The involvement of glycoprotein CD36 and fatty-acid-binding protein (FABP) in cellular growth, differentiation, lipid transport and metabolism led us to examine the possible biochemical and physiological relationship(s) between these two proteins. We investigated three aspects of this relationship. We first attempted to identify any physical complex formed between CD36 and FABP in bovine milk fat globule membranes. These membranes are the product of mammary gland secretory epithelial cells. The second aspect studied was the effect of synthetic peptide analogs to the C-terminus (amino acid residues 121-131) of bovine mammary gland FABP on cell proliferation, as a result of the interaction of these peptides with the ectodomain of CD36. Finally, mammary gland CD36 and FABP coexpression was defined at different stages of lactation and during involution. Immunoprecipitation, Western immunoblotting with anti-FABP and anti-CD36, Northern-blot analysis and a mammary epithelial cell proliferation assay demonstrated that: (a) bovine milk fat globule membranes contain the complex of CD36 and FABP, and that this complex is, most likely, formed as a result of FABP binding to the cytoplasmic segments of CD36; (b) synthetic analog of the C-terminus of FABP with the sequence Val-Thr-Cys, identical to the sequence found in the CD36-binding domain of thrombospondin, was a more potent inhibitor of bovine mammary gland epithelial cell proliferation than a synthetic peptide with the Val-Cys-Thr sequence; (c) the expression of FABP and CD36 is related to the state of mammary cell differentiation, since it reaches its maximum during lactation and declines during the involutionary period.

Bovine growth hormone (51.5 IU/day) and placebo injections were administered for 10 days to four Holstein cows in early lactation (wk 12) and again in late lactation (wk 35). Milk productions in the last 5 days of each period were compared. In early lactation, growth hormone increased milk yield by 15%, fat yield by 17%, protein yield by 14%, and lactose yield by 21%. In late lactation the respective increases were 31, 42, 18, and 35%. For responses of early and late lactation to growth hormone on a quantitative basis, increases for milk yield (4.3 versus 3.9 kg/day) and milk energy secretion (3.3 versus 3.4 Mcal/day) were similar. Concurrent with these increased milk yields, ad libitum intakes of a complete mixed diet declined during the period of growth hormone treatment by 3% in early lactation and 16% in late lactation. During the 6 h immediately following injections of growth hormone, blood plasma concentrations of growth hormone were elevated about 400% in early lactation and 700% in late lactation. Concentrations in plasma of free fatty acids were also higher during growth hormone treatment in late lactation but not in early lactation. Treatments did not affect plasma concentrations of glucose, insulin, glucagon, prolactin, tri-iodothyronine, thyroxine, or cortisol in either early or late lactation. Daily administration of growth hormone in early or late lactation resulted in similar and substantial increases of milk yield and efficiency of milk production.

The objectives of the present study were 1) to determine the heterogeneity of the MAC-T cell line; 2) to examine whether homogeneous clones could be derived from MAC-T cells; and 3) to examine cell morphology, cytoskeletal characteristics, size, colony-forming ability, growth characteristics, beta-casein production, response to oxytocin, and cytogenetic properties of the clones. Three clonal cells, designated CU-1, CU-2, and CU-3, were derived from MAC-T cells. CU-1 and CU-2 cells were morphologically homogeneous. CU-3 cells were heterogeneous and contained two distinct subtypes. All clones contained cytokeratin 14 and 18. CU-2 and CU-3 cells were 30 and 18% larger, respectively, than CU-1 cells. CU-1 cells did not grow in serum-free medium. Doubling times for MAC-T, CU-2, and CU-3 were 46, 48, and 78 h, respectively, in serum-free medium. MAC-T cells and clones constitutively expressed beta-casein in culture ranging from .1 to .3 micrograms/ml per 24 h. Cytogenetic analyses revealed Robertsonian translocations and isochromosomes in the clonal lines. We conclude that parental MAC-T cells are heterogeneous in morphology, growth, and cytogenetic characteristics.

Bovine growth hormone (bGH) was administered to high-yielding Holstein cows fed a complete mixed ration ad libitum. Commencing on day 74 of lactation, 10 cows averaging 34.4 kg milk per day were divided into two groups and received a daily subcutaneous injection of bGH (51.5 IU/day) or a placebo. Injections were continued for an 11-day period and differences in lactational performance, nitrogen balance and estimated energy balance between the two groups were compared for the last 5 days of the preinjection and injection periods. Growth hormone resulted in increases of 9.5% in milk yield, 22.7% in milk fat yield, 14.5% in milk lactose yield and a 17.1% increase in milk energy secretion. Feed intake was slightly reduced (-4.3%, nonsignificant) while milk protein secretion and nitrogen balance were unchanged. Serum growth hormone levels in the bGH group were maintained at the higher concentrations of the normal physiological range during the injection period. By 48 hours following the last injection, declining bGH concentrations approached control values, and milk production decreased to preinjection values. Serum prolactin levels and plasma concentrations of free-fatty acids were slightly increased during the injection period in the bGH group. Growth hormone clearly enhances milk synthesis in the high-yielding dairy cow.

The relative amounts of immunoreactive plasminogen and active plasmin in different fractions of bovine milk were examined. Raw milk was centrifuged to separate skim, cream, and a somatic cell pellet. Skim milk was centrifuged to separate milk serum and casein micelles. Milk fat globule membranes were isolated from the cream fraction of bovine milk. Proteins from somatic cells were isolated following sonication of the cells. Western blot analysis showed the presence of several forms of plasminogen in bovine milk. The predominant forms of plasminogen identified following electrophoresis under nonreducing conditions were proteins with approximate molecular weights of 88,000, 152,000, and 160,000. The predominant forms of plasminogen identified after electrophoresis under reducing conditions were two proteins with approximate molecular weights of 88,000 and 50,000. The highest amount (82% of the total plasminogen), as determined by an ELISA, was associated with the casein fraction. Lower plasminogen concentrations were associated with the serum, cream fractions, and milk fat globule membranes. The SDS-PAGE of the cream and milk fat globule membranes indicated that some casein was present in both fractions. Thus, the low plasminogen concentrations in these fractions may be associated with the caseins there. No immunoreactive plasminogen was present in the somatic cells. Active plasmin was present in the same milk fractions in which plasminogen was detected: casein, serum, and cream.