Fatty acid-binding protein in bovine longissimus dorsi muscle.

Experiments were conducted to purify and obtain amino acid sequence data for fatty acid-binding protein from bovine longissimus dorsi muscle. 1. Separation of sarcoplasmic proteins by fractionation on Sephadex and DEAE cellulose columns indicated that a low abundance fatty acid-binding protein exists in bovine l. dorsi muscle. 2. Fatty acid-binding protein in bovine l. dorsi muscle cannot be separated from myoglobin by standard protein purification procedures. However, partial sequencing of a partially purified protein fraction indicated the presence of an NH2-terminally blocked protein. 3. Myoglobin bound palmitate under the in vitro conditions of these experiments.

Ruminal biohydrogenation of fatty acids from high-oleate sunflower seeds.

The objective of these experiments was to examine methods of modifying the fatty acid composition of bovine tissues. In the first experiment, four steers were fitted with duodenal fistulas and were assigned to four diets in a Latin square design. The steers were fed a control diet or the same diet containing 10% high-oleate partially crushed sunflower seeds, serum-coated sunflower seeds, and heat-treated, serum-coated sunflower seeds for 5 d. Samples of digesta and feces were collected on d 5. The inclusion of sunflower seeds (plain or serum-coated) in the diet increased (P less than .05) the digesta concentration of stearate. The percentage of stearate in the digesta and feces was increased (P less than .05) from 51 to 67% and from 64 to 74%, respectively, when steers were fed the untreated sunflower seed. The fecal concentration of oleate was increased (P less than .05) by dietary sunflower seeds in steers that were fed the serum-coated, unheated sunflower seeds. In a second experiment, heifers (four per group) were fed a corn-based control diet or diets containing 10% of high-oleate sunflower oil encapsulated with calcium alginate, either plain, coated with blood meal, or with blood meal integrated into the pellet. After 50 d on treatment, samples of perianal adipose tissue were obtained by biopsy. The fatty acid composition of the adipose tissue was not modified by the inclusion of the encapsulated oleate in the diet. In summary, limited ruminal bypass of sunflower seed oleate was accomplished with sunflower seed but not with encapsulated oleate.

Relationship between fatty acid-binding protein activity and marbling scores in bovine longissimus muscle.

Studies were conducted in an attempt to establish a relationship between fatty acid-binding protein (FABP) activity and marbling score in bovine longissimus muscle. Longissimus muscle was obtained from four 20-mo-old Charolais-Hereford crossbred heifers, three 16-mo-old Angus steers, and four 18-mo-old Angus steers. Immediately after slaughter, longissimus muscles were removed for the extraction of FABP. Supernatant (S104) fractions containing 41.3 to 144 mg of protein (depending on animal group) were eluted over Sephadex columns, and elution fractions were analyzed for the binding of radiolabeled palmitoyl-coenzyme A (CoA). Specific activities of FABP were 23, 32, and 101 nmol palmitoyl-CoA bound/mg protein for the Charolais-Hereford, 16-mo-old Angus, and 18-mo-old Angus cattle, respectively. These preliminary results suggested that longissimus muscle FABP activity was positively correlated with marbling score. To test specifically for this possibility, longissimus muscle was obtained at slaughter from each of four Wagyu steers, Angus heifers and Braford heifers. Marbling scores taken at the 12th-13th rib junction were Sm45, Sm43, and SI50 for the Wagyu, Angus, and Braford cattle, respectively. Interfascicular adipose tissue was exhaustively removed from sections of the 5th to 8th thoracic region of the longissimus muscle to eliminate any contribution of adipose tissue to FABP activity. For each animal, 300 mg of the S104 were eluted over Sephadex columns. Specific activities for the Wagyu, Angus, and Braford longissimus muscle FABP were 3.1, 3.8, and 3.9 pmol palmitoyl-CoA bound/mg protein, respectively, and were not different (P greater than .05) among the three animal groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Lipid synthesis and adipocyte growth in adipose tissue from sheep chronically fed a beta-adrenergic agent.

The effects of the chronic ingestion of the beta-adrenergic agonist clenbuterol on ovine sc adipose tissue were investigated. Three groups of 10 wether lambs with an average initial weight of 22.7 kg were used as experimental animals. After culling 2 to 3 animals per group, one group of eight sheep was slaughtered (initial). The remaining two groups of sheep (control, n = 7 and clenbuterol-fed, n = 8) were fed either a control, high-energy diet or one containing 2 ppm clenbuterol for 40 to 44 d. At slaughter, sc adipose tissue was obtained from all animals for assays in vitro. Subcutaneous fat accretion observed over time in the control sheep was due primarily to an increase in the number of lipid-filled adipocytes. This phenomenon was not observed in the clenbuterol-fed sheep. The incorporation of acetate into lipid increased in the clenbuterol-fed group relative to the initial group and was numerically greater than the rate observed for the control group. Similar results were observed for lipogenic enzyme activities and fatty acid-binding protein activity. Palmitate esterification in vitro tended to be elevated in the clenbuterol-fed group, relative to the control group, suggesting increased triacylglycerol turnover. The in vitro data indicate that clenbuterol did not decrease sc fat accretion in sheep by inhibiting lipogenesis.

Adipose tissue, longissimus muscle and anterior pituitary growth and function in clenbuterol-fed heifers.

The present study was conducted to determine the effects of feeding clenbuterol on adipose tissue and longissimus muscle growth in heifers. For 50 d, 14 heifers were fed either a sucrose-based, clenbuterol supplement or a placebo in which the clenbuterol had been omitted. The heifers were slaughtered in two groups, based on initial weight. Adipose tissue from several anatomical sites and longissimus muscle (depending on slaughter group) were obtained fresh at slaughter. Changes in carcass characteristics elicited by clenbuterol were similar to those reported by others for steers and sheep. Subcutaneous (sc) and intramuscular (im), but not perirenal, adipocytes were smaller and there were more cells per g tissue in the adipose tissue depots of the clenbuterol-fed heifers. Clenbuterol decreased lipogenic enzyme activities, fatty acid-binding protein activity, basal lipolysis and acetate incorporation into glyceride-fatty acids (P less than .05) in sc adipose tissue, but had no effect (P greater than .05) on lipogenesis or lipolysis in im adipose tissue. Clenbuterol elicited a 20% increase in type II myofiber diameters (P less than .05) but had no effect on type I myofiber diameters. In vitro growth hormone release by perifused anterior pituitaries was not affected significantly by long-term in vivo exposure to clenbuterol. These data indicate that a depression in lipogenesis is the mechanism by which clenbuterol decreases subcutaneous fat accretion in cattle.

Lipogenesis in adipose tissue from ovariectomized and intact heifers immunized against estradiol and (or) implanted with trenbolone acetate.

Forty-two heifers were allotted randomly to six treatment groups: intact controls, intact heifers implanted with trenbolone acetate, ovariectomized heifers, ovariectomized heifers implanted with trenbolone acetate, intact heifers immunized against estradiol and intact heifers immunized against estradiol and implanted with trenbolone acetate. Blood titers of estradiol-17 beta were increased over 100-fold in heifers immunized against estradiol in Freund's complete adjuvant or saline:squalene/arlacel containing Mycobacterium. Lipogenic enzyme activities and acetate incorporation into fatty acids were increased in subcutaneous adipose tissue obtained at slaughter from heifers receiving immunization or the combination of immunization and trenbolone acetate. The increased lipogenic capacity was not reflected in either cell diameter or cells per gram adipose tissue. Ovariectomy in combination with trenbolone acetate caused the lowest activities for all enzymes measured. This treatments also caused the greatest decrease in cell diameter, which resulted in the largest number of cells per gram of adipose tissue. Trenbolone acetate alone had no detectable effect on lipogenesis in the intact heifer, but the combination of ovariectomy and trenbolone acetate caused substantial decreases in enzyme activities, in most cases a significant decrease as compared with ovariectomized heifers. The data suggest that trenbolone acetate is able to depress lipogenesis only when not competing with the effects of circulating estradiol.

Fatty acid-binding protein activities in bovine muscle, liver and adipose tissue.

Subcutaneous adipose tissue, sternomandibularis muscle and liver were obtained from steers immediately postmortem. Muscle strips and adipose tissue snips were incubated with 0.75 mM [1-14C]palmitate and 5 mM glucose. Muscle strips esterified palmitate at the rate of 2.5 nmol/min per gram tissue, which was 30% of the rate observed for adipose tissue. Fatty acid-binding protein activity was measured in 104,000 x g supernatant fractions of liver, muscle and adipose tissue homogenates. Muscle and adipose tissue fractions bound 840 and 140 pmol [1-14C]palmitoyl-CoA per gram tissue, respectively. Fatty acid-binding protein activity was greater in adipose tissue than in muscle when data were expressed per milligram protein (35 vs. 13 pmol palmitoyl-CoA bound per milligram of soluble protein, respectively). Fatty acid binding-protein activity was correlated with the rate of palmitate esterification within each tissue. Liver contained the highest fatty acid-binding protein activity (13,000 pmol palmitoyl-CoA bound per gram tissue and 215 pmol palmitoyl-CoA bound per milligrams soluble protein).