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SUMMARY
The effects of intravenous injection of synthetic luteinizing hormone releasing hormone (LH-RH) on the release of LH and FSH have been studied in the adult guinea-pig. In all the experiments the secretion of FSH was apparently unaffected by administration of LH-RH. The release of LH was log dose-dependent over the range 0·5–50 μg LH-RH.
During the female cycle, the maximum increment in the concentration of LH after a single i.v. injection of 0·5 μg LH-RH decreased progressively from day 1 to day 13. Double or triple injections of 1 μg LH-RH at 1 h intervals produced no potentiation on day 3 but progressively greater LH responses occurred on days 7, 10 or 13 of the oestrous cycle. Ovariectomy immediately before the first injection of LH-RH on day 7 blocked the potentiated response to subsequent injections, whereas ovariectomy immediately before the second injection still permitted this potentiation. These results suggest that there is direct ovarian involvement in the potentiated response to LH-RH observed during the later part of the guinea-pig oestrous cycle.
Infusion of LH-RH (1 μg over 200 min) produced a potentiated release of LH in female guinea-pigs on day 7 (but not on day 3) of the oestrous cycle after a delay of 1·5 h. It is proposed that there are two 'pools' of LH in the pituitary gland of the female guinea-pig and that the second pool is 'activated' consequent upon previous hypophysial stimulation of secretion from an ovary containing adequately developed follicles.
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A.R.C. Institute of Animal Physiology, Babraham, Cambridge, CB2 4AT
(Received 1 November 1977)
Thyrotrophin releasing hormone (TRH) can have a stimulatory effect on the release of both prolactin and thyrotrophin (TSH; Deis & Alonso, 1973), although in the rat, supraphysiological doses of TRH are required to affect the secretion of prolactin (Burnet & Wakerley, 1976). A more important factor in the control of the release of prolactin is considered to be prolactin release inhibiting factor (PIF), which is thought to act through the catecholamine, dopamine (MacLeod, 1976). Stimuli which cause the concomitant release of TSH and prolactin are thought to have a direct effect at the hypothalamic level such that neurones releasing TRH are excited, whereas those releasing PIF are inhibited. In the present work, we have tested this hypothesis using the suckling stimulus to elicit the simultaneous release of prolactin and TSH (Blake, 1974; Burnet & Wakerley, 1976). If
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The changes in FSH and LH secretion after placement of lesions in the hypothalamus were traced in ferrets serially bled at 15 min intervals. Passage of the lesioning current through platinum electrodes in anoestrous females was associated with an immediate surge in LH and FSH output. Damage to the hypothalamus of oestrous females also caused an immediate surge of LH secretion, but then a long-lasting second rise in blood LH content set in. High concentrations of LH were never sustained overnight. The response of long-term spayed females to the placement of hypothalamic lesions was similar to that of anoestrous ferrets, while that of anoestrous or oestrous ferrets was not altered by acute removal of the ovaries. Manipulation of the ovaries appeared to facilitate FSH and LH secretion. The response of males was similar to that of anoestrous females. Marked increases in FSH and LH release were also seen in females when lesions were made with steel electrodes, but had subsided on the following day.
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SUMMARY
Heterologous radioimmunoassays for FSH and LH were employed to examine the effect of synthetic LH-RH upon gonadotrophin secretion in the ferret. Intravenous injection of 4 μg LH-RH induced a surge of FSH and of LH secretion in male and in female animals. In intact and in castrated males, the rise of LH was much more marked than that of FSH. The gonadotrophin response to LH-RH was greater in anoestrous than in oestrous females; FSH secretion was not enhanced during oestrus. Ovariectomized females behaved as anoestrous females with respect to LH secretion, while FSH secretion remained unchanged. Treatment of ovariectomized females with progesterone did not alter the pattern of response to LH-RH, but oestradiol treatment depressed the reaction to match that seen in oestrous females. Repetitive injections of LH-RH induced repetitive surges of FSH and LH in anoestrous females, but only of LH during oestrus: slow i.v. infusion of LH-RH induced a sustained elevation of plasma LH levels both in oestrous and in anoestrous females; again FSH levels rose only in anoestrous females. Injection of synthetic TRH did not alter gonadotrophin secretion in corresponding groups of male or female ferrets.
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SUMMARY
The possible occurrence of long-term changes in gonadotrophin control mechanisms following the administration of oestrogen to adult female rats has been studied. Administration of 2·5 mg oestradiol benzoate (OB) to normal female rats at 60 days of age did not result in failure of ovulation at 120 days of age but significant impairment of the LH and FSH responses to progesterone after ovariectomy and oestrogen priming was observed at 160–180 days of age. Treatment with the same dose of OB at 60 days of rats injected with 10 μg testosterone propionate on Day 4 of postnatal life resulted in an increased incidence of failure of ovulation at 120 though not at 150 days of age but did not further impair the already reduced gonadotrophin response to progesterone at 160–180 days of age. Removal of the ovaries at 60 days of age did not modify the effects of oestrogen given at 60 days of age in either group nor did ovariectomy at 60 days improve the response of neonatally androgen-treated rats to progesterone at 160–180 days of age. The increases in plasma prolactin and TSH levels in response to oestrogen priming after ovariectomy were not affected in any of the experimental groups.
The administration of a long-acting oestrogen preparation (oestradiol cyclopentyl propionate, 2·5 mg at 60 days of age) to normal female rats suppressed ovulation and depressed plasma LH and FSH concentrations for at least 90 days; anterior pituitary weights were greatly increased and plasma prolactin concentrations were very high.
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*A.R.C. Institute of Animal Physiology, Babraham, Cambridge, CB2 4AT, and ‡Department of Physiology, Royal Veterinary College, London, NW1 OTU
(Received 28 March 1978)
Ovulation can be induced precociously in the prepubertal female rat by the administration of pregnant mare serum gonadotrophin (PMSG), provided that the animal weighs over 60 g (Wilson, Endersby & McDonald, 1974). The hormonal changes brought about by this treatment have been studied (J. C. Buckingham, M. B. ter Haar, A. S. McNeilly & C. A. Wilson, data to be published) and it has been found that the preovulatory concentrations of radioimmunoassayable luteinizing hormone (LH) varied according to the antiserum used. These findings are described in the present communication.
Female Sprague–Dawley rats (Tuck & Sons, Rayleigh, Essex) were brought into the department on day 21 of life, kept under conditions of controlled lighting (lights on 05.00–19.00 h) and provided with pelleted rat diet (No. 86, Dixon &
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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.
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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.
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The effects of changes in the intensity of the suckling stimulus on the reflex release of oxytocin and prolactin were compared in urethane-anaesthetized lactating rats. Mothers which had previously suckled 12 pups (Group 1) showed a graded increase in the amount of oxytocin released during a 3 h suckling test when the number of pups applied to the nipples was increased from six to eight or ten. Mothers which had suckled six pups during their lactation (Group 2) appeared to show a maximum frequency of milk ejection whether six, eight or ten pups were applied to the nipples.
The release of prolactin was not elicited from either Group 1 or Group 2 mothers when six pups were applied to their nipples. With eight pups suckling, the Group 1 mothers again showed no evidence of prolactin release. In contrast, the Group 2 mothers showed a significant increase in the level of prolactin in the plasma during the 3 h suckling test. With ten pups suckling the release of prolactin was evident in both groups of mothers, although the response was earlier and more pronounced in Group 2 than Group 1.
These results suggest that in the urethane-anaesthetized rat, the threshold for the suckling-induced reflex release of oxytocin is distinct from the threshold for the release of prolactin and that these thresholds are, at least in part, set by the preceding suckling experience of the mothers. In those animals which showed both reflex milk ejection and prolactin release there was a linear relationship between the magnitude of the two endocrine responses.
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SUMMARY
The incorporation of [35S]methionine into protein of the anterior pituitary and discrete brain areas was measured following the administration of antibodies to oestrogen, ovariectomy, or adrenalectomy on the afternoon of dioestrus. The antibody to oestrogen deleted the circadian rhythms of methionine incorporation normally observed in the various brain areas together with the peaks of incorporation normally observed in the median eminence area and anterior pituitary on the evening of pro-oestrus. The peaks of incorporative activity normally observed in the preoptic area and amygdala (relative to the putamen) at 03.00 h on the day of pro-oestrus were also deleted. Administration of the antiserum on the morning of pro-oestrus failed to alter the pattern of methionine incorporation normally observed on the evening of pro-oestrus.
Ovariectomy performed at 16.00 h of dioestrus blocked the preovulatory rise of luteinizing hormone (LH) (as did the antibody to oestrogen) and inhibited the peak of methionine incorporation normally observed in the anterior pituitary on the evening of pro-oestrus. However, for the peak in the median eminence to be inhibited, ovariectomy had to be performed on the morning of the preceding oestrus. Adrenalectomy alone, or adrenalectomy with ovariectomy, performed on the afternoon of dioestrus did not affect the levels of methionine incorporation in the brain or anterior pituitary at 18.00 h on the day of prooestrus.
Animals which had been ovariectomized and injected with 2·5 μg oestradiol benzoate on the morning of oestrus showed significantly increased levels of methionine incorporation in all the brain areas and the anterior pituitary at 18.00 h of expected pro-oestrus. The administration of antibody to oestrogen to a similar group of animals on the afternoon of expected dioestrus inhibited the rise at 18.00 h of expected pro-oestrus. The apparent discrepancy between the results obtained with ovariectomy and the antiserum appeared to be due to the ability of the antiserum to neutralize the activity of oestrogens retained by the tissues.
The present results suggest that the changes in incorporation of methionine into protein in the brain and anterior pituitary are brought about by the action of endogenous oestrogen: there appears to be a steady summative effect on the median eminence throughout the oestrous cycle together with a short-lived effect occurring during pro-oestrus and affecting the anterior pituitary.
