An analysis of specific stimuli causing the release of prolactin and growth hormone at milking in the goat.

Experiments to investigate the relative importance of the tactile, conditioned and possible metabolic components of the milking stimulus on the release of prolactin and growth hormone (GH) from the anterior pituitary of the goat are described. A comparison of the hormonal responses to milking the auto-transplanted mammary gland (i.e. denervated gland) with that obtained by milking the intact mammary gland of the same goat showed that the concentration of prolactin in the plasma increased only after milking the intact gland, whereas in two out of four goats an increase in plasma GH was detected several minutes after milking the transplanted gland. In the intact animal significantly more prolactin (P less than 0-01) was released in response to milking both teats for 6 min as compared with that released by milking only one teat for the same time. No significant difference (P less than 0-1) was found for GH. Similar quantities of prolactin and GH were released by goats milked when conscious or under anaesthesia. A comparison of the hormonal responses to teat stimulation in the same anaesthetized goats with and without the removal of milk from the mammary gland showed a significant reduction (P less than 0-001) in the quantity of prolactin and GH released in the absence of milk removal. The significance of these results is discussed.

The secretion of citrate into milk.

1. The time course of changes in specific activities of citrate, lactose and fatty acids in milk during frequent milking, following the I.V. administration of labelled glucose, acetate and chylomicrons in goats has been studied. Peak specific activities of lactose and citrate in milk were reached at 2-3 hr, while peak specific activites of fatty acids were reached at 5-7 hr. 2. Following short I.A. infusions of 24Na, 36Cl, and 42K, peak specific activities in milk were reached in 1 hr or less. 3. The mammary epithelium of lactating goats was found to be virtually impermeable to labelled citrate in both directions. 4. Labelled citrate had an apparent volume of distribution in lactating guinea-pigs mammary slices in vitro similar to that of extracellular space markers. 5. Treatment of goats with large doses of oxytocin markedly increased the permeability of the secretory epithelium to labelled citrate. 6. In the goat mammary gland, citrate, protein and calcium failed to enter milk which had been diluted with isosmotic lactose by intraductal injection, whereas Na, K and Cl did enter, thus tending to restore the concentrations of these ions to normal. 7. It is suggested that citrate, which is formed within the sucretory cell, enters milk not by passage across the apical cell membrane but, in common with lactose and milk protein, by exocytosis of Golgi vesicles. It appears that citrate is held at high concentrations in milk by virtue of the impermeability of the mammary epithelium to the forms in which it occurs in milk.

The effects of prolactin and oxytocin on milk secretion and on the permeability of the mammary epithelium in the rabbit.

The effects of prolactin or oxytocin on milk secretion and the permeability of the mammary epithelium have been investigated in rabbits. 2. Milk yield was increased by prolactin treatment in late (25-28 days) but not in established (11-14 days) lactation. 3. Prolactin treatment increased milk [lactose] and [K] and decreased [Na] and [Cl] in late lactation, and thus reversed the normal changes in late lactation, but had no significant effect in established lactation. 4. [14C]sucrose movements from blood to milk were significantly decreased to levels characteristic of established lactation, following prolactin treatment in late lactation. No significant effect was evident with treatment in established lactation. Na and Cl movements showed similar trends. 5. It is suggested that prolactin in some way affects paracellular movements of ions and small molecules like lactose across the mammary epithelium, and that this mechanism is responsible for the changes in the composition of the aqueous phase of milk. 6. Immediately following a single dose of 100 m-u. oxytocin no significant effects on milk composition were evident but after 1 u. milk [Na] and [cl] were significantly increased. 7. Twenty-four hr after 1 u. oxytocin, milk [Na] and [cl] were decreased while [K], [lactose], [fat] and [protein] were increased. 8. During an I.V. infusion of oxytocin milk [Na] and [Cl] increased while [K] and [lactose] decreased. The passage of [(14)C]sucrose, 24Na and (36)Cl from blood to milk also increased. 9. These effects of oxytocin are discussed in relation to the permeability of the mammary epithelium and the pathways for ion movements, and to other studies on milk composition in the rabbit involving the administration of oxytocin to aid in the evacuation of milk.

Secretory activity of goat mammary glands during pregnancy and the onset of lactation.

1. The volume of the udder and the composition of the secretion have been followed in five goats through pregnancy to the onset of lactation. 2. During the middle of pregnancy udder volume was minimal and there was little or no fluid in the teats. 3. Two stages of commencing secretory activity (lactogenesis) were defined. In the first, starting up to ten weeks pre-partum, udder volume increased and the fluid in the teats changed from an extracellular-fluid-like to a milk-like composition and acquired a high concentration of immunoglobulins. Four goats accumulated several litres of a pre-colostral fluid with a high [lactose] 6-7 weeks pre-partum. 4. Comparison of the rate of increase in udder volume with previous data for the rate of increase in empty udder volume in pregnant goats showed that the rate of secretion, even in the last few days of pregnancy, was only a few per cent of the rate immediately after parturition. 5. In the last 2-3 days of pregnancy there was a three to elevenfold increase in [citrate] in the secretion; this heralded the onset of copious secretion at about the time of parturition. 6. The changes in mammary gland activities are discussed in relation to changes in plasma hormone concentrations during pregnancy.

Metabolic clearance rate, production rate, and mammary uptake and metabolism of progesterone in cows.

Tracer kinetic techniques have been used to measure the production rate, metabolic clearance rate and mammary uptake of progesterone in six experiments on two Jersey cowsmthe cows were surgically prepared so that the carotid artery, jugular vein and mammary vein concentrations of progesterone, and udder blood flow, could be determined in conscious animals without anaesthesia or stress. The mean production rate of progesterone was 173 +/- 23-3 (S.ET) mug/min, with values ranging from 80 to 276 mug/min in pregnancy. The metabolic clearance rate was 22-5 +/- 2-0 1/min, or 0-21 +/- 0-025 1/min/kg metabolic body weight. The mammary uptake of progesterone was low, 3-1 +/- mug/min, and udder uptake accounted for about 3% of progesterone production rate. During [3H]A1progesterone infusion, radioactivity was transferred from blood to milk, probably by diffusion down a concentration gradient. Progesterone accounted for more than 88% of the ether-soluble radioactivity recovered from milk.

Utilization of amino acids by the isolated perfused sheep liver.

To test Elwyn's suggestion [1970], that high concentrations of amino acids supplied to the liver from the hepatic artery do not stimulate protein synthesis, substrates containing amino acids have been infused into either the hepatic artery or portal vein of isolated sheep livers. The livers received highly oxygenated blood from the hepatic artery and partly deoxygenated blood from the portal vein. There were no significant differences in amino acid uptake (81 +/- 4% of input), urea output (69 +/- 6% of uptake) or 'protein synthesis' as assessed by N accumulation in the liver. Amino acids were actively extracted from the plasma by both routes of infusion and , when the concentrations simul-ted those occurring in the portal vein of fed sheep, the uptake was very similar to that in vivo. When an amino acid mixture based on that absorbed in dogs was infused, the extraction of individual amino acids was similar except for a negligible uptake of valine, leucine and isoleucine. In these experiments protein synthesis was also very low.

Metabolic clearance rate, production rate and mammary uptake of progesterone in the goat.

The dynamics of progesterone uptake and metabolism in the mammary gland of the goat have been measured and related to the metabolic clearance rate and production rate of the hormone determined by tracer kinetic techniques. The metabolic clearance rate of progesterone from blood was 3-13 plus or minus 0-35 (S.E.M.) 1/min in ten experiments on six goats; values tended to be slightly higher in pregnant than in non-pregnant goats. The production rate of progesterone at oestrus, and at day 3 of the normal cycle, was less than 0-01 mug/min. During the luteal phase of the oestrous cycle the production rate was 8-5 and 14-6 mug/min in 2 animals, and in the second half of pregnancy, 15-3 plus or minus 0-6 mug/min (5 animals). Progesterone was extracted from the circulation by the mammary gland of conscious goats with an efficiency of 49-4 plus or minus 11-3% in non-pregnant, and 51-7 plus or minus 11-5% in pregnant aniamals. The mean clearance rate of progesterone by the udder was 0-279 1/min, 8-8% of the metabolic clearance rate. Mammary uptake of progesterone in goats with an actively secreting corpus luteum was 0-64 plus or minus 0-29 mug/min, which gave an estimated value of 0-11-1-88 ng/min/g mammary gland. The mammary extraction of progesterone was investigated in a goat 3 days after oestrus when any high affinity receptor sites would presumably be unoccupied. During the infusion of progesterone into a mammary artery, tissue sample were taken from various organs, including the mammary gland, and the concentration of labelled compounds at steady state was determined. A high mammary extraction of progesterone was found to be determined. A high mammary extraction of progesterone was found to be attributable principally to progesterone metabolism. The metabolites of progesterone were removed from the gland in venous blood and were not stored to any appreciable extent in mammary tissue. Experiments in vitro confirmed the findings in vivo that mammary tissue metabolized labelled progesterone and also pregnenolone and androstenedione; metabolism of dehydroepiandrosterone, oestradiol-17 beta, oestrone and cortisol was relatively small. Confirmation of our previous finding that the mammary gland of the goat can synthesize progesterone from labelled pregnenolone infused into the gland in vivo, further implicated this organ as an active site of metabolism of certain steroids. The physiological role of steroid metabolism in the mammary gland is discussed.

The distribution and movements of carbon dioxide, carbonic acid and bicarbonate between blood and milk in the goat.

1. A-V differences and milk concentrations of respiratory gases, pH, HCO3 and H2CO3 have been measured in lactating goats and cows. 2. The pH and [HCO3 minus] of milk were significantly lower than those of plasma while milk PCO2 was virtually identical to that of mammary venous blood. [H2CO3+ dissolved CO2] was similar in milk and blood. 3. 14-C (from injected [14-C]HCO3 minus was found to cross the mammary epithelium in both directions. 14-C also passed across the duct epithelium and since this epithelium has previously been shown to be impermeable to ions it is argued that 14-C crossed in an unionized form, i.e. as CO2 and/or H2CO3. 4. Hourly milking with the aid of oxytocin raised milk pH, [HCO3 minus], [H2CO3], [Na] and E1Cl], and lowered [K], [lactose] and [phosphate]. These effects are discussed in relation to the hypothesis proposed previously for the action of oxytocin on milk composition. 5. A scheme for the distribution and movements of CO2, H2CO3 and HCO3 minus between extracellular fluid and milk is suggested, and discussed in relation to Cl minus transport.

Changes in colostrum composition and in the permeability of the mammary epithelium at about the time of parturition in the goat.

1. Changes in milk composition and in mammary permeability to labelled sucrose and monovalent ions have been studied in late pregnancy and at the time of parturition. These data have been compared with those obtained previously in lactating goats.2. Colostrum contained more sodium, chloride, protein, immunoglobulins and less potassium and lactose than milk. The composition of the aqueous phase changed near term and normal milk composition was reached either several days before or several days after parturition. The main decrease in the immunoglobulin concentration occurred when the animal was first milked or suckled.3. In contrast to lactating animals, [(14)C]sucrose passed from blood to milk and [(14)C]lactose from milk to blood. There was a significant correlation between the entry of labelled sucrose into milk and the sodium concentration.4. The rate of passage of (24)Na and (36)Cl into milk from blood was higher in late pregnancy than in lactation.5. In late pregnancy, the concentrations of both sodium and chloride were inversely correlated with the concentration of lactose while there was a positive correlation between potassium and lactose.6. Hydrostatic pressure in the lumen of the mammary gland increased during late pregnancy but there was no significant correlation with the composition of the aqueous phase of milk.7. When one gland of 4 animals was milked twice-daily starting 9-19 days before parturition, the composition of the secretion changed to that of normal milk only in the gland that was being milked. Similarly, [(14)C]sucrose, (24)Na and (36)Cl entry from blood decreased. Milk yield also increased but a further increase occurred post-partum.8. Comparison of the passage of (3)HOH, (24)Na and (36)Cl into different fractions of milk during I.A. infusions indicates that the mammary ducts are less permeable to ions than the alveoli, but it was not possible to decide whether the ducts are as impermeable as during lactation.9. It is suggested that ion and lactose movements occur via a para-cellular route and these results are discussed in relation to transport mechanisms and in terms of local and systemic controlling mechanisms.