Evidence for a spinal site at which opioids may act to inhibit the milk-ejection reflex

Morphine was injected into the lumbar subarachnoid space of anaesthetized lactating rats (7-10 days post partum) to examine its effect on the milk-ejection reflex at a spinal level. Although the delay until the first milk-ejection response to the suckling of hungry pups was variable (3-60 min) the subsequent responses occurred at regular intervals of 7.5 +/- 0.4 min; milk-ejection responses were detected by measurement of intramammary pressure and by the characteristic behaviour of the pups. Injection of morphine (4-50 micrograms) via a cannula inserted into the spinal subarachnoid space inhibited reflex milk ejection in a dose-related manner without affecting the sensitivity of the mammary gland to exogenous oxytocin (1 mu., i.v.); injection extradurally was without effect. The opiate antagonist naloxone (10 micrograms), when injected intrathecally, did not significantly alter the pattern of reflex milk ejection or the amplitude of the intramammary pressure response, but prevented the inhibitory effect of morphine when administered with the opiate. Pethidine (250 and 400 micrograms) also inhibited the milk-ejection reflex. It is unlikely that the effect of spinal administration of morphine occurred as the result of the transportation to a supraspinal site since release of oxytocin evoked by intraventricular injection of hypertonic sodium chloride (3 mol/l) was blocked by intraventricular injection of morphine (4 micrograms) but not by a much larger dose (40 micrograms) injected intrathecally.     

Plasma levels of vasoactive intestinal polypeptide and oxytocin in response to suckling, electrical stimulation of the mammary nerve and oxytocin infusion in rats

The aim of the present study was to measure plasma levels of vasoactive intestinal polypeptide (VIP) and oxytocin following suckling, electrical stimulation of the mammary nerve and oxytocin infusion in lactating rats. Trunk blood was collected by decapitation after 5 and 20 min of suckling in conscious lactating rats. Repeated blood samples were drawn from the carotid artery in anesthetized rats, in connection with suckling, oxytocin infusion (0.22 nmol/l/kg/h) and electrical stimulation of the mammary nerve (5 V, 5 Hz, 2 ms). VIP and oxytocin levels were measured by radioimmunoassay. In conscious rats, VIP levels rose significantly from 18 +/- 5 to 102 +/- 30 pM after 5 min of suckling and to 123 +/- 25 pM after 20 min of suckling when milk ejection occurred. Oxytocin levels rose significantly from 90 +/- 24 to 269 +/- 45 pM during milk ejection. Suckling, oxytocin infusion and mammary nerve stimulation in anesthetized rats raised VIP levels significantly from 13 +/- 2, 18 +/- 5 and 10 +/- 2 to 43 +/- 8, 45 +/- 16 and 53 +/- 22 pM, respectively, whereas oxytocin levels rose from 111 +/- 34 to 294 +/- 66 pM after 20 min of suckling and to a peak value of 500 +/- 70 pM after oxytocin infusion. This study shows that VIP is elevated in plasma in lactating rats when the pups are suckling. The results showing that VIP levels rise following mammary nerve stimulation and oxytocin infusions indicate that both neurogenic and hormonal mechanisms can contribute to the regulation of VIP levels in plasma.     

Reappraisal of the influence of mammary distension on the frequency of milk ejection in the rat

Experiments were performed to reinvestigate the importance of mammary engorgement for activation of the milk-ejection reflex in the rat. Reflex milk ejection (measured by intramammary pressure recordings during a 2-h suckling test under anaesthesia) was compared in rats with engorged mammary glands (15-h separation from the pups, followed by sham-removal of milk) and in rats with drained mammary glands (15-h separation, followed by milk removal using a foster litter and exogenous oxytocin). In experiment 1, multiple small (2 mu.) doses of oxytocin were used for milk removal: these were effective in emptying the mammary glands and caused no subsequent impairment or change in sensitivity of the mammary response to oxytocin. Using this draining procedure, no significant differences were observed in either the number or relative amplitude of the milk ejections, or the occurrence of pup stretch reactions between engorged and drained rats. Similar results were seen in experiment 2, where an identical draining protocol was used, but the rats were pretreated with propranolol before the suckling test. In experiment 3, large (250 mu.) oxytocin doses were used for milk removal, as in previous studies. Again mammary draining had no effect on milk ejection in a subsequent suckling test (with propranolol pretreatment). However, the number of stretch reactions shown by the pups was significantly (P less than 0.001) reduced from 8.6 +/- 1.4/2 h to 1.9 +/- 0.6/2 h. This effect probably related to long-term impairment of the oxytocin response of the mammary glands following the draining procedure, and could not be attributed to the draining per se.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of L-dopa on milk ejection and prolactin release in lactating rats.

The effect of L-DOPA on milk ejection and on prolactin release during 30 min of suckling was studied in lactating rats. Various doses of L-DOPA (1-25, 2-5, 5 and 10 mg/100 g body wt) were injected i.p. 30 min before the suckling period. Control rats were injected with 0-9% NaCl solution only. An inhibition of milk ejection proportional to the dose of drug administered was obtained. The dose of 10 mg completely blocked milk ejection but 1-25 mg had no effect. A normal milk-ejection response was obtained with a small dose of oxytocin injected immediately before nursing into mothers treated with 10 mg L-DOPA, indicating that the blocking effect was not due to a lack of mammary gland response. In control mothers, serum prolactin levels increased from 67-2 +/- 25-9 (S.E.M.) to 950-3 +/- 118-7 ng/ml after a 30 min suckling period. L-DOPA (5 and 10 mg) prevented the release of prolactin induced by suckling, but 1-25 and 2-5 mg L-DOPA had no effect. The results indicate that oxytocin and prolactin release induced by suckling in lactating rats is inhibited by an increase of catecholamines at the hypothalamic-hypophysial axis.     

Interruption of parturition in rats by morphine: a result of inhibition of oxytocin secretion

Oxytocin secretion is inhibited by opioids, and oxytocin is important in parturition. The effects on parturition of morphine, a relatively selective mu-opioid receptor agonist, were studied in the rat. Morphine or vehicle with or without the opiate antagonist naloxone were administered immediately after the birth of the second pup and the subsequent course of parturition was recorded in a total of 80 rats. Both s.c. morphine (10 mg/kg) and intracerebroventricular (i.c.v.) morphine (18 micrograms through a previously implanted cannula) interrupted parturition, delaying the birth of the sixth pup after treatment to 187.3 +/- 35.9 (S.E.M.) min and 195.4 +/- 19.5 min respectively, compared with 46.4 +/- 3.7 and 66.1 +/- 17.5 min after vehicle alone. The dose of morphine given i.c.v. had no effect when given s.c. Naloxone given concurrently prevented the effects of morphine. Eventually the rate of parturition in the morphine-treated groups recovered. Perinatal pup mortality rate was not increased when morphine was given to the mothers, but it did inhibit the expression of normal intrapartum maternal behaviour. Pup mortality was increased 48 h post partum by morphine given during parturition, and it reduced the proportion of rats with normal maternal behaviour 24 h post partum. Morphine did not affect spontaneous or oxytocin-stimulated contractile activity of the parturient uterus in vitro. The concentration of oxytocin in trunk blood plasma was decreased 40 min after i.c.v. morphine (24.3 +/- 3.9 vs 39.3 +/- 6.5 pmol/l in controls), as was vasopressin (7.2 +/- 1.5 vs 19.7 +/- 4.5 pmol/l in controls). Intravenous infusion of oxytocin (2-5 mU/min for 144.3 +/- 8.2 min; total infused 448.5 +/- 61.9 mU) after i.c.v. morphine re-started parturition; all pups were born to these rats (mean time to pup 6, 110.3 +/- 12.7 min) before the i.v. vehicle-infused rats given i.c.v. morphine re-started (mean time to pup 6, 406.3 +/- 125.2 min). It is concluded that morphine given during parturition acts centrally through opioid receptors to inhibit oxytocin secretion, and impairs the expression of maternal behaviour. Reversal of the effects of morphine on parturition by i.v. oxytocin demonstrates the important role of oxytocin in fetus ejection and expulsion.     

Ablation of the region anterior and ventral to the third ventricle (AV3V region) does not impede parturition in rats

The region anterior and ventral to the third ventricle (AV3V) region is a major source of excitatory afferents to the magnocellular neuroendocrine system, and is essential for the osmotically regulated release of oxytocin. We investigated whether this input has a similarly essential role in parturition. Rats were implanted with a guide cannula in the AV3V region on days 9-18 of pregnancy. Following the birth of the third pup, rats were anaesthetized briefly with ether and either given an electrolytic AV3V lesion or a sham procedure was carried out. In eight AV3V-lesioned rats the mean (+/- S.E.M.) median interbirth interval following the lesion was 6.3 +/- 1.2 min compared with 5.2 +/- 0.6 min in 11 sham-lesioned rats. All rats completed delivery of their litters. The mean plasma concentration of oxytocin was unchanged following the sham procedure (pre-sham 17.1 +/- 2.8 pmol/l, n = 8; 15 min post-sham 18.1 +/- 2.7 pmol/l, n = 8; 30 min post-sham 19.2 +/- 3.5 pmol/l, n = 8). In AV3V-lesioned rats, plasma oxytocin was significantly raised following the lesion (pre-lesion 14.6 +/- 1.3 pmol/l, n = 7; post-lesion 58.3 +/- 9.8 pmol/l, n = 7) and was still higher than the sham-treated group after 30 min (55.8 +/- 9.9 pmol/l). Thus there was no significant difference in the time-course of parturition between AV3V-lesioned rats and sham-lesioned rats, and no evidence that the lesion impaired the release of oxytocin. Furthermore, in rats given an AV3V lesion on the morning of the expected day of delivery, parturition was neither delayed nor disrupted, suggesting that the AV3V region does not contribute to the mechanisms controlling the onset of parturition.     

Degradation of 125I-labelled prolactin in the rabbit: effect of nephrectomy and prolactin infusion

The concentrations of progesterone and oestradiol-17 beta in the maternal plasma of Bennett's wallaby, Macropus rufogriseus rufogriseus, were measured daily throughout gestation after reactivation of the diapausing corpus luteum by removal of the suckling pouch young (RPY). Progesterone increased from mean concentrations of 382-424 pmol/l (120-133 pg/ml) during lactation to reach peak concentrations of 908 +/- 172 (S.E.M.) pmol/l (285 +/- 54 pg/ml) (n = 8) 4 days after RPY and 971 +/- 220 and 971 +/- 229 pmol/l (305 +/- 69 and 305 +/- 72 pg/ml) (n = 7) 24 and 25 days after RPY respectively. The mean gestation length (RPY to birth) was 26.8 +/- 0.6 (S.D.) days (n = 6, range 25.75-27.50 days). Immediately after birth the plasma progesterone concentration declined to 299 +/- 51 (S.E.M.) pmol/l (94 +/- 16 pg/ml) (n = 6). Oestradiol-17 beta increased from mean concentrations of 291-553 pmol/l (80-152 pg/ml) during lactation to reach a peak concentration of 967 +/- 331 pmol/l (266 +/- 91 pg/ml) (n = 9) 1 day after RPY. The concentration declined from 7 days after RPY and fluctuated between mean concentrations of 273 and 480 pmol/l before reaching a minimum of 207 +/- 69 pmol/l (57 +/- 19 pg/ml) (n = 6) 19 days after RPY. A transient increase to 542 +/- 207 pmol/l (n = 7) occurred at 22 days after RPY. Plasma concentrations declined to a low of 156 +/- 55 pmol/l (43 +/- 15 pg/ml) (n = 6) 5 days after parturition. The mean concentration of plasma 13,14-dihydro-15-oxo-prostaglandin F2 alpha was less than 2.8 nmol/l (1 ng/ml) for all samples from 13 days after RPY until 4 days after parturition. The results suggest that oestradiol-17 beta may be important in the early stages of blastocyst reactivation to synergize with progesterone in stimulating uterine secretions. 13,14-Dihydro-15-oxo-prostaglandin F2 alpha is unlikely to be involved in the birth process and any luteolytic effect is likely to be from a local production of PGF2 alpha.     

Characteristics of milk ejection, associated intramammary pressure changes and oxytocin release in the mare

The neuroendocrine reflex theory of milk ejection was investigated in the horse under natural suckling conditions. To this end 12 lactating mares were provided with acute jugular catheters and with intramammary pressure (IMP) recording catheters. The foal had free access to the contralateral mammary complex. Intramammary pressure could thus be recorded while blood was drawn simultaneously for oxytocin analysis from the undisturbed animal. Suckling periods associated with a characteristic increase in IMP lasted significantly longer than unsuccessful nursing attempts. Elements of successful sucklings involved physical stimulation of the mammary gland, a quiet phase and a sudden increase in IMP. Successful suckling took place at about 20-min intervals with a wide range from less than 5 min to greater than 100 min. Between 5 and 10 mU oxytocin i.v. were sufficient to evoke an increase in IMP identical in shape and duration to a naturally induced increase in IMP. Mean peak oxytocin levels reached 15.8 pmol/l plasma, with a maximal release of 39 pmol/l. In the majority of cases (greater than 80%) peak oxytocin release did not occur until after the increase in IMP; in some cases an oxytocin surge was not detectable at all, despite a milk ejection-associated increase in IMP. In three cases increase in IMP could be observed while the foals were away from the mother with no signs of any intention to suckle. The data indicate that in the horse some elements of the neuroendocrine reflex, such as tactile stimulation of the teat and a surge of oxytocin before an increase in IMP, are facultative and not essential for normal milk ejection.     

Milk ejection reflex linked to slow wave sleep in nursing rats.

Correlations between cerebral activity of nursing rats and the milk ejection reflex were studied in Sprague-Dawley rats with 15- to 17-day-old litters. The stretch reaction of the pups, which expresses the onset of milk ejection, was closely correlated with the slow sleep epochs of the mother. Once the litter started suckling, milk ejection only took place when the mother fell asleep and electroencephalographic features of slow wave sleep appeared. Milk ejection was never found during paradoxical sleep nor when the mother was awake. Sleep deprivation for 30 min impaired milk ejection in spite of continuous suckling of the nipples by the pups. If the mother was allowed to sleep immediately afterwards, ejection of milk occurred. A 24-h sleep-wakefulness pattern did not show differences between nursing and controls. Our results show that suckling, although necessary, is not enough to set off milk ejection. This reflex only appears when the mother falls asleep, suggesting that oxytocin release is linked to suckling and slow wave sleep.     

Relationship between oxytocin release and amplitude of oxytocin cell neurosecretory bursts during suckling in the rat

Plasma concentrations of oxytocin and vasopressin were measured in relationship to oxytocin cell firing during suckling in urethane-anaesthetized rats. Preliminary experiments showed that plasma concentrations of oxytocin and vasopressin, which were increased immediately after anaesthesia, reverted to basal concentrations 3 h later. Moreover, it was found that exogenous oxytocin had entirely disappeared 5 min after i.v. bolus injections of known doses of oxytocin. Suckling did not modify the basal plasma concentration of oxytocin (14.6 +/- 2.9 compared with 14.6 +/- 1.5 pmol/l before suckling) except during a brief period immediately after neurosecretory bursts on oxytocin cells (37.8 +/- 5.2 pmol/l; P less than 0.001, n = 11). The plasma concentration of oxytocin did not differ significantly from the basal concentration 1.5 min later. The plasma concentration of vasopressin never varied. After two neurosecretory bursts of similar amplitude (total number of spikes during the burst) recorded on the same oxytocin cell, the variations in plasma concentration of oxytocin were also similar. When, for a given cell, the amplitude of neurosecretory bursts increased or decreased, the amount of oxytocin released changed in the same way. These data demonstrate (1) that suckling induces pulsatile release of oxytocin without vasopressin, and (2) a direct relationship between the amounts of oxytocin released and the amplitude of oxytocin cell neurosecretory bursts which argue in favour of simultaneous increases or decreases in the neurosecretory burst amplitudes on all oxytocin cells.     

Oxytocin secretion during milking in dairy cows with regard to the variation and importance of a threshold level for milk removal

Oxytocin secretion in dairy cows was measured during milking by means of a specific radioimmunoassay. Basal oxytocin concentrations before milking recorded from 147 milkings from 21 cows, where an assay method with enhanced sensitivity was employed, were 1.5 +/- 0.6 pmol/l and there was no evidence for any conditioned release of oxytocin. A 1-min manual stimulation before milking evoked a variable but distinct increase in oxytocin concentrations in 188 out of 195 milkings performed using 29 cows. Despite the high variation in absolute concentrations five types of typical secretion pattern could be distinguished. There was no obvious relationship between patterns or absolute concentrations of oxytocin and milk-flow characteristics. Evidence is given that milk ejection seems to follow the threshold principle in that small releases of oxytocin up to a range of 3-5 pmol/l plasma are sufficient to evoke maximum milk ejection.     

Maternal modulation of growth hormone secretion in the neonatal rat: involvement of mother-offspring interactions

Serum GH levels increased in 2- or 8-day-old rat pups when sucking mammary glands whose main milk ducts were ligated. Although intragastric administration of rat milk has been shown to increase serum GH levels in neonatal rats, ingestion of milk during suckling did not increase serum GH values further. In another experiment, 2-day-old pups obtained no milk when they were suckled by anaesthetized mothers, and in this instance the serum GH concentration of the pups decreased. This decrease was prevented if the mothers were injected with oxytocin to counteract the depressant effect of the anaesthesia on milk ejection; nevertheless, GH levels in neonatal animals failed to increase following suckling. Thus some aspect of maternal activity appears to be involved in the suckling-induced increase of serum GH in rat pups. To elucidate which components of maternal activity might be involved, the effects of manipulations of ambient temperature as well as stimulation of the oral or anogenital regions were examined. Exposing rat pups to 37 degrees C (nest temperature) during the 6-h separation period before suckling prevented the separation-induced decrease in serum GH levels of 2-day-old pups. Moreover, exposure to 37 degrees C for 30 min following a 6-h separation at room temperature (22 degrees C) mimicked the effect of suckling in increasing serum GH levels in the pups. Suckling following separation at 37 degrees C was unable to increase serum GH values further.(ABSTRACT TRUNCATED AT 250 WORDS)     

Posterior pituitary lobectomy abolishes the suckling-induced rise in prolactin (PRL): evidence for a PRL-releasing factor in the posterior pituitary

The aim of this study was to determine the role of the posterior pituitary in the regulation of PRL release during suckling. Lactating rats were subjected to posterior pituitary lobectomy (LOBEX) or sham surgery (SHAM) and separation from pups in the evening; experimental manipulations and blood collection were performed the next morning. In the first experiment rats were divided into three groups: SHAM, LOBEX, and LOBEX treated with a vasopressin analog, 1-desamino-8-D-arginine vasopressin and oxytocin. Plasma PRL levels in SHAM rats increased 20- to 25-fold upon introduction of pups and remained elevated for the duration of suckling. In contrast, basal plasma PRL levels in LOBEX rats were 3- to 4-fold higher than in SHAM but suckling failed to induce a further increase. Treatment of LOBEX rats with 1-desamino-8-D-arginine vasopressin and oxytocin reduced water consumption and allowed for milk ejection and milk intake by the pups but did not restore the suckling-induced rise in PRL. The second experiment tested the functional integrity of the hypothalamic dopamine (DA) and serotonergic systems after LOBEX and the ability of LOBEX-lactating rats to respond to PRL-releasing stimuli other than suckling. Injections of alpha-methyl-para tyrosine, an inhibitor of tyrosine hydroxylase, and 5-hydroxytryptophan, a precursor of serotonin, caused 20- to 30-fold rises in plasma PRL levels in both LOBEX and SHAM rats. Exposure to ether elicited a 3- to 4-fold rise in PRL which was higher in magnitude and of longer duration in LOBEX than in SHAM rats. Conclusions: Removal of the posterior pituitary from lactating rats results in an increase in basal PRL levels and a complete abolishment of the suckling-induced rise. Vasopressin and oxytocin restore water balance and milk ejection in the LOBEX rat but fail to affect PRL secretion. The LOBEX-lactating rat is not refractory to PRL-releasing stimuli other than suckling and its hypothalamic DA and serotonergic systems are functionally intact. In addition to DA, the posterior pituitary appears to contain a PRL-releasing factor(s) which mediates the suckling-induced rise in PRL.     

Inhibition of suckling-induced milk ejections in the lactating rat by delta 9-tetrahydrocannabinol

The effect of delta 9-tetrahydrocannabinol (THC) on suckling-induced oxytocin release was investigated by recording intramammary pressure changes in suckled rats treated iv with THC (0.5 mg/kg BW) or vehicle. Latency to the first posttreatment milk ejection and posttreatment milk ejection intervals and pressure wave amplitudes were compared between THC- and vehicle-treated rats. Before treatment, intervals between milk ejections averaged 6.5 +/- 1.3 (+/- SE) and 7.0 +/- 0.7 min for vehicle- and THC-treated groups, respectively. Vehicle injections did not alter the frequency of milk ejections, which continued at an overall mean interval of 7.6 +/- 0.7 min after treatment. In contrast, THC treatment was followed by a transient suspension of milk ejections, with a latency of 59.3 +/- 7.4 min before the first posttreatment milk ejection was recorded (P less than 0.001). Intervals between subsequent ejections averaged 15.3 +/- 2.0 to 16.1 +/- 1.3 min and were lengthened relative to corresponding intervals in vehicle-treated animals (P less than 0.05). The amplitudes of pressure waves were not significantly affected by treatment. Oxytocin (0.5 mU) injections 10 or 30 min after THC treatment evoked abrupt increases in intramammary pressure, indicating continued responsiveness of the mammary gland to oxytocin stimulation. These data suggest that THC interferes with the release of oxytocin in response to suckling. To our knowledge, this provides the first evidence that THC inhibits posterior pituitary function.     

Role of the paraventricular nucleus in controlling the frequency of milk ejection and the facilitatory effect of centrally administered oxytocin in the suckled rat

The milk-ejection reflex was studied in anaesthetized, lactating Wistar rats in order to evaluate the contribution of the paraventricular nucleus (PVN) to the patterning of milk ejection and the facilitatory action of centrally administered oxytocin. In the first series of experiments, radiofrequency lesions were performed and centred: (1) antero-dorsal to the PVN, damaging parts of the medial septum and anterior hypothalamus; (2) in the PVN, such that much of the parvocellular division was destroyed, but parts of the magnocellular division remained intact; or (3) in the PVN, destroying both parvocellular and magnocellular divisions. Suckling tests performed before and after lesioning showed that the milk-ejection interval was significantly increased (decreased frequency) after lesioning in groups 2 and 3, but that milk-ejection amplitude was significantly decreased only in group 3. These results suggest that damage to the parvocellular division of the PVN affects milk-ejection frequency, but that damage to the magnocellular PVN only affects amplitude. Subsequent tests on rats injected into the PVN with the neurotoxin N-methyl-D,L-aspartate revealed a fall in the amplitude and frequency of milk ejection, similar to that after complete radiofrequency lesions of the PVN. In the second series of experiments, the facilitatory action of centrally administered oxytocin (1 mU, 2.2 ng) was examined in animals bearing either sham or complete PVN lesions. In both groups, intracerebroventricular injection of oxytocin was able to increase the frequency of milk ejections, although the incidence of milk ejection was lower in the pre- and post-injection period in the PVN-lesioned animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship between the suckling-induced release of oxytocin and prolactin in the urethane-anaesthetized lactating rat

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.     

Influence of mating and intra-uterine oestradiol infusion on peripheral oxytocin concentrations in the sow

Oxytocin concentrations were measured radioimmunologically in sows on the day of standing oestrus over a 6-h period (controls, n = 6) or 1 h before and 5 h after mating (n = 5) or transcervical infusion of either 100 ml saline (0.9% (w/v) NaCl, n = 7) or saline plus 10 micrograms oestradiol (simulation of seminal oestrogens, n = 5). In the controls, oxytocin was low, at around 1.0 pmol/l, throughout the investigation period. Similarly, saline infusion did not lead to a noticeable change in oxytocin concentrations in six out of seven sows. In one sow, however, infusion led to a maximum of 86 pmol/l at 1 min after infusion. Oestradiol led to no immediate increase in oxytocin concentrations. Later in the post-treatment period (2-5 h) they were only slightly increased (1 pmol/l vs 3 pmol/l). All mated sows reacted with a rapid and clear increase in oxytocin. Maximal concentrations (42.0 +/- 5.1 pmol/l; mean +/- S.E.M.) appeared 2 min after the onset of ejaculation. Clearly increased concentrations were found for 40 min. It was concluded that mating specifically leads to a rise in oxytocin, probably due to both mechanical and pheromonal stimuli provided by the boar.     

Secretory leukocyte protease inhibitor concentration increases in amniotic fluid with the onset of labour in women: characterization of sites of release within the uterus.

Secretory leukocyte protease inhibitor is a potent inhibitor of neutrophil function, a mediator of mucosal immunity and an inhibitor of NF|gkB regulated inflammatory responses. However, its source, function and regulation within the uterus during pregnancy and at parturition are not well defined. In amniotic fluid, the concentration of secretory leukocyte protease inhibitor increased significantly from 2nd trimester (24+/-3 ng/ml; mean+/-s.e.m.; n=20) to term (751+/-53 ng/ml; P<0.05; n=15) with a further profound increase (P<0.005) with the onset of labour (3929+/-1076 ng/ml; n=15). To establish the intra-uterine sites of secretion, explants (n=6 different patients per tissue) were collected at term after elective caesarean section. High levels of secretory leukocyte protease inhibitor were released by decidua (135.2+/-12.4 pg/mg; mean+/-s.e.m.) and chorio-decidua (325.1+/-26.4 pg/mg) with less by amnion (55.6+/-6.0 pg/mg) and placenta (9.2+/-1.9 pg/mg). Intense immunoreactivity for secretory leukocyte protease inhibitor was detected predominantly in decidua parietalis cells adherent to the chorion laeve and myometrium, and also in decidua basalis. We propose that, within the pregnant uterus, secretory leukocyte protease inhibitor is released by decidua, fetal membranes and potentially the fetal lung. The increase in secretory leukocyte protease inhibitor may act to modulate pro-inflammatory paracrine interactions for the maintenance of pregnancy and limit those occurring at parturition within the uterus.     

The physiological release of specific individual neurophysins into the circulation of pigs.

Specific homologous radioimmunoassays for the two major porcine neurophysins have been developed and utilized to measure plasma neurophysins during events known to release vasopressin (dehydration and hemorrhage) and oxytocin (parturition and suckling). During hemorrhage plasma neurophysin I increased 2-25 times and decreased toward basal levels after reinfusion of the blood while plasma neurophysin II was low and showed only minor fluctuations. Neurophysin II was released during parturition and suckling in a pattern similar to that reported for oxytocin release during these events. A rise in plasma neurophysin II occurred towards the end of parturition and spurt release occurred in suckling. The function of neurophysins in the plasma is unknown but porcine neurophysin I has been shown to be released independently into the circulation in response to hemorrhage. Independent release of neurophysin II during parturition and suckling was not demonstrated. In the pig, release of neurophysin I may be associated with vasopressin release and neurophysin II associated with oxytocin release.     

Emergence of the diurnal rhythm in plasma melatonin concentrations in newborn lambs delivered to intact or pinealectomized ewes

We have monitored the 24-h profiles of plasma melatonin concentrations between birth and 10 weeks of age, in lambs which were delivered to, and suckled, either pineal-intact (control group) or pinealectomized (pinealectomized group) ewes. Between 0 and 2 weeks of age, plasma concentrations of melatonin in lambs suckling either intact or pinealectomized ewes were highest at 01.00 h. At this age, however, there was no significant difference in the mean plasma concentrations of melatonin between the entire dark and light phases in lambs in either the control group (dark, 39.7 +/- 6.0 (S.E.M.) pmol/l; light, 39.5 +/- 8.1 pmol/l) or the pinealectomized group (dark, 79.8 +/- 43.3 pmol/l; light, 60.9 +/- 8.7 pmol/l). Between 3 and 4 weeks of age, however, a diurnal rhythm in plasma melatonin concentrations was clearly present in the lambs in both the control and pinealectomized groups (control group: dark, 164.1 +/- 5.6 pmol/l; light 26.2 +/- 2.5 pmol/l; pinealectomized group: dark, 52.7 +/- 8.0 pmol/l; light, 19.1 +/- 5.3 pmol/l; P less than 0.001). Between 3 and 10 weeks of age, plasma concentrations of melatonin in the dark phase were significantly (P less than 0.05) lower in the lambs suckling pinealectomized ewes than in the control group. In both the control lambs and lambs suckling pinealectomized ewes, the mean plasma concentrations of melatonin in the dark and light phases increased significantly (P less than 0.05) between 3 and 6 weeks after birth. In conclusion, we have demonstrated that a clear diurnal plasma rhythm in melatonin concentrations does not emerge until 3-4 weeks of age in lambs suckling either pinealectomized or intact ewes.(ABSTRACT TRUNCATED AT 250 WORDS)