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The aim of this study was to compare the plasma concentration profile, mammary blood flow response and transfer into milk of intact IGF-I with that of its truncated analogue, des(1-3)IGF-I (des-IGF-I). Each peptide was infused for 24 h into the pudic artery supplying one mammary gland of lactating goats (n = 5). Concentrations of IGF-I in plasma (from the jugular vein) rose rapidly during infusion of IGF-I or des-IGF-I to reach 510 +/- 62 and 640 +/- 32 ng/ml (mean +/- S.E.M.) respectively, compared with 262 +/- 35 ng/ml after a similar infusion of saline. Ligand blotting analysis indicated a significant increase in the intensity of [125I]IGF-I binding to the 40-43 kDa doublet (binding protein-3 (BP-3), P < 0.01) and the band at 31 kDa (P < 0.05) during infusion of either IGF-I or des-IGF-I, as compared with saline. Furthermore des-IGF-I induced a significant increase in intensity of binding to the 35 and 24 kDa bands, but IGF-I did not. Whereas [125I]IGF-I was distributed between BP-3 and the other binding proteins, [125I]des-IGF-I bound exclusively to BP-3. Mammary blood flow (MBF) increased 48 +/- 6% after 12 h of infusion of des-IGF-I, compared with an increase of 22 +/- 6% during IGF-I. The difference in response was significant at P < 0.05. In addition, more IGF-I was secreted into the milk of the infused than the non-infused gland during either infusion of IGF-I or des-IGF-I.(ABSTRACT TRUNCATED AT 250 WORDS)

Five lactating goats were infused, via an external pudic arterial catheter, directly into the mammary gland with 0.9% (w/v) NaCl (20 ml/h), recombinant human insulin-like growth factor-I (IGF-I; 80 nmol/h), recombinant human IGF-II (133 nmol/h) or IGF-I and IGF-II combined. The infusion was for 6 h and milk yield was determined every 2 h. The ratio of milk yield in the infused relative to the non-infused gland was changed only slightly by saline (2%), but increased to 9% (P < 0.05) in response to IGF-I and 8% (P < 0.05) in response to IGF-II. When combined, both peptides increased this ratio by 6%. These effects were elicited within 2-4 h of the beginning of infusion. Mammary blood flow increased 50-80% (P < 0.05) during all IGF infusions, but only 28% during saline treatment. Plasma insulin decreased 50% (P < 0.01) during the infusion of IGF-I alone or in combination with IGF-II and 25% in response to IGF-II alone. Whereas plasma glucose increased by approximately 10% during infusion of IGF-I alone or with IGF-II, it was not altered by infusion of IGF-II only. The rapidity and unilateral nature of the milk-yield response to IGF-I and IGF-II is consistent with their acting directly on mammary tissue itself. Thus, the present results demonstrate similar local and systemic actions induced by intramammary infusion of IGF-II and IGF-I, although the magnitude of the response to IGF-II tends to be less than that to IGF-I.

The milk yield and mammary blood flow responses to close-arterial, intra-mammary infusion of IGF-I were investigated in five Saanen goats milked frequently or normally the day before. Animals were infused for 6 h with recombinant human IGF-I (1.3 nmol/min) and milked hourly following i.v. injection of oxytocin beginning 2 h before infusion and then every 2 h. On one occasion animals were milked five times (after i.v. injection of oxytocin) on the day before infusion and on the other they were milked twice, without oxytocin. The ratio of milk yield from the infused to that from non-infused gland increased by 17 +/- 4% (mean +/- S.E.M.) in goats milked twice the day before infusion and by 6 +/- 2% when the infusion was preceded by frequent milking. Maximal responses were obtained 4 h after the start of the infusion and differed significantly (P < 0.05), according to pretreatment milking. Blood flow through the infused gland rose in parallel to the milk yield response. At 5 h, when maximal levels were achieved, blood flow was 182 +/- 23% of the pre-infusion flow rate following twice-daily milking and 139 +/- 3% of the pre-infusion flow rate following more frequent milk removal. Thus, more frequent milk removal on the day before close-arterial infusion of IGF-I attenuated both the milk yield and mammary blood-flow response to the infusion of IGF-I.

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 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)

During pregnancy in goats the concentration of endogenous oestrone sulphate in milk increased more than twofold, and that in arterial and mammary venous plasma 10- and 20-fold respectively. The concentration in milk was higher than that in arterial plasma, particularly in lactating goats during mid-gestation. This was partly related to mammary production of oestrone sulphate (or of a closely related steroid which cross-reacted in the radioimmunoassay) since in tracer infusion studies the specific activity of oestrone sulphate in milk was significantly lower than that in arterial or mammary venous plasma. It was also related to the existence of a mechanism within the gland which concentrates oestrone sulphate in milk since when infused close-arterially into the mammary gland of a non-pregnant goat with undetectable levels of the endogenous compound in the circulation, a concentration ratio of 7.4:1.0 was reached for oestrone sulphate in milk:arterial plasma. Tracer kinetic studies showed that mammary extraction of [3H]oestrone sulphate was variable (up to 41.3 +/- 30.6%, mean +/- S.E.M.). During intravenous or close-arterial infusion, radioactivity in arterial and mammary venous plasma at steady state was mainly in the form of [3H]oestrone sulphate (range, 64 +/- 10.6 to 80.2 +/- 5.9% of total radioactivity in plasma). The remainder was in the form of compounds chromatographically similar to oestradiol-17 beta-3-monosulphate, oestradiol-17 alpha-3-monosulphate and unconjugated oestrogens. The distribution of radioactivity between these different steriods was similar in arterial mammary venous plasma indicating a low level of selective mammary metabolism or extraction. The amount of labelled oestrone sulphate transferred into milk was low, and it was significantly less in pregnant (range, 0.11 +/- 0.07 to 0.27 +/- 0.16% of total infusate) than in non-pregnant animals (3.23 +/- 0.50%). Studies of the rate of transfer of [3H]oestrone sulphate from blood to milk indicated the presence of a transcellular route with peak activity in milk occurring about 110 min after the start of the infusion.

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.

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.