Effect of photoperiod and heat stress in the third trimester of gestation on milk production and circulating hormones in dairy goats.

The influence of photoperiod manipulation in the dry period was examined in dairy goats experiencing environmental heat stress. Multiparous Israeli Saanen goats were blocked at dry off (∼60 d prepartum) into 2 groups of 4 goats each based on body weight, previous milk production, and detected embryo number. Treatments consisted of long-day (16 h light:8 h dark) and short-day (8 h light:16 h dark) photoperiods (LDPP and SDPP, respectively). Heat-stress conditions were applied by manipulating the environment of metabolic rooms to reach a maximum temperature of 37°C between 1000 and 2200 h, and a minimum of 23°C and 70.3% relative humidity. All goats were returned to ambient photoperiod after kidding, milked twice daily, and milk yield was automatically recorded. Dry matter intake during the dry period was similar between treatments, averaging 1,114 g/d. Milk production was significantly higher in the SDPP than the LDPP group (2,172 vs. 1,550 g/d) during the 12-wk experimental period. Milk protein and fat contents were similar in both groups and averaged 3.63 and 4.34%, respectively, whereas milk lactose was higher in the LDPP group (4.77 vs. 4.67%). Heart rates were similar between treatments and averaged 112.6 beats per minute (BPM). Respiration rates were lower in the morning (58.4 BPM) compared with the afternoon (91.2 BPM) and were not influenced by photoperiod. Rectal temperature was higher for the LDPP than the SDPP group (40.4 vs. 39.6°C). The thyroid hormone level (mean ± SE) was similar in both groups during the dry period, but higher during lactation in the LDPP goats up to 40 d postpartum (110±6.59 vs. 156±8.76 ng/mL). Plasma IGF-1 (mean ± SE) was higher in the LDPP group (279±62 vs. 162±27 ng/mL in SDPP) during the dry period but was similar postkidding, averaging 132±24 ng/mL. Plasma prolactin level (mean ± SE) was higher in the LDPP than the SDPP group during the dry period (17.2±1.6 vs. 10.6±0.99 ng/mL), whereas it was similar throughout lactation (0.61±0.28 ng/mL). These data support the idea that SDPP manipulation during heat load in dry goats can be used as an abatement strategy to reduce the carryover effect of heat stress observed during the subsequent lactation. The higher milk production in SDPP goats is explained by changes in circulating prolactin profile rather than differences in feed intake or secretion of insulin-like growth factor 1.

Hyperinsulinemic clamp modulates milk fat globule lipid composition in goats.

We determined the effect of insulin on milk fatty acid (FA) and lipid composition in goats. Four dairy goats, 150 d in milk, were subjected to hyperinsulinemic clamp (treatment) or saline (control) infusion for 4d in a crossover design study. Composition and concentration of plasma and milk FA, triglycerides, phospholipids, sphingolipids, and cholesterol were determined. Mammary gland biopsies were taken at the end of each experimental period and lipogenic gene expression was determined. Plasma insulin was elevated 3.5-fold, whereas plasma glucose remained constant during the treatment period. Feed intake decreased by 26% and fat yield decreased by 17% relative to controls. No change in nonesterified FA concentration was found between controls and treatment. Compared with controls, insulin decreased yield of long-chain saturated FA by 14%. Milk concentration of long-chain FA was reduced by 3%, whereas that of medium-chain FA increased by 5% during the treatment compared with controls. Hyperinsulinemic clamps increased the yields of milk phospholipids by 9% and cholesterol by 16%, whereas it only tended to decrease triglyceride yields (by 11%). Hyperinsulinemic treatment resulted in compositional changes in the milk fat globule membrane, as reflected by 15 and 9% decreases in phosphatidylethanolamine and phosphatidylcholine concentrations, respectively. Lipogenic gene expression of acyl coenzyme A carboxylase, stearoyl coenzyme A desaturase, and FA synthase did not change, whereas lipoprotein lipase gene expression tended toward an increase in the treatment period compared with controls. Hyperinsulinemic clamps reduce the availability of long-chain FA, which are considered to originate from the diet and adipose lipolysis for milk lipid synthesis by the mammary gland of goats. Under these conditions, long-chain FA might be preferentially channeled to phospholipid rather than triglyceride synthesis, hence increasing phospholipid yields. Mechanisms determining FA distribution among milk lipid components and the consequences of altered milk fat globule membrane lipid composition remained to be elucidated.

Long-term effects of ad libitum whole milk prior to weaning and prepubertal protein supplementation on skeletal growth rate and first-lactation milk production.

Our objectives were to determine the effects of rapid growth rate during the preweaning period and prepubertal protein supplementation on long-term growth pattern and milk production during the first lactation. Forty-six Israeli Holstein heifer calves were fed either milk replacer (MR) or whole milk (WM) from 4 to 60 d age. Calves had free access to WM or MR for 30 min twice daily and free-choice water and starter mix for the entire day. From weaning until 150 d of age, all heifers were fed the same ration. At 150 d of age the heifers were divided into 2 subgroups, with one subgroup supplemented with an additional 2% protein until 320 d of age. Thereafter, all heifers were housed and fed together until calving. Another cluster of 20 heifers was raised on MR and WM treatments and 3 animals from each nursery treatment were slaughtered at 60 d and 10 mo age to determine effects of nursery treatment on organ and adipose tissue mass. Prior to weaning, the MR heifers consumed 0.12 kg/d more DM than the WM heifers, but metabolizable energy intake was not different. Body weight at weaning and average daily gain during the preweaning period were 3.1 kg and 0.074 kg/d higher, respectively, in the WM treatment than in the MR treatment, with no differences in other measurements. Nursery feeding treatment and added protein had no effect on growth rate in the prepubertal period, but the postweaning difference in BW between the WM and MR heifers remained throughout the entire rearing period. The age at first insemination was 23 d earlier and age at pregnancy and first calving was numerically lower for the WM heifers than for the MR heifers. Adipose tissue weights at weaning were doubled in the WM calves. First-lactation milk production and 4% fat-corrected milk were 10.3 and 7.1% higher, respectively, for WM heifers than for MR heifers, whereas prepubertal added protein tended to increase milk yield. In conclusion, preweaning WM at high feeding rates appears to have long-term effects that are beneficial to future milk production. The positive long-term effects of feeding WM on first-lactation milk production were independent of their effects on skeletal growth. Enhanced milk production observed with WM treatment may be related to the milk supply, paracrine or endocrine effects of fat tissues on mammary parenchyma, or a combination of both factors.

Leptin up-regulates the lactogenic effect of prolactin in the bovine mammary gland in vitro.

The ability of leptin to up-regulate prolactin action in the mammary gland is well established. We examined the effect of leptin and prolactin on traits associated with lactation. Leptin and prolactin enhanced proliferation (thymidine incorporation) of the mammary gland cells, elevated the cells' proliferation in a dose-responsive manner, and synergized to elevate the expression of amino acid metabolism via a 90% increase in aminopeptidase N expression. Leptin and prolactin decreased apoptosis (decreased caspase-3 expression by 60%) in the same manner. Leptin enhanced the effect of prolactin on all of these processes in bovine mammary explants. Leptin and prolactin regulated mTOR (mammalian target of rapamycin) by increasing expression by 66%, which is one of the signal-transduction junctions involved in the regulation of proliferation, apoptosis, and protein synthesis. These findings support the hypothesis that leptin up-regulates prolactin action in the bovine mammary gland.

Heat production and retained energy in lactating cows held under hot summer conditions with evaporative cooling and fed two rations differing in roughage content and in vitro digestibility.

The objective of this study was to measure the effect of feeding two total mixed rations (TMRs), differing in their roughage content and in vitro dry matter (DM) digestibility, on the physiological response and energy balance of lactating cows. The partitioning of metabolizable energy intake (MEI) between heat production (HP) and retained energy (RE) of cows held under hot weather conditions and external evaporative cooling was measured. In all, 42 lactating cows were divided into two similar sub-groups, each of 21 animals, and were fed either a control (CON) ration containing 18% roughage neutral detergent fiber (NDF) or an experimental (EXP) TMR containing 12% roughage NDF and used soy hulls as partial wheat silage replacer. The in vitro DM digestibility of the CON and EXP TMR was 75.3% and 78.6%, respectively (P < 0.05). All cows were cooled by evaporative cooling for 2 adaptation weeks plus 6 experimental weeks under hot weather conditions. The EXP diet reduced rectal temperature and respiratory rate of the cows while increasing their DM intake (DMI) from 23.1 to 24.7 kg/cow per day, milk yield from 41.9 to 44.2 kg and yield of energy-corrected milk from 38.7 to 39.7 kg, as compared with the CON group. Cows fed the EXP TMR had increased RE in milk and body tissue, as compared with the CON group, but the diets had no effect on the measured HP that was maintained constant (130.4 v. 130.8 MJ/cow per day) in the two groups. The measured MEI (MEI = RE + HP) and the efficiency of MEI utilization for RE production were also similar in the two dietary groups.

Effect of photoperiod in the third trimester of gestation on milk production and circulating hormones in dairy goats.

Multiparous Israeli Saanen goats (n = 8) were blocked at dry off (approximately 45 d prepartum) into 2 treatments of 4 goats each based on body weight (BW), previous milk production, and the number of detected embryos in utero. Treatments consisted of long-day (16 h light:8 h dark) and short-day (8 h light:16 h dark) photoperiods at normothermic ambient temperature (22 degrees C, 72% relative humidity). All goats were returned to ambient photoperiod after kidding, milked twice daily, and milk yield was automatically recorded. Dry matter intake was similar between treatments and averaged 980 g/d. Milk production was greater in the short-day than in the long-day treatment (2,932 vs. 2,320 g/d) during the 12-wk experimental period. Milk protein and lactose contents were similar in both treatments and averaged 3.61 and 4.88%, respectively, whereas milk fat was greater in the long-day treatment (4.80 vs. 4.22%). Plasma insulin-like growth factor 1 was greater in the long-day treatment (149 vs. 73 ng/mL) during the dry period than in the short-day treatment, but was similar postkidding, averaging 76 ng/mL. Concentrations of triiodothyronine in plasma were similar in both treatments during the dry period, but greater during lactation in the short-day treatment (122.1 vs. 94.1 ng/mL). Plasma prolactin was greater in the long-day than in the short-day treatment during the dry period (28.0 vs. 17.5 ng/mL), whereas it was similar throughout lactation (11.7 ng/mL). These data support the idea that greater milk production in goats exposed to short days during the dry period is not explained by differences in feed intake or increased secretion of insulin-like growth factor 1.

Prolactin affects leptin action in the bovine mammary gland via the mammary fat pad.

One of the roles of the endocrine system is to synchronize mammary function. Hormones, such as estrogen, progesterone, and prolactin act directly on the mammary gland. Metabolic hormones, such as GH, glucocorticoids, insulin, and leptin are responsible for coordinating the body's response to metabolic homeostasis. Leptin has been shown to be an important factor in regulating the metabolic adaptation of nutrient partitioning during the energy-consuming processes of lactation. In the present study, we show that leptin is secreted from the mammary fat, and is regulated by prolactin. The expression of alpha-casein in a co-culture of epithelial cells and fat explants was enhanced by prolactin compared with that in epithelial cells cultured alone. Leptin antagonist abolished the effect of leptin on alpha-casein expression in mammary gland explants when exogenous leptin was not present in the medium. This finding supports our hypothesis that the antagonist abolishes the action of endogenous leptin secreted by the mammary adipocytes. These results lead us to the hypothesis that prolactin and leptin act in the bovine mammary gland, via mammary fat pad/adipocytes.

Aminopeptidase N gene expression and abundance in caprine mammary gland is influenced by circulating plasma peptide.

This study examined the localization and the effect of circulating peptides on the expression of aminopeptidase N (EC 3.4.11.2) in caprine mammary gland. Four lactating goats in mid to late lactation were used in a crossover design and were subjected to 2 dietary treatments. Abomasal infusion of casein hydrolysate was used to increase the concentration of peptide-bound amino acid in the circulation. Samples of mammary gland tissue from each goat were taken by biopsy at the end of each treatment period to measure gene and protein expression of aminopeptidase N in the tissue. There were no measurable effects on feed intake and milk production for any of the treatments. Western blot analysis showed that aminopeptidase N is located on the basolateral side of parenchymal cells and not on the apical membranes. Abomasal infusion of casein hydrolysate caused a marked change in the profile of arterial blood free amino acids and peptide-bound amino acids smaller than 1500 Da. Abundance of aminopeptidase N mRNA and protein increased by 51 and 58%, respectively, in casein hydrolysate-infused goats compared with the control treatment. It was concluded that aminopeptidase N is one candidate actively involved in the mammary gland to support protein synthesis and milk production. In accordance with the nutritional conditions in the current experiment, it is suggested that aminopeptidase N expression is partly controlled by the metabolic requirements of the gland and postabsorptive forms of amino acids in the circulation.

Effect of nursing management and skeletal size at weaning on puberty, skeletal growth rate, and milk production during first lactation of dairy heifers.

Forty Israeli-Holstein 5-d-old calves were used to determine the effect of increasing calf body weight (BW) and skeletal size during the nursing period on age and skeletal size at puberty and on skeletal size and performance during first lactation. The calves were randomly allotted to 2 experimental groups as follows: milk replacer (MR) [calves were given 0.450 kg/d dry matter of milk replacer for the first 50 d of life] and milk-fed (MF) [calves had free access to milk in two 30-min meals/d]. From weaning to 180 d of age, all calves were fed the same diet. At 180 d of age, the MR and MF calves were each divided into 2 equal subgroups: one subgroup from each treatment was given only growing ration, and the other was given the same ration supplemented with fish meal to supply 2% crude protein (CP) (treatments MR + CP and MF + CP, respectively). Finally, at 270 d of age, all calves were housed together and fed a growing heifer's ration until first calving. During the entire nursing period, the MF calves consumed 9.8% more DM, 39.7% more CP, and 52.4% more metabolizable energy than the MR calves. At 60 d of age, BW and all skeletal parameters were higher in the MF calves than in the MR calves. During the entire rearing period (60 to 550 d), the average BW of the MF calves was greater by 16 kg than the BW of the MR calves. Nursing management did not affect differences in skeletal parameters at calving. Average age at puberty onset was 272 +/- 26.8 d; MF calves reached puberty 23 d earlier than MR calves. Yields of milk (kg/305 d) and fat-corrected milk (FCM, kg/d) were greater for the MF + CP heifers than for the MR heifers. It was concluded that nursing by ad libitum milk, as compared with milk replacer, affected BW but not skeletal size of the adult animal, decreased age of puberty onset, and increased FCM yield at first lactation. Supplementing the diet with 2% CP during the prepubertal period increased BW but not skeletal size of the adult animal and 305-d milk and FCM yields during first lactation.

Expression of albumin in nonhepatic tissues and its synthesis by the bovine mammary gland.

Albumin is a well-characterized product of the liver. In the present study, objectives were to determine if the albumin gene is also expressed in various nonhepatic tissues in the bovine; whether mammary gland epithelial cells synthesize albumin; and how its synthesis is affected by bovine mastitis. Albumin expression was monitored using reverse transcription-polymerase chain reaction. Tissues examined were: liver, mammary gland, tongue, intestine, lymph gland, testicle, ovary, and uterus. All tissues except the ovary expressed the albumin gene, albeit less so than the liver. The highest level of expression (other than liver) was found in the lymph nodes but expression was also found in the mammary gland. Incubation of mammary gland explants with the labeled amino acid L-[(35)S] methionine resulted in formation of labeled immunoprecipitable albumin, newly synthesized in the explant. Immunoprecipitable albumin in the medium verified that newly synthesized albumin was also secreted into the medium. This shows that the gland itself is a source of milk albumin. Albumin mRNA expression was approximately 4 times higher in mammary gland tissue from 6 mastitic cows compared with expression in mammary tissue from 6 healthy glands. Further, secretion of albumin was increased 3.5-fold from explants of mastitic mammary glands compared with secretion from explants of healthy mammary glands. Addition of lipopolysaccharide increased the synthesis and secretion of albumin in mammary gland cells in a dose-dependent manner. Exposure to lipopolysaccharide accelerated albumin synthesis in a time-dependent manner up to 48 h. These results lead us to suggest that the secretion of albumin by the mammary gland is part of the innate nonspecific defense system.

Leptin affects prolactin action on milk protein and fat synthesis in the bovine mammary gland.

Leptin, a protein hormone produced and secreted predominantly by white adipose tissue, has a critical role in the regulation and coordination of energy metabolism. Identification of leptin in the milk of several mammals, including humans, led us to investigate its presence and regulatory effect in the cow mammary gland. The expression of leptin receptor in tissue culture of lactating mammary gland was augmented approximately 25 times by prolactin, but had no effect on virgin calf mammary tissue. Expression of leptin in tissue culture from mammary glands of lactating cows was enhanced 2.2-fold by prolactin. No effect of prolactin on leptin and leptin receptor expression was found in mammary gland tissue culture from calves. Leptin-enhanced fatty acid synthesis in the presence of prolactin, but had no effect without presence of prolactin. A similar pattern was found in the expression of alpha-casein and beta-lactoglobulin in mammary gland explants from a lactating cow. Our findings indicate that leptin plays an important role in mammary gland lactogenesis, and that the expression of leptin requires the presence of prolactin.

Changes in milk composition as affected by subclinical mastitis in sheep.

The mechanism of the effects of glandular-level subclinical mastitis in dairy sheep on milk yield and on its composition as expressed in curd yield was studied. Thirty-six Israeli-Assaf dairy sheep with one udder half infected with identified coagulase-negative staphylococci and the contralateral gland free of bacteria were chosen. The milk yield of the infected halves was significantly lower than that of the uninfected ones (0.36 vs. 0.76 kg/milking). The somatic cell count and N-acetyl-beta-D-glucosaminidase activity were significantly higher in the infected halves than in the uninfected ones. The plasminogen activator and plasmin (PL) activities were significantly higher in the infected glands than in the uninfected ones, whereas plasminogen (PLG) activity and the ratio PLG:PL were significantly lower in the infected glands. Concentrations of Ca2+ did not differ, whereas Ca2+ activity was significantly lower and proteose peptone concentration was 2.4 times as high in the infected glands than in the uninfected ones. Curd yield was significantly lower in the infected glands than in the uninfected ones.

Infusions of casein hydrolyzates into the mammary gland disrupt tight junction integrity and induce involution in cows.

Milk stasis triggers local stimuli, which make the tight junctions leak and trigger involution. The aim of the study was to test the hypothesis that casein hydrolyzates compromise tight junction integrity and dry-off milk secretion in dairy cows. Six repeated doses of casein hydrolyzates after each milking during 3 d caused drastic changes in mammary secretion and composition, which were associated with irreversible cessation of milk secretion. No such changes were recorded in the control glands that had been treated with nonhydrolyzed casein. Treatment with casein hydrolyzates disturbed tight junction integrity within 8 h (as indicated by changes in Na+ and K+ concentrations), reduced the concentrations of lactose precipitously, activated the plasmin activator-plasminogen-plasmin system, and induced the secretion of immunoglobulin type G and lactoferrin. At the end of the 3-d treatments, we stopped milking the experimental and control glands. Milk composition 19 d later was similar in the experimental and control glands and was consistent with the composition expected in fully involuted glands. We conclude that casein hydrolyzates are among the milk-borne factors that cause the disruption of tight junction integrity and induce involution in cows. The process induced by casein hydrolyzate was more rapid and synchronized than the involution induced at drying-off.

Interrelationships among ambient temperature, day length, and milk yield in dairy cows under a Mediterranean climate.

We examined the effect of calving month (CM) on the production of milk and milk protein by Israeli Holstein dairy cows located in the main climatic zone of Israel during their third and fourth lactations, and found it to be significant. Cows that calved in December produced the highest milk and milk protein yields, and those that calved in June produced the lowest, 92.8% of the maximum. The combined effect of the environmental average temperature and day length accounted for 0.96 of the variability in average milk production during lactation and 0.93 of that in average protein production during lactation. Average milk production was reduced by 0.38 kg/degree C and average protein production was reduced by 0.01 kg/degree C. Elongation of daylight increased average milk production by 1.2 kg/h and average protein production by 0.02 kg/h of daylight. Analysis of the temperature pattern effect on milk and protein yield during lactation indicated that cows at the second month (the pike of their milk yield) are more vulnerable to the negative temperature effect than cows on the ninth month of lactation.

Adrenocorticotrophic hormone and dexamethasone failed to affect milk yield in dairy goats: comparative aspects.

The ability of adrenocorticotrophic hormone (ACTH; single i.v. injection of 2.5IU/kg BW) and dexamethasone (single i.m. injection of 36mg/kg BW) to affect milk production was studied in mid-lactating Israeli Saanen goats. None of these treatments produced changes in milk yield and composition of the goats. The effects of ACTH on blood cortisol levels, and the effects of ACTH and dexamethasone on blood plasma concentrations of glucose, however, were consistent with previous reports in goats and cows. These responses suggest that ACTH and dexamethasone treatments produced their expected glucocorticoid effects. It is suggested that obstructing the axis: stress-ACTH-glucocorticoid-down regulation of milk yield, which was demonstrated in dairy cows, reflects the adaptation of goats to harsh conditions, and the selection pressure to produce milk under conditions which are considered stressful for other ruminants.

Stress down regulates milk yield in cows by plasmin induced beta-casein product that blocks K+ channels on the apical membranes.

Stress and stress related hormones such as glucocorticoids inhibit lactation in cows. In the present study we propose a novel mechanism connecting stress with plasminogen-plasmin system (PPS) (an enzymatic mechanism in milk, which leads to the breakdown of the major milk protein casein). We show that stress activates the PPS leading to an increase in plasmin activity, and that a distinct plasmin-induced beta-casein breakdown product (fraction 1-28) is a potent blocker of potassium channels in mammary epithelia apical membranes. The reduction in milk production due to dehydration stress or glucocorticoid (dexamethsone) was correlated with the activities of plasmin and channel blocking activity in the milk of the tested cows. The notion that the axis Stress-PPS-beta-casein fraction 1-28 is responsible for the reduction in milk yield is supported by the results of experiments showing that injecting solution composed of casein digest enriched with beta-casein fraction 1-28 to the udder lumen leads to a transient reduction in milk production. Furthermore, injecting a pure beta-casein fraction 1-28 to the udder lumen of goat's lead also to a transient reduction in milk production with kinetics that was similar to the kinetics observed in cows.

Effect of acquisition of improved thermotolerance on the induction of heat shock proteins in broiler chickens.

The role of heat shock proteins (HSP) in the protection of cells from heat stress is well established. However, very little is known about their contribution to thermotolerance in the complexity of a whole homeotherm animal. Here we report on the analysis of protein synthesis in lung and heart muscle tissues of broiler chickens following exposure to high ambient temperature. Half of the flock was treated by an early age exposure to heat (conditioning), to improve thermotolerance. In contrast to what has been expected, lower levels of HSP induction was observed in the treated chickens. We suggest that 1) the induction of HSP in the heart and lung tissues of the whole animal correlates with the body temperature and 2) HSP response does not represent a part of the long-term mechanism that is evoked by the early age conditioning.

Osteocalcin (BGP), gene expression, and protein production by marrow stromal adipocytes.

This study was designed to demonstrate the expression and production of osteocalcin, a bone Gla-protein (BGP), by marrow stromal cells. We were able to accomplish this by using a series of marrow stromal cell lines (MBA cells). A unique expression of the osteocalcin was detected by the adipocyte 14F1.1 cells. This was at the mRNA level by Northern blot and by RT-PCR analysis. The secreted protein was quantitated by radioimmunoassay (RIA), in conditioned medium (CM) harvested from these cultured cells. These findings offer the first evidence that marrow adipocyte 14F1.1 derived cells express mRNA for osteocalcin and produce the protein.

Large-scale preparation and characterization of recombinant ovine placental lactogen.

To clone ovine placental lactogen (oPL) cDNA, total RNA from sheep placental cotyledon was reverse transcribed and the single-stranded cDNA was PCR-amplified with 5' and 3' primers containing, respectively, NcoI and PstI sites. The oPL cDNA fragment amplified between these two primers extended from A(-1) to the natural stop codon. The PCR product was gel-purified and subcloned into a Puc vector and the insert was sequenced on both strands, revealing several differences relative to the published sequence: S19N, S69N, D129E and R165Q. We assume that these differences can be accounted for by the high level of individual polymorphism, which has been described in detail for PLs of different species. The insert was subcloned into NcoI/ PstI-digested pTrc99A procaryotic expression plasmid and protein expression was induced by isopropyl-1-thio-beta-D-galactopyranoside. Because of low expression, oPL's cDNA was further subcloned into pET8 procaryotic expression plasmid. Its expression in E. coli strain BL21 transformed with this vector yielded 30-40 mg/l. The expressed protein, found in the inclusion bodies, was refolded into a monomer and purified on a Q-Sepharose column to homogeneity. Structural analysis using circular dichroism revealed a spectrum similar to that of human GH (hGH) thereby indicating proper refolding. Gel filtration and binding experiments, including real-time kinetic measurements using the surface plasmon resonance method revealed that oPL forms transient homodimeric complexes with extracellular domains of prolactin receptors from rabbit, rat and bovine and with hGH receptor. The purified oPL was biologically active in an Nb2-11C cell proliferation bioassay, in its ability to stimulate beta-casein synthesis in explants of ovine and rabbit mammary gland and fat synthesis in explants of bovine mammary gland, and in a proliferation assay using FDC-P1 cells transfected with rabbit or hGH receptors.

Inhibition of lactogenic activities of bovine mammary gland explants by the whey fraction of bovine milk.

The regulation of milk constituents, synthesis and secretion in tissue cultures of the bovine mammary gland was altered by a whey fraction of bovine milk. alpha-Casein gene expression, casein secretion and fatty acid synthesis were inhibited by the whey fraction in a dose-dependent manner. The whey fraction inhibited the enhancement activity of prolactin on alpha-casein gene expression and fatty acid synthesis, and also inhibited casein secretion to the medium, in explants cultured in a medium with or without prolactin. No effect on the expression of the beta-lactoglobulin gene was found.