Rise In Intramammary Pressure Research Articles

Atosiban, an oxytocin receptor blocking agent: pharmacokinetics and inhibition of milk ejection in dairy cows.

Plasma concentrations of the oxytocin receptor blocking agent Atosiban were measured at 2, 4, 10, 15 and 20 min after injection of 5, 10, 20 and 50 micrograms Atosiban/kg body weight in six dairy cows. The half life of Atosiban was 18 min and the total body clearance was 3301 ml/min. Intramammary pressure (IMP) within the teat cistern was measured in six cows before and after i.v. injection of 0 or 20 micrograms Atosiban/kg body weight and repeated injections of 0.2 or 0.5 i.u. oxytocin. IMP was also measured in eleven cows after injection of 0, 10 or 50 micrograms Atosiban/kg body weight: in seven during oxytocin infusions, in four after oxytocin injections in successively increasing dosages (0.05, 0.1, 0.2, 0.5, 1 and 10 i.u.). The occurrence of milk ejection was indicated by a rise in IMP. After injection of 20 micrograms Atosiban/kg body weight, 0.2 i.u. oxytocin did not induce an IMP rise before 48 min, whereas 0.5 i.u. oxytocin induced an IMP rise within 4 min. The time from the start of infusion until the beginning of the IMP rise and the duration of IMP rise during oxytocin infusions both increased, whereas the IMP rise itself was diminished by increasing Atosiban dosages. The amount of injected oxytocin necessary to induce an IMP response increased with increasing Atosiban dosages. Atosiban was shown to have a powerful effect in inhibiting milk ejection in dairy cows.

Relaxin inhibits the pulsatile release of oxytocin but increases basal concentrations of hormone in lactating rats.

The effects of porcine relaxin were examined in urethane-anesthetized, lactating rats to clarify the actions of relaxin on basal levels and the pulsatile release of oxytocin during suckling. Baseline plasma oxytocin concentrations were 27.6+/-2.9 pM in unsuckled rats. They were significantly increased in rats after the onset of suckling (34.5+/-2.4 pM) compared with unsuckled animals. Oxytocin concentrations were increased further during reflex milk ejection (41.2+/-2.7 pM). Treatment with porcine relaxin (5 microg in 0.1 ml saline) caused a significant (p < 0.05) increase in plasma oxytocin compared with pretreatment concentrations or saline injection in all three groups. Frequent blood samples were taken before and during milk ejection to confirm pulsatile oxytocin associated with reflex milk ejection. Short-lived (5-10-sec) pulses with peak concentrations of oxytocin 392.4+/-122.1 pM were observed shortly before or at the time of the peak rise in intramammary pressure associated with reflex milk ejection. Relaxin (5 microg i.v.) completely suppressed the pulsatile release of oxytocin. These data suggest that relaxin increases basal secretion of oxytocin but inhibits the pulsatile secretion associated with reflex milk ejection.

Integrity of the secretory epithelium of the lactating mouse mammary gland during extended periods after suckling.

The transepithelial potential difference (TEPD) and resistance (TER) in the alveoli, and the pressure in the mammary gland were measured in lactating mice at different times after they had finished suckling their young. Oxytocin administration caused a rise in TEPD, the amplitude of which decreased with time after cessation of the pups feeding. At 2.5 h after cessation, oxytocin administration caused a short-term decrease in TER, but at 5 h there was a two phase reaction: a fall in TER followed by a rise. At 20 h there was a one phase reaction, a rise in TER. TEPD in the mammary gland alveoli increased from its initial value of 18 +/- 1 mV to 25 +/- 1 mV at 2.5 h, and thereafter fell to zero by 15 h after suckling. The initial value of TER, 114 +/- 1 k omega. increased to 131 +/- 4 k omega at 2.5 h and then decreased by 5 h after suckling, but unlike TEPD it did not reach zero. These experiments show that extending the period after cessation of suckling does not cause a rise in intramammary pressure and that oxytocin injections cause a short-term rise in milk pressure. From these results, we propose a hypothesis of reducing ionic concentration gradients between intracellular fluid, cytoplasm of the secretory cell and milk at extended periods after cessation of suckling starting at 2.5 h.

Central inhibitory effects of muscimol and bicuculline on the milk ejection reflex in the anaesthetized rat.

1. In order to determine whether GABAergic mechanisms are involved in the control of the milk ejection reflex in the rat, we examined the effects of central administration of a GABAA receptor agonist (muscimol) and antagonist (bicuculline) on the milk ejection reflex in the urethane-anaesthetized rat. 2. Intracerebroventricular (i.c.v.) injection of both muscimol (n = 17), at doses of 5, 10 and 20 ng, and bicuculline (n = 15), at doses of 0.01, 0.1 and 0.3 microgram, inhibited the milk ejection reflex in a dose-dependent manner. The bicuculline-induced inhibition was accompanied by desynchronization of the electroencephalogram and, at the highest dose, by alteration in the sensitivity of the mammary gland to oxytocin. No significant effect on the milk ejection reflex was seen with i.c.v. isotonic saline (n = 5). 3. Injection of 20 (n = 5) or 40 ng (n = 2) muscimol or 0.1 microgram bicuculline (n = 5) i.c.v. did not significantly alter the rise in intramammary pressure evoked by electrical stimulation of the neurohypophysis. 4. Bilateral 400 nl microinfusions directly into the supraoptic nuclei of either muscimol (20-100 ng microliter(-1); n = 10) or bicuculline (0.15 micrograms microliter(-1); n = 5) [corrected] resulted in an inhibition of the milk ejection reflex, which was not accompanied by desynchronization of the electroencephalogram. 5. The effects of i.c.v. injections of muscimol (15 and 20 ng) and bicuculline (0.01, 0.12 and 0.3 microgram) on the electrical activity of twenty-seven antidromically identified supraoptic magnocellular neurones were examined. Both compounds resulted in an inhibition of the background firing of oxytocinergic and vasopressinergic cells, and delayed the occurrence of high frequency bursts in oxytocin neurones. In five supraoptic neurones, bicuculline induced a transient activation before inhibition. 6. The powerful inhibitory action on the milk ejection reflex of both muscimol and bicuculline provides evidence for the importance of GABA neurones in maintaining the functional integrity of the mechanisms which allow the intermittent and pulsatile release of oxytocin during suckling.

Transepithelial potential difference in the goat mammary gland and its change during hand milking, and administration of oxytocin and catecholamines.

The reaction of secretory epithelium and myoepithelial cells in the alveoli to hand milking and i.v. injection of oxytocin and catecholamines was studied in lactating goats. The reaction of secretory cells was assessed by changes in the transepithelial (blood-milk) potential difference (PD), and the contractile reaction of myoepithelial cells by the growth of intramammary pressure (IP). The initial value of PD was 24.6 +/- 0.6 mV, that of IP 3.32 +/- 0.08 kPa (24.9 +/- 0.6 mmHg). Milking and oxytocin administration caused a rise in PD and an increase in IP. After noradrenaline and adrenaline injections two-phase PD changes and a short-term rise in IP were recorded. Isoproterenol, a beta-agonist, caused a rise in PD but did not affect IP. Phenylephrine, an alpha-agonist, caused two-phase and one-phase changes in PD. Simultaneously, a rise in IP was recorded. The results show that the reaction of the mammary gland to the substances administered is complex. Myoepithelial and secretory cells respond differently to short-term rises in the level of mediators and hormones in the blood.

Oxytocin release evoked by electrical stimulation of the medial forebrain in the rat: Analysis of stimulus parameters and supraoptic neuronal activity

The role of the medial forebrain area (vertical limb of the diagonal band, medial septum and medial nucleus accumbens) in the control of oxytocin secretion in lactating rats was investigated. Electrical stimulation of the medial forebrain evoked a reproducible rise in intramammary pressure, equivalent to that caused by i.v. injection of 1 mU oxytocin. No presser effect accompanied this response. Radio-immunoassay of plasma samples showed that stimulation caused a significant rise in the concentration of circulating oxytocin. The effects of changing the parameters of stimulation to the medial forebrain were compared with those evoked by stimulation of the neural stalk. The optimal frequency for stimulation of the forebrain was found to be four-fold lower (10–20 Hz) than that for stimulation of the neural stalk (50 Hz). During continuous prolonged stimulation of the forebrain (20 Hz; 2 min) only a single transient response was obtained, whereas a protracted response was obtained as a result of prolonged stimulation of the stalk. Recordings were made from antidromically identified neurosecretory cells in the supraoptic nucleus. Electrophysiological responses to electrical stimulation of the medial forebrain were characterized by two main features. (1) Single-pulse stimulation produced only a small excitation (one or two action potentials), while high-frequency trains produced a profound facilitation of this response, with each pulse evoking short-duration ‘bursting’ behaviour in the supraoptic neurons. (2) During long trains of stimulation this frequency-dependent facilitation declined and could only be renewed after a period of rest. The study indicates that the medial forebrain provides a major excitatory afferent input to the oxytocin-secreting neurons of the supraoptic nucleus, and that the pulse-like secretion of oxytocin following medial forebrain stimulation results from the complex facilitatory and adaptive characteristics of this input.

Effects of chronic infusion of porcine relaxin on oxytocin release in lactating rats.

The effects of chronic infusion of porcine relaxin on oxytocin release were studied in lactating rats. Infusion of relaxin (4.2 micrograms/h for either 4 or 6 days) suppressed reflex milk ejection and reduced litter weight gain for 48 h compared with saline-infused controls. After 2 days, neither the rate of growth nor the frequency of milk ejection were significantly different from controls. For 24 h after the infusion of relaxin ended, litters gained weight more quickly than controls but there was no difference seen in the frequency of milk ejection. The effects on oxytocin release of stopping an infusion of relaxin after 3 days were investigated. There was a significant (P less than 0.01) rise in plasma oxytocin (up to 90 pmol/l) 30 min after the infusion was stopped, followed by a sustained rise in intramammary pressure. Treatment of relaxin-infused rats with naloxone (0.1 mg/kg) when the infusion was halted caused a more rapid release of oxytocin (within 2 min), a greater release of oxytocin (up to 140 pmol/l) and a prolonged rise in intramammary pressure.

Somatosensory systems and the milk-ejection reflex in the rat. I. Lesions of the mesencephalic lateral tegmentum disrupt the reflex and damage mesencephalic somatosensory connections

Bilateral electrolytic lesions and unilateral tracer injections were performed in lactating rats in order to study the participation of the mesencephalic lateral tegmentum in the milk-ejection reflex. The release of oxytocin was detected as a rise in intramammary pressure during each milk ejection. In animals with lesions, the lateral part of the deep grey layers of the superior colliculus, the intercollicular area and the rostromedial portion of the external nucleus of the inferior colliculus were destroyed. The mesencephalic lateral tegmentum of animals in which the milk-ejection reflex was blocked sustained a larger damage than in rats where the frequency of the milk-ejection response was only slowed down. Solutions of True Blue, horseradish peroxidase or horseradish peroxidase coupled to wheat germ agglutinin were injected in the mesencephalic lateral tegmentum of rats with and without lesions. Retrogradely labelled cells were found in several nuclei of the somatosensory pathways: the principal sensory and spinal parts of the trigeminal complex, the cuneate and gracile nuclei, the lateral cervical nucleus and the nucleus proprius of the spinal cord. Labelled cells were also found in the ventral nucleus of the lateral lemniscus. the ventral parabrachial nucleus, the gigantocellular reticular nucleus, the lateral nucleus of the substantia nigra, the prerubral nucleus of the thalamus, the hypothalamic ventromedial nucleus, the zona incerta and in the anterior and lateral hypothalamic areas. Labelled fibres and “terminal-like” labelling were found in the anterior pretectal area, in the thalamic parafascicular nucleus, in the posterior nucleus and the ventroposterior complex, in the zona incerta and in the fields of Forel, but none were observed in the supraoptic or paraventricular nuclei. Injections made in the area of the lateral cervical nucleus and in the cuneate and gracile nuclei labelled fibres and “terminal-like” fields in the external nucleus of the inferior colliculus. the intercollicular area, the deep grey layers of the superior colliculus and in the mesencephalic lateral tegmentum. After injections in the posterior nucleus and ventroposterior complex of the thalamus, retrogradely labelled cells were found in the lateral tegmentum, the intercollicular area and the external nucleus of the inferior colliculus. These results indicate that bilateral lesioning of the mesencephalic lateral tegmentum, which disrupts the milk-ejection response, could damage somatosensory projections originating from the dorsal horn of the spinal cord, the lateral cervical nucleus, the dorsal column nuclei and the sensory and spinal trigeminal nuclei. The importance of the intercollicular area and of the external nucleus of the inferior colliculus in the processing of the somatosensory informations is further suggested by the projection of those two areas onto the ventroposterior complex of the thalamus.

The milk ejection reflex in the pig.

1. The milk ejection reflex in response to suckling was studied in conscious sows by continuous recording of intramammary pressure, radioimmunoassay of plasma concentrations of neurohypophysial hormones, and observation of the behaviour of the sows and piglets.2. A regular pattern of nursing, suckling and milk ejection was observed. The mean duration of the suckling period was 6.3 min. Over 144 suckling periods, 113 milk ejections were recorded. Each milk ejection was characterized by a sudden rise in intramammary pressure reaching 20-49 mmHg, and lasting 8-41 sec. Milk ejections occurred only once per suckling period, at a mean interval of 44.3 min.3. Each milk ejection occurred with a mean latency of 2.4 min from the onset of a period of initial massage of the udders by the piglets, and was coincident with a period of quiet suckling when the piglets were consuming milk. The onset of nursing was signalled by the sows grunting in a rhythmic manner. In most cases, the frequency of grunts, at first low, increased suddenly 23 sec before milk ejection.4. During eighteen suckling periods leading to milk ejection, neurohypophysial hormone assays performed on serial blood samples showed an increase in plasma concentration of oxytocin up to 30 sec before milk ejection. The concentration of lysine-vasopressin did not rise above basal levels.5. In 21.4% of the suckling periods, no rise in intramammary pressure was observed. In these ;incomplete sucklings', the sow usually failed to grunt rapidly, and the piglets obtained no milk. For three of these periods, hormone assay showed no increase in oxytocin or vasopressin concentrations in blood.6. Oxytocin given intravenously produced variations in intramammary pressure which depended on the dose and the rate of injection. Rapid injections of 25-50 m-u. oxytocin, caused milk ejections similar to those induced by suckling. When oxytocin was administered at different rates, the faster the injection, the shorter the latency and the higher the amplitude of the response. Plasma concentrations of oxytocin after injection of 25 m-u. were similar to those observed during reflex milk ejection.7. Trains of electrical pulses were applied to the posterior pituitary of four anaesthetized sows. At frequencies of stimulation above 10 Hz, a rise in intramammary pressure and an increase in plasma oxytocin and vasopressin concentrations were observed. At frequencies of stimulation of 30-50 Hz, the response of the mammary gland and the time course of the variations in oxytocin plasma concentrations were similar to those observed during natural reflex milk ejection.8. It is concluded that reflex milk ejections during suckling in the pig are caused by the intermittent and spurt-like release of about 25 m-u. oxytocin, without concomitant vasopressin release. It is postulated that the release of oxytocin is probably precipitated by a brief and massive activation of oxytocin-secreting neurones in the hypothalamus. Central mechanisms controlling the intermittent release of oxytocin are discussed.

Electrophysiological recordings from oxytocinergic neurones during suckling in the unanaesthetized lactating rat.

A method is described for making extracellular recordings of the spontaneous activity of single hypothalamic neurones in unanaesthetized, freely moving, lactating rats using chronically implanted micro-wire electrodes. Extracellular recordings taken from individual neurones were maintained for periods of between 1 and 12 days. These records were not affected by any normal movement of the animal. As several micro-wires were implanted into each animal it was possible to make simultaneous recordings from several different hypothalamic sites in the same animal. Some recordings were identified as those from magnocellular neurones in the paraventricular nucleus on the basis of antidromic invasion after electrical stimulation of the neurohypophysis. Milk ejection in response to the prolonged sucking of ten or more pups was intermittent, and individual milk ejections recurred at intervals of 2-10 min throughout each period of nursing. The rise in intramammary pressure at milk ejection was associated with a vigorous extensor response from the pups. This was monitored by radar to provide an index of milk ejection in the unanaesthetized rat. Eleven antidromically identified neurones were recorded through 321 milk ejections. Eight of these neurones displayed a transient (2-6s) and very substantial acceleration in discharge at the time predicted for oxytocin release, i.e. 10-12s before milk ejection. The background discharge of these cells was 0.1-2.6 action potentials/s; this increased to 16-50 action potentials/s during the brief period of accelerated activity. Twenty-five neurones were studied during 365 milk ejections in rats which did not have a stimulating electrode implanted in the neurohypophysis. Thirteen of these neurones displayed a burst of high frequency discharge before each milk ejection, similar to that described for the antidromically identified neurones. Two of the non-responsive cells displayed a phasic pattern of discharge, characteristic of vasopressinergic neurone discharge recorded in anaesthetized rats. These observations of putative oxytocinergic neurones in unanaesthetized, freely moving rats are identical with those previously made on anaesthetized rats, and establish that the high frequency burst of electrical activity displayed by magnocellular neurones some 10-12s before milk ejection is responsible for oxytocin release under normal physiological circumstances.

Sleep is not a prerequisite for the milk ejection reflex in the pig.

Electroencephalographic and electromyographic activities were recorded together with variations in intramammary pressure in unanaesthetized lactating sows during suckling. During each suckling period, milk ejection resulted in a sudden and brief rise in intramammary pressure. From the onset of suckling to the beginning of milk ejection, polygraphic recordings as well as observations of behaviour revealed that the sow was invariably in a state of arousal. This suggests that, unlike what has been proposed for the rat, sleep is not a necessary component of the milk ejection reflex in the pig.

Electrochemical deposition of iron onto rat oxytocinergic neurones does not result in milk-ejection.

Exposure of the preoptic-tuberoinfundibular system to iron ions causes ovulation in the pro-oestrous rat. This electrochemical treatment has been used frequently to study the way that the hypothalamus triggers secretion of gonadotrophic hormone and most investigators have assumed that the effect is mediated by the direct excitation of those neurones exposed to the cation. The present paper reports experiments designed to confirm that iron directly excites hypothalamic neurones to the firing frequencies essential for ovulation. Because of the difficulty of measuring input-output relationships in the diffuse preoptic-tuberoinfundibular system the experiments were performed on the anatomically distinct oxytocinergic neurones of the paraventricular-hypophysial tract in lactating rats. These neurones were simulated electrically (from 10 to 50 Hz for between 1 and 300 s) and subjected to the electrochemical treatment by depositing iron ions from the tip of the stimulating electrode (up to 250 muA anodal current for between 60 and 180 s). Changes in intramammary pressure were used to indicate oxytocin release and mammary glands were calibrated for sensitivity by intravenous injection of oxytocin. Electrical stimulation in excess of about 15 Hz invariably caused release of sufficient oxytocin to cause a rise in intramammary pressure. In contrast, no changes in pressure were observed during or after the electrochemical deposition of iron. The results suggest that the response of hypothalamic neurones to electrochemical treatment is not the same as their response to electrical stimulation.

Excitation of phasically firing hypothalamic supraoptic neurones by carotid occlusion in rats.

1. The activity of supraoptic neurones has been recorded extracellularly during bilateral occlusion of the common carotid arteries in anaesthetized rats. 2. Experiments in lactating rats showed that occlusion liberated sufficient amounts of neurohypophysial hormones to cause a rise in intramammary pressure 15-25 s after the onset of occlusion. 3. Ninety-one percent of the phasic neurones (defined as those showing bursts of activity alternating with periods of silence) were activated by carotid occlusion less than 10 s after the onset of occlusion. Most randomly firing neurones were inhibited or were unaffected. 4. The activation of phasic neurones is unlikely to be just a nonspecific effect, because in the same animals, phasic neurones were excited whilst random neurones were not. 5. Moreover, in phasic neurones, statistical analysis shows (a) that the intervals during which an occlusion was performed were significantly shorter than the intervals between spontaneously occurring bursts, and (b) that this activation was followed by a period of reduced firing probability. 6. The results are discussed with reference to the correlation of supraoptic neuronal activity with hormone release. The possibility is considered of relating the tendency of some supraoptic neurones to fire in bursts with the secretion of vasopressin.

Isolation of kallikrein from mammary gland of cows.

Small amounts of kallikrein were isolated from bovine mammary tissue. Immediately after slaughter, the udder was removed, perfused with ice-cold xylocaine-Rheomacrodex solution, and homogenized in ice-cold acetic acid solution. Kallikrein-like substances were adsorbed on DEAE-cellulose, eluted with ammonium formate and fractionated by acetone precipitation. The 40-70% acetone fraction was separated on a Shephadex-G 75 column and aliquots of each fraction were incubated for 1 or 2 hr with kininogen. After incubation, extracts were tested for kinin activity on the isolated rat uterus, the rat duodenum, blood flow through the femoral artery of dogs and by intramammary pressure assay in sheep. Release of kinins was demonstrated by all these techniques, highest kallikrein activity being detected in the molecular weight range 50,000 to 60,000. Direct injection of the enzyme into the udder artery of sheep, without prior incubation with kinninogen, resulted in vasodilation and a rise in intramammary pressure. The kallikrein activity was strongly inhibited by aprotinin, but not by soybean trypsin inhibitor or ovomucoid. Kallikrein activity was always associated with esterase activity.

Factors governing the periodic activation of supraoptic and paraventricular neurosecretory cells during suckling in the rat.

1. One hundred and thirty-two antidromically identified paraventricular (PV) and supraoptic (SO) neurones were studied during milk ejection evoked by the suckling of the young, in thirty-four lactating rats anaesthetized with urethane (1.1 g/kg I.P.). The spike activity of these neurones was analysed to determine some of the factors governing their orthodromic activation. Changes in spike activity were compared with the amount of oxytocin released from the neurohypophysis, as measured by the rise in intramammary pressure at milk ejection. 2. Sixty-five units, from both the PV and SO nuclei, displayed a characteristic burst of accelerated activity 12-18 sec before milk ejection. With a suckling stimulus of nine or ten pups, these neurosecretory responses were both regular in their occurrence (recurring evey 4-8 min) and uniform in character, i.e. all responsive cells accelerated to 30-80 spikes/sec for 1-4 sec. Two units deviated from this pattern and both were recorded in animals which failed to display a detectable milk ejection...

Electrophysiological evidence for the activation of supraoptic neurones during the release of oxytocin.

1. Antidromically identified supraoptic (SO) units were recorded in lactating rats anaesthetized with urethane (1.1 g/kg), and their activity was studied during milk ejection evoked by the suckling of the young. Fifty-eight SO units were recorded through 174 milk ejections. Each milk ejection was the result of a neurohypophysial release of an oxytocin pulse of 0.5-1.5 m-u.2. Fifty-five of the SO units displayed background activity and three were silent. The firing rates ranged from 0 to 15.4 spikes/sec, the distribution was exponential with 26% of the units firing at < 1 spike/sec. Sixteen (28%) of the SO units displayed a phasic pattern of activity characterized by periods of activity (6-132 sec) and silence (4-71 sec).3. Twenty-five of the non-phasic units displayed a large and stereotyped acceleration in spike activity some 10.5-17.4 sec before the rise in intramammary pressure at milk ejection. Units accelerated to rates between 9-66 spikes/sec, an increase of about thirtyfold (median) on background activity. The response was brief (0.9-4.7 sec) and was followed by a period of after-inhibition.4. It was concluded from studies of double recordings and observations of multi-unit activity that all the responsive units were synchronously activated. The mean latency of 13.3 sec between the onset of the neurosecretory response and milk ejection was similar to that observed following the experimental release of endogenous oxytocin by electrical stimulation of the neurohypophysis (50 pulses/sec for 2-4 sec).5. Four of the phasically active units were correlated with the oxytocin release for milk ejection. Three of these units displayed a burst of activity superimposed on the terminal portion of an active phase, some 10.2-14.7 sec before milk ejection. The fourth unit, recorded in conjunction with a responsive non-phasic unit, consistently switched from silence to activity coincident with the onset of the SO activation.6. The remaining SO units and a further ten units that were not antidromically activated by neurohypophysial stimulation displayed no change in activity during either the period of neurosecretory activation or the period of after-inhibition.7. This activation of the SO neurones, in the formulation of oxytocin release and milk ejection, is the same as that we have previously observed in recordings from the paraventricular (PV) region, and the proportion of neurones displaying the response is similar: 48% in the SO nuclei, 58% in the PV nuclei. We conclude, since the SO nuclei contain 80% of the neurosecretory cells that project to the neurohypophysis, that the SO nuclei are as important, if not more so, than the PV nuclei in the control of oxytocin release.

Effect of locally implanted oestrogen on the response of the rat's lactating mammary gland to oxytocin.

ABSTRACT On the day following delivery (day 1 of lactation) one abdominal mammary gland was implanted with oestrogen and the contralateral gland received an empty needle. At 2, 5 or 10 days of lactation the rats were anaesthetized with pentobarbital and the nipples of both abdominal glands were cannulated and their pressures recorded by means of transducers coupled to an amplifier and recording system. The normal mammary glands of 5-day lactating rats responded to very low doses of oxytocin (Syntocinon®, Sandoz) (5× 10−8 mU) with a rhythmic elevation in pressure. However, saline infusion also evoked a small rise in intra-mammary pressure. Earlier (2 days) and later (10 days) in lactation the responses were smaller. Oestrogen decreases significantly the milk ejection response to oxytocin, and the effect was maximal at day 10 of lactation. Histological observations confirmed the diminished reaction of the gland to oxytocin, since the milk was retained in the alveoli of rats bearing a mammary-oestrogen implant. A paradoxical rise in pressure was detected in normal as well as in oestrogen-implanted glands when the lowest dose of oxytocin was injected in lactating rats which had previously received a high dose of oxytocin (50 mU or 500 mU). These results reinforce the hypothesis that oestrogen alters the milk ejection response to oxytocin and that the mechanism is probably related to changes in the contractility of the myoepithelial cells.

The milk-ejection reflex of the rat: a 20- to 40-fold acceleration in the firing of paraventricular neurones during oxytocin release.

SUMMARY Antidromically-identified paraventricular (PV) neurones were studied in the anaesthetized rat during milk ejection (ME) evoked by the natural stimulus provided by the suckling young. An intermittent pattern of ME was observed, though the ME interval was slightly longer (10–20 min) in the doe during unit recording than in unoperated or conscious animals (5–15 min). PV neurones displayed a steady background of spike discharge in both the non-lactating rat and lactating rat during suckling. The firing rates ranged from 0 to 9·4 spikes/s and were exponentially distributed with 32% of the units firing at &lt;1 spike/s. Phasic patterns of discharge were seen in 18% of the units. Fifty-eight per cent of the PV units displayed a stereotyped and explosive acceleration in spike activity some 15–20 s before the rise in intramammary pressure at ME. The peak firing rate during this brief (2–4 s) response was in the range of 24–84 spikes/s. A prominent after-inhibition was then displayed by these responsive units, though this second component of the response was variable in duration (7–56 s). The electrical activity of the remaining units, including all the cells with phasic discharge, was not correlated with ME. There was no apparent change in the intensity of the suckling stimulus at the time of PV activation, i.e. 15–20 s before ME. Secondly, the activity of the PV neurones was not influenced by the rise in the intramammary pressure and the increased sucking of the pups at the time of ME. Simulation of an individual ME was obtained with both 1 mu. exogenous oxytocin, i.v., and electrical stimulation of the neurohypophysis (50 pulses/s for 4 s). The latencies to ME ranged from 7–14 s and 11–23 s, respectively. In conclusion, an explosive increase in the electrical activity of PV neurones precipitates the release of oxytocin at ME.

The milk-ejection reflex of the rat: an intermittent function not abolished by surgical levels of anaesthesia.

SUMMARY Milk ejection (ME) was studied in the lactating rat by the measurement of intramammary pressure, milk yield and behaviour. An intermittent pattern of ME was observed, both in conscious rats and in rats anaesthetized with tribromoethanol, pentobarbitone sodium and ethyl carbamate. Milk ejection was never seen as a reflex response with the onset of the suckling stimulus; the latency to the first ME varied from 10 min in the conscious rat to &gt; 60 min during deep anaesthesia. Thereafter, ME occurred at regular intervals ranging from 3 to 5 min under optimal conditions to &gt; 15 min during less favourable circumstances, though the young were content to suckle continuously for several hours irrespective of whether they obtained milk. Each ME was associated with an abrupt and uniform rise in intramammary pressure and simulation of the individual ME was obtained by a rapid i.v. injection of 1 mu. oxytocin. The pups reacted immediately to the rise in pressure with a synchronized extensor reaction and a vigorous increase in suckling which lasted for about 10 s. There was, however, no apparent change in the suckling stimulus to account for the release of endogenous oxytocin some 10–20 s prior to these events. Milk ejection was a reflex phenomenon, nonetheless, for the removal of the pups abolished the intermittent release of oxytocin. The rise in intramammary pressure was associated directly with the ejection of the milk and the pups abruptly increased in weight. Yields of 10–12 g were obtained from both conscious and anaesthetized rats after 18 h of separation from their young. Most of this milk was expressed in the first ten milk ejections. Mammary distension and the size of the litter were important parameters determining the pattern of ME. The incidence of ME was appreciably lower in unseparated rats whether anaesthetized or not and in separated rats after prolonged suckling or artificial removal of the milk. A similar reduction was encountered when the number of suckling pups was changed from ten to five. The physiological principles of the ME reflex in the rat are discussed and a theoretical model is presented.

The blood levels of oxytocin during machine milking in cows with some observations on its half-life in the circulation.

SUMMARY Blood oxytocin was assayed on the lactating guinea-pig mammary gland. Transient release of oxytocin was detected in the range of 9–889 μu./ml. plasma in the external jugular vein blood of cows during machine milking. In 24 experiments oxytocin was detected in 17; in nine of these, hormone release by conditional reflex was obtained in response to auditory or visual stimuli associated with the milking routine. All stimuli associated with the milking routine caused oxytocin release. There was no difference in the plasma concentrations detected in response to conditional and nonconditional stimuli. In this and previous work no release of oxytocin was detected in 32% of experimental machine milkings although in most cases milk yields were normal. Milk ejection was accompanied by increases in intramammary pressure from 12·5–24 mm. to 23–44 mm. Hg; the pressure after ejection was maintained virtually constant for over 1 hr. There was no initial drop in pressure, associated with a neural segmental reflex. Rise in intramammary pressure accompanied the detection of oxytocin in jugular blood in four out of seven experiments. The pressure changes and latency could be simulated by rapid intravenous injection of 100 m-u. synthetic oxytocin. The half-life of synthetic oxytocin in the circulation was 127 ± 9·3 sec. (range 85–164 sec.).