Lipogenesis and adipose tissue cellularity in steers switched from alfalfa hay to high concentrate diets

An experiment was conducted to 1) evaluate the effects of diet (alfalfa hay vs high concentrate) on adipose tissue cellularity and rates of in vitro lipogenesis and 2) determine if there was a relationship between in vitro lipogenic rates from acetate and lactate and rates of L- or D-lactate disappearance from plasma. Number of adipose cells/g of tissue decreased with time on experiment; however, hay-fed steers had fewer, but larger cells/g of subcutaneous adipose tissue compared with concentrate-fed steers (.78 +/- .04 vs 1.20 +/- .13 X 10(-6)/g, respectively). These results, however, are likely due to a higher (approximately 25%) intake of dry matter and metabolizable energy by the hay-fed steers. Carcass data obtained at slaughter (460 kg) indicated that the concentrate-fed steers had as much or more adipose tissue compared with the hay-fed steers. Characteristics describing D- or L-lactate disappearance from plasma were not highly correlated with lactate utilization for fatty acid synthesis. Utilization of acetate as a substrate for fatty acid synthesis in vitro was correlated (r = .64) with the rate of lactate utilization for fatty acid synthesis.     

Effect of suppression of prolactin secretion on adipose lipogenesis in the postpartum beef cow

Effects of inhibiting prolactin secretion and of calf removal at 3 d postpartum on the lipogenic capacity of s.c. adipose tissue were investigated in postpartum beef cows. The rate of fatty acid synthesis (SYN) from [1-14C]acetate and the activity of fatty acid synthetase (FAS) were assessed on adipose tissue obtained by biopsy at 1, 2, 4, 8 and 10 wk postpartum. Administration of bromocriptine (BR; a drug that suppresses prolactin secretion in rats) between d 7 and 42 postpartum decreased average serum prolactin concentrations nearly 90%, but BR had no effect on lipogenic rates at any week compared to control (CO) cows. Rates of SYN (nmol acetate.min-1.g-1 tissue) increased linearly in CO and BR cows from a nadir of 3.1 at wk 1 to 19.3 by wk 8. Within CO and BR, cows with the greater energy intake relative to requirements for lactation (energy balance) had the faster rates of recovery of SYN. Cows whose calves were weaned early (3 d) showed rapid early increases in SYN, reaching an average maximum rate of 46.2 by wk 2. Activity of FAS generally followed a pattern similar to that of SYN for all groups. Results indicate that prolactin is not responsible for low rates of postpartum lipogenesis in s.c. adipose tissue and that energy intake influences the rate of recovery.     

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.     

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.     

Copper and lipid metabolism in beef cattle: a review

Results from experiments investigating Cu metabolism in ruminants indicate that Cu is involved in lipid metabolism. Copper supplementation ranging from 10 to 40 mg of Cu/kg of DM to high-concentrate finishing diets decreased subcutaneous adipose tissue deposition and decreased cholesterol concentrations but increased unsaturated fatty acid composition of LM. Serum norepinephrine tended to be greater in Cu-supplemented steers after a 24-h feed withdrawal and at 2 h postfeed consumption, potentially explaining the reduction in subcutaneous adipose tissue deposition. However, when exogenous norepinephrine was administrated, serum NEFA concentrations were less in Cu-supplemented steers, possibly due to the nonsupplemented control steers having a greater subcutaneous adipose tissue depth at slaughter relative to Cu-supplemented steers. Furthermore, in vitro basal and epinephrine-stimulated lipolytic rates of subcutaneous adipose tissue were greater in Cu-supplemented steers relative to controls. These data indicate that that Cu may influence lipogenic or lipolytic function in subcutaneous adipose tissue. In an attempt to further investigate the effect of Cu on lipid metabolism, lipogenic, lipolytic, and homeostatic mechanisms related to Cu are currently being studied. Recent data indicate that genes involved in bovine liver Cu homeostasis are correlated with Cu transporter protein gene expression in the bovine liver. Therefore, the overall intent of this review is to discuss possible mechanisms whereby Cu may affect lipid metabolism in ruminants.     

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.     

Somatotropin and adipose tissue metabolism: substrate and temporal effects

The purpose of these studies was to determine the time course for changes in feed intake, blood metabolites, and lipogenic activity in adipose tissue in response to the initiation of porcine somatotropin (pST) treatment and following withdrawal from treatment in barrows. An initial study was conducted to determine the impact of chronic pST treatment (4 wk of daily injection; 0 vs 4 mg/d) on adipose tissue lipid metabolism in barrows (initial weight 67 kg). Feed efficiency was improved 27%, backfat thickness was decreased 43%, and glucose and lactate oxidation and incorporation into lipid in adipose tissue was reduced 70 to 86% in pST-treated pigs. Palmitate esterification was decreased 44%, whereas palmitate oxidation was unaffected. In vitro metabolism of lactate, glucose, and palmitate in liver slices was not affected by pST treatment. The time-course for changes in intake and adipose tissue metabolism in response to 7 d of pST (0 vs 4 mg/d) treatment and 7 d of withdrawal was examined in subsequent studies in barrows (initial weight 75 kg). Feed intake during pST treatment was significantly (P < .05) less than in control pigs within 24 h of the initiation of treatment and remained low through 3 d after withdrawal. Adipose tissue biopsies were obtained on d 0, 1, 2, 4, and 7 of the treatment phase and on d 2, 4, and 7 after withdrawal from 7 d of treatment. Maximal inhibition of lipogenesis by pST treatment in adipose tissue in vitro was observed on d 4 (-68%) and d 7 (-69%). Similarly, fatty acid synthase activity declined during the treatment period, with the greatest change noted on d 7 (-26%). After withdrawal from treatment, lipogenesis gradually increased, returning to control values 7 d after withdrawal. Levels of IGF-I began to increase from d 1 to d 7 of treatment, continually decreased during withdrawal, and were normalized by the end of the withdrawal period. Plasma urea nitrogen concentrations decreased during treatment, increased during the withdrawal phase, and were normalized 4 d after the last pST treatment. Overall results indicate that most of the metabolic changes in response to pST occur within 1 wk of treatment and return to pretreatment values after 7 d of withdrawal from treatment.     

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.     

The relationship between mitochondrial function and residual feed intake in Angus steers

The objective of this study was to examine the relationship between mitochondrial function and residual feed intake in Angus steers. Individual feed intakes were recorded for a contemporary group of 40 steers via the GrowSafe feed intake system. Intakes were then used to calculate residual feed intake (RFI), a measure of efficiency. Based on these calculations, 9 low (RFI = -0.83) and 8 high (RFI = 0.78) RFI animals were selected for further study. Blood samples were collected via jugular venipuncture 1 wk before slaughter for the determination of plasma glucose and insulin concentrations. Tissue samples were taken from the LM from both the high and low RFI animals and mitochondria were isolated for measurement of oxygen consumption and hydrogen peroxide production. Average daily gain and carcass composition were not different between the high and low RFI steers; however, ADFI by the high RFI animals was 1.54 kg/d greater (P < 0.001) than for the low RFI animals. Low RFI steers exhibited a greater (P < 0.05) rate of state 2 and 3 respiration, respiratory control ratio, and hydrogen peroxide production than high RFI steers when provided with glutamate or succinate as a respiratory substrate. The acceptor control and adenosine diphosphate:oxygen ratios were not different between the 2 groups for either substrate. When hydrogen peroxide production was expressed as a ratio to respiration rate there was no difference between groups, signifying that electron leak was similar for both groups. Plasma glucose concentration was greater (P < 0.05) in the high RFI steers than in the low RFI steers; however, plasma insulin concentration was not different (P = 0.22) between the 2 groups. The ratio between plasma glucose and insulin concentration was similar (P = 0.88) between the 2 groups indicating no difference in glucose metabolism. The increased plasma glucose concentration observed in the high RFI steers was presumed to be the result of a greater feed intake by these animals. It seems that mitochondrial function is not different between the high and low RFI groups but rather the rate of mitochondrial respiration is increased in low RFI steers compared with high RFI steers.     

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.     

Circulating ghrelin and leptin concentrations and growth hormone secretagogue receptor abundance in liver, muscle, and adipose tissue of beef cattle exhibiting differences in composition of gain

Data from species other than cattle indicate that ghrelin and GH secretagogue receptor (GHS-R) could play a key role in fat deposition, energy homeostasis, or glucose metabolism by directly affecting liver and adipose tissue metabolism. Beef steers (n = 72) were used to test the hypothesis that plasma ghrelin and leptin concentrations and abundance of the GHS-R in liver, muscle, and adipose tissues differ in steers exhibiting differences in composition of gain. At trial initiation (d 0), 8 steers were slaughtered for initial carcass composition. The remaining 64 steers were stratified by BW, allotted to pen, and treatment was assigned randomly to pen. Steers were not implanted with anabolic steroids. Treatments were 1) a low-energy (LE) diet fed during the growing period (0 to 111 d) followed by a high-energy (HE) diet during the finishing period (112 to 209 d; LE-HE) or 2) the HE diet for the duration of the trial (1 to 209 d; HE-HE). Eight steers per treatment were slaughtered on d 88, 111, 160, and 209. Carcass ninth, tenth, and eleventh rib sections were dissected for chemical composition and regression equations were developed to predict compositional gain. Liver, muscle, and subcutaneous adipose tissues were frozen in liquid nitrogen for subsequent Western blotting for GHS-R. Replicate blood samples collected before each slaughter were assayed for ghrelin and leptin concentrations. When compared at a common compositional fat end-point, the rate of carcass fat accretion (g·kg of shrunk BW(-1)) was greater (P < 0.001) in HE-HE steers whereas the rate of carcass protein accretion (g·kg of shrunk BW(-1)) was less (P < 0.001) compared with LE-HE steers. When compared at a common compositional fat end-point, plasma leptin, ghrelin, and insulin concentrations were greater (P < 0.05) for HE-HE compared with LE-HE steers. Abundance of the GHS-R, to which ghrelin binds, increased over time in liver and adipose tissue but did not differ as a result of treatment. Plasma ghrelin concentrations were increased for cattle continuously fed the HE diet as they became increasingly fatter; however, abundance of the GHS-R in liver, muscle, and subcutaneous adipose tissue was not different between treatment groups. The role of ghrelin in cattle metabolism warrants further investigation as it could have a significant effect on composition of BW gain, feed efficiency, and metabolic disorders such as ketosis and fatty liver.     

Stimulation of lipogenesis in bovine adipose tissue by insulin and insulin-like growth factor

The present study was undertaken to determine if insulin and insulin-like growth factor 1 (IGF-1) stimulated lipogenesis in bovine adipose tissue and determine the effects of insulin on lipogenic capacity in adipose tissue cultured for 48 h. In contrast to previous studies, insulin markedly stimulated lipogenesis in short-term (2 h) incubations. The stimulation of lipogenesis by insulin was dependent upon the source of bovine serum albumin used in the buffer. Insulin-like growth factor 1 also stimulated lipogenesis; however, the potency was 80- to 100-fold lower than for insulin. Lipogenic capacity was decreased approximately 75% after 48 h of culture in the absence of insulin. When insulin was present in the culture medium, the reduction in lipogenic capacity was attenuated in a dose-dependent manner. However, insulin alone did not totally maintain lipogenic capacity after 48 h. In contrast, inclusion of hydrocortisone (HC; 50 ng/ml) and insulin (10 ng/ml) in the medium completely prevented the decline in lipogenic capacity of cultured bovine adipose tissue. In summary, these results indicate that bovine adipocytes are quite sensitive to insulin in short-term in vitro incubations and that insulin plays a predominant role in maintenance of lipogenic capacity of bovine adipose tissue during culture. Furthermore, the marked potentiation of insulin's effects of lipogenesis after 48 h of culture by HC suggests that the glucocorticoid is involved in regulation of insulin receptor number and(or) other cellular proteins (e.g., enzymes) which are important for lipogenesis to occur.     

Effect of sex on lipogenic activity in swine adipose tissue

Adipose tissue was obtained from male, female and male pigs castrated either at birth, 2 or 4 mo of age. Pigs were biopsied at 11 and 20 wk of age to obtain adipose tissue samples. Lipogenic capacity was assessed by measurement of in vitro glucose incorporation into lipids of adipose tissue slices. At 20 wk of age, males were less fat than females or males castrated either at birth or 20 mo of age. Adipocyte volume at 20 wk of age was smaller in males, females and recently castrated males (4 mo) than males castrated at birth or 2 mo of age. Lipogenic rate at 20 wk of age was lower in males than in castrated pigs; females had intermediate lipogenic rates. The results provide a partial metabolic explanation for the difference in subcutaneous fat deposition in the sexually diverse groups. There was no effect of estradiol-17 beta or testosterone in vitro on adipose tissue lipogenesis, suggesting that sex hormone effects in vivo may not be involved in short-term regulation of lipogenesis.     

Influence of abomasal carbohydrates on subcutaneous, omental, and mesenteric adipose lipogenic and lipolytic rates in growing beef steers

To determine the response to alteration in site and form of carbohydrate delivery to the digestive tract, in vitro rates of lipogenesis and lipolysis in mesenteric (MESA), omental (OMA), and subcutaneous (SQA) adipose depots were compared. Forty crossbred beef steers (243 +/- 2 kg of BW) were fed 161 (LI) or 214 (HI) kcal of ME/(kg of BW(0.75) x d) or they were fed LI and infused for 35 d into the rumen (R) or abomasum (A) with starch hydrolysate (SH) or into the abomasum with glucose (G). Jugular blood samples were collected, steers were slaughtered, and adipose depots were sampled and prepared for assessment of lipogenesis and lipolysis in vitro. Blood concentrations of glucagon were increased (P = 0.04) in HI-H2O compared with LI-H2O steers, whereas A-SH tended to increase (P = 0.08) circulating IGF-I relative to R-SH, and A-G tended to have elevated (P = 0.09) T3 compared with A-SH. Lipolysis, as assessed by NEFA release, was unaffected by treatment. Glycerol release by the MESA and SQA was increased or tended to be increased (P < or = 0.08) in HI-H2O compared with LI-H2O steers. In A-G compared with A-SH steers, glycerol release from OMA increased (P = 0.008) and from SQA tended to be increased (P = 0.08). Acetate incorporation into total neutral lipids (TNL) increased or tended to increase with ME intake and SH infusion (P < or = 0.09) across all depots. Rates of acetate incorporation into fatty acids (FA) also increased or tended to be increased (P < or = 0.1) by SH infusion across all depots, but only that of SQA was increased with ME intake (HI-H2O vs. LI-H2O; P = 0.02). Rates of acetate incorporation into FA and TNL in MESA were increased (P < or = 0.03) by A-SH compared with R-SH, but site of SH infusion did not affect the rates in SQA or OMA. Glucose incorporation into TNL for MESA and SQA increased or tended to be increased (P < or = 0.1) by dietary and infused energy, whereas for OMA they tended to be increased (P = 0.1) only by SH infusion. In contrast, glucose incorporation into FA was unaffected by energy supply but tended to be increased (P = 0.07) by SH in MESA and tended to be greater (P = 0.08) for A-G than A-SH in OMA. The general across-depot pattern of acetate incorporation rate into FA and TNL was SQA > OMA > MESA, whereas, for glucose incorporation, rates across depots were equivalent. These data provide evidence that the postruminal supply of energy, specifically carbohydrate, stimulates lipogenesis from acetate and glucose and is more pronounced in abdominal depots relative to the subcutaneous depot.     

Lipogenesis and stearoyl-CoA desaturase gene expression and enzyme activity in adipose tissue of short- and long-fed Angus and Wagyu steers fed corn- or hay-based diets

Angus and Wagyu steers consuming high-roughage diets exhibit large differences in adipose tissue fatty acid composition, but there are no differences in terminal measures of stearoyl-CoA desaturase (SCD) activity or gene expression. Also, adipose tissue lipids of cattle fed corn-based diets have greater MUFA:SFA ratios than cattle fed hay-based diets. We hypothesized that any changes in SCD gene expression and activity would precede similar changes in adipose tissue lipogenesis between short- and long-fed endpoints. Furthermore, changes in SCD activity and gene expression between production endpoints would differ between corn- and hay-fed steers and between Wagyu and Angus steers. Angus (n = 8) and Wagyu (n = 8) steers were fed a corn-based diet for 8 mo (short-fed; 16 mo of age) or 16 mo (long-fed; 24 mo of age), whereas another group of Angus (n = 8) and Wagyu (n = 8) steers was fed a hay-based diet for 12 mo (short-fed; 20 mo of age) or 20 mo (long-fed; 28 mo of age) to match the end point BW of the corn-fed steers. Acetate incorporation into lipids in vitro was greater (P < 0.01) in corn-fed steers than in hay-fed steers and tended (P = 0.06) to be greater in Wagyu than in Angus s.c. adipose tissue because the rate in Wagyu was twice that of Angus adipose tissue in the corn-fed, short-fed steers. There were diet x end point interactions for lipogenesis in i.m. and s.c. adipose tissues (both P < 0.01) because lipogenesis was 60 to 90% lower in the long-fed cattle than in short-fed cattle fed the corn-based diet. The greatest SCD enzyme activity in Angus s.c. adipose tissue was observed at 24 mo of age (corn-based diet), but activity in Wagyu adipose tissue was greatest at 28 mo of age (hay-based diet; breed x diet x end point interaction, P = 0.08). For short- vs. long-fed endpoints in Angus, s.c. adipose tissue SCD activity was less (hay diet) or the same (corn diet). Conversely, SCD gene expression was greatest in long-fed Wagyu steers fed the hay- or corn-based diets (breed x end point interaction; P < 0.01). Contrary to our hypotheses, SCD activity increased over time, whereas lipogenesis from acetate decreased. However, the developmental pattern of SCD gene expression and activity differed markedly between hay-fed Angus and Wagyu adipose tissues, which may explain the differences in the MUFA:SFA ratios observed in adipose tissues from these cattle.     

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.     

Response to metabolic challenges in early lactation dairy cows during treatment with bovine somatotropin

Milk production is increased in lactating cows treated with bovine somatotropin (bST) because a greater portion of absorbed nutrients are partitioned for milk synthesis. This homeorhetic action may be caused by alterations in response of key tissues to homeostatic signals. To examine this theory, acute metabolic challenges were administered to 8 multiparous Holstein cows (61 +/- 2 days postpartum) receiving daily subcutaneous injections of pituitary-derived bST (26.3 mg) or excipient during two 14-day treatment periods (crossover experimental design). Treatment with bST increased milk yield 12%. Feed intake did not change so that net energy balance decreased (+ .5 vs. -4.3 Mcal/day). Plasma concentrations of nonesterified fatty acids (NEFA) were chronically elevated in bST-treated cows, consistent with energy balance differences. However, baseline concentrations of glucose, insulin, and glucagon in plasma did not differ. On the last 3 days of treatment, individual metabolic challenges were administered via jugular cannulas: epinephrine (700 ng/kg BW), glucose (250 mg/kg BW), insulin (1.0 micrograms/kg BW), and glucagon (175 ng/kg BW). Plasma glucose was reduced after the insulin challenge to a lesser extent during bST treatment. In bST-treated cows, the increase in plasma NEFA in response to epinephrine was greater, and NEFA concentrations were lowered to a greater extent after insulin and glucose challenges. Glucose, insulin, and glucagon removal rates were not altered, nor was plasma glucose response to epinephrine or glucagon challenges. Treatment of lactating cows with bST primarily altered the response of adipose tissue to homeostatic signals which affect lipid metabolism.     

Gene expression of resistin and TNF-α in adipose tissue of Japanese Black steers and Holstein steers

The aim of the present study was to compare the expression of adipose tissue mRNA related to glucose metabolism between Japanese Black steers (n = 5) and Holstein steers (n = 5). We examined the expression of the resistin, tumor necrosis factor‐α (TNF‐α), glucose transporter 1 (GLUT1) and growth hormone receptor (GHR) genes using real‐time polymerase chain reaction of cDNA in adipose tissue. The cDNA sequence identified by 5′/3′‐rapid amplification of cDNA and the deduced amino acid sequence were highly conserved in human, porcine and murine resistin. Expression of resistin mRNA was significantly greater in Holstein steers than in Japanese Black steers. In contrast, expression of TNF‐α mRNA was slightly greater in Japanese Black steers. Expression of GHR mRNA was significantly greater in Japanese Black steers compared with the Holstein steers, although there was no significant difference in the expression of GLUT1 mRNA. However, the plasma non‐esterified fatty acid (NEFA), glucose, insulin and growth hormone concentrations did not differ between Japanese Black and Holstein steers. The present results show that there is a difference in the expression level of mRNA related to glucose metabolism between Japanese Black steers and Holstein steers.     

Exogenous pituitary and recombinant growth hormones induce insulin and insulin-like growth factor 1 resistance in pig adipose tissue

In the present study, pigs were treated daily for 7 days with exogenous porcine growth hormone (pGH; 70 micrograms/kg BW) in order to determine whether pGH induced insulin and insulin-like growth factor 1 (1GF-1) resistance in pig adipose tissue. In the first experiment, pituitary-derived pGH (ppGH) decreased basal and insulin-stimulated lipogenesis by 50%. Insulin sensitivity decreased more than 90% as the result of pGH treatment. Sensitivity and responsiveness to IGF-1 were decreased 50% by ppGH. In a second experiment, pigs were treated daily (70 micrograms/kg BW) with exogenous pituitary pGH (ppGH) or recombinant pGH (rpGH) for 7 days in order to determine if the effects of pGH were intrinsic properties of the hormone. Both rpGH and ppGH caused similar decreases in basal rates of lipogenesis, insulin- and IGF-1-stimulated lipogenesis, and insulin and IGF-1 responsiveness in pig adipose tissue. In summary, the decrease in adipose tissue growth of pigs treated chronically with pGH is due in large part to the suppression of fatty acid synthesis and a decrease in the ability of insulin to stimulate lipid synthesis in pig adipocytes. These responses are intrinsic properties of pGH since the effects of rpGH mimicked those of ppGH. The role and importance of a decrease in IGF-1 responsiveness remains to be resolved.