remained at baseline levels on days 2 and 3 with the exception of noradrenaline at 17.00 h on day 3 when levels appeared higher (P<0·05) than at either 15.00 or 16.00 h. Subsequent measurements of PNMT activity in oestrogen-treated 22-day-old rats at 4-hourly intervals throughout days 2 and 3 showed the presence of a clear circadian rhythm with peak levels occurring at 16.00 h. In conclusion, a temporal relationship (not necessarily specific) exists between increased adrenergic activity in the MBH of oestrogen-treated 22-day-old rats and a surge of gonadotrophins (LH and/or prolactin) 2 h later. This relationship apparently depends on an oestrogen-stimulated circadian rhythm of PNMT activity.

J. Endocr. (1986) 109, 45–51

SUMMARY

Serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels were measured by radioimmunoassay in groups of Wistar rats at 6-hourly intervals during the course of a 4-day oestrous cycle. In addition to a surge at about 18.00 h on the day of pro-oestrus, a circadian rhythm in LH levels was observed which was accentuated during metoestrus. FSH levels showed a protracted periovulatory rise which reached peak levels at about 03.00 h on the day of oestrus. A daily rhythm was not observed.

The incorporation of [³⁵S]methionine (injected subcutaneously 1 h before death) into the trichloroacetic acid-precipitable proteins of the anterior pituitary and of discrete brain areas implicated in the control of gonadotrophin release, was also measured. An increase in protein synthetic activity was observed in the anterior pituitary and the area of the median eminence on the evening of pro-oestrus with peak levels coinciding with the LH surge at 18.00 h on the day of pro-oestrus. An increase in protein synthetic activity relative to that in a 'control' area (the putamen) was observed in the preoptic area and in the amygdala 15–18 h before the LH surge. It is suggested that these changes in protein synthetic activity are associated with the train of neural and humoral events leading to ovulation.

After ovariectomy, protein synthetic activity in all areas investigated was at the low levels observed during the oestrous cycle. Daily injections of 20 μg oestradiol benzoate into intact females led to levels of protein synthesis as great as, or greater than, the maximal levels observed during the oestrous cycle.

Measurements of variations in total protein concentration/unit wet weight of pituitary or brain tissue showed that they were apparently unrelated to variations in protein synthetic activity. Protein concentration appeared to be high in all the brain samples taken at oestrus and low in those taken at metoestrus. Furthermore, superimposed upon a marked daily rhythm, there was a peak of protein concentration on the evening of dioestrus both in the preoptic area and in the area of the median eminence.

ABSTRACT

To investigate the role of suprachiasmatic efferent connections in the expression of diurnal hormone rhythms, the efferent pathway from the suprachiasmatic nucleus (the putative circadian generator in the rat) to the sub-paraventricular zone (the main terminal area of suprachiasmatic efferents) was disrupted using bilateral horizontal knife cuts in ovariectomized oestrogen-treated rats. The position of the knife cut was assessed by observing its effect on vasoactive intestinal polypeptide immunoreactivity (a marker for suprachiasmatic efferents into the sub-paraventricular zone). The size of both the diurnal plasma LH and prolactin surges was markedly and consistently reduced over the 3-week period following the lesion in animals with a total deafferentation of the sub-paraventricular zone, compared with sham-operated animals or lesioned animals with an intact sub-paraventricular zone. When lesioned animals were grouped according to the presence or absence of damage to the preoptic area, no significant differences were found in the sizes of the plasma hormone surges. When similar knife cuts were given to animals whose activity cycles were observed, no significant effects were noted in the ability of the animals to synchronize to a light/dark regime or to free-run in constant light conditions. These results suggest that the suprachiasmatic nucleus influences the diurnal surges of plasma LH and prolactin in oestrogen-treated ovariectomized rats, initially by an interaction with the sub-paraventricular zone, and not by a direct influence on gonadotrophin-releasing hormone neurones or other more rostral structures.

Journal of Endocrinology (1989) 122, 593–604

ABSTRACT

Flow cytometric analysis of nuclei stained with propidium iodide (PI) has been used to study the distribution of cells throughout the different phases of the cell cycle in the anterior pituitary gland of adult male Sprague–Dawley rats at different times of the day. According to PI fluorescence intensity the relative numbers of cells in S phase (cells with a DNA content between that of somatic cells in interphase (2n) and that of somatic cells after duplication of the DNA prior to cell division (4n)) and G₂/M phase (4n) were calculated. A significant circadian rhythm was found for cells in both the S phase (P < 0·05) and the G₂/M phase (P < 0·01). The wave of cells in S phase with a peak at the middle of the light period (14.00 h) precedes by about 6 h the wave of cells in G₂/M phase (peak at 20.00 h). Most of the DNA-replicating cells were found during the early S phase at 11.00 h, advancing further up to the middle of this phase at 14.00 h. Cells were distributed homogeneously throughout the S phase at 17.00 h. These data strongly suggest that the beginning of the light period triggers a wave of cells to leave G₀/G₁ into S phase.

Journal of Endocrinology (1991) 129, 329–333

ABSTRACT

Androgenized, oestrogenized and control female and male rats were used to establish possible differences in the alteration of the prolactin control system. Neonatal treatment involved administration of oestradiol benzoate or testosterone propionate (TP) on days 1 and 5 to the males and on day 5 to the females. Oil-treated animals were used as controls. Plasma prolactin levels were measured in these animals during adulthood (a) before gonadectomy, performed on day 80, and 27 days after gonadectomy and (b) on the 2 days (at 10.00 and 17.00 h) after administration of a single dose of TP to gonadectomized animals. Oestrogenized rats had the highest plasma prolactin concentrations just before and after gonadectomy. Testosterone propionate increased plasma prolactin levels in all groups. This response was more notable in the female than in the male groups, and was highest in the oestrogenized animals. Temporal rhythms of the prolactin response to TP were daily, perhaps circadian, with increased levels in the afternoon compared with those in the morning. This pattern was not present in oestrogenized females and androgenized males. Results suggest (a) that oestrogens and androgens given neonatally differ in their ability to alter the prolactin control system, and (b) that females seem to be more sensitive than males to changes in hypothalamic differentiation induced by neonatal steroid treatment.

J. Endocr. (1985) 105, 429–433

SUMMARY

The concentration of ovine placental lactogen (oPL) was measured by radioimmunoassay in plasma samples from chronically catheterized ewes and their fetuses from day 110 of gestation to term (about day 145).

Concentrations of oPL in the plasma of the mother and fetus were raised after surgery, and remained raised for 3–5 days after the operation. Concentrations of oPL were greatest in the fetus at days 120–124 of gestation, and then declined until delivery. Mean concentrations of oPL in the fetus in late pregnancy for single, twin and triplet pregnancies were 101±6 (s.e.m.), 100±11 and 117±59 ng/ml respectively and were not significantly different.

Mean concentrations of oPL in the mother in late pregnancy for single, twin and triplet pregnancies were 718±227, 1387±160 and 1510±459 ng/ml respectively; the difference between these means was significant (P <0·05). Peak concentrations were noted at days 130–139 of gestation after which concentrations fell and were significantly lower on the day of delivery (P <0·01). Concentrations of oPL in the mother showed no circadian rhythm. The mean concentrations of oPL in maternal plasma during late pregnancy was significantly correlated to the combined fetal weight at birth (r = 0·624, P <0·01).

SUMMARY

The physiological regulation of the plasma corticosteroid concentration, measured by competitive protein-binding, was studied in female rhesus monkeys (M. mulatta) sedated with phencyclidine hydrochloride. Morning basal levels of plasma corticosteroids were found to be in the range 8·0–25·2 μg/100 ml, which is lower than that previously reported in this species. A circadian rhythm in plasma cortisol concentration was demonstrated. Prolonged sedation with phencyclidine was associated with a gradual increase in the plasma cortisol concentration. Synthetic α^1–24 adrenocorticotrophic hormone given intravenously caused a rapid rise in plasma cortisol, the minimum effective dose was between 1 and 10 ng/kg body weight and the response was maximal after 1000 ng/kg. The administration of lysinevasopressin and the induction of hypoglycaemia by insulin were both followed by an increase in the plasma corticosteroid concentration. Metyrapone caused a decline in plasma 11-hydroxycorticosteroids and a concomitant increase in total corticosteroids measured by competitive protein-binding. It is concluded that the hypothalamic-pituitary-adrenal system in the rhesus monkey functions in a manner which is qualitatively and quantitatively similar to that of man.

SUMMARY

Ovulation was delayed for 24 h after the administration of sodium pentobarbitone (Nembutal, 35 mg/kg body weight) at 14.00 h, before the critical period on the afternoon of prooestrus. The expected preovulatory surge of serum LH at 18.00 h of pro-oestrus was also delayed until 21.00 h on the following day; however, increased levels (> 12 ng/ml) were observed in 14 out of 23 animals (killed by decapitation) at 21.00 h on the day of Nembutal administration. The serum FSH rise observed on the morning of expected oestrus was extended after Nembutal treatment, and a further rise was noted 24 h later.

Peak levels of incorporation of ³⁵S from methionine into protein of the median eminence area (ME) and of the anterior pituitary (AP) which normally occur about the time of the preovulatory LH surge, were also delayed until 21.00 h on the day following Nembutal administration.

Neither ovulation nor the preovulatory gonadotrophin rises with their accompanying changes in incorporation in the ME and the AP, were altered by Nembutal administered after the pro-oestrous critical period.

Thus Nembutal, while blocking ovulation, inhibits the circadian rhythm of incorporation of ³⁵S from methionine in the brain as well as the peaks of incorporation in the median eminence and the anterior pituitary which accompany the normal preovulatory surges of gonadotrophin.