Mammary uptake and excretion of prostanoids in relation to mammary blood flow and milk yield during pregnancy-lactation and somatotropin treatment in dairy goats.

Mammary arterious-venous differences (A-V) and excretion into milk of four prostanoids were related to changes in milk yield and milk vein blood velocity (MBV) in goats at different stages of pregnancy and lactation, and during somatotropin (ST) treatment in mid-lactation. Arterial concentrations and mammary A-V for the vasodilators prostacyclin (PGI(2)) and prostaglandin (PG) E(2) (measured as 6-keto-PGF(1 alpha) and bicyclic PGE(2), respectively) decreased from late pregnancy to lactation. A-V were negatively correlated to MBV (r = -0.32 to -0.34). Arterial concentrations of the vasoconstrictors PGF(2 alpha) and TXA(2) (measured as TXB(2)) changed similarly, but no A-V across the mammary gland were found. The vasodilator to vasoconstrictor ratio in plasma was around 1:1, and in skimmed milk around 0.29-0.49 due to significantly higher TXB(2) levels in milk compared to plasma. Close linear correlations were established between milk yield and excretion of TXB(2) into milk (r = 0.80, P < 0.001), and between MBV and PGE(2) excretion into milk (r = 0.69, P < 0.001). ST treatment stimulated MBV and mammary prostanoid supply, and decreased prostanoid concentration in milk vein plasma. The high arterial levels of prostaglandins during pregnancy most likely reflected uterine synthesis. Our results support a role for PGI(2) and PGE(2) in local mammary blood flow regulation during lactation. Increased mammary uptake of these two prostanoids may be involved in the mammary blood flow response to ST. TXA(2) may be synthesized by mammary epithelial as well as vascular cells, and TXA(2) may be an important factor in regulation of mammary function.

The effects of relaxin on the response of intramammary pressure and mammary blood flow to exogenous oxytocin in the goat.

Lactating goats were given relaxin (50 micrograms) by close-arterial infusion into one mammary gland. The increase in intramammary pressure and in mammary blood flow elicited by exogenous oxytocin i.v. was attenuated by relaxin in goats during pregnancy and early lactation but not in a group studied during the oestrous cycle. Intramammary pressure in both mammary glands was affected at the dose of relaxin used. It is concluded that a change in responsiveness of the mammary myoepithelium to oxytocin is one possible effect of relaxin, acting directly or indirectly, circulating systemically during pregnancy and produced locally by the mammary gland at the onset of and during lactation.

Blood prolactin concentrations affect prolactin transfer into goat milk: implications for maintenance of lactation.

125I-Labelled ovine prolactin was infused for 15 min into a pudic artery supplying one mammary gland of lactating goats (n = 17). Between 0 and 4.25 h significantly more total (P < 0.01) and trichloroacetic acid (TCA)-precipitable (P < 0.001) radioactivity appeared in the milk of the infused compared with the non-infused gland. Gel chromatography and antibody precipitation indicated the presence of undegraded 125I-labelled prolactin in milk whey. Maximum transfer occurred 60-80 min after the end of infusion suggesting passage via a transcellular route. High plasma prolactin concentrations, resulting from infusion of cold prolactin with labelled prolactin in late lactation or from seasonally elevated prolactin at peak lactation, reduced the specific activity of infused prolactin and depressed the difference in secretion of 125I-labelled prolactin into milk of infused and non-infused glands. This suggests the operation of a competitive and saturable mechanism. Together with the increase in the milk to blood ratio of prolactin in goats given long-term (3 week) bromocriptine treatment, the results suggest that the goat mammary gland has a high avidity for prolactin especially when circulating prolactin is low. There was also evidence from TCA precipitation that prolactin may be protected from degradation in these circumstances. These mechanisms may contribute to the resistance of ruminant lactation to reduction in plasma prolactin and protect lactation from seasonal prolactin fluctuations.

The local differential effect of prostacyclin, prostaglandin E2 and prostaglandin F2 alpha on mammary blood flow of lactating goats.

Mammary blood flow (MBF) and milk yield are closely related in dairy ruminants, but little is known about the regulation of MBF in vivo. The local effects on MBF of injections or continuous infusions into the mammary artery of prostaglandins (PG) or indomethacin (an inhibitor of prostaglandins) respectively, were investigated in surgically prepared conscious goats. Prostacyclin (PGI2) was found to be a potent stimulator of MBF which increased linearly over the dose range 50-1000 ng. PGE2 was almost as potent as PGI2 at low doses, but tachyphylaxis occurred at doses at and above 100 ng. The response to repeated injections of PGE2 quickly declined depending on the dose. PGF2 alpha had no effect on MBF. During infusion of indomethacin into the mammary artery MBF was reduced markedly, showing that endogenous mammary prostaglandins are involved in the regulation of vasodilatation. The results indicate that PGI2 (and to a lesser extent PGE2) has an important role in the local regulation of vascular tone in the mammary gland.

Unilateral control of ovarian oxytocin release and the facilitatory effects of insulin-like growth factor-I in sheep.

Prostaglandin F2 alpha (PGF2 alpha)-induced release of ovarian oxytocin was investigated to determine whether the effect in vivo was local. [3H]PGF2 alpha infused downstream into a single ovarian lymphatic was transferred into the adjacent ovarian vasculature (estimated transfer 1.1 and 1.7%, two experiments). When unlabelled PGF2 alpha was infused in a similar manner (76 pmol min-1), there was a prompt eightfold increase in ovarian oxytocin release from the adjacent ovary containing a corpus luteum, but no effect on the opposite corpus luteum, showing that the effect was local. Instillation of 2% lignocaine into the ovarian vascular pedicle did not affect PGF2 alpha-induced oxytocin release, supporting the idea that neural mechanisms are not involved. Repeated doses of PGF2 alpha given close-arterially produced a successive reduction in oxytocin release. This effect was prevented by a prior infusion of insulin-like growth factor-I (IGF-I), which itself gave a small, but significant, increase in oxytocin release. The results show that PGF2 alpha in ovarian lymphatics acts locally and directly to stimulate ovarian oxytocin secretion, that repeated exposure of the corpus luteum to pulses of PGF2 alpha can result in tachyphylaxis, and that this latter effect can be ameliorated by IGF-I infused in vivo.

Transfer of insulin-like growth factors I and II from plasma to lymph in young goats.

The plasma clearance of intravenously injected 125I-labelled insulin-like growth factor I (IGF-I, n = 13) and IGF-II (n = 12) and their transfer into lymph draining the foreleg of 3.5- to 8-week-old British Saanen goats was studied. Both peptides were initially distributed into a volume equivalent to the plasma volume, while the final distribution spaces for IGF-I and IGF-II were 90 +/- 9.8 and 125 +/- 12 ml/kg live weight respectively. There were two phases to the plasma clearance of both IGF-I and IGF-II, with the half-lives of both phases for IGF-I (9.6 +/- 0.9 and 287 +/- 23 min, first and second phase respectively) being significantly (P less than 0.001) longer than those of IGF-II (4.2 +/- 0.6 and 172 +/- 18 min, respectively). The radioactivity transferred into lymph originated from intact IGF-I and IGF-II as well as degraded products of these compounds, as assessed by precipitation with trichloroacetic acid and gel filtration. Levels of undegraded IGF-I in lymph were 50% greater than IGF-II. While more than 90% of either peptide was bound to specific IGF-binding proteins in plasma, in lymph 34 +/- 2% of IGF-I and 23 +/- 3% of IGF-II remained in the free form 60-80 min after injection. The plasma: lymph ratio for free IGF-I was 2:1 and for bound IGF-I, 8:1. The corresponding values for IGF-II were 3:2 and 8:1 respectively. These results provide direct experimental evidence for transfer of undegraded IGF-I and IGF-II from blood into lymph of the foreleg, consistent with the ability of these factors to act in an endocrine role in growing tissues. Differences between plasma clearance and transfer of IGF-II into lymph compared with IGF-I may be due to its greater cellular uptake and/or degradation in vivo.

Secretion of insulin-like growth factor II into milk.

125I-labeled insulin-like growth factor II (IGF-II) was infused directly into the pudic artery supplying one gland of lactating goats (n = 4). Maximum specific activity for [125I]IGF-II transferred into milk from the infused gland was reached 60 min after that in plasma and was 2.5 fold greater than in milk from the non-infused gland. Inclusion of either 67.5 nmoles unlabeled IGF-II or IGF-I had no influence on the amount or pattern of secretion of [125I]IGF-II into milk from either gland. While the temporal pattern of secretion of [125I]IGF-II into milk was consistent with a transcellular mechanism of transfer, the lack of competition by unlabeled IGF-II or IGF-I suggests a non-specific mechanism is operable, which contrasts to secretion of IGF-I.

Mechanism of secretion of plasma insulin-like growth factor-I into milk of lactating goats.

125I-Labelled insulin-like growth factor-I (IGF-I) was infused as the free form directly into the pudic artery supplying one gland of lactating goats (n = 6). The infusion was for 60 min and 0.4 +/- 0.09% (S.E.M.) of the infusate was secreted into milk from the infused gland during its first passage through that gland. A large proportion of the 125I-labelled IGF-I escaped into the systematic circulation and was secreted into milk of both glands. A total of 5.2 +/- 0.4% of infused radioactivity was recovered in milk from both glands from 0 to 720 min. Radioactivity consisted of trichloroacetic acid (TCA)-precipitable and -soluble counts which were shown by gel filtration to be authentic IGF-I and degraded products of the peptide. The amount and time course of TCA-soluble radioactivity in milk from both glands was similar, suggesting degradation of 125I-labelled IGF-I at extramammary sites. Maximum specific activity for 125I-labelled IGF-I in milk from the infused gland was reached 80-120 min after the start of infusion and was 2.5-fold greater than milk from the non-infused gland. The time course of appearance of 125I-labelled IGF-I in milk suggests that transfer was via the transcellular pathway and this was further supported by comparing the pattern of transfer of [14C]sucrose and [14C]amino acids. When excess unlabelled IGF-I was included in the infusate, specific activity in milk from the infused gland was reduced to that of the non-infused gland, indicating a competitive and saturable mechanism of secretion for 125I-labelled IGF-I.(ABSTRACT TRUNCATED AT 250 WORDS)

The galactopoietic effect of bovine growth hormone in goats is associated with increased concentrations of insulin-like growth factor-I in milk and mammary tissue.

Lactating goats exhibiting widely divergent responses to short-term (4 days) treatment with bovine GH (bGH) were retrospectively divided into two groups based on the magnitude of this response. There was no difference between groups in terms of the pretreatment milk yield, but by day 4 of treatment milk secretion had increased by 4.99 +/- 2.5 (S.E.M.) ml/h (P greater than 0.05 compared with pretreatment) for group 1 and 22.9 +/- 2.4 ml/h (P less than 0.001) for group 2. Plasma GH increased in both groups, but concentrations were significantly higher both before and during treatment in group 1 compared with group 2. Plasma concentrations of insulin-like growth factor-I (IGF-I) increased significantly during bGH treatment for both groups and there was no significant difference between the two until day 4 of treatment when levels of IGF-I in group 1 began to decline, whereas those from group 2 were maintained. Concentrations of IGF-I in milk from goats in group 1 were not significantly altered by GH administration, whereas those in goats in group 2 were increased by 40% (P less than 0.01 compared with pretreatment). Levels of IGF-I in mammary secretory tissue from four animals from group 1 were not altered by bGH (2.8 +/- 0.2 and 2.77 +/- 0.08 nmol/kg tissue before and after treatment respectively), but were significantly (P less than 0.05) increased in four animals from group 2 (2.80 +/- 0.2 and 9.9 +/- 1.1 nmol/kg tissue).(ABSTRACT TRUNCATED AT 250 WORDS)

Increase in milk secretion and mammary blood flow by intra-arterial infusion of insulin-like growth factor-I into the mammary gland of the goat.

The close-arterial infusion of free insulin-like growth factor-I (IGF-I; 1.1 nmol/min) for 6 h into the pudic artery supplying one mammary gland of lactating goats caused a 25 +/- 6% (mean +/- S.E.M., n = 6) increase in the rate of milk secretion of that gland. The increase in the rate of milk secretion in the adjacent noninfused gland (14 +/- 4%) was not significantly different from that observed during saline infusion (4 +/- 5%). Blood flow to the infused gland was increased from 378 +/- 26 ml/min 1 h before to 487 +/- 56 ml/min approximately 5 h after the start of the infusion of IGF-I, declining to 420 +/- 44 ml/min approximately 2 h after the end of the infusion. The total concentration of IGF-I (free and bound) in milk of the infused gland was significantly higher than that of the non-infused gland. The concentrations of IGF-I in carotid arterial plasma samples increased during IGF-I infusion from a mean value of 32 +/- 2 nmol/l before to a maximum of 49 +/- 3 nmol/l 5 h after the infusion commenced. Circulating concentrations of total IGF-I declined slowly after the infusion with an estimated half-life of 5 h. Infusion of saline alone did not alter mammary blood flow or the concentration of total IGF-I in milk or plasma. The results indicate that the infusion of free IGF-I into the mammary arterial supply enhances milk secretion and mammary blood flow in intact, conscious goats.(ABSTRACT TRUNCATED AT 250 WORDS)

Neurotransmitters and lymphatic-vascular transfer of prostaglandin F2 alpha stimulate ovarian oxytocin output in sheep.

The mechanisms of lymphatic-vascular transfer across the ovarian vascular pedicle were studied in anaesthetized sheep 8-15 days after ovulation. [3H]Prostaglandin F2 alpha (PGF2 alpha), [14C]mannitol and [36Cl]Na were infused continuously into either a uterine lymphatic or a uterine vein and the kinetics of transfer into the adjacent utero-ovarian vein or ovarian plasma were studied. Transfer occurred according to the sequence [36Cl] greater than [14C] greater than [3H] indicating that PGF2 alpha is not transferred by rapid diffusion, as with [36Cl]Na, nor by a paracellular route, as with [14C]mannitol, but by a slower process probably involving facilitated diffusion. Transfer into the adjacent utero-ovarian vein or ovarian blood was greater when compounds were infused into a uterine lymphatic than into a uterine vein. Substantially more [3H]PGF2 alpha occurred in the adjacent corpus luteum than either of the other compounds after a lymphatic infusion. Intra-lymphatic infusion of PGF2 alpha stimulated the release of ovarian oxytocin but the effect was not confined to the adjacent ovary. Intravenous (jugular) infusion of PGF2 alpha failed to stimulate ovarian oxytocin secretion whereas close-arterial infusion into the ovaries was effective, and the possibility was investigated that any systemic effect of PGF2 alpha was mediated through neural mechanisms. Noradrenaline and acetylcholine were both effective in causing the release of ovarian oxytocin when infused close-arterially into the ovary. With infusions of acetylcholine, ovarian oxytocin secretion rate was increased over fivefold without any change in posterior pituitary release. Noradrenaline and acetylcholine produced a concomitant fall in ovarian blood flow, and neurotransmitter-induced ischaemia may have played a role in ovarian oxytocin release. The finding that PGF2 alpha infused into a uterine lymphatic stimulates ovarian secretion of oxytocin, and that the effect is bilateral whereas PGF2 alpha accumulation in ovarian tissue is unilateral, implies that its mechanism of action may not be solely directed at the luteal cell.

Increased secretion of insulin-like growth factor I into milk of cows treated with recombinantly derived bovine growth hormone.

Six lactating, non-pregnant Jersey cows were given subcutaneous injections of recombinantly derived bovine growth hormone for 7 d. Milk yield was increased by 4.5 kg/d on d 7, compared with the average yield of 10.7 +/- 0.4 kg/d (mean +/- s.e.m.) for the 7 d preceding treatment. Concentrations of insulin-like growth factor I (IGF-I) in the milk increased from 0.44 +/- 0.04 nmol/l (mean +/- s.e.m.) during the 7 d preceding treatment to 1.6 +/- 0.2 nmol/l on d 7 of treatment. Taking the increase in milk yield into account the total increase in the secretion of IGF-I into milk of one udder half was 6-fold. Plasma concentrations of total IGF-I rose from 15.5 +/- 1.3 nmol/l (mean +/- s.e.m.) on the day preceding treatment to 56.9 +/- 3.6 nmol/l (mean +/- s.e.m.) on d 7 of treatment. Mammary plasma flow increased from 1.6 +/- 0.09 to 2.2 +/- 0.06 l/min.udder half over the same time. Estimates of the amount of IGF-I that reached the mammary gland gave values of 24 and 116 nmol/min.udder half before and during treatment respectively. IGF-I in milk of treated cows was associated predominantly with proteins ranging from 40,000 to 150,000 mol.wt, but a significant proportion (19%) of the total IGF-I was present in the free unbound form. IGF-I crosslinking studies revealed the presence in milk of one specifically labelled band at 31,000 mol.wt.

Cardiovascular responses and mammary substrate uptake in Jersey cows treated with pituitary-derived growth hormone during late lactation.

Pituitary-derived bovine growth hormone (bGH) was administered to Jersey cows during late lactation for 7 d. Milk yield increased significantly during treatment and by a maximum of 49.6% on d 7. The magnitude of the increase was similar to that of mammary plasma flow (47.8 +/- 18.3%) over the same period. By 15-21 d after treatment, both variables had returned to pretreatment values. With respect to milk composition, bGH had negligible effect on lactose and fat concentrations but there were significant decreases in protein, sodium and chloride. Arterial plasma concentrations of bGH increased substantially during treatment, but the associated rise in insulin was not statistically significant. Haematocrit decreased significantly, the lowest value being recorded 3 d after bGH treatment ceased. Mammary respiratory quotient fell progressively after the start of bGH treatment and reached the lowest recorded value 3 d after treatment ceased (62.2 +/- 7.3% of pretreatment value). Glucose and acetate uptake by the mammary gland increased significantly during treatment, increase in glucose uptake being due both to a greater arterio-venous difference and to mammary plasma flow. There was strong evidence that the acute response in increased milk yield was associated with multiple effects in terms of mammary plasma flow and metabolism, as well as haematocrit changes indicative of increased plasma volume.

From animal to molecule: aspects of the biology of insulin-like growth factors.

The synthesis of IGF-II mRNA in sheep foetal tissues is considerably higher than IGF-I. IGF-II probably has a paracrine role in the foetus; however it is likely that IGF-I originates mainly from the foetal liver and has an endocrine function. Although in the adult system IGF-I is tightly bound to serum carrier proteins it is potentially biologically active. Galactopoiesis in the goat mammary gland provides a useful model for demonstrating the importance of circulating IGF-I as a mediator of GH action. Ligand-receptor interactions involved in the stimulation of Swiss 3T3 fibroblasts by IGF-I, II and insulin were examined. It was found that the potency of binding to type I receptors was IGF-I greater than IGF-II much greater than insulin by competitive binding assays and chemical cross-linking studies, and that some cell lines secrete an IGF binding protein which is specific for IGF-I and II and which acts as an inhibitor in cellular binding assays. Maximal stimulation of DNA synthesis induced by IGF-I, II and insulin in the presence of synergising mitogens were similar. While the actions of the IGFs were consistent with type I receptor binding insulin appeared to act through its own receptor. The reduction of EGF receptor affinity following the addition of IGF-I and insulin to 3T3 cells may involve a protein kinase that is not sensitive to phorbol esters. 3T3 cell nuclei contain endogenous inositol phospholipids and their corresponding kinases and monoesterases.(ABSTRACT TRUNCATED AT 250 WORDS)

Cardiovascular and metabolic responses during growth hormone treatment of lactating sheep.

Pituitary-derived bovine growth hormone (bGH) was administered to six lactating Friesland ewes for 7 d. There was no consistent galactopoietic response, with changes in milk yield varying from 0 to 33% during treatment compared with the pretreatment period. The major effect of bGH on the concentration of milk constituents was to increase fat by 14.2% (P less than 0.05). Treatment resulted in significant increases in arterial plasma concentrations of growth hormone, insulin-like growth factor I and glucose, with decreases in the plasma arterial concentrations of acetate and certain amino acids. There was a marked reduction in haematocrit and in haemoglobin concentration which took at least 3 d to recover. The arterio-venous difference across the mammary gland decreased for O2 during treatment and the veno-arterial difference for CO2 decreased after treatment. Mammary respiratory quotient therefore decreased significantly after bGH treatment. The results suggest that bGH exerts effects at a number of separate loci.

Protein synthesis and degradation in the mammary gland of lactating goats.

Lactating goats were given a close arterial infusion of [1-14C]leucine and [4,5-3H]4-methyl-2-oxopentanoic acid into one half of the mammary gland at 2-3 weeks and 34-39 weeks after kidding. Rates of protein synthesis, degradation and net output were determined from measurements of arteriovenous difference and blood flow using a model of leucine metabolism previously developed for muscle (Oddy & Lindsay, 1986). Protein leucine output in milk (Y mumol/min) correlated well with the difference between synthesis and degradation (X mumol/min) derived from the model: Y = 1.30 + 1.24X (r2 = 0.9; n = 9, P less than 0.01). There was substantial synthesis and degradation of protein within the mammary gland. Although only an approximate value could be obtained for the partitioning of protein synthesis and degradation between tissue and milk proteins, there was evidence of appreciable turnover of both. There was no significant difference between mammary leucine and protein metabolism in early and late lactation other than that imparted by a greater mass of mammary tissue in early lactation, although there was a tendency for greater oxidation of leucine in late lactation.

Suppression by perchlorate of technetium-99m and iodine-123 secretion in milk of lactating goats.

Lactating goats were infused with either technetium-99m (99mTc) or iodine-123 (123I) together with chlorine-36 (36Cl) through an indwelling catheter previously placed in an external pudic mammary artery. The radioisotope infusions were repeated together with 100 mg of sodium perchlorate. There was a rapid transfer of 99mTc and 123I into milk, reaching a peak concentration 30 min after a 15-min infusion. The fractional secretion of 99mTc and 123I in milk was reduced by 70%-80% and 60%-66%, respectively, by perchlorate. The fractional secretion of 36Cl was not affected by perchlorate, and the shape of the 36Cl secretion curve differed from those of 99mTc and 123I, which were similar. It is probable, therefore, that the latter nuclides were secreted by a transport route different from that of chloride. Available data describing the secretion of 99mTc in human milk after pertechnetate administration was reviewed, and it was concluded that perchlorate pretreatment significantly reduced the secretion of 99mTc in human breast milk.

Mechanisms of transfer of steroid hormones and growth factors into milk.

In this paper we examine the ability of the mammary gland to remove from circulating blood three compounds which differ in their physico-chemical and structural properties. Mammary extraction of progesterone, oestrone sulphate and epidermal growth factor (EGF) is similar at peak lactation in goats, but the proportion of labelled infusate that is transferred into milk is greater for oestrone sulphate and EGF than progesterone which is rapidly metabolised by mammary tissue. The kinetics of transfer of progesterone, oestrone sulphate and EGF from blood into milk show that transcellular processes are involved, and on the basis of earlier hypotheses and new information reported here the results indicate the probable importance of simple and facilitated diffusion pathways for progesterone and oestrone sulphate, and secretory mechanisms for oestrone sulphate and EGF. Although evidence is lacking for a direct effect of hormones in milk on mammary function, their concentration in milk may reflect changes in local regulation of mammary secretion. Considerable practical value is attached to the immunodiagnostic use of milk hormone concentrations to determine ovarian and placental endocrine activity during pregnancy in domestic ruminants.