Glycerolipid biosynthesis in porcine adipose tissue in vitro: effect of adiposity and depot site

To compare genetic differences in glycerolipid biosynthesis, rates were determined in s.c. adipose tissue of lean and obese pigs at 28, 60 and 110 d of age. To compare depot-specific differences, glycerolipid biosynthetic rates were determined in outer s.c., middle s.c., perirenal and omental adipose tissues obtained from 105-kg contemporary pigs. Rates were determined with a 700 x g infranatant fraction of an adipose tissue homogenate by measuring glycerophosphate incorporation into total lipids (mostly phosphatidic acid) during 4 min. This assay represents entrance of substrates into the glycerolipid synthesis pathway or glycerophosphate acyltransferase (GPAT) activity. Rates measured for 60 min represent maximal synthesis of glycerolipid (more triacylglycerol than phosphatidic acid) or lipid synthesis capacity (LSC). Adipocyte diameter and volume were greater for adipose tissue of obese than of lean pigs both at 60 and 110 d. When expressed per cell, activity of GPAT and LSC were similar for lean and obese pigs at 28 d. At 60 d and 110 d, LSC was greater for obese than for lean pigs; GPAT activity was greater at 60 but not at 110 d in obese than in lean pigs. Expressed on a cell basis, GPAT activity was highest in omental and outer s.c., intermediate in perirenal and lowest in middle s.c. adipose tissue depots. Lipid synthesis capacity was highest in perirenal and lowest in outer and middle s.c. depots. Our results indicate that the LSC assay was more closely related to the accretion of fat in vivo than to GPAT activity.     

Glycerolipid biosynthesis in porcine adipose tissue in vitro. II. Synthesis by various types of cellular preparations

Glycerolipid biosynthesis by porcine adipose tissue homogenates did not yield the 90+% triacylglycerol observed in situ. Consequently, we compared intact tissue slices and various subcellular fractions to characterize the usefulness of such systems to assess glycerolipid biosynthesis in vitro. Glycerolipid biosynthesis by porcine adipose tissue homogenates was measured in vitro using either [14C]-fatty acid or [14C]-glycerol-3-phosphate (G3P) as a radiolabelled substrate. Removal of residual 14C-labelled fatty acid from lipid extracts was difficult. Because G3P is soluble in water, residual [14C]-G3P separated easily from the glycerolipid-containing organic phase and, thus, was the preferred radiolabelled substrate. With tissue slices, glycerol and G3P were minimally incorporated into lipid so that [14C]-fatty acid was the preferred radiolabelled tracer. A washing procedure followed by thin layer chromatography was devised to separate residual [14C]-fatty acid from glycerolipids, including phospholipids. Fatty acid esterification into glycerolipids in tissue slices yielded about 4% phospholipids, whereas with homogenates, esterification yielded up to 50% phospholipids. Comparison of several subcellular fractions indicated that microsomes contained most of the glycerolipid biosynthetic activity when activity was expressed on a protein basis. However, when activities were expressed on a tissue wet weight basis, the 700 x g infranate and the 10,000 x g supernate had about equal activity that was far greater than the microsomes. The 700 x g infranate was the preferred enzyme preparation for assay of the entrance of G3P into the pathway as well as the capacity to synthesize triacylglycerol. Several methods of freezing and storing tissue or 700 x g infranates were not acceptable. Freezing of the 700 x g infranate in liquid N2 with storage at -80 degrees C may be an acceptable procedure.     

Effects of adrenergic agonists and insulin on porcine adipose tissue lipid metabolism in vitro

Because this laboratory has been able to demonstrate only a small and somewhat inconsistent stimulation of glucose metabolism by insulin in porcine adipose tissue in vitro, the tissue was preincubated with insulin to attempt to enhance the hormone effect. Preincubation with or without insulin did not increase insulin stimulation. Furthermore, insulin did not stimulate triacylglycerol biosynthesis. Adrenergic hormones stimulated lipolysis in porcine adipose tissue in vitro. Several analogs of norepinephrine incubated with porcine adipose tissue in vitro did not inhibit glucose incorporation into CO2 or total lipids, in contrast to inhibition observed in adipose tissue from other species. Isoproterenol inhibited glycerol-3-phosphate incorporation into lipids; the maximal inhibition was 50% for the initial stages of the pathway. Palmitate incorporation into lipids also was inhibited 50% by isoproterenol but this may have been an artifact. Preincubation of adipose tissue, with no exogenous hormone, might decrease the concentration of endogenous adrenergic hormones and thus make the tissue more responsive to exogenous adrenergic hormones. Preincubation of porcine adipose tissue did not consistently lower the basal lipolytic rate but enhanced the stimulated lipolytic rate; the mechanism is not known. These experiments provide no evidence that preincubation is beneficial to measurement of lipolysis or glucose metabolism in porcine adipose tissue in vitro.     

Effect of bovine growth hormone on incorporation of [14C]acetate into lipids by co-cultures of bovine mammary, liver, and adipose tissue explants

Incorporation of [14C]acetate into lipids was measured in 24 hr co-cultures of mammary, liver and adipose tissue from Holstein cows at 53, 210 and 318 d of lactation in the presence or absence of bovine growth hormone. Little (less than 1%) of the labeled lipids appeared in the media relative to that incorporated into the tissue. In mammary tissue, incorporation of [14C]acetate was highest into triglycerides (16,298 cpm/mg mammary tissue), followed by phospholipids (1,887 cpm), free fatty acids (1,252 cpm), diglycerides (708 cpm), free cholesterol (360 cpm) and monoglycerides (93 cpm). Bovine growth hormone did not increase incorporation of [14C]acetate when mammary or adipose tissue were incubated separately. However, in the presence of liver and adipose tissue, bovine growth hormone significantly increased the incorporation of [14C]acetate into triglycerides, diglycerides, free fatty acids and free cholesterol by mammary tissue. These results suggest that bovine growth hormone acts on mammary tissue indirectly through liver and adipose tissue to increase lipid synthesis. This mechanism may play a role in the action of bovine growth hormone in vivo to increase milk and milk fat production.     

Human acylation-stimulating protein and lipid biosynthesis in bovine adipose tissue explants

Human acylation-stimulating protein (hASP) up-regulates triacylglycerol synthesis in human adipocytes. The objectives of this research were 1) to determine the effect of hASP on triacylglycerol synthesis in bovine adipose explants and 2) to determine whether nutritional status influences the sensitivity of adipose tissue to hASP. Fresh s.c. adipose tissue was sectioned into 20- to 30-mg explants and incubated for 1 to 6 h in M199 media containing 3% BSA and either 0.75 mM [1-14C]palmitate, 0.75 mM [9, 10-3H]oleate, or 2.5 mM [1-14C] acetate, as well as hASP and(or) insulin. The explants were extracted, and lipid fractions were separated by TLC and quantified by liquid scintillation. Acetate incorporation into lipids increased 15 to 30%, and palmitate or oleate incorporation increased 10 to 25%, when explants were exposed to hASP, although this response was not significant in every experiment. Insulin increased triacylglycerol synthesis in some experiments, but not in others. Our interpretation is that acylation-stimulating protein (ASP) can mildly enhance triacylglycerol synthesis in bovine adipose tissue. To fulfill the second objective, nine 9-mo-old steers were housed individually for two periods of 3 wk each. During the first period, four of the nine steers were fed to 50% of NEm requirement and the other five consumed the same diet ad libitum. After the first period, all steers consumed feed ad libitum for 2 wk and were assigned the opposite ration for the second period. Steers gained 40.5 kg BW when allowed ad libitum access to feed but lost 30.2 kg BW when feed intake was restricted (SE = 7.84; P < 0.01). At the end of each period, s.c. adipose tissue was sectioned into explants and incubated as described above. Four explants per steer per period were used to test effects of insulin (0 and 1 nM) and hASP (0, 0.01, 0.1, and 1 microM). Insulin did not influence incorporation of acetate or oleate. Acetate incorporation (P < 0.32) was 0.99, 1.03, 1.04, and 1.10 nmol x mg(-1) h(-1) (SE = 0.13) and oleate incorporation (P < 0.01) was 0.347, 0.357, 0.353, and 0.420 nmol x mg(-1)h(-1) (SE = 0.022) for 0, 0.01, 0.1, and 1 microM hASP, respectively. Feed restriction reduced (P < 0.01) acetate and oleate incorporation by 95 and 40%, respectively. No interactions among feed intake, insulin, and hASP were detected. In conclusion, the effect of hASP on fatty acid esterification is not influenced by feed restriction.     

Stimulation of lipogenesis by insulin in swine adipose tissue: antagonism by porcine growth hormone

The effects of physiological (1, 10 ng/ml) and pharmacological (1,000 ng/ml) concentrations of insulin (INS) and porcine growth hormone (pGH) on lipid metabolism were determined in short-term (2 h) and long-term (26, 50 h) incubations of swine adipose tissue. The short-term effects of three different commercial sources of bovine serum albumin (BSA) on adipose tissue metabolism were also evaluated. Two of the three BSA preparations were found to be unsuitable for inclusion in the short-term incubation buffer because they caused a stimulation of lipid synthesis in adipose tissue and masked the stimulatory effects of insulin. Physiological concentrations of insulin stimulated glucose metabolism in 2-h incubations by 100% in adipose tissue from 80-kg swine. After a 26-h incubation period, INS maintained rates of glucose metabolism at levels comparable to maximally stimulated rates in fresh tissue. Insulin also enhanced glucose metabolism following 50-h incubations; however, rates were less than for 2- or 26-h incubations. Glucose metabolism was also stimulated in adipose tissue from 127-kg swine when incubated for 2 h with INS; however, INS responsiveness declined with increasing body weight. Lipogenesis and glucose oxidation were partially maintained by INS using tissue from the heavier swine. A pharmacological but not physiological concentration of pGH stimulated glucose metabolism in short-term incubations by 50% in adipose tissue from 80-kg swine, and by 10% in adipose tissue from 127-kg swine. Long-term culture of adipose tissue in the presence of pGH had no effect on glucose metabolism. Physiological levels of pGH directly antagonized the stimulation of glucose metabolism by INS in short- and long-term incubations. In summary, these results are the first to establish that swine adipose tissue is quite sensitive to insulin and that pGH directly antagonizes insulin action.     

Lipid metabolism and low molecular weight solute uptake in Ascaridia galli

Adult Ascaridia galli, an intestinal nematode parasite of fowl, reveals a large variety of complex lipids such as phospholipids containing choline, ethanolamine, inositol, serine and glycerol. Lysophospholipid species, vinyl ether phospholipid (plasmalogen), neutral acylglycerols, cholesterol and non-esterified fatty acids are also present. Sugar-containing lipids, such as cerebrosides, sulphatides and gangliosides are abundantly present. Female parasites contain more lipids, particularly acylglycerols and phospholipids. Acylglycerols, phosphatidyl choline, phosphatidyl ethanolamine and glycolipids incorporate a large amount of radiolabelled precursor substrate in A. galli. The presence of important enzymes of lipid biosynthesis like glucose-6-phosphate dehydrogenase, malate dehydrogenase and hydroxymethyl glutaryl-CoA reductase as well as an enzyme of lipid ester hydrolysis, triacylglycerol lipase is detected in the parasite. These enzymes show subcellular distribution patterns and Michaelis-Menten kinetic characteristics comparable with that from rat liver homogenate. Studies on the uptake of labelled precursor molecules for lipid biosynthesis, glucose, acetate and palmitate show that the parasites can take up the isotopes readily in a time-dependent manner, showing substrate saturation kinetics, dependence upon Na ions, and can be inhibited by the presence of the bile salts sodium cholate and sodium deoxycholate. The substrate affinity constant (Kt) and maximum apparent velocity of glucose uptake in A. galli were found to be 9.09 mM and 26.67 mM per 100 mg tissue dry weight per min at 37 degrees C.     

Lipogenesis in acute and 48-hour cultures of bovine intramuscular and subcutaneous adipose tissue explants

In this study, the interactions among breed of cattle, adipose tissue site and specific incubation conditions were investigated. Subcutaneous and i.m. adipose tissues were obtained from 10 Angus and 9 Santa Gertrudis steers immediately postmortem. Adipose tissue explants were incubated acutely for 2 h immediately at slaughter or after being cultured 48 h with or without 1 mU/ml insulin and 30 mg/ml bovine serum albumin; the incorporation of 14C-labeled acetate and glucose (5 mM, plus 5 mM unlabeled lactate) into lipid fractions was measured. AT the same chronological age, Angus steers had a more youthful lean maturity score, higher USDA marbling score and higher USDA quality grade (P less than .05) than did carcasses from Santa Gertrudis steers. The lower marbling score of the Santa Gertrudis steers was paralleled by smaller i.m. adipocytes (P less than .05) relative to Angus steers. Pentose cycle reductase and NADP-malate dehydrogenase activities were greater in Angus i.m. adipose tissue than in Santa Gertrudis i.m. adipose tissue, which would provide more reducing equivalents (NADPH) and glycerol for fatty acid biosynthesis and triacylglycerol esterification. Correspondingly, Angus i.m. adipose tissue exhibited a greater rate of lipogenesis from acetate and glucose (P less than .05) than did Santa Gertrudis i.m. adipose tissue in acute incubations. The presence of insulin resulted in higher rates of lipogenesis from acetate in Angus s.c. adipose tissue than in Santa Gertrudis s.c. adipose tissue after 48 h of explant culture. These data indicate that i.m. and s.c. adipose tissues exhibit aspects of lipid metabolism unique to each tissue and suggest that breed-related differences in adipose tissues may explain the divergent responses to insulin observed in different laboratories.     

Adiposity of calf- and yearling-fed Brangus steers raised to constant-age and constant-body weight endpoints

We tested the hypothesis that fatty acid biosynthesis and adipocyte diameter and volume would be greater in s.c. and i.m. adipose tissues of calf-fed steers than in yearling-fed steers at a constant BW, due to the greater time on feed for the calf-fed steers. Conversely, we predicted that the capacity for s.c. and i.m. preadipocytes to divide, as estimated by 3H-thymidine incorporation into DNA, would be greater in the less mature adipose tissues of calf-fed steers and in yearling-fed steers at 16 mo of age than in yearling-fed steers fed to 18 mo of age. Brangus steers were fed a corn-based finishing diet as calves (calf-fed; n = 9) or yearlings (n = 4) to 16 mo of age (CA yearling-fed); another group of yearlings (n = 5) was fed to a constant-BW end point of 530 kg (CW yearling-fed). Both groups of yearling-fed steers had free access to native pasture until 12 mo of age. At slaughter, the fifth to eighth thoracic rib section of the LM was removed, and fresh s.c. and i.m. adipose tissues were removed for in vitro incubations. There were no differences in the number of s.c. adipocytes/g or mean peak volumes of adipocytes across production groups (P > or = 0.14). However, s.c. adipose tissue of CA yearling-fed steers contained greater proportions of smaller adipocytes (<1,500 pL) than calffed or CW yearling-fed steers, and similar results were observed for i.m. adipose tissue. Acetate incorporation into total lipids was greater (P = 0.02) in s.c. adipose tissue of CA yearling-fed steers than in calf-fed or CW yearling-fed steers, and tended to be different (P = 0.10) across production groups in i.m. adipose tissue. The production system x cell fraction interaction was significant (P = 0.03) for s.c. adipose tissue DNA synthesis, which was greatest in adipocytes from CA yearling-fed steers, whereas there were no differences across production system in stromal vascular (SV) DNA synthesis. For i.m. adipose tissue, DNA synthesis was greatest in adipocytes and SV cells from CA yearling-fed calves, and was greater in SV cells than in adipocytes (both P = 0.01). Therefore, stage of adipose tissue development more strongly influenced fatty acid synthesis, adipocyte volume, and DNA synthesis than age at sampling, final BW, or time on the finishing diet.     

Influence of the sex-linked dwarfing gene (dw) on the contribution of dietary lipid to yolk lipid synthesis

1. The efficiency and time course of dietary fatty acid incorporation into lipids of egg yolk and abdominal adipose tissue was compared in "White Leghorn", normal (Dw) and dwarf (dw) laying hens at 56 weeks of age, using 14C labelled linoleic acid. 2. The sex-linked dwarfing gene, dw, was shown to reduce not only body weight and abdominal fat pad deposition, but also yolk production and the average clutch size. 3. Higher peak incorporation and total recovery of the linoleic acid radioactivity into yolk lipids, but lower label recovery into adipose tissue triglycerides were found in dwarf hens. 4. The higher esterification of the dietary linoleic acid in its native form into dwarf yolk triglycerides indicates that dwarf hens use more dietary lipids to synthesise yolk lipids but these results also suggest that the dw allele might reduce the lipogenic capacities of the liver and adipose tissue in laying hens.     

In vitro lipogenesis in the liver and adipose tissues of the female Aylesbury duck at different ages

The in vitro incorporation of [U — ¹⁴C]‐glucose into total lipids was uniformly high in liver slices from birds aged 2, 4 and 10 weeks. The separate addition of prolactin, oestrogen or insulin in vitro always failed to increase the rate of lipogenesis.

The rate of incorporation of [U — ¹⁴C] glucose in vitro into total lipids of subcutaneous, peritoneal and inguinal adipose tissues decreased with age and was considerably lower than that in the liver. There was no difference between the rates of lipid synthesis in adipose tissues from different sites.

Whereas 75% of the glucose converted into total lipids by the liver was recovered as fatty acids, the proportion was only 9 to 15% for adipose tissues. These results suggest that adipose tissue is not an important site for the synthesis of lipid de novo in the growing duck. Hepatic lipogenesis did not reflect the previously reported changes in the rate of fat deposition during growth in the duck.     

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.     

Technical note: adipose tissue blood flow in miniature swine (Sus scrofa) using the 133xenon washout technique

The purpose of this study was to examine the 133xenon washout technique as a viable method for measuring adipose tissue blood flow (ATBF) in swine. Using a total of 32 female Yucatan miniature swine (Sus scrofa), the partition coefficient for 133xenon in swine subcutaneous adipose tissue was determined and ATBF was measured at rest and under various physiological conditions. These conditions included feeding, anesthesia, epinephrine infusion, and acute exercise. The effects of epinephrine and acute exercise were examined in both sedentary and exercise-trained swine. The partition coefficient value for 133xenon in swine subcutaneous adipose tissue was 9.23+/-0.26 mL/g (mean +/- SD, n = 10). The average value for resting ATBF in swine was 3.98+/-2.72 mL/(100 g tissue-min) (n = 19). Feeding increased ATBF by approximately fivefold over fasting values, and isoflurane anesthesia significantly decreased ATBF compared to rest (1.64+/-1.12 vs 3.92+/-4.22 mL/[100 g x min], n = 10). A 30-min epinephrine infusion (1 microg/[kg BW x min]) significantly increased ATBF from a resting value of 3.13+/-2.61 to 10.35+/-5.31 mL/(100 g x min) (n = 12). Epinephrine infusion into exercise-trained swine increased ATBF to the same extent as when infused into sedentary swine. An acute, 20-min bout of exercise significantly increased ATBF in swine, and the sedentary swine showed a larger increase in ATBF than their exercise-trained littermates relative to rest: 7.83 vs 2.98 mL/(100 g x min). In conclusion, the 133xenon washout technique appears to be a viable method for measuring ATBF in swine; our findings are comparable to swine ATBF values reported using the microsphere method and are consistent with values reported in animal and human studies.     

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.     

Technical Note: In vivo estimation of lipogenesis using a bolus injection of [U-13C]glucose in pigs

The use of radioactive isotopes to measure de novo lipogenesis in pigs has been well established. Different from radioactive isotopes, stable isotopes present little or no risk to human and animal subjects. Therefore, the objective of this study was to adapt the method of bolus injection of radioactive glucose (¹⁴C) to use ¹³C-labeled glucose to estimate de novo lipogenesis in finishing pigs. Five vein-catheterized gilts received 3.0 kg/d of a commercial diet for 2 wk. On the last day, the pigs received a bolus injection of [U-¹³C]glucose (12 mg/kg body weight). A serial of blood samples was taken for 4 h to determine the glucose rate of disappearance (R_d) from plasma glucose isotopic enrichment (IE). The ¹³C IE of lipids was determined from adipose tissue biopsies collected at 1, 2, and 3 h after the bolus injection and from adipose tissue collected after pig euthanasia 4 h after the bolus. Lipogenesis was estimated from the incorporation of ¹³C from glucose into adipose tissue lipids. Glucose R_d, estimated using a double-exponential function, averaged 5.4 ± 1.4 mmol/min. The IE of lipids increased linearly during the 4 h following the bolus injection (P < 0.05). The rate of incorporation of glucose into lipids, estimating lipogenesis, averaged 9.0 µg glucose/(min × g of lipids) 4 h after the bolus injection. In conclusion, the in vivo method using a bolus injection of [U-¹³C]glucose allows a successful estimation of de novo lipogenesis in finishing pigs.     

Effect of porcine somatotropin on in vivo glucose kinetics and lipogenesis in growing pigs.

Crossbred barrows were used for in vivo studies investigating hormonal regulation of lipogenesis. The first experiment examined an in vivo method for determining rates of lipogenesis. Three barrows were infused with [U-14C]glucose and incorporation of radioactivity into triglycerides was determined in up to five biopsies of subcutaneous adipose tissue obtained over 7 h. Incorporation was linear after blood glucose specific radioactivity had reached a plateau and was constant over the entire infusion. For the second experiment, eight pigs (71 +/- 2.5 kg) were allocated to one of two treatments involving daily injections of excipient (control) or porcine somatotropin (pST; 120 micrograms/kg of BW). On d 10, beginning 15 h after injection, glucose incorporation into adipose tissue lipid was determined under both basal and hyperinsulinemic/euglycemic conditions. Basal glucose incorporation into lipid, particularly fatty acids, was markedly reduced (greater than 90%) during pST treatment. Although glucose incorporation was increased to a similar extent in both groups by hyperinsulinemia, the pST-treated pigs still exhibited markedly lower rates. Based on kinetic data, the decrease in lipid accretion of pST-treated pigs was primarily the result of a decrease in the rate of de novo synthesis. Furthermore, the reductions in glucose incorporation into fatty acids, glucose irreversible loss rate, and feed intake that occur with pST treatment were quantitatively similar.     

Effect of a high-linolenic acid diet on lipogenic enzyme activities,fatty acid composition,and meat quality in the growing pig

Forty-eight Duroc-cross gilts (40 kg initial BW) were fed a control or a linseed diet containing 60 g of whole crushed linseed/kg. Both diets were supplemented with 150 mg of vitamin E/kg. Eight pigs from each dietary treatment were slaughtered at 20, 60, or 100 d after the start of the experiment. There was no effect (P > 0.05) of diet on growth, carcass characteristics, or foreloin tissue composition. Feeding the linseed diet increased (P < 0.05) the content of n-3 PUFA in plasma, muscle, and adipose tissue, but docosahexaenoic acid was not (P > 0.05) altered by diet. The proportions of n-3 PUFA were highest (P < 0.01) in pigs fed the linseed-diet for 60 d, regardless of tissue (plasma, muscle, or adipose tissue) or lipid (neutral lipids and phospholipids) class. The linseed diet produced a PUFA:saturated fatty acid ratio > or = 0.4 in all groups and tissues, which is close to the recommended value for the entire diet of humans, as well as a robust decrease in the n-6:n-3 ratio. The decrease (P < 0.01) in the percentage of oleic acid in adipose tissue of pigs fed the linseed diet for 60 d could be attributed to a 40% decrease (P < 0.001) in stearoyl-CoA-desaturase activity. Diet did not (P > 0.05) affect the activities of acetyl-CoA-carboxylase, malic enzyme, or glucose-6-phosphate-dehydrogenase in any tissues. Muscle vitamin E content was decreased (P < 0.001) 30% in pigs fed crushed linseed for 60 d, whereas lower (P < 0.001) concentrations of skatole in pork fat were observed in linseed-fed pigs at all slaughter times. Inclusion of linseed (flaxseed) in swine diets is a valid method of improving the nutritional value of pork without deleteriously affecting organoleptic characteristics, oxidation, or color stability.     

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.     

Acute and long-term lipogenic response to insulin and clenbuterol in bovine intramuscular and subcutaneous adipose tissues

The present study was conducted to determine whether insulin and clenbuterol affected either short-term (2-h) incubations or long-term (48-h) tissue cultures of i.m. and s.c. adipose tissue explants. Samples were taken from control steers and steers fed 7 mg.head-1.d-1 clenbuterol for 50 d, after which time the drug was withdrawn from the diet for 90 d prior to slaughter. Neither short-term incubations nor long-term explant cultures contained bovine serum albumin (BSA). Insulin (6.67 x 10(-9) M) had no effect (P greater than .05) on lipogenesis in s.c. and i.m. adipose tissue in 2-h tissue incubations of fresh adipose tissue. There was a substantial decrease in activity during the culture period, which was ameliorated somewhat in s.c. adipose tissue by the presence of insulin in the culture media. Clenbuterol exposure for 48 h in vitro decreased the production of lipids from acetate in both adipose tissue depots but had no effect in short-term adipose tissue incubations. Results from the present study confirm that omitting BSA from incubation media does not enhance the responsiveness of bovine s.c. adipose tissue or the less mature i.m. adipose tissue to insulin. Insulin may maintain greater cell viability in 48-h explant cultures.     

Incorporation in vitro of 14C fatty acids into bovine sebaceous gland and dermal lipids

A study has been made of the incorporation in vitro of 14C palmitic, oleic and linoleic acids into the skin lipids of cattle. Linoleic acid was incorporated into the triglyceride (triacylglycerol) fraction of the sebaceous gland lipid at a greater rate than palmitic and oleic acids. Its incorporation was much greater when presented as a free acid than when presented either as cholesteryl linoleate or linoleoyl lecithin. However, the ability of cholesteryl ester and phospholipid to make a substantial indirect contribution of linoleic acid to sebum triglyceride synthesis by hydrolytic release of fatty acid was indicated. The association between the observed preferential incorporation of linoleic acid into the sebum triglycerides and the uniquely unsaturated triglycerides found on the skin surface of cattle is discussed.