Acetyl-CoA carboxylase of sheep adipose tissue: problems of the assay and adaptation during fetal development

The rate of fatty acid synthesis of perirenal adipose tissue of fetal lambs decreased by 90% during the last month of gestation. There was also a 90% decrease in the activity of fatty acid synthetase during this period, but the activity of this enzyme exceeded lipogenic flux by a factor of 10. The activity of acetyl CoA carboxylase in the active state (initial activity) was very similar to the lipogenic flux in adipose tissue from lambs at 120 d of gestation; although activity decreased towards term, the decline was insufficient to account for the fall in rate of fatty acid synthesis. The study also shows that assay of acetyl CoA carboxylase in the active state of ovine adipose tissue and of caprine mammary gland requires the presence of citrate, thus differing from that for rat adipose tissue. Evidence that pyruvate carboxylase can interfere in the assay of acetyl CoA carboxylase also is presented.     

Dietary L-carnitine suppresses mitochondrial branched-chain keto acid dehydrogenase activity and enhances protein accretion and carcass characteristics of swine.

A trial was conducted to biochemically explain the decreased lipid deposition and increased protein accretion observed in pigs fed carnitine. Our hypothesis was that an increase in the ratio of acetyl CoA:CoA-SH produced by stimulation of fatty acid oxidation by supplemental L-carnitine may decrease branched-chain alpha-keto acid dehydrogenase activity and increase pyruvate carboxylase activity. Such changes could reduce oxidative loss of branched-chain amino acids and provide more carbons for amino acid biosynthesis. Yorkshire gilts (n = 36; 12 per treatment) were fed a control diet or diets containing either 50 or 125 ppm of added L-carnitine during growth from 56 to 120 kg. After slaughter, the semitendinosus muscle and liver were collected for isolation of mitochondria and hepatocytes. Increasing dietary L-carnitine did not influence growth performance (P > 0.10) but linearly decreased (P < 0.05) 10th rib backfat thickness and increased (linear, P < 0.05) percentages of lean and muscle. The rates of [1-(14)G]palmitate oxidation in isolated hepatocytes and isolated mitochondria, and incorporation of [35S]methionine into the acid insoluble fraction of isolated hepatocytes were increased (linear, P < 0.01) in pigs fed L-carnitine. Flux through branched-chain alpha-keto acid dehydrogenase linearly decreased (P < 0.01) in isolated liver and muscle mitochondria with increasing dietary carnitine. Flux through pyruvate carboxylase was increased (linear, P < 0.01) in isolated mitochondria from liver of pigs fed carnitine, and assays with particle-free extracts indicated that the amount of mitochondrial pyruvate carboxylase was tripled by feeding carnitine (linear, P < 0.01). The association of increased protein accretion and reduced backfat thickness with greater rates of palmitate oxidation, more rapid flux through pyruvate carboxylase, and reduced flux through branched-chain alpha-keto acid dehydrogenase suggests pigs fed carnitine are more able to use fat for energy, divert carbon toward synthesis of amino acids, and spare branched-chain amino acids for protein synthesis.     

Changes in activities of lipogenic enzymes in adipose tissue and liver of growing goats.

The lipogenic capacity of omental adipose tissue and liver was measured in vitro from samples obtained at slaughter from 33 young male goats. The animals were slaughtered either on the day of weaning (d 0) or 2, 14, or 56 d after weaning. Ages at weaning were 4 wk (early weaning) or 6 or 8 wk (late weaning). Blood samples from the jugular vein were taken before slaughter to measure the concentrations of plasma glucose and nonesterified fatty acids. There was a 30% decrease in glucose concentration after weaning. Nonesterified fatty acid concentration increased fourfold between d 0 and 2 after weaning. By d 14 after weaning, nonesterified fatty acids returned to basal concentration. The lipoprotein lipase (LPL) activity of adipose tissue declined markedly (90%) on d 2 after weaning. Lipoprotein lipase activity returned to preweaning values by d 56 after weaning in those goats that had ad libitum access to feed. In adipose tissue, nicotinamide adenine dinucleotide phosphate (NADP)-malate dehydrogenase activity fell by only 17% by d 2 after weaning and to 63% by d 14 after weaning. Lipoprotein lipase activity was closely related to metabolizable energy intake the day before slaughter. Acetyl-coenzyme A carboxylase activity was low in adipose tissue and it increased only slightly by d 56 after weaning. The data indicated that LPL played a preponderant role in the restoration of lipid stores after weaning. High NADP-malate dehydrogenase activity together with a high concentration of plasma glucose by d 56 after weaning suggested that this enzyme activity could be enhanced by high glucose availability in goat kids. Activities of lipase, acetyl-coenzyme A carboxylase, NADP-malate dehydrogenase, and glucose-6-phosphate dehydrogenase in liver were essentially unaffected by weaning. The extent and rapidity of change of lipogenic enzymes of goat kids was much more pronounced in adipose tissue than in liver.     

Expression of fat deposition and fat removal genes is associated with intramuscular fat content in longissimus dorsi muscle of Korean cattle steers

Intramuscular fat (IMF) in cattle is an important component of traits that influence meat quality. We measured carcass characteristics and gene expression in Korean steers to clarify the molecular mechanism(s) underlying IMF deposition in LM tissue by determining the correlation between IMF content and gene expression abundance and by developing models to predict IMF content using gene expression abundance. The deposition of IMF is determined by a balance between fat deposition and fat removal in the LM. We measured mRNA abundance of lipid metabolic genes including lipogenesis [acetyl CoA carboxylase (ACC), fatty acid synthase (FASN)], fatty lipid uptake [lipoprotein lipase (LPL), fatty acid translocase (CD36), fatty acid transport protein 1 (FATP1)], fatty acid esterification [glycerol-3-phosphate acyltransferase 1 (GPAT1), acylglycerol phosphate acyltransferase 1 (AGPAT1), diacylglycerol acyltransferase 1 (DGAT1), DGAT2], lipolysis [adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL), monoglyceride lipase (MGL)], and fatty acid oxidation [carnitine palmitoyl transferase 1B, very long-chain acyl-CoA dehydrogenase (VLCAD), medium-chain acyl-CoA dehydrogenase (MCAD)] in the LM. The mRNA abundance of the GPAT1 gene showed the greatest correlation (r = 0.74; P < 0.001) with IMF content among 9 fat deposition genes. The gene expression abundance of other fat deposition genes including ACC, FASN, LPL, CD36, FATP1, AGPAT1, DGAT1, and DGAT2 also exhibited significant positive correlations (P < 0.05) with IMF content in the LM. Conversely, ATGL mRNA abundance showed the greatest negative correlation (r = -0.68; P < 0.001) with IMF content in the LM among 6 fat removal genes. The expression of other fat removal genes including MGL, VLCAD, and MCAD showed significant negative correlations (P < 0.05) with IMF content. Our findings show that the combined effects of increases in lipogenesis, fatty acid uptake, fatty acid esterification, and of decreases in lipolysis and fatty acid oxidation contribute to increasing IMF deposition in Korean steers. The multiple regression analysis revealed that the mRNA abundance of the GPAT1 gene in the LM was the first major variable predicting IMF content (54%) among 15 lipid metabolic genes. The second was mRNA abundance of ATGL (11%). In conclusion, these results suggest that GPAT1 and ATGL genes could be used as genetic markers to predict IMF deposition in the LM.     

Glucose-6-phosphate dehydrogenase and leptin are related to marbling differences among Limousin and Angus or Japanese Black x Angus steers

This work investigated the metabolic basis for the variability of carcass and i.m. adiposity in cattle. Our hypothesis was that the comparison of extreme breeds for adiposity might allow for the identification of some metabolic pathways determinant for carcass and i.m. adiposity. Thus, 23- to 28-mo-old steers of 3 breeds, 2 with high [Angus or Japanese Black x Angus (J. Black cross)] and 1 with low (Limousin) i.m. and carcass adiposity, were used to measure activities or mRNA levels, or both, of enzymes involved in de novo lipogenesis [acetyl-coA carboxylase, fatty acid synthase (FAS), glucose-6-phosphate dehydrogenase (G6PDH), malic enzyme], circulating triacylglycerol (TAG) uptake (lipoprotein lipase), and fatty acid esterification (glycerol-3-phosphate dehydrogenase), as well as the mRNA level of leptin, an adiposity-related factor. In a first study, enzyme activities were assayed in the s.c. adipose tissue (AT), the oxidative rectus abdominis, and the glycolytic semitendinosus muscles from steers finished for 6 mo. Compared with Angus or J. Black cross, Limousin steers had a 27% less (P = 0.003) rib fat thickness, and 23 and 29% less (P < or = 0.02) FAS and G6PDH activities in s.c. AT. In rectus abdominis and semitendinosus, the 75% less (P < 0.001) TAG content was concomitant with 50% less (P < 0.001) G6PDH activity. In a second study, enzyme activities plus mRNA levels were assayed in an oxido-glycolytic muscle, the longissimus thoracis (LT), in the i.m. AT dissected from LT, and in s.c. AT from the same Limousin steers and from Angus steers finished for 10 mo. Compared with Angus, the 50% less (P < 0.001) rib fat thickness in Limousin contrasted with the 1.1- to 5.8-fold greater (P < or = 0.02) mRNA levels or activities, or both, of acetyl-coA carboxylase, G6PDH, lipoprotein lipase, and glycerol-3-phosphate dehydrogenase in s.c. AT. Conversely, the 90% less (P < 0.001) TAG content in Limousin LT was concomitant to the 79 and 83% less (P < or = 0.002) G6PDH activity and leptin mRNA level. Such differences could arise from a greater number of adipocytes in LT from Angus steers because no difference was found between Limousin and Angus for G6PDH activity and leptin mRNA in i.m. AT. We conclude that FAS and G6PDH in s.c. AT could be involved in differences in carcass adiposity, but this relationship disappeared when the fatness increased strongly. Leptin and G6PDH are related to the expression of marbling whatever the body condition and thus could be relevant indicators of marbling in beef cattle.     

Activities of certain cytoplasmic enzymes in bovine T lymphoma.

Activities of certain cytoplasmic enzymes were measured in bovine T lymphoma (BTL-PC3 cells). The activities of hexokinase, pyruvate kinase and glucose-6-phosphate dehydrogenase in PC3 cells were elevated as much as 2 or 3-fold of those in bovine thymocyte. The high activities of these enzymes derived from activation of glucose metabolism may reflect the high growth potential of PC3 cells.     

Subcutaneous and intramuscular adipose tissue stearoyl-coenzyme A desaturase gene expression and fatty acid composition in calf- and yearling-fed Angus steers

We proposed that stearoyl-CoA desaturase (SCD) activity dictates fatty acid composition of adipose tissue and muscle in beef cattle, regardless of ruminal or hepatic fatty acid hydrogenation or desaturation. Twelve Angus steers were assigned to a calf-fed (CF) group and slaughtered at weaning (8 mo of age; n=4), 12 mo of age (n=4), or 16 mo of age (n=4). Twelve steers were assigned to a yearling-fed (YF) group and slaughtered at 12 mo of age (n=4), 16 mo of age (n=4), and 17.5 mo of age (n=4; 525 kg, market weight). Data were analyzed based on time on the corn-based finishing diet, with terminal age as a covariate, and orthogonal polynomial contrasts were tested on the main effects of treatment group and time on the finishing diet. Fatty acids from duodenal digesta, plasma, liver, LM, and subcutaneous and intramuscular adipose tissue were measured, and SCD gene expression was measured in intramuscular and subcutaneous adipose tissues. In duodenal digesta, palmitic and linoleic acids increased by 100% over the sampling period, α-linolenic acid decreased over the sampling period, and trans-vaccenic acid was greater in YF than in CF steers (all P < 0.01). The proportion of α-linolenic acid decreased over time in all tissues, including liver. The SCD index (ratio of SCD fatty acid products to SCD fatty acid substrates) increased over time in LM and in intramuscular and subcutaneous adipose tissues. The SCD:glyceraldehyde 3-phosphate dehydrogenase mRNA ratio was virtually undetectable at the initial sampling periods in subcutaneous adipose tissue of YF and CF steers, and it increased over time (P < 0.01). The SCD index and SCD:glyceraldehyde 3-phosphate dehydrogenase ratio were greater in intramuscular adipose tissue of CF steers than in that of YF steers. The SCD index did not change over time in liver and decreased over time in duodenal digesta. We conclude that, unlike essential fatty acids, the SFA and MUFA composition of adipose tissue is regulated by adipose tissue fatty acid desaturation, with little contribution from hepatic or duodenal fatty acids.     

Glucose transport and glycolytic enzyme activities in erythrocytes of two-year-old thoroughbreds undergoing training exercise

d-Glucose transport and cytosolic enzyme activities were measured in erythrocytes from 2-year-old thoroughbreds under continuous training exercise (race horses) and compared with those from untrained horses of various ages (sires, mares and untrained 2-year-old thoroughbreds). The activities of the glucose transport and glycolytic enzymes, hexokinase and pyruvate kinase, in the race horses' erythrocytes were elevated to 2–3.5 times above those of untrained horses. There were no significant differences in plasma glucose, triglyceride or IRI concentrations between the horses in training and untrained horses. The increases in glucose transport and glycolytic enzyme activities in their erythrocytes are considered to reflect an increased metabolic activity in the race horses resulting from the training exercises.Abbreviations D-GT d-glucose transport - EDTA ethylenediaminetetraacetic acid - ELISA enzyme-linked immunosorbent assay - G6PD glucose-6-phosphate dehydrogenase - HK hexokinase - IRI immunoreactive insulin - PBS phosphate-buffered saline - PK pyruvate kinase     

Effect of dietary energy source on in vitro substrate utilization and insulin sensitivity of muscle and adipose tissues of Angus and Wagyu steers

Angus (n = 8; 210 kg of BW) and 7/8 Wagyu (n = 8; 174 kg of BW) steers were used to evaluate the effects of dietary energy source on muscle and adipose tissue metabolism and insulin sensitivity. Steers were assigned to either a grain-based (corn) or hay-based (hay) diet and fed to similar final BW. At slaughter, LM and s.c. and i.m. adipose tissue samples were collected. Portions of the LM and adipose tissues were placed immediately in liquid N for later measurement of glycolytic intermediates. Fresh LM and s.c. and i.m. adipose tissues were incubated with [U-(14)C]glucose to assess glucose metabolism in vitro. All in vitro measures were in the presence of 0 or 500 ng/mL of insulin. Also, s.c. and i.m. adipose tissues were incubated with [1-(14)C]acetate to quantify lipid synthesis in vitro. Glucose-6-phosphate and fructose-6-phosphate concentrations were 12.6- and 2.4-fold greater in muscle than in s.c. and i.m. adipose tissues, respectively. Diet did not affect acetate incorporation into fatty acids (P = 0.86). Insulin did not increase conversion of glucose to CO(2), lactate, or total lipid in steers fed hay but caused an increase (per cell) of 97 to 110% in glucose conversion to CO(2), 46 to 54% in glucose conversion to lactate, and 65 to 160% in glucose conversion to total lipid content in adipose tissue from steers fed corn. On a per-cell basis, s.c. adipose tissue had 37% greater glucose oxidation than i.m. adipose (P = 0.04) and 290% greater acetate incorporation into fatty acids than i.m. adipose (P = 0.04). Insulin addition to s.c. adipose tissue from corn-fed steers failed to stimulate glucose incorporation into fatty acids, but exposing i.m. adipose tissue from corn-fed steers to insulin resulted in a 165% increase in glucose incorporation into fatty acids. These results suggest that feeding hay limited both glucose supply and tissue capacity to increase glucose utilization in response to insulin without altering acetate conversion to fatty acids. Because s.c. adipose tissue consistently utilized more acetate and oxidized more glucose than did i.m. adipose, these results suggest that hay-based diets may alter i.m. adipose tissue metabolism with less effect on s.c. adipose tissue.     

Coordinate regulation of ovine adipose tissue gene expression by propionate

The current study examined the acute effects of intravenous propionate infusion on plasma hormones and metabolites and the expression of adipose tissue lipogenic genes. Four yearling rams were assigned to one oftwo groups (saline or propionate infusion) in a crossover design. All sheep were cannulated in both jugular veins and infused with 1.2 M propionate at a rate of 64 micromol x mix(-1) x kg BW(-1) for 30 min. Blood samples were collected at -10, 0, 5, 10, 20, 30, 60, and 120 min after initiation of infusion. Subcutaneous adipose tissue biopsies were obtained from the tailhead at 0 and 2 h after propionate infusion and analyzed for gene expressions of lipoprotein lipase, acetyl CoA carboxylase, fatty acid synthase, peroxisome proliferator-activated receptor gamma, leptin, and uncoupling protein-2 using a nonisotopic ribonuclease protection assay. The partial cDNA of the enoyl reductase region of ovine fatty acid synthase was cloned and sequenced from s.c. adipose tissue of sheep. The deduced amino acid sequence (210 amino acids) was 86% identical to human, 88% identical to rat, 88% identical to mouse, and 72% identical to chicken. Plasma glucose and insulin concentrations abruptly increased 5 min after beginning propionate infusion and further increased up until 30 min but were unaffected in saline-infused sheep (P < 0.05). Plasma concentration of NEFA decreased (P < 0.05) during propionate infusion, whereas IGF-I levels were unaltered. The amounts of lipoprotein lipase, acetyl CoA carboxylase, fatty acid synthase, peroxisome proliferator-activated receptor gamma, and leptin mRNA increased (P < 0.05) in s.c. adipose tissue of propionate-infused sheep compared with those of saline-infused sheep. However, uncoupling protein-2 mRNA decreased (P < 0.05) in propionate-infused sheep. This study demonstrates that an acute nutrient challenge, in the form of i.v. propionate, can stimulate or inhibit the expression of various adipose tissue genes involved with lipogenesis and adipose tissue metabolism.     

Effects of in vitro ronnel on metabolic activity in subcutaneous adipose tissue and skeletal muscle from steers

Ronnel [0,0-dimethyl 0-(2,4,5-trichlorophenyl) phosphorothioate] is an organophosphate pesticide with growth-promoting properties. Experiments were conducted to determine effects of ronnel on oxidation of and fatty acid synthesis from acetate and glucose as indices of metabolic activity in subcutaneous adipose tissue and skeletal muscle from 6-, 12- and 18-mo-old steers. Ronnel depressed metabolic activity in adipose tissue from 6- and 12-mo-old steers without concomitantly decreasing metabolic activity in skeletal muscle. Production of CO2 and fatty acids from acetate and glucose in tissues from 18-mo-old steers was influenced less by ronnel than in tissues from younger steers. Interactions of ronnel with thyroxine or growth hormone on acetate oxidation and conversion to fatty acids in adipose tissue also were investigated. Thyroxine increased acetate oxidation and decreased fatty acid synthesis. Ronnel interfered with the metabolic effects of thyroxine. Growth hormone, with or without ronnel, did not affect metabolic activity of adipose tissue. Ronnel seemingly alters the partitioning of acetate and glucose between major metabolic processes in adipose tissue and skeletal muscle.     

Effects of epigallocatechin gallate on lipid metabolism and its underlying molecular mechanism in broiler chickens

The objective of this study was to investigate the effects of epigallocatechin gallate (EGCG) on fat metabolism and to establish the molecular mechanism of these effects in broilers. Seventy‐two 28‐day‐old male Ross 308 broiler chickens were divided into three groups with different levels of EGCG supplementation for 4 weeks: normal control (NC) group, L‐EGCG (a low‐level supplement of EGCG, 40 mg/kg body weight daily) and H‐EGCG (a high‐level supplement of EGCG, 80 mg/kg body weight daily). After 4 weeks of oral administration, EGCG significantly reduced the level of abdominal fat deposition in broilers. The serum triglycerides and low‐density lipoprotein cholesterol of chickens in H‐EGCG group were also significantly decreased compared with the NC group, and the high‐density lipoprotein cholesterol was notably increased at the same time. Moreover, the vital role of the liver and abdominal adipose tissue in lipid metabolism of poultry animals was examined through gene expression and enzyme activities related to fat anabolism and catabolism in these organs. Our data show that EGCG supplementation for 2 weeks significantly downregulated the expression of fatty acid synthesis and fat deposition‐related genes, and upregulated the expression of genes involved in fatty acid β‐oxidation and lipolysis genes. Simultaneously, the activities of hepatic fatty acid synthesis enzymes (fatty acid synthase and acetyl CoA carboxylase) were significantly decreased, and the activity of carnitine palmitoyl transferase‐1 was notably elevated. The results suggest that EGCG could alleviate fat deposition in broilers through inhibiting fat anabolism and stimulating lipid catabolism in broilers.     

Effects of body condition score at parturition and postpartum supplemental fat on adipose tissue lipogenic activity of lactating beef cows

Three-year-old Angus x Gelbvieh beef cows nutritionally managed to achieve a BCS of 4 +/- 0.07 (479.3 +/- 36.3 kg of initial BW) or 6 +/- 0.07 (579.6 +/- 53.1 kg of initial BW) at parturition were used in a 2-yr experiment (n = 36/yr) to determine the effects of BCS at parturition and postpartum lipid supplementation on cow adipose tissue lipogenesis. Beginning 3 d postpartum, cows within each BCS were randomly assigned to be fed hay and a low-fat control supplement or supplements with either cracked high-linoleate safflower seeds or cracked high-oleate safflower seeds until d 60 of lactation. Diets were formulated to be isonitrogenous and isocaloric, and safflower seed diets provided 5% DMI as fat. Adipose tissue biopsies were collected near the tail-head region of cows on d 30 and 60 of lactation. Dietary treatment did not affect (P > or = 0.43) adipose tissue lipogenesis. Body condition score at parturition did not affect acetate incorporation into lipid (P = 0.53) or activity of acetyl CoA carboxylase (P = 0.77) or fatty acid synthase (P = 0.18). Lipoprotein lipase activity and palmitate incorporation into triacyl-glycerol tended to be greater (P = 0.06), and palmitate esterification into total acylglycerols was greater (P = 0.01) in cows with a BCS of 4 at parturition. Mean activity of acetyl-CoA carboxylase (P < 0.001), lipoprotein lipase (P = 0.01), and rate of palmitate incorporation into monoacylglycerol (P = 0.02), diacylglycerol (P = 0.001), triacylglycerol (P = 0.003), and total acylglycerols (P = 0.002) were greater at d 30 than d 60, suggesting a greater proclivity for fatty acid biosynthesis and esterification by adipose tissue at d 30 of lactation. Although dietary lipid supplementation did not affect adipose tissue lipogenesis, results suggest that cows with a BCS of 4 at parturition have a greater propensity to deliver exogenously derived fatty acids to the adipocyte surface and incorporate preformed fatty acids into acylglycerols as stored adipocyte lipid. Additionally, cows in early lactation seemed to be able to synthesize and incorporate more fatty acids into stored lipid than cows during peak lactation.     

Adipocyte metabolism and cellularity are related to differences in adipose tissue maturity between Holstein and Charolais or Blond d'Aquitaine fetuses

This paper reports the metabolic and morphological characteristics of bovine adipose tissue (AT) at end of fetal life and its variability with breed and anatomical site of AT. Our hypothesis was that, in cattle, end-of-fetal-life differences in adipocyte number, size, and histology may account for differences in AT maturity. To address this question, perirenal and intermuscular AT were sampled from Charolais, Blond d'Aquitaine, and Holstein fetuses at 260 d postconception. Holstein fetuses showed greater leptin mRNA abundance, which is consistent with the greater perirenal AT weight (P = 0.03) than Blond d'Aquitaine fetuses. Compared with Blond d'Aquitaine or Charolais fetuses, Holstein fetuses had larger (P < 0.001) adipocytes, greater (P < 0.05) activities of enzymes involved in de novo fatty acid (FA) synthesis (FA synthase, glucose-6-phosphate dehydrogenase, malic enzyme) and FA esterification (glycerol-3-phosphate dehydrogenase), and greater (P = 0.06, P = 0.10, P < 0.001) mRNA abundance for lipolytic enzymes (hormone-sensitive lipase and adipose triglyceride lipase) and uncoupling protein 1 in both perirenal and intermuscular AT. This indicates increased FA turnover in Holstein adipocytes through FA storage, mobilization, and oxidation pathways. Whatever the breed, adipocytes were smaller in perirenal AT than intermuscular AT. Whatever the breed or anatomical site, bovine AT at 260 d postconception contained predominantly unilocular adipocytes believed to be white adipocytes together with a few multilocular brown adipocytes. We conclude that the greater metabolic and morphologic maturity of adipocytes from Holstein than Blond d'Aquitaine and Charolais fetuses may contribute to the greater thermogenic aptitude of Holstein newborns. Moreover, the presence of both white and brown adipocytes at the end of fetal life highlights the complexity of AT structure and may indicate that the cellular and functional heterogeneity of AT repeatedly observed postnatally has a developmental origin.     

Antibiotics promote abdominal fat accumulation in broilers

Antibiotics stimulate the growth of animals but result in drug residues and bacterial resistance. In this study, the negative effect of antibiotics on abdominal fat deposition was evaluated in broilers. The results showed that adding both chlortetracycline (50 g/1,000 kg) and tylosin (50 g/1,000 kg) significantly increased abdominal fat weight, abdominal fat percentage (p < .05), and triglyceride and cholesterol levels (p < .05) in blood. Also, both products synchronously stimulated intestinal absorption and synthesis of liver fat. The expression levels of the peroxisome proliferator‐activated receptor γ (PPARγ), diacylgycerol acyltransferase 2 (DGAT2), lipoprotein lipase (LPL), and fatty acid‐binding protein (FABP4) genes in abdominal fat tissue significantly increased (p < .05 or 0.01) when antibiotics were added to the feed. However, no significant difference was found in expression of the fatty acid synthesis (FAS) or acetyl CoA carboxylase (ACC) genes. Further in vitro study results revealed that antibiotics had no effect on fat content or the related gene expression levels in preadipocytes. In summary, the antibiotics induced fat deposition in adipose tissues by activating extracellular absorption of fatty acids from intestinal absorption and synthesis of liver fat. However, it shows no direct regulation by adipose tissue.     

Effects of dietary energy level on lipid metabolism‐related gene expression in subcutaneous adipose tissue of Yellow breed × Simmental cattle

This study was conducted to estimate the effect of dietary energy level on lipid metabolism‐related gene expression of subcutaneous adipose tissue in Yellow breed × Simmental cattle. The experiment was conducted for 60 days. The results showed that final weight, average daily gain, average backfat thickness, (testicles + kidney + pelvic) fat percentage and subcutaneous fat percentage in the high and medium energy groups were significantly higher than in the low‐energy group but that the feed conversion ratio was significantly lower. The glucose, triglycerides, cholesterol, high‐density lipoprotein and low‐density lipoprotein in the high‐energy group were significantly higher than in the low‐energy group. With dietary energy increasing the activities of lipoprotein lipase (LPL), fatty acid synthase (FAS) and acetyl‐CoA carboxylase (ACC) significantly increased, whereas hormone‐sensitive lipase (HSL) and carnitine palmitoyltransferase‐1 (CPT‐1) significantly diminished. Peroxisome proliferator‐activated receptor γ (PPARγ), LPL, FAS, sterol regulatory element binding protein 1 (SREBP‐1), ACC, stearoyl‐CoA desaturase (SCD) and adipocyte‐fatty acid binding proteins (A‐FABP) gene expression were significantly increased by dietary energy increasing, and HSL and CPT‐1 gene expression were significantly decreased. These results indicated that with dietary energy increasing, the subcutaneous fat accumulation mainly increased due to adipose tissue lipogenic gene expression and decreased lipolytic gene expression.     

Acetyl-CoA carboxylase and fatty acid synthase activity and immunodetectable protein in adipose tissues of ruminants: effect of temperature and feeding level

To gain insights into the regulation of fat synthesis, we have investigated the effect of cold environmental exposure and feed restriction of sheep on activity and immunodetectable protein content of acetyl-CoA carboxylase (ACC) and fatty acid synthase in adipose tissue. Subcutaneous and mesenteric adipose tissues were collected at slaughter from sheep exposed to either cold (0+/-2 degrees C) or warm (23+/-2 degrees C) environment, and given either ad libitum or restricted access to feed for three 5-wk periods. Acetyl-CoA carboxylase was isolated from frozen adipose tissue samples and activity determined as the rate of incorporation of H14CO3- into acid stable malonyl-CoA. Cold exposure and feed restriction reduced (P < .05) ACC activity in the two adipose tissue depots. Western blot analysis with peroxidase-conjugated streptavidin showed that both adipose tissue depots express a single isoform of ACC. In s.c. adipose tissue, cold exposure increased (P < .05) ACC protein abundance, which is opposite to the change in activity. However, feed restriction reduced immunodetectable ACC protein. There was no significant effect of environment or feeding level on ACC protein abundance in mesenteric tissue. Fatty acid synthase activity determined in ammonium sulfate extract by measuring the malonyl-CoA- and acetyl-CoA-dependent oxidation of NADPH was decreased (P < .05) by feed restriction in both s.c. and mesenteric tissues. Cold exposure reduced fatty acid synthase activity in s.c. but not in mesenteric tissue. There was no effect of environment on fatty acid synthase protein abundance in either adipose tissue depot. However, feed restriction significantly reduced fatty acid synthase protein abundance in the two depots. The data suggest that feed restriction and exposure of ruminants to cold environmental conditions may significantly down-regulate the activity of key lipogenic enzymes.     

Fatty acid elongation and desaturation enzyme activities of bovine liver and subcutaneous adipose tissue microsomes

Assay conditions were established for the fatty acid elongation and the delta 9 desaturase enzyme systems of bovine liver and adipose tissue microsomes; rat liver microsomes were used as a reference. Overall fatty acid elongation was determined by measuring the incorporation of [2-14C]malonyl-coenzyme A (CoA) to 14C-labeled stearate. Rat liver elongation activity was .50 +/- .02 nmol.min-1.mg protein-1; bovine liver microsomal elongation activity was substantially lower (P less than .05), with a mean value of .15 +/- .02 nmol.min-1.mg protein-1. The elongation activity of bovine s.c. adipose tissue microsomes (.42 +/- .10 nmol.min-1.mg protein-1) was not different (P greater than .05) from the activity observed in rat liver microsomes. To determine the fatty acid delta 9 desaturase activity, microsomes were incubated in the presence of [1-14C]stearoyl-CoA and nicotinamide adenine dinucleotide (reduced form) (NADH), and the production of radioactively labeled oleate was quantified. Microsomal delta 9 desaturase activity was similar in rat liver and bovine s.c. adipose tissue microsomes with rates of .15 +/- .04 and .21 +/- .05 nmol.min-1.mg protein-1, respectively. However, no desaturase activity was detected in bovine liver microsomes, indicating that the liver is not a major site of oleate synthesis in this species. To investigate differences in fatty acid metabolism relative to breed type, eight Angus and seven Braford heifers were slaughtered at approximately 12 mo of age. Subcutaneous fat thickness over the 12th-13th thoracic vertebrae was greater in the Angus heifers than in the Braford heifers. However, no differences (P greater than .05) were observed in mean adipocyte size or number of cells per gram of adipose tissue between the Angus and Braford heifers. Similarly, there were no significant differences between the Angus and Braford s.c. adipose tissues for microsomal fatty acid elongation or delta 9 desaturation, or for nicotinamide adenine dinucleotide phosphate (NADP)-malate dehydrogenase, fatty acid synthetase, or the pentose cycle reductases. The inability of bovine liver to convert stearate to oleate was in agreement with the fatty acid composition of the liver lipid, which had a smaller percentage of oleate and a higher percentage of stearate than s.c. adipose tissue.